routine impurity analysis with uplc/ms and upc2/ms
TRANSCRIPT
©2014 Waters Corporation 1
Routine Impurity Analysis with UPLC/MS,
UPC2 /MS
©2014 Waters Corporation 2
Outline
Sources of Impurities – Why is it important to be able to: o Separate o Detect o Identify o Quantify o Isolate impurities
Chemical Materials Workflow
Waters Technologies
Application Examples
©2014 Waters Corporation 3
Sources of Impurities
Either naturally occurring or purposely, accidentally, inevitably, or incidentally added into the substance – Raw materials – Contamination / Degradation o During production /
synthesis o Packaging / storage / use
The presence of impurities, even in small amounts, could affect the efficacy, performance, quality, lifetime and safety of the final product
©2014 Waters Corporation 4
Chemical Materials Workflow
IDENTIFICATION – UNKNOWN IMPURITIES Structural Elucidation – Exact mass, elemental composition, fragments, MS/MS confirmation
IDENTIFICATION – KNOWN IMPURITIES Calibration against standards User library/database UV, Mass, RT, Fragment
DETECTION
PDA / ELSD / FL MS / MRM / SIR MSE / MS/MS NMR
SEPERATION
HPLC UPLC UPC2 APGC
©2014 Waters Corporation 5
Separation Technology Overview
Gas Chromatography
Liquid Chromatography
Convergence Chromatography
Separation achieved by a temperature gradient
•High efficiency [N] • Virtually no limitation on column length
•Limited selectivity [α] • Limited stationary phase options
Separation achieved by a solvent gradient
•High efficiency [N] • Limited to pressure drop across column
•Moderate selectivity [α] • Different modes: reversed-phase, normal-phase, SEC, IEX, affinity, ion pair, HILIC, GPC…etc.
Separation achieved by density/solvent gradient
•High efficiency [N] • Very low viscosity enables longer columns and smaller particles
•High selectivity [α] • Wide variety of stationary phase and mobile phase co-solvent and modifier options
GC
LC
CC
APGC
UPLC
UPC2
HPLC
©2014 Waters Corporation 6
Ultra Performance Liquid Chromatography is a variant of HPLC using columns with particle size <2 µm (typically, 1.8 µm, which provides significantly better separation than the traditional (5 µm ) columns and enables much faster analysis
Higher efficiencies with a much wider range of linear velocities,
flow rates, and back pressures
More resolution and sensitivity
While increasing throughput (faster run times)
Optional elevated temperatures (column oven)
Highly robust, dependable, and reproducible system
ACQUITY UPLC
©2014 Waters Corporation 7
Convergence Chromatography (UPC2)
Convergence Chromatography (CC) is a normal phase separation technique – Uses carbon dioxide as the
primary mobile phase – Choice of adding an co-solvent
such as methanol or acetonitrile
The Waters UltraPerformance Convergence Chromatography (UPC2) builds upon the potential of CC while using proven and robust Waters UPLC technology
©2014 Waters Corporation 8
ACQUITY UPC2 Flow Path and Components
Inject valve
Auxiliary Inject valve
Column Manager
PDA detector
Back Pressure Regulator (Dynamic and Static)
Waste Modifier CO2 Supply CO2
Pump Modifier
Pump
mixer Thermo-electric heat exchanger
Make-up Pump
Mass Spec
Splitter
©2014 Waters Corporation 9
The Key Benefits of UPC²
Simplify the workflow with UPC2
– Combine multiple techniques (LC & GC into CC) – Access robust normal phase separations – Eliminate solvent exchange steps for organic extracts
Deal with compound Similarity challenges – Chiral Separations (enantiomers & diastereomers) – Positional isomers (differ in location of functional groups)
Deliver Orthogonal separations
– Different relative retention helps ensure full characterization – Check method specificity by comparison to a second
procedure – Reveal “hidden” impurity or degradation peaks – Increase confidence in characterization of complex samples
SIMPLICITY
SIMILARITY
ORTHOGONALITY
©2014 Waters Corporation 10
What are the benefits of using SFC based separations?
Deliver Orthogonal separations – Different relative retention helps ensure full characterization – Check method specificity by comparison to a second
procedure – Reveal “hidden” impurity or degradation peaks – Increase confidence in characterization of complex samples
Simplify the workflow with UPC2
– Combine multiple techniques (LC & GC into CC) – Access robust normal phase separations – Eliminate solvent exchange steps for organic extracts
Deal with compound Similarity challenges – Chiral Separations (enantiomers & diastereomers) – Positional isomers (differ in location of functional groups)
SIMPLICITY
SIMILARITY
ORTHOGONALITY
Built upon proven UPLC Technology – Quantifiable increase in productivity
Robust, Reliable and Reproducible – Modernization of SFC-based technology,
making this technique a viable analytical separations tool
©2014 Waters Corporation 11
Detection
Tandem Quadrupoles
Single Quadrupoles
SQD2
XEVO TQD
XEVO TQ-S
XEVO G2-XS QToF
Time of Flight (ToF)
ACQUITY UPC2 System
ACQUITY UPLC System
Mass Detector
QDa
Point Detectors ELS PDA RI* FLR* TUV* UV/VIS # *UPLC / HPLC only #HPLC only
SYNAPT G2-Si
XEVO TQ-S micro
Alliance HPLC
©2014 Waters Corporation 12
Designed as mass detector integrated with a separations system
A robust mass detector that does not require adjustments and optimization for each sample
Provides orthogonal detection for the range of applications that benefit from the extra information
ACQUITY QDa Detector
©2014 Waters Corporation 13
Automated Calibration Pre-optimized ES Zero Tuning Disposable Sample Cone 50-1250 Da 10,000 Da/s +/- Switching 4 Orders Dynamic Range
Detector Characteristics
PDA
QDa
©2014 Waters Corporation 14
Waters Fraction Manager - Analytical
©2014 Waters Corporation 15
Analytical Scale Fraction Collector System Description:
– Integrates seamlessly with ACQUITY H-Class or Alliance Systems
– Empower Control OR Stand-Alone Control – Flow Rate Range: 0.1 – 2 mL/min
o Optional needle for flow rates up to 5 mL/min – pH Operating Range 2 – 12
Developed on FTN Sample Manager Platform – known for innovative, robust technology.
Fast valve switching and movement between vessels enabling collection of narrow UPLC peaks.
Designed for low carryover and high recovery to accommodate limited sample.
Temperature controlled from 4 to 40OC for thermally labile samples.
Waters Fraction Manager - Analytical
©2014 Waters Corporation 16
Agrochemicals
Application note: 720004824en
PDA
QDa
©2014 Waters Corporation 17
Agrochemicals
The use of agricultural chemicals ensures that there is decreased crop damage resulting in a food supply that is plentiful and of high quality
The detection, characterization and quantitation of the active ingredient/s and all other components in the pesticide formulation is necessary to support product development and product registration
The presence of impurities (>0.1%) could potentially reduce the toxicological properties of the final product, potentially causing health and environmental effects
©2014 Waters Corporation 18
Agrochemicals
Experimental Description: UPLC/PDA/QDa Achiral separation by UPLC, PDA and QDa mass detection were
used to provide a data profile of the pesticide formulations. The ACQUITY QDa mass detector, in combination with PDA,
allowed for low-level components to be detected with increased confidence in the pesticide formulations.
The components were identified as having similar optical and
structural properties to the active ingredients (AI).
Inert formulation components not seen in the UV were readily detected by the ACQUITY QDa Detector.
©2014 Waters Corporation 19
Agrochemicals AU
0.00
0.20
0.40
0.60
0.80
1.00
AU
-0.12
-0.10
-0.08
-0.06
-0.04
-0.02
0.00
0.02
0.04
0.06
Minutes0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00
1 2
4 3
Formulation Sample
Propiconazole Standard
**
UPLC/UV Analysis of a Formulation
©2014 Waters Corporation 20
Agrochemicals AU
0.00
0.20
0.40
0.60
0.80
1.00
QDa 1: MS Scan MS TIC (1: 100.00-1000.00 Da ES+, Continuum, CV=7): not integrated, source of MS spectra
Inte
nsity
0.0
5.0x10 8
1.0x10 9
1.5x10 9
2.0x10 9
2.5x10 9
3.0x10 9
3.5x10 9
4.0x10 9
Minutes0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00
UV at 220 nm
QDa TIC
1
2
3 4
1
2
3 4
(
Minutes6.00 6.20 6.40 6.60
UV
Mass Detector
UPLC/UV and QDa Mass Detection
©2014 Waters Corporation 21
Agrochemicals
UV Chromatogram
Total Ion Chromatogram
Automated Extracted Ion Chromatograms
Combined UV and MS Spectra
1 2 3 4
1 2 3 4
The Empower 3 Mass Analysis Window
©2014 Waters Corporation 22
Agrochemicals
UV Chromatogram
Total Ion Chromatogram
Automated Extracted Ion Chromatograms
Combined UV and MS Spectra
1 2 3 4
1 2 3 4
The Empower 3 Mass Analysis Window
©2014 Waters Corporation 23
Agrochemicals
261.9
261.9
268.0
268.0
nm210.00 220.00 230.00 240.00 250.00 260.00 270.00 280.00 290.00 300.00
342
344
342
344
342
344
342
344
nm260.00 280.00 300.00 320.00 340.00 360.00 380.00 400.00
1
2
3
4
1
2
3
4
UV and Mass Spectra of the Detected Components
©2014 Waters Corporation 24
Agrochemicals
Tebuconazole
Channel Description: PDA 220.0 nm (210-400)nm
2.17
9
4.73
1
5.24
9 6.48
5
AU
-0.20
-0.15
-0.10
-0.05
0.00
0.05
0.10
0.15
QDa 1: MS Scan MS TIC (1: 100.00-600.00 Da ES+, Continuum, CV=5): not integrated, source of MS spectra
Inte
nsity
0
1x108
2x108
3x108
4x108
5x108
6x108
7x108
Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60 4.80 5.00 5.20 5.40 5.60 5.80 6.00 6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40 8.60 8.80 9.00 9.20 9.40 9.60 9.80 10.00
1
2 34
1
23
4
UV
QDa TIC
*UV
QDa TIC
UPLC/PDA/QDa Analysis of a Formulation with two AI’s
©2014 Waters Corporation 25
Channel Description: PDA 220.0 nm (210-400)nm
2.17
9
4.73
1
5.24
9 6.48
5
AU
-0.20
-0.15
-0.10
-0.05
0.00
0.05
0.10
0.15
Channel Description: QDa 1: MS Scan MS Calculated: 256+281+308 (1: 100.00-600.00 Da ES+, Continuum, CV=5)
Inte
nsity
0
1x10 8
2x10 8
3x10 8
4x10 8
Minutes0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00
p ( )
UV at 220 nm
XIC
XIC
2 3
1
4
2 31
4
Agrochemicals
UV
QDa XIC
Extracted Ion Chromatograms
©2014 Waters Corporation 26
Application example – Agrochemicals
2.179 Peak 1
212.4
269.9
4.731 Peak 2220.3
267.4
5.249 Peak 3220.9
269.3
6.485 Peak 4220.9
268.0
nm210.00 220.00 230.00 240.00 250.00 260.00 270.00 280.00 290.00 300.00
256
281
308
308
m/z220.00 240.00 260.00 280.00 300.00 320.00 340.00
1
2
3
4
1
2
3
4
AI
Unknown A
Unknown B
Tebuconazole, AI
UV and Mass Spectra of the Detected Components
©2014 Waters Corporation 27
Application note: 720004426en
Furans in Transfer Oil
©2014 Waters Corporation 28
Transformer oil – Highly-refined mineral oil, stable at high temperatures – Excellent electrical insulating and heat transfer properties – Used in oil-filled transformers, high voltage capacitors,
fluorescent lamp ballasts, high voltage switches, and in circuit breakers
Furans
– Originate from thermal depolymerization of cellulose solid insulation
– Degree of degradation eventually renders transformer ineffective – Periodic analysis for furans can be used to assess the degree of
degradation and does not require taking the unit out of service in order to take a sample
Furans in transformer oil
©2014 Waters Corporation 29
UPLC H-Class System
Mobile phase A: Water (0.1% formic acid)
Mobile phase B: Acetonitrile (0.1% Formic acid)
Inj. volume: 10 µL
Column: ACQUITY BEH C18, 2.1 x 150 mm, 1.7µm
Column temp: 40 °C
Sample temp: 10 °C
Time (min)
Flow Rate (mL/min) %A %B Curve
Initial 0.450 80 20 - 1.50 0.450 80 20 6 2.00 0.450 60 40 6 2.01 0.450 0 100 6 3.00 0.450 0 100 6 3.01 0.450 80 20 6 4.00 0.450 80 20 6
Furans in transformer oil
©2014 Waters Corporation 30
Furans method – ACQUITY PDA Detector
o λ range: 190 – 350 nm o Resolution: 1.2 nm o Sampling rate: 20 points/s
Furans in transformer oil
Furans and additives method – Xevo TQ MS
o Ionisation: APCI+ o Acquisition: MRM o Corona current: 20 µA o Source temp: 150 °C o APCI probe temp: 400 °C o Desolvation gas flow: 1000 L/hr o Cone gas flow: 100 L/hr
©2014 Waters Corporation 31
Furans in transformer oil
Chemical Substance CAS
Number RT
(min) UV absorbance
(nm) 5-hydroxymethyl-2-furaldehyde
5H2F 67-47-0 1.02 284
Furfurylalcohol 2FOL 98-00-0 1.28 216 2-furaldehyde 2FAL 98-01-1 1.45 277 2-furylmethylketone 2ACF 1192-62-7 1.77 274 5-methyl-2-furaldehyde 5M2F 620-02-0 2.13 292
©2014 Waters Corporation 32
Furans in transformer oil
2-furaldehyde (277 nm)
5-methyl-2-furaldehyde (292 nm)
2-furylmethyl-ketone (274 nm)
Furfuryl alcohol (216 nm)
5-hydromethyl-2-furaldehyde (284 nm)
B
A
B
A
B
A
B
A
B
A
MAX Plot
A
UPLC/UV chromatograms
©2014 Waters Corporation 33
Furans in transformer oil
UPLC/UV Empower Results
5-methyl-2-furaldehyde
©2014 Waters Corporation 34
Furans in transformer oil
Compound Replicate injection results
(mg/kg) Average recovery
(%) 1 2
5-hydroxymethyl-2-furaldehyde
Blank ND -- -- 2 mg/kg 1.63 1.64 81.9 10 mg/kg 9.37 9.36 93.6
Furfurylalcohol Blank ND -- --
2 mg/kg 1.89 1.87 93.8 10 mg/kg 9.51 9.38 94.5
2-furaldehyde Blank ND -- --
2 mg/kg 1.90 1.91 95.1 10 mg/kg 9.54 9.55 95.4
2-furylmethylketone Blank ND -- --
2 mg/kg 1.94 1.95 97.3 10 mg/kg 9.59 9.58 95.8
5-methyl-2-furaldehyde Blank ND -- --
2 mg/kg 1.86 1.86 93.2 10 mg/kg 9.96 10.01 99.8
UPLC/UV Recovery Results
©2014 Waters Corporation 35
Furans in transformer oil
Chemical Substance RT
(min)
Cone Voltage
(V)
MRM Transition
Collision energy (eV)
5-hydroxymethyl-2-furaldehyde
5H2F 1.05 20 127.0 > 109.0 15 127.0 > 81.0 20
Furfurylalcohol 2FOL 1.30 25 81.1 > 53.0 15
2-furaldehyde 2FAL 1.46 25 97.1 > 69.0 15 97.1 > 41.1 15
2-furylmethylketone 2ACF 1.79 20 111.1 > 43 20
111.1 > 69.1 15
5-methyl-2-furaldehyde 5M2F 2.15 25 111.1 > 55.0 20 111.1 > 83.0 15
Benzotriazole BTA 1.66 40 120.1 > 65.0 20 120.1 > 92.0 20
©2014 Waters Corporation 36
Furans in transformer oil
Benzotriazole
5-Methyl-2-furaldehyde
2-Furylmethylketone
2-Furaldehyde
Furfuryl alcohol
5-Hydroxymethyl-2-furaldehyde
0
A
A
A
A
A
A
B
B
B
B
B
B
UPLC/MS/MS Chromatogram
©2014 Waters Corporation 37
Impurity profiling of Liquid Crystal intermediates using UltraPerformance Convergence
Chromatography (UPC2) with PDA detection
Application note: 720004743en
©2014 Waters Corporation 38
Background - Liquid Crystals
Properties: o Some properties of liquids:
• Flow, pour like liquids and take the shape of containers o Some optical properties of solids:
• Birefringence • Optical activity
o React predictably to an electric current, enabling the control of light passage
Liquid crystal intermediates: o Building blocks compounds used to prepare liquid crystals
• Used in mixtures (10 to 20 singles used in a typical mixture)
Uses: o Many electronic displays use liquid crystals:
• Watches, calculators, notebooks, personal digital assistant (PDA), mobile phones, projectors, desktops monitors / TVs, viewfinders on cameras / camcorders…..
©2014 Waters Corporation 39
Liquid Crystal Intermediate Compounds
Methods UPC2 conditions Run time: 5.00 min CCM back pressure: 2000 psi Sample temp.: 20 oC Column temp.: 50 oC Flow rate: 2.0 mL/min Column: ACQUITY UPC2 CSH Fluoro-phenyl, 3.0 mm x 100 mm, 1.7 µm Mobile phase A: CO2 Mobile phase B: Methanol (2% Formic Acid + 15 mM ammonium acetate) Injection volume: 1 µL PDA conditions UV system: ACQUITY UPC2 PDA Detector Range : 210 to 450 nm Resolution: 1.2 nm Sampling rate: 20 pts/sec Filter time constant: Slow (0.2 sec)
©2014 Waters Corporation 40
Liquid Crystal Intermediate Compounds
©2014 Waters Corporation 41
Liquid Crystal Intermediate Compounds
Chemical Substance CAS
Number
Retention time
(minutes)
UV optimum absorbance
(nm)
4,4′-Azoxyanisole-d14 39750-11-3 0.69 346
4-Butylbenzoic acid 20651-71-2 1.39 235
4-Octylbenzoic acid 3575-31-3 1.62 235
4-Cyanobenzoic acid 3575-31-3 1.75 252
4-Butoxybenzoic acid 1498-96-0 1.90 252
4-(Octyloxy)benzoic acid 2493-84-7 2.09 235
©2014 Waters Corporation 42
Liquid Crystal Intermediate Compounds
Empower calibration curve
©2014 Waters Corporation 43
Liquid Crystal Intermediate Compounds
UV Chromatograms and UV Spectra
©2014 Waters Corporation 44
Liquid Crystal Intermediate Compounds Impurity Profiling
©2014 Waters Corporation 45
Primary Aromatic Amines
Waters® ACQUITY UPLC H-Class / ACQUITY PDA coupled with SQ Detector 2:
Technology brief: 720004062en Application note: 720004151en
©2014 Waters Corporation 46
Primary Aromatic Amines
Used to produce many commodities – Pharmaceuticals, explosives, epoxy polymers, rubber, aromatic
polyurethane products and azo dyes Found as impurities in the final product
– Due to incomplete reactions, as by-products, or as degradation products PAAs can be produced as by-products of azo-dyes Azo-dyes
– A diverse and widely used group of organic dyes – Wide range of uses
o Specialty paints, printing inks, varnishes and adhesives – Found in many consumer products
o Textiles, cosmetics, plastics Risks to human health
– Proven / suspected carcinogenic nature, highly toxic
©2014 Waters Corporation 47
SQ Detector 2: Ionization mode: ESI+ Acquisition mode: SIR ACQUITY PDA Detector: Wavelength range: 190 – 500 nm
Primary Aromatic Amines
PAA number Primary Aromatic Amines (PAAs) CAS
Number m/z Retention
time (minutes)
Cone Voltage
(V) 1 Aniline 62-53-3 94 2.17 40 2 o-Toluidine 95-53-4 109 3.80 40 3 1,3-Phenylenediamine 108-45-2 109 0.62 40 4 1,4-Phenylenediamine 106-50-3 109 0.41 43 5 2,4-Dimethylaniline 95-68-1 122 5.58 43 6 2,6-Dimethylaniline 87-62-7 122 5.33 43 7 2,4-Toluenediamine 95-80-7 123 1.64 40 8 2,6-Toluenediamine 823-40-5 123 0.85 40 9 o-Anisidine 90-04-0 124 3.74 45 10 4-Chloroaniline 106-47-8 128 4.6 40 11 2,4,5-Trimethylaniline 137-17-7 136 7.06 40 12 2-Methoxy-5-methylaniline 120-71-8 138 5.36 40 13 4-Methoxy-m-phenylenediamine 615-05-4 139 1.51 36 14 2-Naphtylamine 91-59-8 144 6.18 40 15 3-Amino-4-methylbenzamide 19406-86-1 151 2.19 35 16 3-Chloro-4-methoxyaniline 5345-54-0 158 4.00 40 17 5-Chloro-2-methoxyaniline 95-03-4 158 6.06 40 18 1,5-Diaminonaphtalene 2243-62-1 159 2.52 40 19 2-Methoxy-4-nitroaniline 97-52-9 169 4.37 30 20 4-Aminobiphenyl 92-67-1 170 7.57 43 21 2-Aminobiphenyl 90-41-5 170 7.71 50 22 Benzidine 92-87-5 185 4.01 43 23 4-Chloro-2,5-dimethoxyaniline 6358-64-1 188 5.79 40 24 4-Aminoazobenzol 60-09-3 198 7.84 30 25 4,4'-Methylenedianiline 101-77-9 199 5.64 43 26 4,4'-Diaminodiphenylether 101-80-4 201 4.36 45 27 3,3'-Dimethylbenzidine 119-93-7 213 6.01 43 28 4,4'-Thioaniline 139-65-1 217 6.29 43 29 o-Aminoazotoluene 97-56-3 226 8.28 43 30 4,4'- Diamino-3,3'-dimethylbiphenylmethane 838-88-0 227 7.39 40 31 3-Amino-p-anisanilide 120-35-4 243 6.06 40 32 o-Dianisidine 119-90-4 245 6.00 45 33 3,3'-Dichlorobenzidine 91-94-1 253 7.76 45 34 4,4'- Diamino-3,3'-dichlorobiphenylmethane 101-14-4 267 7.90 60
SIR m/z, expected retention times, and cone voltages
©2014 Waters Corporation 48
Primary Aromatic Amines TargetLynx Quantify results browser showing the calibration quantitation results, calibration curve and example SIR chromatogram for Aniline
©2014 Waters Corporation 49
UPLC H-Class System Mobile phase A
10 mL of 1 M aqueous ammonium acetate solution and 990 mL water
Mobile phase B 10 mL of 1 M aqueous ammonium acetate solution and 990 mL methanol
Column: ACQUITY BEH C18, 1.7 mm, 2.1 x 50 mm
Column temperature: 40 oC.
Primary Aromatic Amines SIR chromatograms for 34 PAAs in a mixed 1 µg/mL calibration standard
©2014 Waters Corporation 50
Primary Aromatic Amines Ink analysis Neat ink diluted 1:100 with 5%
methanol / 95% water
Ink spiked at various levels with selected PAAs, and analyzed without any further cleanup or concentration steps
The efficient recoveries obtained (ranging between 83 to 108%)
– minimal signal enhancement / suppression was observed
Ink spiked with PAAs recovery data.
©2014 Waters Corporation 51
Primary Aromatic Amines The advantages of mass spectral detection over PDA (UV) detection
– Improved sensitivity and selectivity – Matrix effects can be greatly reduced by using mass spectral detection
a) UV chromatograms for 2-Aminobiphenyl and 3,3’-Dichlorobenzidine in individual solvent standards
b) In a mixed solvent standard
Example 1 (selectivity) Considering the PAAs,
2-Aminobiphenyl and 3,3’-Dichlorobenzidine Maximum UV absorbance
can be found at 295 and 284 nm respectively (retention times of 7.71 and 7.76 minutes respectively)
The two compounds are not completely resolved.
Which could potential lead to misidentification, poor integration, and false positive results
©2014 Waters Corporation 52
Primary Aromatic Amines
Mass spectral detection The two compounds are
resolved Improved selectivity
Extracted ion chromatograms for 2-Aminobiphenyl and 3,3’-Dichlorobenzidine in fortified ink (containing 4.6 µg/mL PAAs)
©2014 Waters Corporation 53
Primary Aromatic Amines Improvement in selectivity
– The abilities to measure analytes of interest accurately and specifically in the presence of a complex matrix
– Considering the PAA 2,4,5-Trimethylaniline
o When spiked in ink cannot be distinguished due to other UV absorbing compounds present
– However, mass detection is sufficiently sensitive and selective to enable confident detection and quantification of 2,4,5-Trimethylaniline in an ink matrix
UV and extracted ion chromatograms for the PAA 2,4,5-Trimethylaniline spiked in ink (4.6 µg/mL), solvent
standard (5.0 µg/mL) and a blank ink matrix
Blank ink matrix UV:286 nm
Spiked ink UV:286 nm
Solvent standard UV:286 nm
Blank ink matrix m/z:136
Spiked ink m/z:136
Solvent standard m/z:136
©2014 Waters Corporation 54
Primary Aromatic Amines
Improvements in sensitivity – Signal-to-noise (S/N),
(comparing the UV and the mass spectral data)
– The increase in S/N and sensitivity when using mass spectral data
UV and extracted ion chromatograms for 4-4’-Diaminodiphenylether
©2014 Waters Corporation 55
Conclusion
By utilizing Waters technology –Fast, selective, and sensitive methods can be
developed for the analysis of impurities Offering many business benefits using UPLC and UPC2
o Increase in sample throughput o Reduction in toxic solvent usage
Using mass spectral detection over UV detection provides – Improvement in sensitivity and selectivity –Reduced matrix effects
PDA and mass detection provide complementary information for peak assignment and structural confirmation of impurities
©2014 Waters Corporation 56