introduction to chromatography software - mohsin …...2017/03/04 · introduction to...
TRANSCRIPT
![Page 1: Introduction to Chromatography software - Mohsin …...2017/03/04 · Introduction to Chromatography software ©2005 Waters Corporation Agenda • Chromatography Theory • Basic](https://reader034.vdocuments.net/reader034/viewer/2022052518/5f0837f87e708231d420ec56/html5/thumbnails/1.jpg)
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Introduction to Chromatography software
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Agenda
• Chromatography Theory
• Basic Chromatography Sequence
• Data Management
• Data Processing
• Reporting
• Software Option
• Instrument Control
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Chromatography Theory
• Everything is relative– From Area ( Response) Concentration (Amount)– Or Amount = B x (Area)
• But what affects B?– Absorption capabilities of the molecule’s chemistry– UV lamp or other detector qualities (age, efficiency, energy, cell size)– Sensitivity of the recording device– Mobile phase (composition and flow rate)
• Therefore the pharmacopoeia specifies that you should run standards of known concentrations to determine B.– On the same LC– With the same conditions– On the same day– With the same integration algorithms
∝
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Amount (sam) = Amount (std)
Response (sam) Response (std)
Amount (sam) = Amount (std) X Response (sam)
Response (std)
Quantitation Theory
First Principals
This is B:- the slope of a plot
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Quantitation Theory
• Plot amount against response through origin
• For any measured Sample RESPONSE
• We can read off the appropriate Sample AMOUNT
Amount Standard
Res
pons
e
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Identifying StandardsStandard Replicates
• How many standards identified as replicates in the Sample Set Method– No. of vials
Multiple vials of the same standard concentrationMultiple vials with different standard concentrations
– No. of injections
Sample Set Method
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Quantitation Theory
• How do we plot the Std measurements?
• One strength solution multiple injections
• Similar strength single injection
• Multiple strengths
Amount Standard
Res
pons
e
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Position of Standards
• Traditionally placed at the beginning of a run
• Response can change during a long sequence– Particularly with early detectors– With the long run times needed with previous generation
separation columns– Depends on solution stability
• Habit of placing standards through the run was established– Even when up to date columns and equipment removes the
need– Either “every 5 samples” or “every 45 minutes”
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Performing CalibrationBracketing Standards
• No Bracket– Runs all standards first and
– Create Calibration curve
– Run all samples
– Quantitate Samples
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Performing CalibrationBracketing Standards
• Overall Bracket– Standards are injected at the start
and end of the sample set.
– Entire sequence collected
– Standards Calibrated
– Samples Quantitated
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Performing CalibrationBracketing Standards
• Modified Overall Bracket– Standards run AROUND GROUPS of n
samples
– Entire sequence collected
– Standards Calibrated
– Samples Quantitated
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Performing CalibrationBracketing Standards
• Sliding, No Overlap– Standards run AROUND GROUPS of
n samples– One an entire group is collected
Calibrate the Stds of the groupQuantitate the Samples of the group
– Standards are used only once during calibration.
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Performing CalibrationBracketing Standards
• Sliding, with Overlap– Standards run AROUND GROUPS
of n samples– One an entire group is collected
Calibrate the Stds of the groupQuantitate the Samples of the group
– Standards are used multiple times during calibration.
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Performing CalibrationSliding Bracket-No Overlap
Bracket 1
Bracket 2
Bracket 3
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Pre Sequence Checks or System Suitability
• Blank Injection
• Replicate Injections
• Test Quantitation – control samples
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Basic Chromatographic Workflow Process
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1. Login.2. Instrument Method.3. Sequence/Run Samples.4. Process Data.5. Review/Preview6. Print
1.
5.
Basic Sequence
2. 3.
4.
6.
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Single Injection
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Create Sample Set
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Log In
• Log into Empower using your user name– demo user– sdms60
• Be sure you are in the QuickStart Interface
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Methods
• Method Set– Instrument Method– Processing Method– Report Method– Export Method
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Instrument Method
• Method Set– Instrument Method– Processing Method– Report Method– Export Method
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Collect Chromatographic Data
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Processing Method
• Method Set– Instrument Method– Processing Method– Report Method– Export Method
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Process the Data to Create Results
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Report Method
• Method Set– Instrument Method– Processing Method– Report Method– Export Method
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1. Login.2. Instrument Method.3. Sequence/Run Samples.4. Process Data.5. Review/Preview6. Print
1.
5.
Basic Sequence
2. 3.
4.
6.
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Data Management
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Empower 2 Data ViewsQuick Sort - Click on any column heading to sort ascending or descending
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Empower 2 Data ViewsRaw Data = Sample Sets, Injections and Channels
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Empower 2 Data Views
Sample Sets
Injections
Channels
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A Sample Set contains multiple injectionsEach injection may contain multiple channels
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Empower 2 Data ViewsSample Sets -> Injections -> Channels
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Empower 2 Data ViewsMethods = Instrument, Processing, Report, Method Set and Sample Set
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Empower 2 Data Views
• Instrument Method - Specifies instrument control and data acquisition parameters, such as flow rate and wavelength
• Processing Method - Includes a set of instructions defining how Empower processes raw data
• Report Method - Serves as a template to organize the data, results, calibration curves, and method contents in a generated report
• Export Method - Specifies the data file format you want to use to export data.
• Method Set - User-defined collection of methods and data channels– Can include an instrument, processing, report and export method
• Sample Set Method - Contains a set of functions and related information that specifies the parameters to acquire data from each vial (standard or unknown) in a set of vials
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Instrument Method
Wavelengths/Masses
Processing Method
Report Method
Sample Name
Standard or Sample
Vial – Run Time
Method Set to Use
Amounts of Standard
Sequence or Set of Samples to be Run
Empower 2 Data Views
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Empower 2 Data ViewsProcessed Data = Results and Result Sets
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Empower 2 Data Views
Result Sets
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A Result Set contains multiple Results
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View Filters
• View Filter - limits or customizes the types of information available for viewing in the Project window. – You can use View Filters to define the columns of information that appear
and their sequence and sorting. Empower provides several default view filters for project data.
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View Filters
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View Filters
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Find My Results
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Custom Fields
Custom Fields - A custom field is a user-defined field in a table as opposed to a predefined field in Empower.Custom fields can be descriptors and formulas
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Custom Fields
Descriptor Custom Field
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Custom Fields
Formula Custom Field
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Data Processing
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Empower 2 Data Processing
Select a Sample Set, Injection or Channel and Click on Review
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Empower 2 Data Processing
Click Integrate to perform default peak integration
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Empower 2 Data Processing
Modify Integration Parameters or Add Integration Events
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Empower 2 Data Processing
Manually Integration Peaks
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Empower 2 Data Processing
Add Peak Names
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Reporting
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Empower 2 Reporting
Select a Result or Result Set and Click on Preview
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Empower 2 Reporting
Select a Report Method from the list
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Empower 2 Reporting
Edit Report Method
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Empower 2 Reporting
Modify Peak Table
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Empower 2 Options
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Traditional Software Options
• Built In:– Photo Diode Array– Mass Spectrometry– CE/CIA
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Traditional Built-in Option
Check peak purity and see the finest spectral details.
Single wavelength data,multi-wavelength data or complete spectra.
Integrated advance detection to minimize software training.
Photo Diode Array
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Traditional Built-in Option
Data acquisition and instrument control of Mass Detectors.
Greater specificity of analysis.
Easily use MS and MS/MS in QC and Development lab environments.
Mass Detection
1st Chromatography Software to offer Tandem control and data processing in a network or
regulated environment
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©2005 Waters Corporation
Traditional Software Option (Key Disk)
System SuitabilitySystem suitability testing is a means of checking that a chromatographic system and methodology are working within acceptable limits.
System Suitability calculations and limits are selected in the processing method and limits can be set so that a result will be flagged if it is outside the defined limit.
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©2005 Waters Corporation
Integrity of your HPLC dissolution testing methods and results, helping increase lab productivity.
Combined software and hardware solution.
Automated Dissolution
Traditional Software Option (Key Disk)
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©2005 Waters Corporation
Gel Permeation Chromatography (GPC) for Polymer AnalysisGPC is the only proven technique for characterizing the complete molecular weight distribution of a polymer, which is required for predicting material properties and performance.
Polymer analysis includes techniques such as Gel Permeation Chromatography (GPC)
Viscometry (GPC/V) Light Scattering(GPC/LS)
Traditional Software Option (Key Disk)
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©2005 Waters Corporation
Integrated Chemical Structures
Integrate chemical structures to improve data management and reporting.
Perform structure searches, assign structures to components and connect to corporate structure databases.
Traditional Software Option (Key Disk)
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©2005 Waters Corporation
Waters UPLC
• Full control of Waters binary and quaternary pumps.
• Full control of Multiple wavelength, Photodiode array, Fluorescence, and RI detectors.
• Full Control of sample compartment and column oven temperatures
• Full control of autosamplers.
• Full control of Waters Mass Spec detectors.
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©2005 Waters Corporation
Waters HPLC’s
• Full control of Waters binary and quaternary pumps.
• Full control of Multiple wavelength, Photodiode array, Fluorescence, and RI detectors.
• Full Control of sample compartment and column oven temperatures
• Full control of autosamplers.
• Full control of Waters single quad mass detectors.
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©2005 Waters Corporation
Waters Approach
Empower Software puts the right controls in the right hands for efficient, compliant lab work.