automatic vol. abstracts of papers presented at the islar...

Journal of Automatic Chemistry, Vol. 18, No. 2 (March-April 1996), pp. 29-57

Abstracts of papers presented at theISLAR (International Symposium onLaboratory Automation and Robotics) 1995

Thirteen’s a charm for ISLAR ’9513th Annual Symposium Features New Sessions & Record Breaking Attendance. It was a memorable year forthe International Symposium on Laboratory Automation & Robotics (ISLAR ’95). This year’s symposium--the luckynumber 13--enhanced ISLAR’s reputation as the pre-eminent conference on laboratory automation and robotics, withfeatures including: a 25 increase to 500 attendees, 95 podium and poster presentations, plus a first-ever special sessionfocused on combinatorial chemistry.

New Emphasis on Management & Drug Discovery Issues. From keynote speeches and feature presentations tointeractive discussion groups and technical poster presentations, ISLAR ’95 provided a forum for the latest developmentsin the burgeoning field of laboratory automation and robotics. In addition, new emphasis on two of the most pressingissues facing scientists and laboratory technicians today--Managing Laboratory Automation and Automating DrugDiscovery--was instituted. Three comprehensive.sessions were conducted to accommodate presentations on managerialissues, as well as four sessions and a special roundtable discussion on the topic of automated drug discovery.

New Application & Company Luncheons Highly Successful. The three day symposium attracted a world renownedgroup ofscientists that included representatives from 17 countries and 150 different organizations. To facilitate interactionbetween attendees, ISLAR offered two special luncheon programmes--one that grouped participants by applicationarea and a special company-specific luncheon. Both provided colleagues, who share common interests and experience,with a forum to interact and network with peers.

As always, another symposium highligh.t was a reception at Zymark Center, Hopkinton, MA, that featured an introductionto Zymark’s expanded customer service capabilities, including demonstrations of a customer service web-site,video-conferencing support services, plus enhanced validation and field application capabilities. The reception alsofeatured demonstrations of the latest developments in robotic technology--Zymark unveiled a number of exciting newproducts, including the MultiDose Automated Dissolution Testing Workstation, an organic Synthesis System and theRapidTrace SPE Workstation.

The 1996 ISLAR will be held in Boston, MA from 20-23 October, 1996. Paper and Poster submissions are beingaccepted now. For more information on the symposium, contact Chris O’Neil at 508 435 9500, send an E-mail [email protected] or visit the ISLAR pages at http://www.zymark.com

Dr James N. Little

PLENARY SESSIONRe-engineering the laboratory

Francis H. ZenieZymark Corporation, Hopkinton, MA, USA

Our organizations are changing more rapidly than everbefore--and the consequences range from prosperity--tofragile survival--to economic failure. Re-engineering is a

process through which we can initiate and direct changerather than react to changes created by others. Re-engineering business is not new, yet it is timely to meettoday’s challenges.

The two primary forces driving these changes include:

Informed customers--informed customers demand morevalue and stimulate competition. As both individualconsumers and business purchasers, we are bettereducated and more demanding about receiving valuefor our investments. As providers of products andservices, therefore, we must re-earn customers’

business by providing innovative new products andgreater value.

(2) Information Technology--IT is both a primary causeand an enabling technology helping to adapt to

change. IT enables organizations to shift frominefficient hierarchies to highly efficient teams work-ing on high leverage opportunities. Ever increasingglobalization is a strategic opportunity made possiblethrough advances in IT.

Both of these driving forces are irreversible and relentless,that is they will never return to the past. Regulation, onthe other hand, is often a political, and even emotional,driving force. We have seen many examples where thedesired benefits of regulation are better attained throughcustomer education and economic incentives.

Science cannot separate itselffrom business and economics.Science creates the technology which drives our mostrobust businesses. As competitive, consumer and regulatorypressures force business to improve quality and product-ivity, science must lead, or at least enable, the bold stepsahead.

29

Abstracts of papers presented at the International Symposium on Laboratory Automation and Robotics

Our goal is to increase productivity--that is create moreeconomic value for each unit of cost. Only when survivalis at stake, must we place cost or people reduction aheadof increasing productivity. Wherever possible, people andautomation should be teamed for productivity.

Mangement’s role continues to change. In the early phaseof the industrial revolution, managers existed to controlworkers who performed physical tasks. Over time physicalwork was transferred to machines and workers gainedmore education to become knowledge workers. Expertknowledge workers became managers to manage special-ized functions. Organizations were structured as hier-archies to maintain control and facilitate the flow ofinformation. Managers focused on specialized tasks ratherthan whole processes which produced value. It is nowonder that serving customers got lost.Today’s managers lead and facilitate knowledge workers.Effective managers delegate work to individuals andteams and then stimulate innovation and change.

Keynote: The new face of drug discovery withrobotics and screening

Prabhavathi B. FernandesBristol-Myers Squibb Pharmaceutical Research Institute, Princeton,:vj, usThe cloning and identification of new genes as targets fordrug discovery have been enhanced greatly through newmethods in biotechnology and DNA sequencing. Coupledwith this novel target discovery is the urgent need for thepharmaceutical industry to develop innovative drugs fornew therapeutic targets. New approaches to screen designhave resulted in sensitive and rapid screens which aresuited to automation and robotics. Rapid identificationof new leads within weeks of screen design is possiblethrough the screening of synthetic collections, combin-atorial chemistry libraries, as well as diverse naturalproduct samples. The direct delivery of data fromautomated instrumentation to computer data bases hasmade the identification and sorting of leads possible.Through the use ofrobotics and automation, high through-put screening can accommodate thousands ofsamples perday and the identification of leads results within weeks ofscreening. Thus, new targets can now be cycled throughscreens, resulting in rapid lead identification for exploratoryand drug discovery research.

However, the biological matrix cannot be analyseddirectly, and the speed of sample preparation has becomethe rate-limiting step in achieving high throughput assays.The need for automation has now shifted away fromanalytical instrumentation towards the sample preparationstep. In particular, solid-phase extraction has seen a majorincrease as the method of choice for sample preparation.It has been of particular importance in the analysis ofpeptides and biotechnology molecules where solventextraction techniques that are widely used for smallmolecule drug analysis are not always possible.

Commercial automated sample preparation instrumenta-tion for the bioanalytical chemist is not widely available,and the need to design an instrument specifically for thispurpose became apparent. Through a series of meetingsaimed at defining the process of sample preparation, andexploring all of the desirable features for analysis ofbiological matrices, a prototype instrument was con-structed. This prototype was set up in a drug metabolismlaboratory and thoroughly evaluated. Through collabor-ative teamwork, the instrument and its controlled softwarewere modified and ’Rapid Trace’was born.

The initial response to Rapid Trace by the laboratorypersonnel has been very positive, and it has demonstratedthe potential to increase the productivity of the laboratoryfor both methods development and sample analysis. Thesystem has been validated as part ofthe analytical method,and shows superior reproducibility over manual SPE tech-niques for the assays evaluated.

DRUG DISCOVERYHigh throughput screening in a small biotechenvironment

Michelle A. J. PalmerGenetics Institute, Cambridge, MA, USA

High throughput screening in a small biotech group or

company shares many of the hurdles found in a largepharmaceutical screening setting, but there are a numberof issues that are unique. These areas were highlighted inthis presentation and a perspective was given on what isinvolved in implementing a HTS effort.

An automated solution for compound weighing,dissolution and distribution

Keynote: Automation breakthrough for bioana-lytical extractions

R. John StubbsBioanalytical Methods, Drug Metabolism, The R. W. JohnsonPharmaceutical Research Institute, Spring House, PA and Raritan,vj, USA

The explosive growth of analytical technology in recentyears has provided the bioanalytical chemist with thepower to measure drugs and their metabolites in biologicalfluids at extremely low concentrations. With the adventof high specificity assays, there is often little or noendogenous interference from the biological matrix.

John Babiak and Linda HeydtRobotics and Automation Research, Wyeth-Ayerst Research,Princeton, NJ 08543, USA

An integral part of any pharmaceutical automatedscreening effort is the availability of a large number oftest compounds in an assay-ready format. The mosttypical arrangement is to dissolve compounds in acommon solvent, such as DMSO, and distribute theminto microtitre plates for future use. The data generatedby this process include the identity of each compound,the quantity ofeach compound dispensed, its final concen-tration in solution, the ultimate plate and well locationof each compound and the volume of solution distributed

30


into each plate. To meet expected future needs for com-pound sourcing. The authors developed a single automatedsystem, that fits on a 6 x 10 ft laboratory bench, incollaboration with Sagian, Inc.

The primary commercial hardware consists of an HPORCA robotic arm on a 3 m track, Tecan 8051 and 5051diluters, two Zebra 105 bar code printer/applicators, threeSymbol bar code readers, two Hamilton syringe pumpsand a Mettler AT200 balance. Custom componentsengineered by Sagian include a microtitre plate elevator,a vial rack elevator and a microtitre plate sealer. Com-munication is maintained by a Sagian IID; the robot iscontrolled by HP-MDS and data management is handledby MS-ACCESS.

The essence ofthe approach was to develop a single systemto weigh, dilute and dissolve compounds and thendistribute them into microtitre plates with minimalhuman involvement. Three steps are required to generatethe desired microtitre plates of compounds. First, thesystem individually bar codes empty amber vials anddetermines the tare weight of each. Second, an approxi-mate amount of compound is transferred manually intoeach tared vial at a remote log-in station and the identityof each compound is transirred manually in each vial byreading bar codes. Third, the vials containing compoundsare returned to the automated system where compoundweights are determined, solvent is added and compoundsare automatically transferred to ’mother’ and ’daughter’plates which are then bar coded and sealed by thesystem.

The integrated system described will increase the authors’laboratory’s capability to provide dissolved compoundsfor future screens without an increase in the labourinvolved in compound handling. Although many of thesteps involved in compound sourcing can be performedby small workstations, and a few groups have describedlarge integrated systems, the authors believe that theyhave an interesting and flexible integrated system ofmodest size that will have an enormous impact on theirlaboratory’s screening programme.

An integrated laboratory automation system forquantitative high throughput screening (QHTS)with cell-based assays

Mark E. Goldman, Timothy Walton,Lynn J. Ransone and Carla M. SutoSignal Pharmaceuticals, Inc., San Diego, CA, USA

During the past decade, large and small pharmaceuticalcompanies have migrated towards centralized highthroughput screening operations. One of the manyadvantages of this approach is that libraries of screeningsamples can be evaluated rapidly in multiple assays.Combined with the use ofsophisticated data managementtools, the resulting information can be compared toidentify samples (either defined chemicals or naturalproduct extracts) that have selective activity and warrantfurther characterization. In order to properly comparedata across assays, or develop estimates of structure-activity relationships, it is becoming increasingly importantto quantify pharmacological responses. To this end, the

authors have optimized their robotics and data manage-ment systems to conduct quantitative high throughputscreening (QHTS).

Compound are prepared using a BenchMate (ZymarkCorporation) modified to accept 16 x 100 mm test tubesin the 8 x 12 format. Vials of compounds are placed inboxes (8 x 12 format) and logged into the data base.Using spatulas, approximately 5 mg samples are placedin pre-tared test tubes, reweighed and then automaticallysolvated (DMSO) to a concentration of 5 mg/ml. Therack of tubes is transferred to a Packard MultiProbe and500 lal aliquots are placed in deep-well microtitre plates.For the inflammation and osteoporosis targets, cell lineshave been developed which are stably transfected withluciferase reporter genes driven by promoters containingappropriate transcription factor binding sites. Screeningis conducted using a custom Zymate robotics system(Zymark Corporation). Each week, 2500 individual sam-

ples are tested in six assays (15 000 sample-assays/week).This represents a six-fold increase throughput comparedto the semi-automated workstation assay. In addition, theZymate System uses one quarter of the personnel requiredfor the semi-automated assay. The quantity of datagenerated by the Zymate System has exceeded thelaboratory’s ability to use Microsoft Excel spreadsheetsfor data reduction and analysis. An Oracle-based client-server data management system is used to registercompounds (synthesized at Signal as well as thosepurchased from commercial sources) and to reduce/compare data from many assays. This QHTS system hasallowed the authors to significantly increase productivity,reduce screening personnel and accurately compareresults with greater confidence.

Development and implementation of microplateimmunoassays employing Zymark robotics

j. Patrick McCurley and Rose E. SekulovichChiton Biocine, Department of Analytical Viral Immunobiology,Emeryville, CA, USA

The authors’ group is responsible for the developmentand implementation of assays to support Chiron Biocineclinical vaccine trials. With a rapidly expanding vaccineprogramme, and some projects already in phase 111 trials,standard manual assays were not adequate for the requiredthroughput. Therefore several automation steps have beenimplemented to facilitate the receiving, tracking, assaying,data processing and data reporting process for multiplesamples.

Samples labeled with bar codes generated in-house arecompared with corresponding labels on the packing list.The software program identifies discrepancies in theshipment, assigns sample storage locations and recordsrelevant shipment information (date of shipment, shipperand receiver IDs, etc.). A repgrt of storage locations canbe generated by entering the sample IDs manually orfrom a file. Assays on selected samples are then run in a

fully automated system. A Zymark PyTechnology systemhas been adapted to perform microplate immunoassays,

31


and two robots are used full time to run eight differentELISA format assays.

The data processing software matches the sample ID tothe data, generates a graphical analysis and analyses theresults obtained for the standards, controls and samplesto determine if the acceptance criteria have been satisfied.Preliminary results are stored in a table for review by thetechnician; after the review, results are electronicallytransferred to a final database for reporting.

Finally, the authors are evaluating the feasibility ofadapting a Zymark .robot for microplate-based virusneutralization assays, to increase throughput and repro-ducibility relative to conventional plaque assays.

MANAGING LABORATORY AUTOMATIONManaging a high throughput screening groupwithin an international organization

Carol Ann HomonBoehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT, USA

During the last decade, the pharmaceutical industry hasdeveloped high throughput screening (GTS) into a majortool for the discovery of such drugs as Paclitaxel(TAXOL(R) product), and Tacrolimus (FK506). AtBoehringer Ingelheim Pharmaceuticals, Inc. (BIPI), theHTS programme identified compounds which resulted inthe discovery and development of nevirapine and ontazo-last. The probability of finding new structural leadsincreases with the number ofcompounds screened. Hence,successful screening requires a high throughput of com-pounds. HTS within an international company must beorganized on a world-wide basis. BIPI is part of theBoehringer Ingelheim companies, a large internationalpharmaceutical firm. Recently, Boehringer Ingelheim hasorganized three dispensary units at Ridgefield, USA;Ingelheim, Germany; and Biberach, Germany. The entireBoehringer Ingelheim library is available for the HTSprogramme through these three dispensaries. Additionally,natural products and peptide libraries are also screened,as are compounds obtained from outside sources. Screeningcentres for Boehringer Ingelheim have been establishedin Ridgefield, USA, Biberach, Germany, and Vienna,Austria. The activities of the dispensaries and thescreening centres are co-ordinated by an internationalgroup within Boehringer Ingelheim. This is responsiblefor all logistical and technical aspects of the HTSprogramme.

Managing automation for high throughput screening

Derek J. HookBristol-Myers Squibb, Biomolecular Screening, Wallingford, CT,USA

Over the last decade the pressure on new drug discoveryin the pharmaceutical industry has placed increasedemphasis on high throughput screening of chemicalcompound libraries, fermentation extracts, plant samples

and now combinatorial libraries as a source of newchemotypes for novel targets.

In order to accomplish this goal, automation has becomepivotal in achieving the necessary productivity in labor-atories to meet challenges. The key factors critical to

implementing automation in pharmaceutical laboratoriesare closely related to those elements for analyticallaboratories. There are, however, special needs forscreening, and the growth of recent symposia and courseson high throughput drug discovery show how importantautomation is to large pharmaceutical houses and smallerbiotechnology companies.

The experience of the pharmaceutical industry in man-

aging the introduction of this automation shows that thereare common factors that have been experienced by manycompanies. Novel technologies are driving the emergenceofnew types ofinstrumentation in the vendor laboratoriesand show some of the possible directions that might betaken by research laboratories in the pharmaceuticalindustry. These directions will continue to put strain onolder automation concepts and they will continue to

challenge the boundaries of robotics and automationcapability.

Enabling biotechnology research using laboratoryautomation

Steven D. Hamilton, Jason W. Armstrong,Richard A. Stanton and James V. PetersenAmgen Inc., Thousand Oaks, CA, USA

For many years, analytical laboratories have realizedincreased productivity through the use of laboratoryautomation. Similar technology is now becoming an

important tool to enable biotechnology and pharma-ceutical discovery research to progress more quicklyand efficiently. More importantly, automation enablesresearchers to develop discovery processes that go wellbeyond the human capability to manipulate, track andanalyse.

Automation in the research environment involves suchtechnologies as robotics, automatic identification, machinevision, informatics, and image and signal processing. Userfriendly, robust and flexible automation must be developedand/or applied in an environment where project speed iscritical and research directions change rapidly. Theseneeds have led to the creation of the Amgen ResearchAutomation Technologies group, whose focus is to spanthe boundary between automation development andresearch biology and chemistry.

The motivations, problems, rewards and strategy relatedto implementing automation within a biotechnologyresearch organization were discussed, using examples ofthe Amgen automation programme including: automatedcolony imaging and picking; high density spotting,analysis and re-arraying; automatic identification (barcoding) and tracking; high throughput PCR preparationand thermal cycling; automated plasmid preparations;and high throughput compound screening.

32


CHEMICAL ANALYSISThe role of a sample preparation robotics systemat a corporate analytical research and developmentlaboratory

Matthew BenningInternational Specialty Products, Inc., Wayne, NJ, USA

International Specialty Products (ISP) is a manufacturerof high performance specialty chemicals. The productshave diverse applications as ingredients or processing aidsin personal care, pharmaceuticals, agricultural andbeverage formulations. The diversity of product appli-cations is equally matched by their chemical morpho-logical characteristics. Products are available as insolublepowders, soluble powders, highly viscous liquids, andnon-viscous liquids.

The ISP HPLC method development group is responsiblefor methods development and validation in support ofresearch, marketing, quality assurance, and manu-facturing. The corporate effort on continuous productimprovement has placed demands on ISP’s laboratorythat challenge current manpower capability. Routineanalysis in support of research projects and product linesprior to HPLC method implementation and validationat manufacturing facilities, are a significant portion of thelaboratory’s workload.

An application has been developed to allow roboticsample preparation for a variety ofproducts, matrices andprocedures thus freeing the chemists to pursue HPLCmethod development related activities including instru-ment setup and data processing. A Zymate II robot witha System V controller was assembled from standardpysections. Subsequently, hardware and software modi-fications were implemented and validated to enable thesystem to fit into our highly automated laboratory.

The uniqueness of the robotic system described is in thesoftware design that allows a variety of samples and tasksto be performed by users with different sample preparationrequirements. The success of the system is measured bythe increased routine workload handled by a small groupwith increased productivity. Finally, the robotics systemis one part of an automated laboratory. Integration asevidenced by data portability, integrity, and someregulatory concerns is easily met and enhanced with thisimplementation.

(3) Weigh liquid and solid samples in amounts of100-500 mg into different target vessels.

(4) Use different tecnniques for weighing in solid samples.(5) Transfer the weight data to a LIMS.(6) Label the target vessels with barcode and letters by

using an ink jet technique.

The main challenges proved to be handling of a largevariety of sample vessels, finding adequate and robusttechniques for weighing solids of varying consistencyin small amounts, and integrating the robot systeminto an existing on-line data collection system and aLIMS.

Automating the vapour test method for evaluatingchemical protective fabrics

Raymond E. Andreotti, Cyrus Kendrick, and DonaldRivinU.S. Army Natick Research, Development and Engineering Center,Natick, MA, USA

An automated vapour test method that provides a rapidand safe means of evaluating the adsorptive capacity offabrics used in chemical protective garments has beendeveloped utilizing laboratory robotics in conjunctionwith an automated effluent detector. Sequential vapourchallenge analysis of fabric samples can be performed ina pneumatically operated computer-controlled test cellthat was designed and developed not only to facilitate thesample loading and unloading procedure, but also toprovide a test environment for obtaining precise andreliable adsorption kinetics. A laboratory robot wasprogrammed to replace the repetitious and tedious tasksformerly performed manually in the vapour test. Apersonal computer is interfaced to the system and is usedfor programming and control of the robotics as well ascollecting, calculating, and storing effluent concentrationtest data output from the automated detector. Operationalprograms were developed that can easily be updated, toaccommodate changing protocols, such as varying chem-ical concentrations, flow velocities, temperature, andexposure times. Use of this type of technology fordefence- and environment-related applications wouldensure that safe and reliable testing could be conductedin governmental and industrial settings.

Automated procedures for weighing small amountsof samples

H. Speck, M. Kranz, M. Luebke, W. Schmid andE. SametschekBASF AG, Ludwigshafen, GermanySample preparation for ICP-spectrometry consists of twosteps: weighing and disclosure. To automate the first step,the authors designed a laboratory robot (in co-operationwith Zymark), which can do the following tasks:

(1) Identify samples by reading barcode and gettinginformation on the samples from a LIMS.

(2) Open and close different capped (crimp caps, screwcaps, snap caps) and different size sample vessels.

A complete custom automation of a commercialanalytical instrument with a Zymate(R) PyTechnol-ogyTM laboratory robot system and the associatedcomputer interface

Mitchell E. Meyer, Collin B. Hitchcock, H. RobertPinnick, Jr. and Gordon R. StallingsPhillips Petroleum Company, Research & Development, Bartles-ville, OK, USA

The challenge of completely automating an analyticaltechnique is often related to the interface between therobot, the analytical instrument instrument, a LaboratoryInformation Management System (LIMS), and theassociated sharing of data between these systems. Thispaper discussed an approach employed on three Applied

33


Research Laboratories (ARL) Inductively CoupledPlasmas (ICPs) to fully automate these analyticalinstruments utilizing a customized Zymate(R) PyTechnol-ogyTM system, a desktop computer (PC), a LIMS system,and a local area network (LAN). This design can be usedfor future robotization and automation efforts with othercommercially available instrumentation.

Combining robotics with UV-VIS-NIR spectroscopy

Carl G. ZimbaPolaroid Corporation, Cambridge, MA, USA

In the development of new photographic media it isnecessary to evaluate the colorimetry of a large number9g dyes and other components. As these prototypematerials are synthesized, the absorption ofthese materialsin the ultraviolet, visible, and near-infrared spectralregions are measured to verify both their absorbancemaxima and their absorption intensity.

As part ofan ongoing effort to automate routine analyticalspectroscopic measurements, a laboratory robot has beencombined with both a UV-VIS and VIS-NIR spectro-photometer. The result has been the creation of a highlyautomated instrument which prepares solutions frompowder or crystalline samples using a variety of solvents,and measures the absorption spectrum.

Using this instrument, sample characterizations can beaccomplished with more rapid turn-around, more precisionand accuracy, and in some cases with less material.Furthermore, the available staff can be better utilized tomeet the needs of Polaroid’s analysis clients.

The presentation gave an overview of both the hardwareand the software of this automated spectrometer andcompared the performance of this instrument to manualmethods. Potential applications to a more traditionalQA/QC environment were also discussed.

Automation of a NMR measurement of finish-on-fibre

James Rodgers and Joel WeekleyFibers Division of Monsanto, Gonzalez, FL, USA

Finish is applied to the fibre surface primarily to assistthe fibre’s performance during downstream processing.The quantity of finish applied to the fibre surface is calledFinish-On-Fibre (FOF). FOF is an important process andquality control parameter for textile fibres and yarns, andit has historically been measured by solvent extractiontechniques. A Nuclear Magnetic Resonance (NMR)method, using the bench-top Oxford QP20 pulsed NMRanalyser, was developed for the measurement of FOF on

nylon fibre products (carpet, tire, conductive fibre). TheNMR instrumentation yielded rapid, cost effective,accurate, and precise non-destructive measurements ofFOF. Since the NMR analysis requires no solvents,significant environmental and cost-related benefits wererealized. Excellent FOF agreement was obtained betweenthe NMR method and a standard solvent extractiontechnique. A program was initiated to interface the NMR

unit to a laboratory automation system. Several auto-mation vendors were compared using a ComparisonMatrix (technical capabilities, partnership potential,economics). A Zymark Zymate XP system was determinedto be the optimum laboratory automation system for thisapplication. A joint Monsanto-Oxford-Zymark programwas established to implement hardware and softwareintegration of the Zymate XP system to the Oxford NMRunit and to Monsanto’s QC and LIMS systems. Custom-ized Oxford-Zymark software allowed control ofthe NMRand robotic functions by the System V controller.Customized Monsanto-Zymark software resulted in thesuccessful integration of the NMR results into the PlantLIMS system.

Adsorbable organic halogen compounds (AOX):robot-based automated analysis

Helmut Dillenburg and Bernhard KoernerSolvay Alkali GmbH, Rheinberg, Germany

The parameter AOX (DIN 38409 T14 AdsorbableOrganic Halogen Compounds, where X C1, Br, J) is,due to legal regulations and prescriptions, one of the most

’popular’ analytical parameters in Germany, if not inWestern Europe, for the analysis ofwater and wastewater.But it is also very time-consuming and therefore costly.A well-trained laboratory operator can analyse about 8to 10 samples (double estimation) per shift. Thus a lot ofattempts have been made to automate all of the analyticalprocedure or at least major parts of it.

The principle of measurement is as follows: the organiccomponents of the sample are adsorbed on active carbon.Interferences ofinorganic halogen compounds are removedby rinsing the adsorption-columns with an aqueoussolution of sodium nitrate. The loaded active carbon isthen combusted in an oxygen stream. The generated HXare absorbed and the mass of halogen is detected.

This paper described a fully automated, Zymate-robot-based, DIN-compatible procedure, comprising sampleidentification, enrichment step, sample transfer andinterfacing to a commercially available automatedanalysis system, data collection, evaluation and docu-mentation of the results. Starting with the validationprocess, results for calibration standards and real-lifesamples are compared with the results obtained by themanual DIN-procedure in parallel. The comparisonshowed excellent consistency. The sample capacity of thesystem is 10 samples per shift (triple estimation andcalibration) and can be easily adapted to specific needs.

DRUGS IN BIOFLUIDSAn evaluation of automated solid-phase extractionsystems

R. Eric Schmidt, L. A. Kosobud, N. S. Khoshaba,and K. J. MillerG.D. Searle & Company, Skokie, IL, USA

An evaluation ofcommercially available automated solid-phase extraction (SPE) systems to perform the extractionof pharmaceuticals from plasma and urine samples was

34


conducted. The evaluation focused on increased through-put and the creation of automated assays which could betransferred to contract laboratories for routine analyses.The Gilson ASPEC XLTM, Hamilton MicroLab 2200TM,Zymark BenchMateTM, Zymark RapidTraceTM, and theCardinal AutoChem WorkstationTM, were evaluated.The evaluation focused on three components: (1) hard-ware, (2) software/programming, and (3) applicationof an established assay presently using a ZymarkPyTechnology robot. For the hardware and software/programming, the advantages and the disadvantages ofeach system were generated. For the application, theanalysis time, carry-over, precision and accuracy werecompared and conclusions were drawn regarding systemapplicability for automated sample preparation.

Use of a Zymate system for bioanalytical auto-mation as a workstation

Michael Wirth, Mats Svensson andTorbjorn ArvidssonPharmaceutical and Analytical R&D, Analytical Chemistry,Astra Pain Control AB, 151 85 Sodertalje, Sweden

Robotics in bioanalytical work often comprises fullyautomated systems developed for a given method. Thispaper introduced a concept based on automation of thebioanalytical work using different types of generalworkstations. Several such stations are commerciallyavailable--dilutors, solid-phase extraction systems, auto-

injectors etc.

A new type of workstation using a Zymate robotic systemhas been developed for the necessary steps followingextraction (evaporation, reconstitution and transfer to

autosampler vials including capsulation). The develop-ment, test and validation of the robotic system wasdiscussed. The concept has been applied in a bioanalyticalmethod for determination of local anaesthetic drugs inblood plasma using liquid-liquid extraction and gaschromatography.

Flexible automation tools for drug metabolism

Thomas L. Lloyd and Michael G. DoddsDrug Metabolism Department, Glaxo Wellcome, ResearchTriangle Park, NC, USA

Robotics was first introduced into most drug metabolismlaboratories as a means to automate routine assays.Existing methods for biological fluid sample analysis weretransferred to an automated platform to support largeclinical studies. Systems were often dedicated to one

application and the decision for automating a projecthinged on whether the workload for that project justifiedthe investment in equipment and set-up time.

As the flexibility and variety ofautomation tools progressed,the range of application extended upstream in the drugmetabolism process. Robotic systems are still used toautomate analysis for large clinical studies. However, theyare also used for smaller volume pre-clinical and researchsupport applications. The automated systems are flexibleenough to handle up to six different assays in a week and

multiple compounds within an analytical run. Bio-analytical methods are developed and refined directly on

an automated platform in conjunction with experimentaldesign software. By minimizing or eliminating set-up timebetween applications, a steady supply of low volumebioanalytical applications can also be cost effective.

The breadth of automation for these applications hasexpanded as well. Bioanalytical sample preparation tech-niques include solid phase extraction, liquid-liquidextraction, protein precipitation and dialysis. Samplecontainer labels can be generated and applied auto-

matically for a variety of different container dimensions.Standards and quality controls can be prepared from a

concentrated stock and stored in the same labeledcontainers. A number of sample preparation processingoptions are available. Prepared samples can be analysedon-line or stored in autosampler vials. Automationcontinues to be developed for new diversified applicationswithin drug metabolism.

Extraction of morphine and codeine from urinesamples using the Zymark RapidTrace

Francis X. Diamond, William H. Vickery and Johnde KanelNational Medical Services, Inc., Willow Grove, PA, USA

Morphine and codeine are the two opiate drugs designatedby SAMHSA (Substance Abuse Mental Health ServiceAdministration, formerly the National Institute for DrugAbuse, NIDA) for analysis in employment and pre-employment situations to determine if heroin, codeine or

morphine have been used. Numerous methods have beenpublished describing methods to extract morphine andcodeine from urine either by traditional liquid/liquidmeans or by solid phase extraction. Liquid/liquidextractions are time consuming, difficult to automate andrequire great analyst skill if they are to produce repro-ducible and accurate results. Each manufacturer of solidphase extraction cartridges has published methods to

accomplish the extraction of morphine and codeine fromurine. For the purposes of evaluating the ZymarkRapidTraceTM automated extraction workstation theauthors chosen 3 ml Confirm HCX IsoluteTM cartridgeswhich are available in the USA from Jones Chroma-tography, Inc. The method was programmed into theRapidTraceTM software without modification. A set of 10Zymark RapidTrace sample preparation units controlledby one personal computer was capable of extractingcodeine and/or morphine from 50 urine samples per hour.The samples were then evaporated to dryness at 40Cusing a Zymark TurboVap. This step took approximately10 minutes. The samples were then derivatized by reactionwith 50 gl (BSTFA with 19/o TMCS) in 501 gl ethylacetate for 30 minutes at 70C in a sealed tube. Thesamples were then transferred to autosampler vials andanalysed by GC/MS. A technician would previously havespent at least three hours to prepare these extracts

manually--the use of the RapidTrace frees the technicianto work on more productive things such as data reviewfor two of the three hours.

35


Two millilitre urine samples known to be negative forcodeine and morphine were spiked with deuteratedcodeine and morphine at levels of 50 ng/ml as internalstandards and with codeine and morphine at varyinglevels to evaluate the recovery, linearity and carry-over ofthe extraction method when performed by the ZymarkRapidTrace. The results for codeine and morphine areshown in tables and 2. The standards at 150, 300, 600,1200 and 2400 ng/ml were used to determine backcalculated levels for these standards and to quantitatethe controls at 240 and 360 ng/ml, as well as levels invarious blank samples. Urine samples were extractedthrough the procedure after spiking it with 50 000 ng/mlof morphine or 1000.00 ng/ml codeine. Analysis of theblank samples following these spiked samples was done toestimate carry-over. The procedure used was identical tothe routine procedure which would be followed for realsamples. No extra manipulative steps were added in aneffort to minimize carry-over. The extracts from the50 000 ng/ml or 100 000 ng/ml samples were not deriv-atized nor analysed by GC/MS since potential carry-overfrom the GC autosampler could have confused the results.

Under SAMHSA guidelines, an affirmative cut-off of300 ng/ml is used to administratively differentiate positivesamples from negative ones. Irrespective of other data,samples containing codeine or morphine below thisadministrative level are reported to clients as negative.Direct quantitation of the blanks following the 100000ng/ml codeine spike yields 65 ng/ml or 0"07% carry-over.

Table 1. Codeine analysis: Diamond et al.

Calculate Ion Ion IonSpike D ratio ratio rationg/ml level, ng/ml 229/371 356/371 229/356

150 160 0"32 0"13 2"5300 340 0"37 0"12 3"1600 640 0"45 0’ 13 3’51200 1100 0"59 0’ 14 4"2

240 control 220 0"38 0’ 13 2"9360 control 320 0"40 0’ 12 3"3

0 0"56 2"8 1"6 1"80* 65 0"39 0’15 2"6

* Blank extracted after the 100 000 ng/ml spiked sample extraction.

Table 2. Morphine analysis: Diamond et al.

Calculate Ion Ion IonSpike D ratio ratio rationg/ml level, ng/ml 401/429 414/429 401/414

150 150 0"28 0"43 0"65300 330 0"30 0"46 0"65600 560 0"29 0"45 0"641200 900 0" 31 0"47 0"66

240 control 210 0"32 0"53 0"60360 control 330 0"33 0"47 0"70

0 1"1 0"51 2"3 0"220* 16 0"34 0"54 0"63


For morphine, the blank following the extraction of the50 000 ng/ml spike yielded 16 ng/ml or 0"03% carry-over.Some of this level is derived from the background noise.The blanks extracted after relatively low levels ofbenzoylecgonine quantitate to approximately ng/ml, forexample. These levels are all far below the administrativecut off of 300 ng/ml.

Fully automatic determination of the enantiomersof amlodipine in human plasma by robotic samplepreparation and gas chromatography

K. D. Riedel, F. Scharpf, H. Laufen and M. LeitoldDepartment of Pharmacology, Pfizer Mack, Illertissen, Germany

A sensitive and specific gas chromatographic (GC)method using a Zymark II robotic system for completesample preparation was developed for the determinationin plasma of the enantiomers of amlodipine, a calciumchannel blocking therapeutic agent. Plasma samples werealkalinized and extracted with tert.-butyl methyl ether(TBME). Amlodipine and the internal standard (a C1analogue of amlodipine) were then back extracted intocitric acid. After discarding the organic layer the aqueouslayer was alkalinized and again extracted with TBME.The ether phase was then evaporated to dryness under astream of nitrogen. For chiral derivatization the driedextracts were reconstituted into a solution of (+)-(S)--methoxy--trifluoromethylphenylacety chloride. Afterremoving the excess reagent with potassium carbonate thediastereoisomers were then analysed by GC with electroncapture detection.

The limit ofdetection of this method is 0"02 ng/ml plasmafor both enantiomers. The day-to-day %CV ranged from2"6 to 11"7 (both enantiomers). The corresponding withinday %CV ranged from 2"6 to 11"7. The Zymate systemperforms the complete extraction, back extraction,evaporation, derivatization and transfer of the extractsinto autosampler vials for gas chromatography. Thesystem consists of the following components (number ofmodules in brackets): dual-function hand (2), liquid-transfer hand (1), power and event controller (2), rotatingoverhead shaker (1), centrifuge (1), evaporator (1), screwcapper (1), crimp capper (1), vortex mixer (1), dispensingstation (2), pneumatic dosing station (1), as well as varioussample racks.

A total of 50 samples can be automatically prepared bythe robot within 30 hours. This corresponds to a totalsample throughput ofabout one sample per 35 min, whichis similar to the chromatographic time.

PHARMACEUTICAL ANALYSISAnalysis ofwater soluble vitamins in multivitamin-mineral supplements on an automated tabletprocessing-liquid chromatographic system

Andrew L. Deputy and Lionel P. MurrayBayer Corporation, Consumer Care Division, Elkhart, IN, USA

This presentation described a project to develop a ruggedand reliable automated analysis system for ascorbic acid,

36


folic acid, niacinamide, panthothenic acid, pyridoxine,riboflavin and thiamine in multivitamin-mineral supple-ments. In addition, the analytical system must be robustenough to process a large number ofsamples with varyingformulations and physical characteristics. These differ-ences in products can include colour, flavour, coating,tablet shape, tablet hardness, types and amounts of activeingredients, and multiple tablet excipients.

In order to produce such an automated system, a

chromatographic separation was developed which isextremely rugged and insensitive to minor changes in thetablet excipients, including the flavours and colours.During the development of such a rugged separation, thechromatographic separation was optimized for capacityfactor (k’) and selectivity (). Because of the interrelatednature of these two parameters, an ’expert’ approach wasused for the development of the HPLC separation. Inaddition, post-column derivatization was employed forthose vitamins which either do not have an easilyaccessible chromophore or whose chromatographic be-haviour does not yield the desired selectivity overinterfering components of the table matrix. For themultivitamin-mineral supplements, ascorbic acid andpantothenic acid meet these criteria.

The optimization of the aqueous extraction of the watersoluble vitamins was accomplished using a TaguchiMethods(R) Optimization scheme (Lg) to examine theinfluence of various parameters on the extraction of thevitamins from the tablet matrix. Extraction parametersexamined included homogenization time, extractiontemperature, extraction solvent pH, and pre-extractionsoak time. Parameter conditions and ranges were selectedto optimize the precision (S/N) of the extraction process;therefore, providing a rugged automated anlaysis system.In summary, a rugged, precise analysis system is necessaryfor use in the routine release testing of pharmaceuticalproducts. By the use of experimental design techniques,it is possible to build ruggedness into the analytical systemduring development.

The BenchMate II TPWmfrom installation tofinancial success

Simon SmithSmithKline Beecham Pharmaceuticals, Manor Royal, Crawley,West Sussex, UK

The Quality Assurance Department at SmithKlineBeecham performs the testing of samples for stability,process development/new product introduction androutine release for sale. As a consequence of long-termmanagement objectives, the goals are to reduce laboratorytesting time to 24 hours and eliminate waste whilemaintaining or reducing costs. Within the analyticallaboratory, automation can provide the key to thesebusiness challenges. If careful implementation is employed,the automated laboratory will emerge as the successful,cost effective and efficient laboratory of the future. Thisresult will consequently give that certain leading edge to

any product in the aggressive market place and allowgreater flexibility as the automated laboratory will besituated at the end of the production line.

The release for sale and stability testing of a particularfilm coated tablet has been automated within thelaboratory by the successful implementation of a Bench-Mate II Tablet Processing Workstation. Initial justificationfor the purchase of the system was based on the view thatit would replace an analyst and create extra capacity.This view has been realized but many hidden benefitshave emerged, with the main ones being a reduction ininventory costs and the number of samples requiringretesting.

Due to the success of this first automation challenge, asecond system was purchased, validated and implementedin a third of the time of the first system and is now alsoused for the testing of a second product.

Robotic-LC analysis of Benadryl tablets

Fern Smith and Harold ShawParke-Davis/Warner Lambert Company, Brockville, Ontario,Canada

The BenchMate TPW Workstation (Zymark Corporation,Hopkinton) consists of two parts, the BenchMate Work-station and the Tablet Processing Workstation. TheBenchMate TPW Workstation provides a versatile, costeffective alternative to manual sample preparation forcontent uniformity assays, potency (composite) assays,and dissolution testing. These routine tests require samplesto be ground, dissolved, filtered, diluted, and analysed bychromatographic techniques. The BenchMate TPWWorkstation utilizes computer controlled wet-grindinghomogenization, a liquid management system, an internalfour place analytical balance, and a three place toploading balance to consistently provide accurate andprecise results. The workstation also provides an audittrail for all sample weights used and sample volumestransferred.

Fast dissolution sampling and analysis using a

Zymark dissolution robot and on-line UV/visiblespectroscopy

Daniel W. BarrowBristol-Myers Squibb, New Brunswick, NJ, USA

Dissolution testing is a common analytical procedure usedthroughout the pharmaceutical industry to meet legalrequirements for compendial drugs and to provide a

quality control measure for solid dose forms in manu-

facturing and research operations. Fast on-line UV/visibledissolution sampling and analysis have been achievedusing a customized Zymark dissolution robotic system.The system is capable ofsampling and analysing solutionsfrom a single vessel in less than 50 seconds: allowing sixvessels to be tested in under five minutes. It is equippedwith a second six vessel dissolution tester thus enablinganalysis of 12 dissolution samples in under 10 minutes.The dissolution procedure is fully automated includinganalytical standard analysis, system preparation, sampleaddition and analysis, system clean-up, results calculationand print-out. Common productivity gains range from50 when compared to conventional robotic dissolution

37


systems to 300 when compared to manual methods ofanalysis.

This robotic system has been used in the analysis ofseveral.pharmaceutical products. The presentation showedsystem schematics and system validation data.

Using a PyRobotic system to update and automatethe existing analytical methodology in a currentlymarketed product

cation. Instead of programming for specific applications,an expert program interacts with the user to create theapplication program. The installation qualification andoperational qualification protocols were used to demon-strate that the robotic system hardware and expertprogram are working as intended. The performanceprotocols were used to validate the application programsgenerated by the expert program and to demonstrateequivalence to either manual or other automated pro-cedures.

Allan L. Greenberg and Phillip A. LaneAnalytical Research and Development, The R.W. JohnsonPharmaceutical Research Institute, Raritan, NJ, USA

Laboratory robotics have been previously demonstratedto be most efficiently used where conditions make repro-ducibility important and sample throughput primeconsiderations. Most of the previous implementations inthe authors’ laboratories have been in the application ofrobotics to new dosage form sample preparation schemes.However, in the pharmaceutical industry, especially intoday’s climate of having to work faster and moreproductively, it is easy to go backwards and implementautomation into pre-existing analytical procedures ratherthan only considering application of robotics to newchemical entities.

The robotics programs used were taken from a creamapplication, which was presented two years ago at ISLAR.A comparison ofthe flow diagrams ofthe original programand the desired program was used in the presentation toindicate what part of the program could possibly besalvaged. The programs were modified to meet currentanalytical requirements. The new robotic procedures alsoincorporated a system suitability check and preparationof standards.

The results from the assay of robotically prepared creamsamples were compared to manually prepared samples.The evaluation concluded that the results were statisticallyequivalent. The assay reproducibility and sample through-put was presented for both cases.

Using an expert system with laboratory robots toprovide flexibility in the analytical developmentlaboratory

M. E. Hinshaw and D. A. JacksonLilly Research Laboratories, Eli Lilly and Company, Indianapolis,IN, USA

Until recently, robots have been of limited use in theauthors’ pharmaceutical development laboratories. Thetime required to plan, build, and validate systems is notwell accommodated in today’s accelerated developmentenvironment and once the bolus of samples is through thesystem, the robots are generally not flexible enough to beused for other projects without substantial modification.

This problem has been addressed by changing the basicapproach to building robotic systems at Eli Lilly. Eachsystem is now constructed to allow for multiple samplepreparation techniques using various container sizes,regardless of the requirements of the immediate appli-

Installation, validation, and implementation of theZymark MultiDose Automated Dissolution Work-station for profile testing oftablets using the paddleapparatus

John J. Mullen and Timothy J. McCormickThe DuPont Merck Pharmaceutical Company, Wilmington, DE,USA

The MultiDose Automated Dissolution Workstationautomates the paddle method for dissolution testing ofpharmaceutical solid dosage forms. With minimal operatorset-up the system will automatically dispense dissolutionmedia, check the temperature, introduce tablets, andprovide selected profile sampling at intervals as close asfive minutes. The installation process supplied by themanufacturer helped provide for a problem-free start up.Zymark engineers were able to set-up and have the systemoperating in several days.

A thorough system hardware and software validation wasconducted which included the IQ, OO and PQ of thesystem. The Installation Qualification confirmed that theanalytical balance, thermisters, pumps, and all othercomponents were installed correctly and that theyoperated as expected. The Operation Qualificationverified the functional and applicational performance ofthe workstation. All components did operate as they wereintended throughout anticipated ranges. The PerformanceQualification of the system was the actual execution ofan analytical procedure without an error and in thecorrect sequence.

Product/application specific validation was also conductedwhich included: calibration, sample solution carry-overchecks, vessel washing effectiveness and manual versusautomated comparisons.

The results of the above installation/validation processwere presented along with the implementation of thesystem for product stability testing.

Automated dissolution testing in the validation oftablet coating procedures

Kevin K. OlsenESI Lederle Generics, Pearl River, NY, USA

A common medication is manufactured in four strengthsby ESI Lederle Generics at their Pearl River New Yorkfacility. Validating a recent change in the tablet coatingoperation required extensive dissolution testing in whichthe tablets are allowed to dissolve under tightly controlled

38


conditions and the concentration of the active ingredientis measured as a function of time. Such testing is verycommonly used to monitor the release characteristics ofa wide variety of pharmaceutical preparations.

Since two separate pieces of coating machinery were

involved, almost all the analyses were done twice. Eachof the four tablet strengths required testing on uncoatedtablets, dissolution profiles, and testing of tablets takenfrom different steps in the coating process. The estimatedtime to complete the testing by manual methods was over300 hours. In addition, the laboratory had to support itsnormal workload of quality control analyses.

Substantial savings of time were achieved by automatingthe testing procedures using a Zymark MultiDose, an

autosampler on the UV/Vis spectrophotometer and aBeckman LIMS to manage data handling and routinecalculations. The Zymark MultiDose was the criticalelement because it allowed the entire test to be automated,including the very labour intensive steps of cleaning theapparatus and preparing it for the next sample. The finalresult was a documented saving of slightly more than50 in analyst time. Results obtained by the automatedsystem were in good agreement with those from manuallyprepared samples.

DATA HANDLINGAutomation of data handling for trace analysis ofdrug active in on-line cleaning validation samples

Jon P. Sadowitz and Thomas D. GroeschnerSchein Pharmaceutical Inc., Carmel, NY, USA

In response to current concerns a cleaning validationprogram was developed. Within this program, it becamenecessary to automate the analysis of data to increaseproductivity within a small group solely responsible forthe analysis of all cleaning validation samples. Until veryrecently, data analyses were performed by hand, whichconsumed much of the analyst’s time when dealing withthe large number of samples associated with a cleaningvalidation study: a small cleaning validation study of 50tubes would require over 150 individual calculations. Fora small staff of two chemists, automated analysis wasessential.

This presentation included the development and overviewof the final product utilized in the automated transfer ofraw chromatographic data to transform these data intoa workable tbrmat to interpret the results of a cleaningvalidation study.

LABORATORY WORKSTATIONSAutomating the process of removing coating ma-terials from tablet formulations

Kathy DuquetteWyeth-Ayerst, Rouses Point, NY, USAand Chris WernerBohdan Automation, Mundelein, IL, USA

This presentation demonstrated an automated system thatis capable of removing the coating material from the

active inner core of pharmaceutical tablet formulations.In addition to selectively removing the coating material,the instrument also calculates and records the averagetablet core weight..

The initial intention of the automated workstation was tofree the analyst from the time-consuming task of washingthe tablets by hand. The analyst is now able to wash upto 12 batches of coated tablets with a very short amountof time needed to set up the machine. In addition to

improving the use of valuable employees, more consistentresults are achieved using the automated approach.

The tablet washer has the capability of washing, rinsingand drying the tablets. A variety of’run parameters’ areaddressed by the analyst at the beginning of the program.The computer program also stores the weights of thetablets in its memory, as well as a hard copy print-out.This allows the laboratory to have both permanent andelectronic documentation of the sample preparation andsample data report.

To date, the automated workstation has shown resultsthat are equal to or better than manually preparedsamples.

Automation of a tablet assay using a BenchMateTPW

Michele E. Lake, David A. Hollowell,James L. Sabatowski and Mark N. FlairEli Lilly and Company, Lilly Research Laboratories, Indianapolis,IN, USA

A Zymark BenchMate Tablet Processing Workstation(TPW) has been used to successfully automate a potencydetermination for tablets resulting in precise results andsubstantial savings in analyst time. During automationof the potency method, several iterations of the TPWprocedures were examined. TPW results generated withthe various procedures were compared to manual results(as well as the percentage label claim) until both sets ofresults were statistically equivalent. Since validation ofthe automated method, a control sample has been utilizedand a control chart of this data has shown the reproduci-bility of the TPW for the assay of tablets. The TPW datahas been analysed to evaluate the method variability (runto run), as well as tablet variability for the establishedcontrol sample. This presentation described the automatedassay and the results.

Automation of the analysis of an unstable analytein solid dosage forms with the BenchMate TPW

L. A. Cavenaghi and E. GabrieleGuppo Lepetit SpA, Anagni, Italy

With the use of a BenchMate TPW, the authors havebeen able to automate the analysis of Rifampin in soliddosage forms. This antibiotic, which is widely used for thetreatment of difficult infections, is formulated in sugarcoated tablets of different strength ranging from 100 to

600 mg, and up to now could not be analysed automaticallybecause the compound oxidizes in solution.

39


The use of the BenchMate TPW with its capability ofpreparing standards and samples (weigh, dissolve, diluteand inject) at the appropriate moment has solved theproblem.

A new method has been developed to cover all those inuse for the different formulations. The method covers therange 80 to 720 mg/sct (__ 20 of theoretical content),enabling the laboratory to have an enormous increase inflexibility of response. This increase was achieved with noloss of precision and accuracy of the analytical procedure.

An added bonus ofthe use ofthe BenchMate TPW was theease of the validation with automatic tracking of all thedilutions and thus removing the need to validate theanalyses.

Workstation automation of the biochemical oxygendemand method

Lars Lindquist, Sonja Soloway and Donna PociusWMX Environmental Monitoring Laboratories, Inc., Geneva,IL, USA

A fully automated BOD workstation was developed toautomate the Environmental Protection Agency method405.1. This system has the capability to deliver sample,seed, controls, and dilution water to BOD bottles. Thedissolved oxygen is measured with a probe and a waterseal is formed when the cap is placed on the BOD bottles.

The operation of the Zymark workstation with a personalcomputer running WindowsTM is very user friendlybecause it uses Visual Basic images of the samples, racks,and BOD bottles. The user points and clicks on the mouseto screen images of the sample bottles and inputs theinformation to set up the analyses. The BOD bottles thatare analysed are tracked with a bar code reader. Theresults can either be reported from a standard report orcustomized by use of other spreadsheet software.

System validation, method detection limits, samplethroughput, and performance characteristics were pre-sented.

VALIDATING LABORATORY AUTOMATIONThe generic dictionary: a new approach to validation

P. J. Gallant and R. S. WhiteDuPont Merck Pharmaceuticals, Radiopharmaceuticals Division,Process Testing Development, Billerica, MA, USA

In most GLP/GMP environments, the major downtimeto implementing automated systems is the validationprocess. With respect to the Zymark XP system in DMPC’sProcess Testing Group, the major drawback to convertingthe current XP system over to new analyses is validatingthe new configuration on a relatively unchanged system.

The concept of a generic dictionary places emphasis onthe range of functionality versus function for a givenapplication/use. Consider a liquid/liquid transfer station,currently validated and used to aspirate/dispense 2 ml. Ageneric version of this application would be validated foraspirating/dispensing operations with volumes ranging

from 0"5 to 10 ml, as opposed to the fixed volume of 2 ml.When a new sample process is required, any volumebetween 0"5 and 10 ml could be implemented withoutrevalidating the functioning of the liquid/liquid transferstation.

On a broader scale, the entire dictionary can be designedwith this generic focus. The various functions the XPsystem performs can be broken down into functional units.Functional units are similar to laboratory operation units(LUO), although the emphasis for functional units is onthe range of functionality for a given operation versusrepetition of that operation.

This presentation outlined the concepts and the authors’experiences in implementing the generic dictionary on an

existing XP system.

Systematic approach to automation validation

W. J. Ewing and P. J. GallantDuPont Merck Pharmaceutical Company, RadiopharmaceuticalsDivision, Billerica, MA, USA

The trend in today’s laboratories is movement towardautomation. This advancement is accompanied by theneed to validate the automated system. This process canbe very long and time-consuming; however, with theproper knowledge and planning, the validation processcan be short, painless and very successful. Initially, theidea about an automation project is conceived, and thenthe research and investigation begins to find the perfectsystem and design to suit specific needs. A rough layoutor system plan should be generated to ensure that thesystem obtained can perform to the expectations. Thefunctional requirements document should be written inwhich the system will be described in general and theoperational requirements of the system will be stated. Asystem specification document is also required which ismade up of hardware and software design specificationof the system.

A validation plan needs to be established that will includethe documentation necessary to prove that the systemperforms its functions properly and is consistent with thefunctional requirements and system specifications. Thevalidation plan also identifies all the required tasks andresponsibilities needed to have a successful validatedsystem.

The actual validation testing can be broken into threeparts: installation qualification; operational qualification;and performance qualification. The installation qualifi-cation (IQ) test is performed to ensure that all of theequipment is installed according to the manufacturers’specifications and recommendations. The operationalqualification (OQ) tests that the system components eachperform correctly within their operation ranges. Theperformance qualification (PQ) provides proof that thesystem performs all its functions accurately, reliably andin accordance with the functional requirements.

This presentation described the steps involved in validatingan automated system and maintaining that system incompliance once it has been validated.

4O


A generalized performance based approach to theoperation qualification of the Zymark BenchMate/TPW

David A. Hollowell, Mark N. Flair, Jeffrey D. Hofer,and Michele E. LakeEli Lilly and Company, Lilly Research Laboratories, Indianapolis,IN, USA

A generalized approach to the operational qualificationof a Zymark BenchMate/TPW has been defined andexecuted in the Pharmaceutical Analytical DevelopmentDivision at Eli Lilly and Company. The operationalqualification (OQ] .is intended to produce documentedevidence providing a high degree of assurance that theautomated system is capable of reliably performing thedesired analytical functions in the laboratory. Usingvendor specifications and statistical techniques, rationallimits for automated operations have been derived fromNIST Class A specifications for the glassware-equivalentmanual laboratory operations. This approach was under-taken in order to be able to confidently utilize the equip-ment for development of analytical methods as well as

to validate the automation of manual procedures. Thispresentation gave the approach, results, and conclusionsof this operational qualification.

DRUG DISCOVERY RESEARCHThe automation of radioisotopic and luminescenceassays for high throughput screening

Alfred J. KolbPackard Instrument Company, Meriden, CT, USA

The number ofsamples being analysed in high throughputscreening has been growing at an acclerating rate overthe last five years. To keep up with this demand,pharmaceutical companies have been drawing uponinnovations in analytical instruments, microplates, assaytechnology and robotics. The core instruments in most

screening laboratories include liquid handling systems andan array of analytical instruments such as densitometers,luminometers and microplate counters for radioisotopicmeasurements. These instruments can handle a sub-stantial sample throughput when combined with micro-plates and assay technologies that reduce, or eliminate,sample processing steps.

Microplates with integral filters have eliminated the needto handle individual filters for the analysis of receptorbinding or cell proliferation. The harvesting and analysisofsamples occurs in the same microplate, thereby reducingsample preparation time. In-plate assays for immuno-binding, receptor binding and nucleic acid hydribidizationhave been designed that require only a simple aspirateand wash step. Other assay systems completely eliminatethe need to separate bound from free label. Many of theseassays are available using radioisotopic labels or newluminescent reagents with half-lives of hours instead ofminutes. This allows the researcher to choose from a rangeof methods that best suit the individual laboratoryrequirements.

The simplification or elimination of separation steps hasalso simplified the complete robotics automation of

screening assays. Filtration assays for receptor bindingcould not be automated when these assays required theuse offilter disks or sheets offilter paper. The developmentofmicrofiltration plates and non-separation assays offereda practical solution for robotics automation.

The co-operation of Packard Instrument Company andZymark is an example of how companies can combinetheir strengths to offer solutions to the automation of highthroughput screening.

Automated template preparation for high through-put sequencing with BioRobot 9600

Michael CollasiusQIAGEN GmbH, Hilden, Germanyand Alex ZrolkaQIAGEN Inc., Chatsworth, CA, USA

The BioRobot 9600 has been designed specifically for highthroughput production ofultrapure sequencing templates.Templates can be prepared using either the QIAwellUltra or QIAprep M 13 Systems. The QIAGEN kits havebeen optimized in our research facilities for use with theBioRobot 9600. The BioRobot 9600 can meet the through-put requirements of any type of laboratory. Dependingon the type, source and quantity ofDNA templates beingpurified, the workstation can deliver up to 96 templatesin as little as 2 hours.

The BioRobot 9600 hardware can be easily adapted to

meet changing research requirements. The modularplatform allows the option to upgrade as necessary, or asnew technology becomes available. The workstationcombines the novel QIAGEN operating system with theWindows user interface to provide eash set-up andexecution. The point and click operation can be used to

modify existing protocols or even create new applications.

The BioRobot 9600 provides the reliability of provenpurification chemistry with the high throughput require-ments of modern laboratories.

An automated system for preparation of radio-labeled antibody

Jimmy Brunet, Julie Stimmel and Doug WilsonGlaxo Wellcome Inc., Research Triangle Park, NC, USA

A laboratory robot has been developed to perform theradioligand labeling of clinical quantities of a monoclonalantibody. The system was developed in response to a

problem of high radiation exposure to staff using manualmanipulations. A small Zymate XP robotic system wasassembled to perform radiolabel preparation, radioligandbinding and cleanup of the radiolabeled antibody usinga spin, size-exclusion column.

A new 0"1-10 gl Eppendorfpipette hand was implementedoptimized and characterized. The 0"1-10 gl Eppendorfhand (fabricated by Zymark) uses the variable plungermechanism of a standard Zymark pipette hand. Customsoftware was developed to allow variable volume dispensingfrom the Eppendorf with a linear calibration adjustmentmade in the software. Accuracy and precision of 5% and

41


+_59/o were achieved at a lal nominal volume withaccuracy and precision of 2% and 4-1% at 10 gl volume.

The radiation exposure of scientists was reduced signifi-cantly, allowing for more frequent and safer experiments.The robotic method improved reproducibility and overallrecovery of radiolabeled antibody versus the manualmethod.

Motion control of an X-Y stage and autofocus fora microscope using a commercially availablevision and instrument control package

Joe J. Yacobucci, Jeff Guss and Derek HookBristol-Myers Squibb, Biomolecular Research, Wallingford, CT,USA

Many biological responses are accompanied by a shapeor fluorescent change. Q.uantitating these changes by eyeusing a microscope or a television monitor is tedious andprone to error with the added difficulty of controlling foroperator-to-operator variation. The authors report on theapplication ofan X-Y stage and autofocus for a microscopeusing Ludl motion hardware, and software.

Topics discussed included automated X-Y stages, auto-focussing, computer logic commands, image prameters,and the use of a Zymark robot to facilitate unattendedoperation.

This system has processed unattended over 100000microplate wells.

steps. Once this intermediate processing is completed, theoriginal source container is typically returned to itsoriginal location.

Various strategies exist for automating the storage/retrievaland preparation processes. Approaches include attemptingto automate the complete procedure to selectivelyautomating steps in the process. Where full automationhas the attraction of reduced manpower requirements,selective automation will normally increase the likelihoodof success and potentially results in optimum utilizationof manpower with maximum return on investment.

CUSTOM AUTOMATIONCustom automation of polyurethane materialstesting using visual programming tools

Steven E. Robbins and Michael E. RusakAir Products and Chemicals, Inc., Allentown, PA, USA

Polyurethane foam physical test measurements are funda-mental to characterizing end product uses and determineeconomic feasibility. Visual programming tools were usedas the basis for integration of diverse hardware andsoftware necessary in this custom application development.The visual environment improves system operations andminimizes the need for extensive training. This presen-tation described some of the technical approaches used inautomation of data acquisition, instrument and robotcontrol, and system management.

Automated compound storage and retrieval sys-tems-providing a backbone for high throughputscreening programs

Scott C. Atkin and R. Bruce JamiesonSAGIAN, Inc., Indianapolis, IN, USA

As High Throughput Screening technologies are imple-mented in pharmaceutical and agricultural compounddiscovery processes, the rate limiting step in successfulscreening programs becomes the ability to handle the’upstream’preparation of potential compounds. Specific-ally, as HTS processes become robust within an organiz-ation and truly high throughput there comes an everincreasing demand for more novel compounds to screenwith improved access to existing libraries. Combinatorialchemistry and acquisition programs can accelerate theidentification of new compounds. However, withoutincreasing manpower, the preparation ofthese compoundscan only be accomplished with automated systems.

Automated Storage and Retireval Systems (ASRS) allowfor the rapid retrieval of existing compounds for use inHTS programs. A common requirement for manycompound management systems is the retrieval of anarchive compound from a central storage facility. Subse-quent steps may include transferring a sub-sample toanother container for distribution to an internal HTSlaboratory, transfer and dissolution tbr shipment and/orshort-term storage, creation of’daughter’ microplates forimmediate use, storage, or shipment, or other processing

Integration oflaboratory automation for the HumanGenome Project

William Lee, Eric S. Lander, Trevor L. HawkinsWhitehead Institute/MIT, Center for Genome Research, Cam-bridge, MA, USAand Glynn SearlCRS Robotics Corporation, Burlington, Ontario, Canada

There is a major need for the development of a systemwhich can accomplish the integrated tasks of DNAisolation and proceed with purification and the set-up ofsequencing reactions. The authors demonstrated thefeasibility of such a system from both a biochemical andengineering perspective. The authors are collaboratingwith CRS Robotics Corp., Packard Instruments, TecanUS and Techne Include to design and construct afactory-style laboratory system. The major component ofthe system is an articulate CRS 255/A robotic arm whichis track mounted. The deck of the robot contains severalnew/modified XYZ robotic workstations, a novel thermalcycler with automated headed lids, carousels and custombuilt plate feeders.

Biochemically, the authors employed their solid-phasereversible immobilization (SPRI) technique to isolate andmanipulate the DNA throughout the process. The systemis flexible and can be modified relatively easily. In thisway, one system can be used to accomplish many differentbiochemical tasks.

42


An integrated system for automated radio iodina-tion

Andy Chang, Greg Bennett and Randy YenBioAnalytical Technology Department, Genentech, Inc., South SanFrancisco, CA, USA

An automated robotics system was developed to performprotein-labeling experiments involving radioisotopes. Thereason for integrating this system is to allow people tominimize exposure to radiation, not for high throughputnor for running experiments at a high volume. The systemconsists of a m HP ORCA arm and four syringe pumps.It is small enough to reside behind appropriate shieldinginside a chemical hood from which a charcoal filter trapsvolatile radioactive molecules. A Visual Basic softwareprogram was written to control the robot and to providea user interface for scientists or technicians. The user caneasily change parameters and/or create new methodswithout having to know programming for the robot. Thissystem addresses safety concerns by reducing humanexposure to high levels of radioactivity, offers improvedreproducibility and reduces manpower needs.

The robotic automation of the determination ofdoses from metered dose inhalers under a widerange of simulated patient usage conditions

Andrew MonkDepartment of Robotic Automation, Thurnall PLC, Manchester,UK

This system described is an automated system proyidingan repeatable method for handling and firing MeteredDose Inhalers (MDIs) into collection apparatus, measuringloss and determining dosage. The primary elements of thesystem are: a gantry robot system, shaker/stirrer mechanism,analytical balance, dose collection facility with associatedwashdown unit, and an autosampler to provide samplesfor HPLC analysis. The system provides a cost effectiveand flexible alternative to manual testing.

Custom engineered automation for scientific re-search

D. D. McCampbell, M. F. Fischer and C. E. BallMidwest Research Institute, Kansas City, MO, USA

Midwest Research Institute (MRI) has been providingcontract scientific research for over 50 years, in fields suchas energy, environmental, health, and transportation. Theautomation group is a multidisciplinary group with back-grounds in mechanical engineering, engineering design,artificial intelligence, computer programming, machinevision, analytical chemistry, and biology.

MRI’s specialists have developed robotic systems toincrease the throughput and precision of such tasks asnatural pesticide screening, oncogene inhibitor screening,biological matrix extraction, and food product analysis.Some of the custom systems developed by the automationgroup at MRI were discussed in this presentation.

CHEMICAL ANALYSISIntrinsic viscosity measurement using a laboratoryrobot

Philip J. FarrellyHudson Control Group, Springfield, NJ, USA

The presentation described the design and developmentof a robotic system to fully automate the measurement ofintrinsic viscosity of polyester materials for a major petro-chemical manufacturer. The topics covered included:system design overview; identification of technical chal-lenges; required innovations in hardware and software;software development; operational methods; and futureexpansion.

The design, construction, testing and implementa-tion of an automated robotics system

Juan C. Cadavid and Marie SaboClairol, Inc., Stamford, CT, USA

Keeping in line with today’s requirements, the QualityAssurance group at Clairol was faced with the task ofdoing more with less; i.e. they were confronted with thechallenge of making more efficient use of their time. Aftera careful evaluation of possible alternatives, an automatedrobotics system was selected to perform the most timeintensive analyses in QA’s daily workload as well as theones with the most sample throughput. These analyseswere identified to be the determinations of % totalalkalinity, % fatty acid, and Brookfield viscosity.

Both the total alkalinity and the fatty acid analyseswere implemented using existing analytical autotitratormethods. The existing Brookfield viscosity technique (ona dye/developer mixture) was significantly modified dueto physical constraints in automating the method.

The robotics system is capable of handling 130 samplesper day with 10 possible combinations of analysis. Thepossible combinations are determined via bar codescanner from a preprinted list of bar code labels for eachsample. This information is sent to the top level userinterface program Visual BasicTM, an object-orientedprogramming language which operates from the easy-to-use WindowsTM platform. The intelligence and optionalhelp features incorporated into the interface permitoperators to obtain results with minimum supervision.

A description of the automated system was presented,along with the requirements posed to the system, thehardware and software utilized, and the system validationschemes.

ADVANCED TOPICSAn approach to use low-budget-imaging to automizethe detection of liquid-liquid interfaces in labeledtest-tubes

Manfred Jungke, Marius Voehringer, KarlfriedDoerr, Christian Lehmann and Jens LuettgeFachhochschule, Frankfurt am Main, GermanyThe main objective of this work was to investigate whetherlow budget hardware components, combined with a

43


standard PC, could be used to solve a complex task ofliquid-liquid extraction in labeled test-tubes. The goalwas to detect the separation layer of two transparentcolourless liquids, with different refraction coefficients.The tubes may have white labels or light colour labels.The labels may cover up to half of the circumference ofthe test-tube, they may be machine printed or manuallywritten. There is no need to predetermine the position ofthe label. The method was tested by using an example ofplain water and sunflower oil that had been carefullyadded before.

One major task was to set up a solution with low costcommercial consumer components such as a black andwhite camera module, and a consumer PC-video-digitizer,which was used as a frame-grabber. The components weresuccessfully mounted on a Zymate py-plate and wereintegrated in the automation process under control of a

Zymate II System. The method was developed by trainingelectrical engineers in automation tasks in an educationalenvironment.

This work was supported as a research project by theHessian Ministry of Science and Art.

Evolution of machine vision enabled automation:from the factory floor to the laboratory bench

James H. Beyer, Michael E. Dobbs, Paul C. OlsztynVision Instruments, Inc., Ann Arbor, MI, USAand Julie ErbProctor & Gamble Company, Cincinnati, OH, USA

Historically, the application of machine vision to auto-mation problems has been an expensive, one-of-a-kindeffort, requiring special purpose hardware and highlyskilled engineers to support an application. The high costofdeveloping and maintaining a vision enabled automationapplication restricted its deployment to large volume/highmargin manufacturing tasks, such as welding in the auto-motive industry and circuit board soldering in thecomputer industry. The increasing computational powerof commercial personal computers, coupled with theirdecreasing cost, has enabled a new machine visionparadigm. PC hosted image processing software can nowbe combined with robotics and other automation com-ponents for a fraction of the cost of systems created justa decade ago. This decreased cost enables machine visionto move from large scale factory automation, to smallerscale laboratory automation. Vision Instruments Inc. hasintroduced a series ofvision enabled products which allowthe automation of laboratory processes previously notamenable to automation effort. This presentation examinedthe evolution ofimage processing techniques from its DODroots to its application in the modern laboratory. It alsoshowed the implications of this evolution for the auto-mation of laboratory procedures used in high throughputscreening and microbiological counting and classificationtechniques.

A modern robotic automation technology formaterial handling applications

Karl Sharicz, Ed Fournier, David Feindel and MarcO’MaraAsyst Automation Inc., Wilmington, MA, USA

Like most manufacturing environments today, the semi-conductor industry faces many of the same challenges asthose involved with pharmaceutical drug discovery anddevelopment. The common denominator in both industriesis that traditional methods are no longer sufficient insatisfying the demands for increased yields, reduced costs,and accelerated time-to-market. Comprehensive solutionsthat essentially redefine the work process and helporganizations remain competitive into the next centuryare required. At the same time, these solutions must offera total systems approach that allows organizations toretrofit and extend the usefulness of existing facilities, aswell as build new cost-effective facilities. In essence, thesesolutions must provide integrated strategies with afinancial and technological pay-off.

Automation, through the use of robotics technology, hasmade a significant contribution in process optimizationand has led organizations toward realizing the goalsprescribed by these new demands. For the semiconductormanufacturer, the evolution of more advanced chipdesigns, larger wafer diameters, safer and more reliablematerial handling, and needs for reduced particulatecontamination have helped create increased opportunitiesto apply robotic automation.

Since 1981, Asyst Automation engineers have served theneeds for the semiconductor manufacturer by pioneeringthe field of contamination-free robotic material handlingsystems. These systems enable manufacturers to safelystore, retrieve and transport material during the manu-

facturing process. Materials include bare silicon wafers,wafer containers, bare reticles, and reticle containers. Bydesigning the system to work within its own ’mini-cleanroom’ environment, Asyst Automation has helpedsemiconductor manufacturers reduce space requirementsand recover valuable cleanroom floor space.

This presentation, through graphic illustration andvideotape, showed robotic material handling in actionand demonstrated how the material storage, retrieval,transport, and management process applies in a varietyof configurations and applications. These systems providedata links to other aspects of the manufacturing process;the technology is easily learnt and mastered by productionpersonnel through a graphical user interface provided bythe OS/2 Presentation Manager control software.

MANAGING LABORATORY AUTOMATIONA team approach to the transfer of robotics to a

QC environment

j. R. Tricome, N. Muhammad, S. W. Swieck andJ. j. RiceBristol-Myers Squibb Company, Syracuse, NY, USA

In today’s competitive environment in the pharmaceuticalindustry and with increased emphasis on productivity and

44


efficiency, a team approach is essential to successful imple-mentation of a robotic system in a QC environment. AQC initiative was undertaken at Bristol-Myers Squibb inSyracuse to implement a robotic system to analysePenicillin fermentation whole broths on a 24-hour basis.This team effort has resulted in the introduction ofroboticsto the QC laboratories and a smooth transition to a fullyautomated analysis technique.

Approaches to automation for pharmaceuticalanalysis

Alan WickmanG. D. Searle, Skokie, IL, USA

Automation has been an ongoing endeavour within G. D.Searle’s analytical group for the past 12 years. Initialsuccesses were with automation ofdissolution testing. Thiswas followed by not so successful attempts at automatingother sample analysis techniques. With much hard work,lessons learned, and persistence, all major analytical testshave been automated. While automation has meant thatanalysis capacity has been increased, it has also presentednew problems and challenges.

This paper discussed what has been automated at G. D.Searle, what works best and why, the problems, thechallenges and a view on what the next five years willbring.

Regrouping, reforming and re-engineering: applyingrobotics to new challenges

Stephen Scypinski, Theodore Sadlowski andJohn BaianoAnalytical Research and Development, Hoffmann-La Roche, Inc.,Nutley, NJ, USA

At the ISLAR meeting over the past several years, manymanagement session and plenary talks have dealt withthe reality of’ doing much more with less.., faster!’ Mostrecently, Dr Phillip Lane of R. W. Johnson PRI spoke of’The reality of the 90s’. The number of mergers,acquisitions and consolidations in our industry hasintensified. All of these actions always result in more workfor less people. Hammer and Champy’s 1993 bookRe-engineering the Corporation spelt out what will becomereality for a company of the 90s and into the next centuryin that companies will surely improve the way we, asAmerican corporations, conduct business. Most inter-estingly, re-engineering relies heavily on informationtechnology and automation, which is not too surprising.Indeed, many’of the practices spelled out in the casestudies documented in their book could not have evenbeen thought about 15 to 20 years ago.

If one extrapolates the re-engineering fever to the field ofanalytical chemistry, the use of automation will con-comitantly increase over the next several years as well.Such an attitude will require a fresh look at projects andsituations that can be automated. The ability oflaboratorypersonnel to ’think out of the box’ will dictate their riseor fall as successful analysts.

Some of the newest challenges at Roche were discussedin this presentation.

Laboratory automation: a critical tool in com-

petitive, regulated industries

Lane GehrleinSchein Pharmaceuticals, Carmel, NY, USA

Multisource pharmaceutical companies face increasingdemands for fast and efficient research and developmentprogrammes. Large domestic pharmaceutical companiesare developing generic drug programmes and foreignpharmaceutical companies are also entering into thegeneric drug market. In order to stay competitive,companies must increase the number of new productintroductions, since the market share for each product isdivided up among an increasing number of competitors.

Laboratory automation has played an important role inDanbury Pharmacal’s ability to develop an increasingnumber of new generic pharmaceutical products withminimal increases in R&D personnel. Laboratory auto-mation has also been very effective for reducing the impactof increasing government regulatory requirements on

pharmaceutical R&D programs. The laboratories employautomation in eight areas which have demonstrated a

positive impact on productivity: cleaning validation-sample testing; process validation-sample testing; productassay and content uniformity; dissolution; data acquisitionnetwork; LIMS; method validation; and raw materialrelease.

Automation in a highly regulated industry can be veryproductive when properly implemented and validated.Regulation can be equated with consistency which isexactly what is obtained from automation. All ofthe aboveautomated procedures have been designed to deliver datawhich has been validated and reports which directlyreference that data. No additional manipulation of dataor reformatting of results is needed. Auditing of thesereports is easy and fast, because the auditor knows exactlywhat to look for and where to find it. Automation willbe a major contributor towards staying competitive intoday’s highly regulated industries.

Digging out with a robot

Milton LevenbergAbbott Laboratories, Abbot Park, IL, USA

As in every other pharmaceutical company, the chemist/sat Abbott Laboratories are totally dependent on a rapidreturn of spectroscopic data on their intermediates andsynthetic products to ensure that their chemical reactionsare on track. Eight years ago the chemists at Abbott oftenhad to wait three days to a week to receive this data, andhad to start new reactions without this data, hoping theprevious step performed as expected. That this was an

unacceptable situation was clearly recognized by theAbbott chemists and the management of the spectroscopyarea.

45


This presentation discussed the general approach used toanalyse Abbott Laboratories’ needs and to provide totallyautomated sample preparation and running ofboth NMRand mass spectrometric samples. Systems by which thechemists log their own samples into our database havebeen developed. A Zymark robotics system does the entiresample preparation and insertion into the spectrometer.NMR spectra are plotted directly in the client’s lab. Thespectroscopy staff need only provide samples to the robot,fresh supplies (solvents, tubes etc.) and routine main-tenance, but no actual sample handling or spectrometeroperation. The effects of this automation on throughput,sample turnaround time, and cost per spectrum werepresented.

Getting smart with automation

Richard KramerInstrumentation Development Group, Bristol-Myers SquibbPharmaceutical Research Institute, Princeton, NJ, USA

Laboratory productivity and quality issues have givenway to business issues as the driving force behindautomation and in pharmaceutical R&D in recent years.This shift brings with it unprecedented opportunity forautomation where it can be related to meeting businessobjectives. Unfortunately, not all of the driving businessissues are pretty: the budget restrictions, staffing limit-ations, and organizational changes that we live with todayrequire that we thoroughly understand the role automationwill play in the new climate. We have found that we arenot merely charged with engineering the physical auto-mation in the laboratories, but also engineering thestrategy for the evolution of automation. We had to getsmart.

In the author’s R&D environment, getting smart wasachieved through two distinct methods, which werediscussed in depth: getting smart about automation, andgetting smart from automation. The presentation con-sidered the relationship of risk management strategies,and how education, investment and partnerships areingredients ofmanaging automation growth. The chemist’srole as in-house integrator was examined in terms ofthe multitude of hats we have to wear. Also the lessonslearned from implementing automation were discussed.

Additionally, goals for automation technology need to betempered with realistic expectations. This presentationfocused on these issues and provided some operationalsolutions for organizations facing many of these same

problems.

Economic justification of automation systems inthe Dow Chemical Company

Jonathan Zieman, Paul Morabito and RamasamyTamilarasanThe Dow Chemical Company, Midland, MI, USA

Numerous robotic automation systems have been devel-oped and implemented within the Dow Chemical Com-pany. These systems have found homes in productionquality control laboratories, analytical laboratories, andR&D departments. The complexity of procedures auto-mated within the company has varied, ranging fromsample preparation for biological tissue analysis to

polymer sample hot dissolution to polymer physicaltesting. Automation capabilities are currently availablethat can reliably perform most of the common laboratoryprocedures such as weighing, dilution, filtration, vialmanipulations, liquid handling, incubation, and inter-facing to instruments etc. The developed capabilitiesalong with the new emerging technologies have enor-mous automation potential in many areas within thecompany.

As with all capital investments, economic justificationmust be evaluated for the automation impact and returnon investment (ROI). It can be difficult to identify and/orquantify all the major variables needed to adequatelydetermine the automation impact/ROI, especially for thefirst-time user. Lack of suitable information could resultin not funding a project that would save the companymoney and increase productivity. ROI data acquiredfrom several automation systems in operation over thepast couple of years was used to more formally addressfuture justifications. The issues from these systems werepresented to help identify topics to consider if automationplans are in your future.

VALIDATING LABORATORY AUTOMATIONEvaluating the reliability of software

Laboratory automation of the 1990s: goals, expect-ations and business needs

W. Jeffrey Hurst and Robert A. Martin Jr.Hershey Foods Technical Center, Hershey, PA, USA

The laboratory of the 1990s is not immune to the trendsfacing American industry. For many years, laboratoryprofessionals have felt that they could somehow be isolatedfrom the realities of business. Laboratory automation wastargeted as a panacea for a multitude ofissues dealing withlaboratory productivity. There has been a shift inindustrial research and development, requiring labor-atories to refocus their efforts and continuously evaluatethe many facets of laboratory operations includingfinancial objectives along with laboratory automation.

Charles A. SnipesFDA, Rockville, MD, USA

Current computer industry standards for software requirea written validation plan that will include the followingelements:

(1) Design specifications (exactly what the program isintended to do and exactly how it is intended to doit, detailed, both high level and low level, withpredetermined criteria for acceptance of the program,and descriptions of: hardware to be used, algorithms,file structure, limits and parameters to be measured,error and alarm messages, configuration, communi-cation links among subprograms and to equipmentand to other systems, and security measures).

(2) Risk/hazard analysis.

46


(3) Testing plans: Developmental, including structuralanalysis of all individual decision points (in themodules before integration), and testing of worst caseconditions at operational limits, of alarms, of errorroutines, and ofexecutable statements. Installation, forinstallation testing of the software within the environ-ment where it has been designed to function.

(4) Evaluation of test results, particularly as to how theydemonstrate that the design specifications have beenmet.

(5) Change control/revalidation procedures.(6) Documentation about who is responsible for approving

and for executing the validation plan.(7) Archiving the whole of the above, including in

particular all versions of the design specifications andin particular specific test results, rather than pass/fail.

(8) Especially important are the design specifications, asthese are the heart of a validation.

Written development standards (for overall development)and written programming standards (for the work ofindividual programmers) are also of importance.

Development and validation of an automated drugcontent and degradation profile analysis methodfor pharmaceutical tablets

John R. StanleyPharmaceutical Technologies, SmithKline Beecham Pharmaceut-icals, Hertfordshire, UK

An automated bulk tablet drug content assay applicationfor the HPLC analysis of pharmaceutical tablets has beendeveloped and validated using a Zymark BenchMateTablet Processing Workstation (TPW) with EasyFillsample collection module (EZ). The first part of theapplication consisted ofthe development and optimizationof a Zymark TPW method for the homogenization oftablets and the extraction of the drug component ofinterest from the tablet matrix.

The second part of the application was the developmentof a protocol to show equivalence between the automatedmethod and the pre-existing manual method for theextraction of drug for both bulk content drug assay anddegradation profile analysis. This protocol included anassessment ofTPW processed samples that were transferredto vials by the EZ for subsequent off-line HPLC analysis.The third part of the application was performancequalification validation for the equivalence protocol andinterpretation of the data generated. Advantages anddisadvantages of the approaches undertaken for thedevelopment and validation of the automated procedurewere discussed.

Development and validation of a Zymate PyTech-nology potency robot equipped with a tablet pro-cessing workstation and on-line HPLC

Daniel W. BarrowBristol-Myers Squibb, New Brunswick, NJ, USA

A potency and degradant analysis robot has been success-fully developed and validated for use with tablet and

whole encapsulated dosage forms. The system is based ona Zymate PyTechnology robot equipped with a TabletProcessing Workstation PySection and on-line HPLC.Active drug ingredients and degradants are extractedfrom solid dosage forms using a high speed homogenizer.Filtration is performed by a custom filtration PySectionthat uses standard membrane filters and dispenses filtratedirectly into the target test tube. Multiple dilutions are

possible using capped test-tubes. The high capacity, 50 ml,test-tubes allow accurate dilutions and vigorous vortex

mixing, virtually eliminating evaporation through the useof a robotic capping PySection. Final working solutionsare automatically injected onto the on-line HPLC. Totalsample capacity is increased through the use of a customwaste diversion valve which separates hazardous extractionwaste from aqueous wash waste. Productivity gains of thisrobotic method of analysis are approximately 100compared to manual methods of analysis.

This robotic system has been successfully used in theanalysis of whole encapsulated dosage entities. Thepresentation described system schematics and robotic/methodology validation data including accuracy andprecision.

The validation of a Zymark Batch DissolutionSystem for QC and R&D analytical laboratory

Muhammad Alburakeh, Charles DiLiberti andMattio CitardiBarr Laboratories, Pomona, NY, USA

After considerable effort by the Batch Dissolution ProjectTeam, the Zymark PyTechnology-Batch DissolutionRobotics Systems have been validated for use in theauthors’ laboratory as a fully automated dissolutiontesting system. They support samples provided by theQC and R&D areas, as well as process validationsamples. Each system was successfully validated with USPapparatus I & II to perform on-line UV analysis, off-lineHPLC vial filling and storage, single and multi samplepoint profiles, sample weighing and media exchange.These systems were originally installed as serial systemsusing telescoping shafts, filter tips and gravimetric mediadispensing. The upgraded Batch Dissolution Systems are

using volumetric media dispensing, standard 0"45 micronmembrane filters and standard USP dissolution shafts.Some of the authors’ experiences, as well as frustrations,and most importantly, successes in the transfer, pre-validation and validation of the robotics systems were

presented.

DRUG DISCOVERYAn integrated and versatile system to rapidlydetermine potency, selectivity and potential forside-effects: from assay automation and datamanagement to result

j. Elands, C. Widmaier and M. GalvanMarion Merrell Dow Research Institute, Strasbourg, France

In a continuous effort to optimize early drug discoverythe authors set out to automate the following tasks:screening new compounds against a series of receptors,

47


uptake sites and enzymes, to establish selectivity andside-effect potential; development ofHTS assays; secondaryscreening of HTS hits.

Task is run in batch mode: a single receptor/uptakesite per microtitre plate was used. Task 2 requires a

temperature/CO2 controlled incubator, a fluorescenceand UV/Vis plate reader and a luminometer--differentassay conditions are evaluated automatically and assaysare rendered compatible with a fully automated system.Task 3 involves the evaluation of libraries of analoguesaround a hit on the original assay and a series of assaysselected to assess selectivity for the target; assays of thefirst task are also used in this stage.

Assays could not be run during long times. So a flexiblerobotic system with an easy programmable GUI-basedScheduler was needed. Details of the Zymark roboticsystem used and examples ofscheduled assays were shown.Assays are continuously adapted and new assays developedfor every HTS target. The end users need a flexible datamanagement package with powerful protocol managementincorporated. Compatibility with existing corporate data-bases is essential. ActivityBase from idBS was found tohave nearly all these features included, idBS has provento be very flexible by adding those features that wereessential to our company. The architecture ofActivityBasewas also discussed.

A multi-functional microplate workstation as a

strategy for changing priorities

William S. Fillers and Diana K. CohenSandoz Research Institute, Sandoz Pharmaceuticals Corporation,East Hanover, NJ, USA

Shorter project lifetimes, changes in personnel and rapidlyshifting priorities require a flexible approach to highcapacity screening (HCS). A compact microplate roboticsplatform capable of operating concurrent luminescentand spectrophotometric assays was described. The oper-ation schedule of the platform is determined by the abilityto generate plates for cell based, reporter gene assays(RGA), while remaining workstation time is filled wthassays which are less constrained by supply dynamics.Sample preparation efficiency gains and conservation ofthe compound library are made possible by sharingsample dilution plates. Sustained output of 7500 samplesper day creates the opportunity for batch or concurrent

processing of primary and secondary screens. The cost

advantage ofrapid cycle time allows consideration ofmorehigh risk opportunities. Concurrent operation of multiplescreens on a single HCS platform is an effective strategyfor rapidly generating large amounts of data for decisionmaking on competing approaches that require scarceproject resources.

Automating more for less

M. R. KozlowskiDepartment of Screening, Geron Corporation, Menlo Park, CA,USA

A goal oflaboratory automation is to replace human effortwith robotic labour in the performance of predictable,

repetitive laboratory operations. In an ideal world, thisapproach frees up the human element to perform more

sophisticated and rewarding tasks. This goal is onlyrealized, however, when the automation of the task takesless resources over the life of the project than simplyperforming the task manually. Resources include suchthings as human time, financial outlay (which can beconverted to human time via cost per FTE), anddown-time (which can be converted to financial outlayvia ’burn rate’). Resource-intensive aspects ofautomatinga task include training the robot, designing and testing(or purchasing) special robot-operated equipment, andmanaging the robotic system (for example, fixing ’crashes’).Automation of certain tasks is often not undertakenwhen the apparent resource requirement becomes too

great.

This presentation discussed ways to make more tasksamenable to automation by decreasing the resource

output required to perform them with a robot. Two keyelements were focused upon: first, decreasing the need forspecially designed equipment, and the resultant expenseand delays. Many tasks can be performed by a robot usingstandard, or slightly modified, basic laboratory equip-ment. The second focus is decreasing resources lostthrough robot failure. Simple robot ’babysitting’ tech-niques were also presented.

The modular application of automation to highthroughput screening

j. v. Petersen, Jason Armstrong, Steve Hamiltonand Rick StantonAmgen Incorporated, Thousand Oaks, CA, USA

The Research Automation Technologies group at Amgenis charged with development of a coherent and responsiveautomation strategy to accelerate the drug discoveryprocess. Success is measured by the timely application ofthe appropriate technologies to research and developmentapplications. Due to the diversity and quick turnover inresearch projects, the hardware and software needs to bemodular in design to facilitate the ease of development,integration and maintenance.

The cornerstone of the High Throughput Screeningindustry is the microtiter plate. A list of basic modularoperations for an assay are barcoding, liquid handling,transporting, washing, reading and storage. Common toall of these modules are sample interaction, data inter-change, external control and internal status. A variety ofcommercial devices and software provide a core function-ality that requires further customization and enhancementto provide a fully integrated robotic system. Examples ofseveral Amgen automated screening effors were reviewedto illustrate how this module approach can assist in thediscovery of new therapeutic drugs.

48


DISSOLUTION TESTING/VALIDATIONEvolution ofa rugged automated dissolution systemwith high sample throughput

Richard Von CulinAnalytical Research & Development, Bristol-Myers SquibbPharmaceutical Research Institute, New Brunswick, NJ, USA

Automated dissolution testing has become a pharma-ceutical industry standard with companies restructuringand trying to do more with less. Automated dissolutionhas been very successful for the robotics laboratory inAnanalytical R&D, New Brunswick. An on-line UVspectrophotometry robot has processed approximately35 000 samples since 1989 and has accomplished uninter-rupted dissolution runs lasting more than 50 hours. Theevolution of a highly productive and rugged on-linedissolution system can be summarized in three maindevelopmental stages: system flexibility, system relability,and efficient data management. System flexibility wasobtained by programming the robot to run multipledissolution methods. The system can run capsules, tablets,different sample potencies, and different flow cell sizesduring a single dissolution run. The system has proven tobe very reliable over the past six years. The only serviceperformed routinely is preventative maintenance so thesystem rarely fails to complete a dissolution run. Finally,to further increase sample throughout, data managementfunctions were automated. Custom software was writtento electronically input all sample information into theSystem V controller and to lead the final dissolution resultsinto the stability database. The evolution of this on-lineUV dissolution robot into a rugged and productive systemwas presented.

The fibre optic method development was described,together with the testing used to validate this system. Thedissolution throughput has increased considerably forsamples with interfering matrix. The fibre optic endanalysis has provided additional savings to the currenteconomical automated dissolution methods. This systemminimized the handling ofexperimental samples, providingan added measure of safety to laboratory personnel whenhandling very potent or sensitive materials.

Validation of vendor-supplied systems

Bruce Fowler, P. Michael Masterson and ClarenceKemperKemper-Masterson, Inc., Belmont, MA, USA

The purpose of this presentation was to describe somecurrent issues and concepts relative to vendor-suppliedsystems, such as LIMS, laboratory instrumentation, orrobotics. The presentation incorporated practical docu-mentation techniques for optimizing qualification andvalidation of vendor-supplied systems. The presentationfocused on the validation premise that user firms canreduce the amount of effort and expense of qualificationand validation if they rely on vendor-supplied data andinformation to support their systems-related activities.

The conclusions emphasized that vendor and users shouldwork more closely together to minimize the cost and effortrequired to qualify and validate systems used for GLPand GMP functions.

Establishing validation standards

Automated dissolution testing utilizing on-linefibre optic probe UV analysis

Leonard J. Kostek, Paul K. Aldridge, David W.Melvin, Brenda A. Williams, Karl Bratin andSonja SekulicAnalytical Research and Development, Pfizer Central Research,Groton, CT, USA

Automated dissolution testing has become a commonpractice in the pharmaceutical industry. Generally, theend analysis for the dissolution test has been an ultravioletspectrophotometric assay or an HPLC separation withultraviolet detection. The end analyses for both methodshave been performed off-line and on-line depending onthe drug product. For the past 10 years, automateddissolution methodologies have encompassed vessel filling,sample dropping, medium sampling and vessel washingroutines. Recent advances in fibre optic technology havecomplimented the traditional systems by allowing in situanalysis of the dissolution samples. The end analysis isperformed on-line with each dissolution vessel using arobot manipulated fibre optic probe and interfaced to aconventional ultraviolet/visible spectrophotometer. Nosample is removed from the dissolution vessels and nofiltration is required to eliminate background interference.Various computer programs instantaneously calculatethe results and transfer the data to appropriate spread-sheets.

Gregg BellQuality Control--Automation and Technology, Geneva Pharma-ceuticals, Inc., Broomfield, CO, USA

In the more than 10 years that computer system validationhas been an area of concern within the pharmaceuticalindustry, regulators, manufacturers, and system vendorshave been faced with the task of determining how thesesystems should be documented. Although numerousarticles and guidelines on validation principles have beenpublished, the development ofwidely accepted validationstandards has been slow to evolve.

By developing company-level validation standards, signifi-cant benefits can be realized in the areas of regulatorycompliance and business performance. This paper de-scribed the benefits of establishing validation standards;described practical techniques for the development andmaintenance of validation standards; and provided an

example of a validation standard for a company-levelcomputer system validation master plan document.

ADVANCED TOPICS/DATA HANDLING/DATAMANAGEMENTComputer modeling of laboratory automation

Wen-Jeng LiBristol-Myers Squibb Pharmaceutical Research Institute, Prince-ton, NJ, USA

49


Down sizing has made a lot of scientists consider thatautomation will make up the loss of manpower in thelaboratory. Some of them had not thought of laboratoryautomation previously. Some of them had some ideasabout what to automate, but most of them do not knowwhere to start, what to automate, or who to talk to.

Working as internal system integrators, we have theresponsibilities to educate our scientists about laboratoryautomation, help them to identify potential areas, andwork out solutions with them. Describing our suggestionswith words and drawings alone sometimes cannot get themessage across. Most scientists do not envision automationlike engineers. Therefore, we started to use computermodeling software to show the scientists how ideas canwork. If a picture is worth a thousand words, then movingpictures must be worth thousands times more.

With the help of life-like animations, scientists can seeevery step of their processes performed by an automatedsystem. Misunderstandings can be easily corrected in thisdesign phase. This helps to prevent financial losses andminimize wasted time due to rework.

The integration of laboratory automation systemswith LIMS in a pharmaceutical R&D environment

Marie Di Maso, Alain LeBlanc, Carolyn Wells, JudyWybrandts and Sam McClintockMerck Frosst Centrefor Therapeutic Research, Kirkland, Quebec,Canada

To obtain regulatory approval to market a new drugproduct, long term stability studies are required todemonstrate chemical and physical stability during theindicated shelf life of the product. Under the current

changing regulatory environment, more extensive testingand exhaustive documentation is required to support thesestudies. This presentation described the process ofautomating and integrating many of the steps requiredto generate the appropriate data. A LIMS system is usedto set up the study protocols, log in samples for analysis,collate data, produce reports, archive the data andforecast workload. This system receives data from severalsources including an automated chromatography system,balances, KF instruments, a Benchmate Tablet ProcessingWorkstation and a MultiDose dissolution system. Theprocess of validating and maintaining each of the systemsand the associated documentation was presented. Themethods development and optimization processes on therobotic systems and the authors’ approach to issuesencountered were discussed. Emphasis was placed on theintegration of the systems and the flow of informationbetween the system components.

Applications of Visual Basic in laboratory auto-mation

Martin EcholsNycomed Inc., Collegeville, PA, USAand Mark F. RussoIndustrial BioCatalysis, Wynnewood, PA, USA

The Visual Basic programming system for Microsoft(R)

WindowsTM is quickly becoming one of the most promi-nent custom application development tools for theautomated laboratory. Visual Basic combines a simpleprogramming language and an intuitive graphical userinterface development environment with a wide range ofbuilt-in and third-party software libraries. Visual Basicsoftware libraries are available for purposes such as: dataacquisition, serial communications, motion control,scientific graphing, statistics, neural networks, multi-media and even laboratory robot control. This ease-of-useand continually increasing number of pertinent softwarelibraries makes Visual Basic attractive for developingcustom software components for automated laboratorysystems.

The heart of any laboratory automation programmingenvironment is its ability to communicate outside of itsown boundaries. Visual Basic provides many ways to

accomplish this. The authors detailed several of thesecommunication mechanisms by discussing a variety ofreallaboratory automation applications using Visual Basic.These examples covered Serial and IEEE 488 com-

municaions, Dynamic Data Exchange (DDE), DynamicLink Library (DLL) calls, and Object Linking andEmbedding (OLE). The access of databases and localarea networks, as they pertain to laboratory automation,as well as Zymark’s Systems V Controller was alsodiscussed.

Automation of a Houillon viscometer

Katherine S. Bonar, Robert N. Sanders andDonald R. BlevinsAlbemarle Corporation, Baton Rouge, LA 70898, USA

Viscosity measurement is an important part ofAlbemarleCorporation’s R&D lubricating oil programme. Re-searchers can generate more samples in a given timeperiod if sample size is small. An ISL Houillon viscometerwas purchased so that small volume samples could be run.

Using this manual instrument increased the workloadbecause previously fewer large volume samples were beingrun on an automated viscometer. Fast turnaround isneeded to allow efficient progress in research projects. Itwas concluded that automation would make the measure-ment less time-consuming and costly. Robotics would alsoeliminate any potential operator inattention problems dueto boredom.

The Houillon was automated with Zymark PyTechnologyrobotics. Since this application is the first automation ofa Houillon viscometer, custom designs were needed. Aspecial pipet hand for handling small volume pipet tipswas devised. A communications interface between theZymark computer and the Houillon computer was alsodesigned to detect instrument status and to exchangesample identification and results.

The automation has produced substantial savings inoperator time. These savings will return the capitalinvested after running about 6000 samples. It has alsoresulted in the researchers receiving answers sooner,because the robot can operate overnight. Another benefitwas demonstrating a robot’s usefulness and potential inan R&D setting to skeptics in the organization.

5O


Progress toward standard commands for modularanalytical instruments

Gary W. Kramer, Peter J. Grandsard, Torsten A.Staab and Uwe BernhoeftNational Institute of Standards Technology, Chemical Science andTechnology Laboratory, Analytical Chemistry Division, Gaithers-burg, MD, USA

The concept of building automated analysis systems byinterconnecting standard modular instruments offersgreat potential to reduce the costs and complexity ofsystem integration. The Consortium on AutomatedAnalytical Laboratory Systems (CAALS) has beenworking to delineate the requirements for modular instru-ments. Previously, CAALS has described an architecturefor fully automated laboratories, developed the CAALS-ICommunication Specification for instrument-to-controllermessaging, and enumerated behaviours that devicesshould exhibit for good system citizenship. Currently, withthe assistance of CAALS members and other experts inanalytical and clinical chemistry automation, an attemptis being made to define a small, but powerful set ofstandard commands for the remote control ofinstruments.The approach is simple, abstract, and enabling. Sincemany analytical instruments are already computer-controlled and capable of storing externally definedmethods, there is little reason to attempt standardizationat the device command level, rather the approach is todevelop a standard mechanism for invoking such storedmethods. In this way, system control can be achievedwithout requiring instrument manufacturers to give uptheir existing methods of programming their instruments.

A systematic approach to error and exceptionhandling in automated laboratory systems using amachine vision example

Peter J. Grandsard, Torsten A. Staab and Gary W.KramerNational Institute ofStandards Technology, Chemical Science andTechnology Laboratory, Analytical Chemistry Division, Gaithers-burg, MD, USA

Managing errors and exceptions is a system requirementthat is commonly omitted in the initial designs forlaboratory automation applications. Ad hoc approachesfor handling such unexpected events are often addedduring system implementation, often reducing a carefullydesigned control system to ’spaghetti code’. Over thepast five years, the Consortium on Automated AnalyticalLaboratory Systems (CAALS) has developed and pro-moted the concept of modular instruments as build-ing blocks for automated systems. A requirement ofCAAL’s modularity concept is the development of a

systematic, modular approach to the handling of un-planned events.

Classically, unscheduled event handling has been carriedout in three steps: event detection, event reporting, andevent remediation. This process would be more modularifa fourth activity, event classification, was added betweendetection and remediation. In this paper, the authorspresented their recent efforts using machine vision as an

example to manage errors and exceptions in the operationof our Laboratory Automation System Testbed. Thefollowing issues were addressed: the classification ofunplanned events; the level of synchronization betweenerror/exception detection and remediation; and themanner by which an effected module returns to its normaloperating state. Underlying the treatment of all theseissues is compliance to the module behaviour requirementsof the CAALS control model. Some different recoveryprocedures were illustrated with real examples.

Automation integration, and regulationQuo Vadis?

Frank A. SettleVMI Research Laboratories, Virginia Military Institute,Lexington, VA, USA

What is the current state of automation in today’sanalytical laboratories and where is it going? Has the ’fishin-knowledge out’ paradigm been implemented withautomated, integrated, systems, or does automation existonly on isolated islands? This presentation outlineddifferent approaches to laboratory automation in regulatedenvironments, discussed the advantages and limitationsof each, and cited case studies illustrating each strategy.The synergy between the flow of samples and the flow ofdata through automated systems was addressed.

The CAA (Contaminant Analysis Automation, Depart-ment ofEnergy) and CAALS (Consortium for AutomatedAnalytical Laboratory Systems, National Institute forStandards and Technology) hardware and softwarestandards for integration of automated devices intosystems were presented. Advantages of using thesestandards were discussed, as well as problems encounteredin their implementation.

Finally, the path to full laboratory automation via ’plugand play’ standard laboratory modules (SLMS @ Scibus)was projected. The ultimate objective of this strategy isto make laboratory automation as easy as office automationthrough the use of standardized hardware interfaces andsoftware drivers.

DRUG DISCOVERY/HEALTH SCIENCEAutomated compound preparation for high through-put screening

Mark BeggsZENECA Pharmaceuticals, Macclesfield, UK

High compound throughput is a key requirement inempirical screening programmes aimed at pharmaceuticallead identification. The initial compound dilution anddistribution processes that form a common ’front end’ to

many high throughput screens can benefit significantlyfrom effective automation. The primary need was for a

system that could accommodate large numbers of micro-plates and perform the necessary liquid transfer operationsaccurately and reliably. This capability has been providedby an appropriately configured Zymark MicroassaySystem. Deployment of this system at ZENECA Pharma-ceuticals has facilitated a significant increase in compound

51


throughput. The rationale behind automating compounddilution and distribution, the capabilities provided by thesystem and the resulting benefits were described.

Design of track based automated ELISA assay foruse in high throughput screening

determine specific parameters ofa biochemical system (forexample, neurotransmitter receptor availability) importantto normal organ or tissue function. Often, multiple plasmaradiochemical assays including a series of labeled metab-olite correction determinations must be made for everyquantitative PET procedure, requiring additional per-sonnel working in a potentially hazardous environment.

Jeffrey A. Guss, Joe j. Yacobucci, Janet M. Kolband Derek J. HookBristol-Myers Squibb, Biomolecular Research, Wallingford, CT,USA

Drug discovery requires large quantities of samples to beassayed and evaluated in a timely fashion. The screeningprocess contains routine mundane activities which needto be carried out in order to find novel compounds forspecific therapeutic targets. Unfortunately, people are tooexpensive and can introduce variability to the results ofan assay. So most companies interested in drug discoveryautomate to ensure accuracy of the results and to makebetter use of time. A popular assay type is the ELISA.This incorporates liquid handling, shaking, incubating,plate washing, and reading the end point. When creatingan automated platform for this type ofassay, an assortmentofvariables must be addressed if the assay is to be success-fully implemented. Reagents can be light and/or tem-perature sensitive, and volumes of liquid to be handled,sample apacity of the system, and incubation times arejust some of the variables to be considered for the scienceof the assay. The physical layout and overall design ofthe automated system has other needs. Communication,which is both active and has conformation, whether ornot the system is stationary or track based, type of liquidhandling station, peripherals, and space requirements aresome design variables. Special requirements of the systemmay include: reagent cooling and/or heating, filtration,handling radioactive reagents, system waste, and externalreagent pumping stations. There are two other items toconsider when automating an ELISA, or any other typeof assay: development time and cost. If an automationproject such as this one is to be successfully designed andimplemented in a timely fashion, co-operation betweenintegrators and scientists is a necessity.

To help overcome this potential limitation of quantitativeimaging studies, a commercial laboratory robot system(Zymate PyTechnology II Laboratory Automation Sys-tem) was interfaced to standard and custom laboratoryequipment and programmed to perform rapid radio-chemical assays necessary for plasma input functiondetermination in quantitative PET studies in humans andbaboons. A Zymark XP robot arm was used to carry outtwo assays" (1) the determination of total plasma radio-activity concentrations in a series of small-volume wholeblood samples and (2) the determination of unchanged(parent) radiotracer in plasma using only solid phaseextraction methods. Steady steady robotic throughput fordetermination oftotal plasma radioactivity in whole bloodsa,mples (0"350 ml) is 14"3 samples/hour, which includesautomated centrifugation, pipetting, weighing and radio-activity counting. Robotic throughput for the assay ofparent radiotracer in plasma is four to six samples/hourdepending on the radiotracer. Percentage of total radio-activities present as parent radiotracers at 60 minutes postinjection of 25

___5"0 (N 25), 26

__6"8 (N 68), 13

__4"4

(N=30), 32___ 7"2 (N= 18), 16___4"9 (N=20), wereobtained for carbon 11 labeled benztropine, raclopride,methylphenidate, SR 46349B (trans, 4-[(3Z)3-(2-dimethyl-amino-ethyl) oxyimino-3 (2-fluorophenyl)propen-l-yl]-phenol), and cocaine respectively in baboon plasma and84 __+ 6"4 (N= 9), 18

__11 (N= 10), 74

__5"7 (N= 118)

and 16

__3"7 (N 18) for carbon-11 labeled benztropine,

deprenyl, raclopride, and methylphenidate respectively inhuman plasma. The automated system has been used formore than four years for determination of unchangedtracer in plasma for seven different carbon-11 labeledcompounds used routinely in the authors’ laboratory.The robotic parent compound assay runs unattendedand includes automated clean-up procedures that elim-inates all human contact with plasma-contaminatedcontainers.

Automated plasma radiochemical assays for quan-titative functional imaging using a Zymark XPlaboratory robot

Research for this project was supported by USDOE,OHER, and NIH Grants NS- 1538, NS- 15380.

David L. Alexoff, Colleen Shea, Joanna S. Fowler,Payton King, S. John Gatley, David J. Schlyer andAlfred P. WolfChemistry Department, Brookhaven National Laboratory, Upton,3;Y, USA

Positron Emission Tomography (PET) is a nuclearmedicine imaging technique that allows the quantitativeobservation of the time course and spatial distributionof positron-emitting radiopharmaceuticals in an organ(typically the brain or heart) of an awake human subjectafter intravenous injection. Combined with plasma radio-activity measurements and a validated mathematicalmodel, this tissue radioactivity data can be used to

Robot-assisted synthesis of radiopharmaceuticalslabeled with high activity positron emitters

C. Brihaye, C. Lemaire and A. LuxenCylotron Research Center, University of Liege, Sart-Tilman BatB30, B-4000 Liege, Belgium

Positron Emission Tomography (PET) is becoming anincreasingly important tool for studying physiological,biochemical and pharmacological functions at a molecularlevel in living man, whether in health or in disease. Radio-chemical methodology constitutes the most importantbase for successful functioning of a PET group in the

52


routine production and development of radiopharma-ceuticals. Automatic robotic radiosynthesis is more desir-able to avoid excessive radiation exposure to operatives,although the labeling of radiopharmaceuticals with highactivities (>18500 Mbq) of position emitter (laF, IC,13N) by remote control is feasible. The preference forrobotic radiosynthesis exists, because it appears to be moreversatile, and therefore more useful for research. In theCyclotron Research Center of the University of Liege, arobotic system has been developed base on the ZymateTMIILaboratory Automation System (Zymark Corporation).The software control is EasyLab PlusTM, and routines forseveral modules, more specific to the particular radio-synthesis were designed and fabricated in house, includinga column for the separation of [aF]fluoride fromirradiated water, Sep-PakTM and conventional chroma-tography systems, ovens for evaporation equipped withoptical levels probes, a microwave oven to rapidly heatvials. Special sectors were designed for collecting fractionsfrom a chromatographic column and automatic injectionin an HPLC system, as well as for formulating the.injectable solution.

Complete routine productions of[-laF-I Altanserin, 4-[18F]Fluorotropapride, 6-[laF] Fluoro-L-Dopa and 2-[ la]Fluoro-L-Tyrosine are conducted weekly, including thelabeling of the radiopharmaceuticals, the quality controland the formulation of the injectable solution.

Automated sample preparation methods for DNAamplification by PCR

Larry D. Sutton, Werner W. Wilke and Ronald N.ones

Medical Microbiology Division, Department of Pathology,University of Iowa College of Medicine, Iowa City, IA, USA

An automated DNA extraction system was developedwhich was capable ofextracting DNA by several methods.Extraction was accomplished using a Zymate XP roboticsystem with a centrifuge, master laboratory station withremote dispenser, capping station, vortexer, centrifugehand, 200gl eight channel pipetting hand, one mlpipetting hand and a combination syringe-gripping hand.Peripheral equipment included Boekel minirefrigeratorand freezer, and a Hitachi U-2000 double beam, scanningUV-Vis double beam spectrophotometer automated usingZymark’s generic RS-232 interface. The system extractsthe DNA from the sample and quality controls thepreparation by measuring absorbences at 260 and 280 nm.Samples are processed in 1"5 ml screw-capped micro-centrifuge tubes. Only one tube is opened at a time inorder to minimize chances ofcross contamination betweensamples. DNA amplification was accomplished in 96 wellformat using either a Zymate II automated PCR systemwith an MJ thermal controller or by reagent assemblyand sample loading by a Hamilton Micro Lab 2200 andthermal cycling on a Perkin-Elmer 9600. Throughputvaried according to the DNA extraction technique. Theextraction efficiencies were evaluated using severaldifferent sample matrices and DNA analytes.

A liquid handling station to rapidly assemblereagents and sample loading for DNA amplificationby PCR

Werner W. Wilke, Arzu Karabay, Ronald N. Jonesand Larry D. SuttonMedical Microbiology Division, Department of Pathology,University of Iowa College of Medicine, Iowa City, IA, USA

DNA analysis based upon gene amplification by thepolymerase chain reaction (PCR) continues to find new

applications in medicine, agriculture, environmentalanalysis, and the food, pharmaceutical and biotechnologyindustries. Many of these applications require highthroughput and reliability, especially with regard to

prevention of false positives due to carry-over contami-nation. The performance ofthe Hamilton Micro Lab 2200to assemble PCR reagents and load samples into micro-tubes in 96-well format for amplification on a Perkin-Elmer 9600 thermal controller was evaluated. DNA wasextracted from samples using a Zymate XP system.Performance was validated by comparing to a Zymate IIautomated PCR system which has been evaluated overthe past three years. Washing the Teflon-coated steelcannula with 5, sodium hypochlorite followed bycopious rinses with distilled water using the peristalticpump accessory, adequately eliminated sample carry-over. The high flow rate (3 ml over 10 seconds perchannel) achievable with the peristaltic pump wasessential to eliminate carry-over ofboth target and bleach(which inhibited PCR). Up to 50 negative controls wereused to assess aerosolization of target; none was detected.No build up of target DNA overtime was observed. Asconfigured, the system is capable of assembling over 1800PCR reactions in an eight hour shift translating into nearly500 000 reactions per year. At this throughput, use offixedcannula may save from $5000 to $30000 in disposablepipette tips.

Validation of robotic-automation ofthe fluorescentmicrosphere technique for determination of re-

gional blood flow

R. Hecht, S. Raab and K. MessmerInstitute for Surgical Research, LM-University of Munich,GermanyDetermination of regional blood flow (RBF) by means offluorescent-labeled microspheres (FM) has been demon-strated to be an attractive alternative to conventionalradioactive microspheres (RM). However, processing oftissue samples to completely extract fluorescence from thetissue is time consuming because it has to be performedmanually. In addition, it presents a source of error.

The authors designed a new sample processing unit (SPU)which allows the entire processing of the tissue samplesautomatically. After digestion of the tissue samples byexposure to 4 N KOH at 60C for 4 hours and vacuum-filtration, the extraction of the fluorescent dye is accom-plished by adding 2(2-ethoxy-ethoxy)-ethyl-acetate andfollowed by centrifugation. Zymark Corporation (Hop-kinton, MA, USA) provided a modified robotic system

53


for processing the tissue sample to analyse the fluorescenceof multiple colours present in each sample. For validationof the FM-method of the new automated SPU, combi-nations of different RM and FM (0-15 gl) were simul-taneously injected into the left atrium of six pigs. For alltissue samples prepared according to an hierarchicalorgan dissection scheme (N 301/pro animal: heart 201;brain 46; kidney 44; skeletal muscle 10), RBF wascalculated based on radioactivity or fluorescence intensity,respectively.

The RBF determined by FM using the new SPU withautomated measurement corresponded to RM-RBFvalues (r > 0"95).

The automated FM method reported allows reliabledetermination ofRBF with the advantage of automaticallyprocessing 40 samples/hour and without need of radio-isotopes.

LABORATORY WORKSTATIONAnalysis of cleaning validation samples with theZymark BenchMate workstation

Stephen Scypinski, Theodore Sadlowski and JohnBaiano

Hoffmann-La Roche, Inc., Nutley, NJ, USA

The performance ofcleaning validation or cleaning assess-ment studies is necessary to verify that a piece ofpharmaceutical processing equipment is free from con-tamination from both active substance and cleaning agentafter its use. Cleaning validation is especially importantto the development arena, where it is quite common fora single blender, mill or tablet press to be utilized in theproduction of several products. Therefore, the cleaning ofsuch processing equipment has become commonplace,along with the detailed protocols for the proper executionof the cleaning procedures. Because of the rigours of theseprocedures and the complexity of typical equipmentinvolved, cleaning validation typically results in amultitude of samples for a single cleaning. These samplesconsist of either swabs obtained by wiping a prescribedarea of the machine or rinse samples collected during thewashing of its specified parts. While the methods used toanalyse these samples are usually not very complicated,the tedium of analysing such a large amount of samplescan be considerable.

The authors have found that such a procedure was anexcellent candidate for automation via the Zymark Bench-Mate Workstation. BenchMate racks containing 16 x100 mm test tubes with caps are given to the industrialpharmacists who perform the cleaning of the equipmentfollowing a prescribed Standard Operating Procedure(SOP). The tubes are turned to the laboratory containingswabs or rinse solutions. In the case of solutions, the racksare placed in the Zymark TurboVap for concentrationfollowed by placement in the BenchMate for analysis.Racks containing swab samples are placed immediatelyin the BenchMate.

Several products have already been automated by thisprocedure, and the authors anticipate no further need to

perform these analyses manually. The generic method andspecific results for several products were presented.

Automated analysis of oral care products byHPLC using a BenchMate sample preparationworkstation

Anthony J. Wilkes, Gisele Walraven and Jean-MarieTalbotColgate Palmolive Company, Research and Development Inc.,Milmort (Herstal), B 4041 Belgium

Automated methods have been elaborated for the analysesofseveral therapeutic agents of dental creams and mouth-rinses. Sample weighing, dilution, mixing of liquids bycycling (mouthrinses) or vortex agitation ofviscous liquidsor pastes (dental creams), addition of internal standards,filtration of particulate matter, preparation of multi levelcalibration standards and injection onto the HPLCchromatograph was effected using the BenchMate Work-station. The BenchMate was employed in-line, thussamples and multi-level calibration standards were freshlyprepared prior to injection and analysis by HPLC.Operator time was reduced to a minimum and resultswere furnished during unattended overnight or weekendruns. Results obtained using the BenchMate for bothinternal and external standard methods were in excellentagreement with those obtained using the manual pro-cedures; RSDs were typically less than 2.

Automated determination of trace avermectins inwaste streams

Andrew Lange and Mark GrimaldiMerck & Co., Inc., Elkton, VA, USA

Production and environmental demands necessitated thedevelopment ofa quick, sensitive assay for analysing traceavermectins in waste streams. A sensitive, automated assaythat included solid-phase extraction, solvent extraction,evaporation and derivatization, followed by HPLCanalysis using fluorescence detection was developed. Tothis end, several modifications were made to a ZymarkAutoTraceTM Workstation, and custom accessories werefabricated.

To quantify these trace amounts in aqueous solution, theAutoTrace was used to extract the avermectins from wastewater using solid-phase extraction. To quantify avermec-tins adsorbed to solids in waste waters the AutoTraceWorkstation was modified to allow solvent extraction andinjection of samples containing solids. A PTFE adapterwas manufactured to allow stacking of a C-8 solid phaseextraction column and a Florisil clean-up column inseries, combining two steps in the analysis. Specializedglassware was made to accommodate 20-ml of eluate andallow the eluate to be evaporated to a residue in an HPLCvial. This new glassware required modifications to boththe AutoTrace Workstation and TurboVap system. TheTurboVap system was used for drying multiple samples,and a Shimadzu SIL-10A autosampler was used toautomate the derivatization step.

54


The analysis of standards from 20 gg/1 (ppb) to gg/1have been tested thus far with good correlation betweenconcentration and peak area, with a correlation coefficient(R2) of 0"99. The precision ranges from 6-13 (relativestandard deviation) and the detection limit is below10 ng/1 (ppt).

Extraction of benzoylecgonine from urine samplesusing the Zymark RapidTraceTM

William H. Vickery, Francis X. Diamond andJohn de KanelNational Medical Services, Willow Grove, PA, USA

Benzoylecgonine is one of the primary metabolitesexcreted in the urine following cocaine exposure. It is thismetabolite which is designated by SAMHSA (SubstanceAbuse Mental Health Service Administration formerlyNational Institute for Drug Abuse, NIDA) for analysis inemployment and pre-employment situations to determineif cocaine has been used. Numerous methods have beenpublished describing methods to extract benzoylecgoninefrom urine either by traditional liquid/liquid means or

by solid phase extraction. Liquid/liquid extractions aretime consuming, difficult to automate and require greatanalyst skill if they are to produce reproducible andaccurate results. Each manufacturer of solid phaseextraction cartridges has published methods to accomplishthe extraction of benzoylecgonine from urine. For thepurposes of evaluating the Zymark RapidTraceTM

automated extraction workstation, the Applied Separ-ations’ method published for use with their SpeedTM ScanABN cartridges was chosen. The method was programmedinto the RapidTraceTM software without modification. Aset of 10 Zymark RapidTraceTM sample preparation unitscontrolled by one personal computer was capable ofextracting benzoylecgonine from 60 urine samples perhour. The samples were then evaporated to dryness at40C using a Zymark TurboVap. This step took approxi-mately 10 minutes. The samples were then derivatized byreaction with 50 gl /BSTFA with 19/o TMCS) in 50 glethyl acetate for 20 minutes at 70C in a sealed tube. Thesamples were then transferred to autosampler vials andanalysed by GC/MS. A technician would spend at leastthree hours preparing these extracts manually. The useof the RapidTrace frees the technician to work on more

productive things such as data review for two of the threehours.

One millilitre urine samples known to be negative forcocaine metabolites were spiked with deuterated benzoyl-ecgonine at a level of 200 ng/ml as an internal standardand with benzoylecgonine at varying levels to evaluatethe recovery, linearity and carry-over of the extractionmethod when performed by the Zymark RapidTrace. Theresults from this experiment are presented in the table.The standards at 75, 150, 300 and 1000 ng/ml were usedto determine back calculated levels for these standardsand to quantitate the controls at 120 and 180 ng ng/mlas well as levels in various blank samples. A urine samplewas extracted through the procedure after spiking it with100000 ng/ml benzoylecgonine. Analysis of the blanksamples following this procedure was done to estimatecarry-over. The procedure used was identical to the

Ion Ion IonSpike Calculated ratio ratio rationg/ml level, ng/ml 346/240 361/240 361/346

75 78 0"1088 0"2844 2"652150 142 0"1033 0"2761 2"673300 300 0"0992 0"3224 3"2491000 1000 0"0986 0"3088 3"132120 129 0"1145 0"3085 2"694180 175 0’1074 0"3253 3"029

5"5 0"6326 0"9166 1"4490* 15"1 0"0725 0"3483 4"802

5"2 0"1947 0"2934 1"507


routine procedure which would be followed for realsamples. No extra manipulative steps were added in aneffort to minimize carry-over. The extract from the100000 ng/ml sample was not derivatized nor analysedby GC/MS since potential carry-over on the GC auto

sampler could have confused results.

Under SAMHSA guidelines an affirmative cut-off of150 ng/ml is used to administratively differentiate positivesamples from negative ones. Irrespective of other data,samples containing benzoylecgonine below this adminis-trative level are reported to clients as negative. In additionto this quantitative test, three ion ratios must be within

20% of values determined for a standard. Direct quanti-tation of the blank following the 100000 ng/ml spikeyields 15 ng/ml or 0"015% carry-over. Some of this levelis derived from the background noise. The blanksextracted after relatively low levels of benzoylecgoninequantitate to approximately 5 ng/ml, for example. Theselevels are all far below the administrative cut off of150 ng/ml. In addition to this, however, the ion ratio testfails for all the blank runs. Benzoylecgonine cannot evenbe identified in these samples, therefore.

AUTOMATION AND COMBINATORIALCHEMISTRYSecond-generation robotic synthesizer for peptide,pseudopeptide and non-peptide libraries

Jean A. Boutin and Jean-Luc FauchereDivision des Peptides, Institut de Recherches Servier, 92150Suresnes, France

A second-generation peptide library synthesizer has beenconstructed around a Zymark robotic arm. In order to

provide new solutions for the ever-growing field ofcombinatorial libraries, it has been designed to handlegram quantities of resin for the solid phase synthesis ofpotentially any type of library.

To ensure a wide molecular diversity, a series of 24building blocks was used and therefore, the robot synthesisstation is a table hooked with 25 (5 columns of 5) reactors,slaved to individual gas and waste valves. The 25th reactorserves as a control permitting the independent butconcomitant synthesis of a single defined product whichis closely analysed by HPLC, mass spectrometry andnuclear magnetic resonance. Since libraries (i.e. defined

55


mixtures of oligomers made up of at least 24 buildingblocks) are extremely complex mixtures, with up to severalmillions of individual chemicals, a minimum of g (i.e.2 x 10 beads) per reactor vessel was considered theabsolute requirement to ensure the proper and completesynthesis of random tetra- to penta peptide libraries, orof higher oligomer libraries with fixed positions.

Designed to perform the so-called mix-and-divide strategy,the robot comprises a mixing chamber that functionsas a mixer between each step of the combinatorialsynthesis. This chamber is filled and emptied by aCombiTip-based hand that ensures the proper and totaltransfer of the bead suspension. Finally the instrumentcomprises also a section for automatically dissolving thestarting materials (not limited to natural or exotic aminoacids), thus ensuring the maximum stability of thechemicals before coupling. This section comprises five setsof 25 tubes in which the dry building blocks are stockedand five additional devices for large quantities ofcommonreagents. The robot comprises a set of valves and handsthat handle up to 10 different liquids used in the chemicalreactions or as washing solvents.

This instrument is a unique tool for the iterative synthesisof defined libraries. More than a 100 different peptide,pseudopeptide and non-peptide libraries (over 20 millioncompounds altogether) have been synthesized in theauthors’ laboratory in the past two years leading to ligandsof therapeutically relevant targets active in the micro tonanomolar ranges.

reaction blocks are processed by a series of custom-built,task specific workstations organized in an assembly linefashion. Compounds are cleaved from each of the 96individual reaction vessels of the reaction block systemdirectly into the wells of a standard 96-well microtitreplate. These plates, with one compound per well, are

readily accepted onto Ontogen’s automated high through-put screening (HTS) systems. Client/server databasesoftware for inventory control, experiment design, instru-ment control, HTS data analysis and structure activityrelationship (SAR) studies is written in Visual Basic andOracle7. Synthesis of compounds in the OntoBLOCKsystem is driven by instrument control language (ICL)scripts which are generated by the experiment planningsoftware and integrated within the database system. Inthe OntoCODE system, miniature reaction capsules areelectronically coded with a solid state radio frequency tagand undergo a number of split and recombine syntheticsteps. The OntoCODE tag is electronically read after eachstep to maintain a reaction histogram. After the syntheticsteps are completed, the discrete compounds are cleaved.from the coded capsules into standard 96-well microtitreplates for HTS, with one compound per well. In bothsystems, an automated mass spectrometer processes themicrotitre plates and produces spectra for each com-pound.

A totally integrated approach to combinatorialchemistry

Automated preparation and purification ofamides

R. Michael Lawrence, Olga M. Fryszman, MichaelA. Poss, Scott. A. Billet and Harold N. WellerBristol-Myers Squibb Company, Princeton, NJ, USA

With ever-increasing pressures on the pharmaceuticalindustry to bring innovative products to the marketplacemore quickly, Bristol-Myers Squibb Company is exploringautomated organic synthesis as a means to accelerate thedrug discovery process. To this end, we have developedan automated solution phase procedure to rapidly prepareamides. Utilizing a Zymark BenchMate Workstation, upto 100 reactions can be set up and the resulting productspurified by exploiting the solid phase extraction capabilitiesof the BenchMate Workstation. The procedures forreaction set-up, product purification, analytical samplepreparation and electronic data handling were describedin this presentation.

Automated combinatorial chemistry on solid phase

John F. CargillOntogen Corporation, Carlsbad, CA, USA

High speed solid phase synthesis of diverse, non-peptidylsmall organic molecules in a spatially dispersed combina-torial library (SDCL) is accomplished with proprietaryautomation systems called OntoBLOCK and OntoCODE.These systems support reagent introduction and removal,agitation, inert atmosphere, temperature control, pressurecontrol and vacuum drying. In the OntoBLOCK system,

Peter L. MyersCombiChem, Inc., La Jolla, CA, USA

Combinatorial chemistry is emerging as one of a series oftechnology advances, which over the next decade, couldpotentially provide a more efficient and successfulapproach to drug discovery. As with the advent of manynew technologies there has been very significant evolutionfrom the initial creative, but complex, methodologieswhich validated the concept to a growing requirementfor a simplifying strategy to leverage its full potential. Thereal value of the technology to the medicinal chemist, whois ultimately responsible for selecting and optimizing leadstructures, lies in the ability to maximize the informationgained in screening combinatorial libraries of moleculesagainst a wide range of biological targets, and to patentfile composition claims to molecules of most interest, in atraditional manner.

As these are essential requirements, it is obvious that thefocus in library design and synthesis should be on multi-parallel synthesis ofsingle compounds (discretes) in multi-milligram quantities with appropriate QC methods forpurity and confirmation of identity. This is most readilyachieved by making smaller libraries and simultaneouslyeliminates the need for complicated tagging and/ordeconvolution strategies which are not always successfuland may complicate the chemical processes.

However, proportionately larger numbers ofsuch librarieswill now be necessary, to ensure that we exploit thepotential diversity open to us. They must be well designedboth for synthesis and more specifically content. Ideally the

56


synthetic methodology should also be capable of simpleautomation to minimize manpower resource.

Thus CombiChem’s intention as a combinatorial chemistrycompany is to develop an integrated package comprisingstate of the art combinatorial library synthesis incor-porating computational software design and instrumen-tation which together will form the major infrastructuretowards optimal library design and synthesis.

DIVERSOMERTM technology and recent advancesin automated synthesis

S. Hobbs DeWitt’ N. Halim, E. Hogan, A. W.lIzarnikParke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, MI, USAA. MacDonald and R. RamageUniversity of Edinburgh, Department of Chemistry, Edinburgh,UK

Advances in molecular biology and automation haverevolutionized the testing ofcompounds for drug discovery.Biologists at Parke-Davis can now test up to 500000samples in one month. Therefore, traditional methods forchemically synthesizing compounds one-by-one can no

longer meet the demands of mass screening. The ability

to synthesize multiple compounds in a simultaneousfashion is the goal of Parke-Davis’ DIVERSOMERTM

technology. Automation of routine and labour-intensivesteps of chemistry is critical to advancing combinatorialand high throughput synthesis. Although automation hasbeen extensively used in clinical chemistry and highthroughput screening, it has not been integrated intosynthetic chemistry laboratories. This is largely due to thefact that chemistry requires individual and precisemanipulations. The objectives of automation in chemistryare not only speed and productivity but also precision insample transfers and manipulations.

DIVERSOMERTM technology enables the high through-put synthesis of organic compounds using both manualand automated methods. The system design incorporates amodular approach employing robotic automation toachieve parallel processing of40 intermediates or products.Automation and integration has been achieved bymodifying and exploiting commercial software andhardware products. Representative hardware and soft-ware includes the DIVERSOMERTM apparatus, a Tecanrobotic sample processor, Microsoft Excel, and MDLsoftware. The combination of these tools enables a fullrange of operations necessary for the generation andtesting ofcompound libraries for combinatorial chemistryand molecular diversity programs.

57

automatic vol. abstracts of papers presented at the islar...

Documents