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Table of Contents

Cover

Title

Preface

The Structure of HPLC-MS for Practitioners

List of Contributors

Part I: Overview, Pitfalls, Hardware-Requirements

1 State of the Art in the LC/MS
1. 1.1 Introduction
2. 1.2 Ionization Methods at Atmospheric Pressure
3. 1.3 Mass Analyzer
4. 1.4 Future Developments
5. 1.5 What Should You Look for When Buying a Mass Spectrometer?
6. References
2 Technical Aspects and Pitfalls of LC/MS Hyphenation
1. 2.1 Instrumental Requirements for LC/MS Analysis – Configuring the Right System for Your Analytical Challenge
2. 2.2 LC/MS Method Development and HPLC Method Adaptation – How to Make My LC Fit for MS?
3. 2.3 Pitfalls and Error Sources – Sometimes Things Do Go Wrong
4. 2.4 Conclusion
5. 2.5 Abbreviations
6. References
3 Aspects of the Development of Methods in LC/MS Coupling
1. 3.1 Introduction
2. 3.2 From Target to Screening Analysis
3. 3.3 The Optimization of Parameters in Chromatography and Mass Spectrometry
4. References

Part II: Tips, Examples, Trends

4 LC/MS for Everybody/for Everything? – LC/MS Tips
1. 4.1 Introduction
2. 4.2 Tip Number 1
3. 4.3 Tip Number 2
4. 4.4 Tip Number 3
5. 4.5 Tip Number 4
6. 4.6 Tip Number 5
7. 4.7 Tip Number 6
8. 4.8 Tip Number 7
9. 4.9 Tip Number 8
10. 4.10 Need More Help?
11. References

Part III: User Reports

5 LC Coupled to MS – a User Report
1. References
6 Problem Solving with HPLC/MS – a Practical View from Practitioners
1. 6.1 Introduction and Scope
2. 6.2 Case Example 1
3. 6.3 Case Example 2
4. 6.4 Case Example 3
5. 6.5 Case Example 4
7 LC/MS from the Perspective ofaMaintenance Engineer
1. 7.1 Introduction and Historical Summary
2. 7.2 Spray Techniques
3. 7.3 Passage Through the Ion Path
4. 7.4 The Analyzer
5. 7.5 Maintenance
6. References

Part IV: Vendor’s Reports

8 LC/MS – the Past, Present, and Future
9 Vendor’s Report – SCIEX
10 Manufacturer Report – Thermo Fisher Scientific
1. 10.1 Liquid Chromatography for LC/MS
2. 10.2 Mass Spectrometry for LC/MS
3. 10.3 Integrated LC/MS Solutions
4. 10.4 Software
5. References

About the Authors

Index

End User License Agreement

List of Tables

2 Technical Aspects and Pitfalls of LC/MS Hyphenation

Table 2.1 Volumes and backpressure of a 30 inches/750 mm capillary with different inner diameters (I.D.) in the viscosity maximum of water/acetonitrile and water/methanol mixtures.
Table 2.2 Recommended re-equilibration volume for high-throughput and high-resolution columns under typical MS-compatible conditions.
Table 2.3 Suitability and purpose of various mass spectrometer types; + = well-suited, o = moderately suitable, – = inappropriate.
Table 2.4 Applicable and ideal working ranges for selected ionization processes.
Table 2.5 Common gas phase adducts at positive (left) and negative (right) polarity. Mass differences refer to the difference between [M + H]⁺ (left) or [M − H]⁻ (right), respectively, and the related gas phase adduct.

3 Aspects of the Development of Methods in LC/MS Coupling

Table 3.1 Overview of the different LC/MS workflows.
Table 3.2 A list of the target compounds with important physicochemical parameters [9].
Table 3.3 A list of the stationary phases selected for the screening.
Table 3.4 Comparative overview of the effective column volumes depending on the length and diameter of HPLC columns.
Table 3.5 Resulting source parameters depending on the evaluation strategy used.

List of Illustrations

1 State of the Art in the LC/MS

Figure 1.1 Analysis of saffron using direct-inlet probe-APCI with high-resolution QTOF-MS. (a) TIC of the toal analysis. (b) mass spectrum at the time of 2.7 min.
Figure 1.2 Polarity range of analytes for ionization with various atmospheric pressure ionization (API) techniques. Note: The extended mass range of APLI against APPI and APCI results from the ionization of nonpolar aromatic analytes in an electrospray Reproduced with kind permission of O. J. Schmitz, T. Benter, Advances in LC-MS Instrumentation: Atmospheric pressure laser ionization, Journal of Chomatography Libary, Vol 72 (2007), Chapter 6, Pages 89-113.
Figure 1.3 Reduction of the droplet size.
Figure 1.4 Reaction mechanism in APCI.
Figure 1.5 Ion suppression in APCI-MS of PAH in urine.
Figure 1.6 Analysis of a mixture of glucose and fructose with IM-qTOF-MS.

2 Technical Aspects and Pitfalls of LC/MS Hyphenation

Figure 2.1 Illustration of the gradient delay volume (GDV) and the extra-column volume (ECV) of an LC system.
Figure 2.2 Descending top-down (a) and ascending bottom-up (b) flow path for minimized connection tubing length between LC column outlet and MS inlet (ion source).
Figure 2.3 LC/MS chromatogram of two isomers, m/z = 240.10; (a) PEEK bulk capillary behind the column (0.13 mm I.D.), PEEK fingertight fittings; (b) SST capillary with virtually zero-dead volume connection behind the column (Viper™ fingertight fitting technology, 0.13 mm I.D.)
Figure 2.4 Reduction of re-equilibration time and throughput enhancement by using a second separation column and alternating sample injection (Tandem LC); (a) flow scheme, (b) injection interlacing.
Figure 2.5 Distinction of isobaric compounds by RP chromatography and UV detection for a reaction control analysis (N-arylation of an E/Z acrylic ester mixture.)
Figure 2.6 Signal intensity of Leu-enkephalin dissolved in various common LC/MS solvents; data acquired in ESI(+) mode during syringe pump infusion [12].
Figure 2.7 Separation of a cytochrome C digest by adding 0.05% TFA (a) or 0.1% FA (b) under identical chromatographic conditions
Figure 2.8 Influence of the dry gas temperature on MS signal quality using FIA of astemizol in 10 mM aqueous ammonium acetate/methanol 20/80 (v/v) at 50 μL/min on a triple quad instrument; (a) extracted ion chromatogram of astemizol ([M + H]⁺), m/z 459.3; (b) reconstructed total ion chromatogram.
Figure 2.9 Mass spectrum of two co-eluting compounds m₁ and m₂, once measured with a low-resolving (a) and a high-resolving (b) mass spectrometer.
Figure 2.10 Mass spectrum of a single (a, b) and fourfold charged (c, d) compound, once measured with a low-resolving (a, c) and a high-resolving (b, d) mass spectrometer.

3 Aspects of the Development of Methods in LC/MS Coupling

Figure 3.1 Structural formulae of (a) cyclophosphamide (CP) and (b) ifosfamide (IF).
Figure 3.2 Structural formulae of (a) epirubicin and (b) doxorubicin.
Figure 3.3 Matrix effect chromatogram of a house dust extract. The selected mass transitions of different mycotoxins (nonsmoothed raw data) are presented. For further explanations, see the text.
Figure 3.4 Structural formulae of the target analytes 5-fluorouracil (1), gemcitabine (2), methotrexate (3), topotecan (4), irinotecan (5), ifosfamide (6), cyclophosphamide (7), doxorubicin (8), epirubicin (9), etoposide (10), paclitaxel (11), and docetaxel (12).
Figure 3.5 Graphic representation of the experimental design for determining the appropriate phase system.
Figure 3.6 Schematic layout of the HPLC system used for partially automated screening.
Figure 3.7 Comparative separation of 12 cytostatic drugs on (a) Supelco Ascentis Express C-8 (50 × 2.1 mm, 2.7 μm), (b) Restek Raptor ARC-18 (50 × 2.1 mm, 2.7 μm), (c) ChromaNik SunShell RP-Aqua C-28 (50 × 2.1 mm, 2.6 μm) columns; chromatographic parameters: temperature: 30 °C injection volume: 2 μL; mobile phase: A = water + 0.1% formic acid, B = acetonitrile + 0.1% formic acid; flow rate: 350 μL min^–1; detection: UV at 200 nm and 254 nm.
Figure 3.8 Comparative separation of 12 cytostatic drugs on (a) Supelco Ascentis Express RP-amide (50 × 2.1 mm, 2.7 μm), (b) Restek Raptor biphenyl (50 × 2.1 mm, 2.7 μm), (c) TCI Kaseisorb LC ODS-SAX Super (50 × 2.0 mm, 3.0 μm) columns; chromatographic parameters: temperature: 30 °C; injection volume: 2 μL; mobile phase: A = water + 0.1% formic acid, B = acetonitrile + 0.1% formic acid; flow rate: 350 μL min^–1; detection: UV at 200 nm and 254 nm.
Figure 3.9 Comparative separation of 12 cytostatic drugs on (a) Supelco Ascentis Express C-8 (50 × 2.1 mm, 2.7 μm), (b) Restek Raptor ARC-18 (50 × 2.1 mm, 2.7 μm), (c) ChromaNik SunShell RP-Aqua C-28 (50 × 2.1 mm, 2.6 μm) columns; chromatographic parameters: temperature: 30 °C; injection volume: 2 μL; mobile phase: A = water + 0.1% formic acid, B = methanol + 0.1% formic acid; flow rate: 350 μL min^–1; detection: UV at 200 nm and 254 nm.
Figure 3.10 Comparative separation of 12 cytostatic drugs on (a) Supelco Ascentis Express RP-amide (50 × 2.1 mm, 2.7 μm), (b) Restek Raptor biphenyl (50 × 2.1 mm, 2.7 μm), (c) TCI Kaseisorb LC ODS-SAX Super (50 × 2.0 mm, 3.0 μm) columns; chromatographic parameters: temperature: 30 °C; injection volume: 2 μL; mobile phase: A = water + 0.1% formic acid, B = methanol + 0.1% formic acid; flow rate: 350 μL min^–1; detection: UV at 200 nm and 254 nm.
Figure 3.11 Comparative separation of 12 cytostatic drugs on an Supelco Ascentis Express C-8 (50 × 2.1 mm, 2.7 μm) column; chromatographic parameters: (a, b) temperature: 30 °C, 50 °C; injection volume: 2 μL; mobile phase: A = water + 0.1% formic acid, B = acetonitrile + 0.1% formic B = methanol + 0.1% formic acid; flow rate: 350 μL min^–1; detection: UV a 200 nm and 254 nm; (c, d) temperature: 30 °C, 50 °C; injection volume: 2 μL; mobile phase: A = water + 0.1% formic acid, B=methanol+0.1% formic acid; flow rate: 350 μL min^–1; detection: UV at 200 nm and 254 nm.
Figure 3.12 Comparative separation of 12 cytostatic drugs on an Agilent Zorbax SB C-18 (50 × 2.1 mm, 1.8 μm) column; chromatographic parameters: (a) solvent gradient: in 30 min from 1 to 99% B, (b) solvent gradient: in 10 min from 1 to 99% B; temperature: 30 °C; injection volume: 5 μL; mobile phase: A = water + 0.1% formic acid, B = acetonitrile + 0.1% formic acid; flow rate: 350 μL min^–1; detection: UV at 200 nm and 254 nm.
Figure 3.13 Comparative separation of 12 cytostatic drugs on a ChromaNik SunShell RP-Aqua C-28 (50 × 2.1 mm, 2.6 μm) column; chromatographic parameters: (a) temperature: 40 °C; injection volume: 100 μL; mobile phase: A = water + 0.1% acetic acid, B = acetonitrile + 0.1% acetic acid; flow rate: 350 μL min^–1; (b) temperature: 40 °C; injection volume: 100 μL; mobile phase: A = water + 0.1% formic acid, B = acetonitrile + 0.1% formic acid; flow rate: 350 μL min^–1; detection: MS – multiple reaction monitoring.
Figure 3.14 Typical system set-up in LC/MS coupling.
Figure 3.15 Optimized system configuration for coupling a flexible HPLC system with the mass spectrometer.
Figure 3.16 The inner diameter of the column plotted against the linear velocity of the mobile phase for a constant flow rate of 0.5 mL min^–1.
Figure 3.17 UV spectra of (a) cyclophosphamide and (b) ifosfamide.
Figure 3.18 Schematic representation for the “normal” injection: (a) HPLC column, (b) chromatogram.
Figure 3.19 Schematic representation of a large volume direct injection for (a) injection from aqueous phase and (b) injection from organic phase. The injection plug takes up approximately half of the volume available to the mobile phase in the column.
Figure 3.20 Separation of 12 cytostatic drugs on a ChromaNik SunShell RP-Aqua C-28 (50 × 2.1 mm, 2.6 μm) column; chromatographic parameters: temperature: 40 °C; injection volume: 5 μL; mobile phase: A = water + 0.1% acetic acid, B = acetonitrile + 0.1% acetic acid; flow rate: 350 μL min^–1; detection: MS – multiple reaction monitoring; composition of the injection solution: 30/70 (v/v) water/isopropanol. The dark area is the elution band of irinotecan.
Figure 3.21 Separation of 12 cytostatic drugs on a ChromaNik SunShell RP-Aqua C-28 (50 × 2.1 mm, 2.6 μm) column; chromatographic parameters: temperature: 40 °C; injection volume: 10 μL; mobile phase: A = water + 0.1% acetic acid, B = acetonitrile + 0.1% acetic acid; flow rate: 350 μL min^–1; detection: MS – multiple reaction monitoring; composition of the injection solution: 30/70 (v/v) water/isopropanol. The dark area is the elution band of irinotecan.
Figure 3.22 Separation of 12 cytostatic drugs on a ChromaNik SunShell RP-Aqua C-28 (50 x 2,1 mm, 2,6 μm) column; chromatographic parameters: temperature: 40 °C; injection volume: 100 μL; mobile phase: A = water + 0.1% acetic acid, B = acetonitrile + 0.1% acetic acid; flow rate: 350 μL min^–1; detection: MS – Multiple Reaction Monitoring; composition of the injection solution: 93/7 (v/v) water/isopropanol.
Figure 3.23 Separation of four cytostatic drugs (a) without and (b) with online enrichment. Chromatographic conditions: stationary phase: (a) Waters XBridge C-18 (50 × 2.1 mm, 3.5 μm), (b) Thermo Hypercarb (10 × 2.1 mm, 5 μm) coupled with a Waters XBridge C-18 (50 × 2.1 mm, 3.5 μm) column; mobile phases: A) water with 0.1% trifluoroacetic acid, B) acetonitrile with 0.1% trifluoroacetic acid; flow rate: 0.5 mL min^–1; gradient: 0 to 90% B in 10 min, 10–20 min, 100% B; injection volume: 1000 μL; temperature: 35 °C; detection: MS. analytes: 1) gemcitabine, 2) ifosfamide, 3) cyclophosphamide, 4) fenofibrate.
Figure 3.24 Separation of four cytostatic drugs. Chromatographic conditions: stationary phase: Thermo Hypercarb (10 × 2.1 mm, 5 μm) coupled with a Waters XBridge C-18 (50 × 2.1 mm, 3.5 μm) column; mobile phase: A) Water with 0.1% trifluoroacetic acid, B) acetonitrile with 0.1% trifluoroacetic acid; flow rate: 0.5 mL min^–1; gradient: 0 to 90% B in 10 min, 10–20 min, 100% B; injection volume: see diagram; temperature: 35 °C; detection: MS. Absolute amount of substance on column: 25 ng. Analytes: 1) gemcitabine, 2) ifosfamide, 3) cyclophosphamide, 4) fenofibrate.
Figure 3.25 System configuration of the online SPE/LC/MS/MS coupling: 1) HPLC pumps, 2) PAL RTC autosampler, 3) automatic cartridge exchanger, 4) HPLC oven, 5) tandem mass spectrometer.
Figure 3.26 Comparison of the chromatograms of an enriched sample on (a) resin SH cartridge and (b) C-18 cartridge.
Figure 3.27 Comparison of the recovery rates of online SPE (C-18 cartridge, n = 3) and offline SPE (Waters HLB cartridge, n = 5) for a waste water-multianalyte method with 25 pharmaceuticals and an injection volume of 10 mL in the online SPE.
Figure 3.28 Time schedule for sample preparation in manual offline SPE and the automated online SPE.
Figure 3.29 Resulting intensities for the two mass transitions of each analyte depending on the set source temperature.
Figure 3.30 Resulting intensities for the two mass transitions of each analyte depending on the setting of the volume stream of the nebulizing gas with source temperature of (a) 450 °C and (b) 250 °C.
Figure 3.31 LC/MS/MS chromatogram of 182 mass transitions on a 50 × 2 mm Merck Chromolith Fast Gradient RP18 HPLC column. Chromatographic parameters: temperature: 40 °C; injection volume: 20 μL; mobile phases: A = water + 0.1% formic acid, B = methanol + 0.1% formic acid; gradient: 5 to 95% B in 20 min; flow rate: 400 μL min^–1; mass spectrometric parameters: pause time: 5 ms; dwell time: 100 ms (unsmoothed raw data).
Figure 3.32 LC/MS/MS chromatogram of 182 mass transitions on a 50 × 2 mm Merck Chromolith Fast Gradient RP18 HPLC column. Chromatographic parameters: temperature: 40 °C; injection volume: 20 μL; mobile phases: A = water + 0.1% formic acid, B = methanol + 0.1% formic acid; gradient: 5 to 95% B in 20 min; flow rate: 400 μL min^–1; mass spectrometric parameters: pause time: 5 ms, dwell time: 10 ms (unsmoothed raw data).
Figure 3.33 Extracted LC/MS/MS chromatogram of 182 mass transitions on a 50 × 2 mm Merck Chromolith Fast Gradient RP18 HPLC column. Chromatographic parameters: temperature: 40 °C; injection volume: 20 μL; mobile phases: A = water + 0.1% formic acid, B = methanol + 0.1% formic acid; gradient: 5 to 95% B in 20 min; flow rate: 400 μL min^–1; mass spectrometric parameters: pause time: 5 ms, MRM detection window: 60 s, target scan time: 2 s (unsmoothed raw data).
Figure 3.34 Comparative representation of the principle of different MRM modes. (a) Retention-time-independent MRM mode; (b) retention-time-independent MRM mode with the division of the chromatogram in periods; (c) retention-time-independent MRM mode with variable detection windows.
Figure 3.35 General workflow for an LC/MS and MS/MS screening analysis.
Figure 3.36 Comparison of (a) the obtained peak width using a conventional (black line) and highly efficient (red line) LC separation. In addition, the corresponding cycle times of combined MS full scan and MS/MS acquisition data are shown for (b) four and (c) eight information dependent MS/MS experiments.
Figure 3.37 Comparison of commercially available emitter tips. (a) Classical emitter tip with an inner diameter of 100 μm, compatible with 1/16^′′ fittings; (b) miniaturized emitter tip with an inner diameter of 25 μm, compatible with 1/32^′′ fittings.
Figure 3.38 Experimental setup for the separation of pharmaceuticals with a monolithic nano-HPLC column (Merck CapROD 150 × 0.1 mm).
Figure 3.39 LC/MS/MS chromatogram of a separation of 50 pharmaceuticals on (a) a 50 × 2 mm Merck Chromolith Fast Gradient RP18 column; chromatographic parameters: temperature: 40 °C injection volume: 10 μL; mobile phases: A = water + 0.1% formic acid, B = acetonitrile + 0.1% formic acid; flow rate: 500 μL min^–1; mass spectrometric parameters: pause time: 5 ms; dwell time: 20 ms; (b) a 150 x 0.1 mm Merck Chromolith CapROD C-18 column; chromatographic parameters: temperature: room temperature; injection volume: 100 nL; mobile phases: A = water + 0.1% formic acid, B = acetonitrile + 0.1% formic acid; flow rate: 5 μL min^–1; mass spectrometric parameters: pause time: 5 ms; dwell time: 20 ms.

4 LC/MS for Everybody/for Everything? – LC/MS Tips

Figure 4.1 Application ranges of API techniques.
Figure 4.2 Schematic representation of an electrospray ion source.
Figure 4.3 Electrospray mass spectrum of myoglobin.
Figure 4.4 Schematics of an APCI ion source.
Figure 4.5 Schematics of an APPI ion source.
Figure 4.6 Influence of trifluoroacetic acid (TFA) on LSD (lysergic acid diethyl amide) and LAMPA (lysergic acid methylpropyl amide).
Figure 4.7 Adduct formation in ESI.
Figure 4.8 APCI adducts of prednisolone.
Figure 4.9 ESI calibration curves of o-toluidine and 3,3^′-dimethylbenzidine.
Figure 4.10 APCI calibration curves of o-toluidine and 3,3^′-dimethylbenzidine.
Figure 4.11 Signal/noise as a function of MSⁿ stages.
Figure 4.12 Ionization principle at atmospheric pressure.
Figure 4.13 Drop of signal intensity after extreme matrix stress of an ion source.
Figure 4.14 Comparison of solvent standard with matrix spike.
Figure 4.15 Extended evaluation of the matrix effect.
Figure 4.16 Method optimization by analyte infusion.
Figure 4.17 Evaluation of the matrix effect by analyte infusion.

5 LC Coupled to MS – a User Report

Figure 5.1 (a) Schematic representation of the quadrupoles of a triple quadrupole instrument. Only two quadrupole rods of the four quadrupole rods are shown for each quadrupole. The first quadrupole (Q1) filters for the precursor ion, the precursor ion is fragmented in Q2 by collision induced dissociation and Q3 filters for a specific daughter ion. (b) Isomers of hydroxyeicosatetraenoic acid (HETE). (c) Schematic representation of ion traces by measuring just the precursor ions or daughter ions of HETE isomers. Reproduced with kind permission of Novartis.
Figure 5.2 Chromatographic separation of four hydrophilic molecules by IC and detection with a triple quadrupole MS instrument. Reproduced with kind permission of Novartis.

6 Problem Solving with HPLC/MS – a Practical View from Practitioners

Figure 6.1 Wrong choice of ionization mode can lead to incorrect conclusions in stability tests, for details see text. Reproduced with kind permission of MCC.
Figure 6.2 The comparison of chromatograms with MS, DAD, and ELSD detection increases the amount of information and, thereby, the accuracy of statements regarding structure and purity. Here: Methohexital Imp. 236. Reproduced with kind permission of MCC.
Figure 6.3 The comparison of chromatograms with MS, DAD, and ELSD detection increases the amount of information and, thereby, the accuracy of statements regarding structure and purity. Here: Tribenuron Impurity 1. Reproduced with kind permission of MCC.
Figure 6.4 Chromatogram of a component after synthesis optimization; comparison between three different detector traces (DAD, ELSD, MS-TIC). Reproduced with kind permission of MCC.
Figure 6.5 Structures of the dopamine impurities.

7 LC/MS from the Perspective ofaMaintenance Engineer

Figure 7.1 Micromass Quattro Ultima Pt, pollution after the first ion tunnel.
Figure 7.2 Micromass Quattro Ultima Pt, contamination after the second ion tunnel.
Figure 7.3 Black shadows show contamination of the components of the ion block.
Figure 7.4 Micromass Quattro Premier XE, extremely corroded ion source.
Figure 7.5 Agilent MSD, ion burn on a hyperbolic quartz quadrupole.
Figure 7.6 Micromass Quattro Micro ion source.
Figure 7.7 Rusty oil–water mixture from the backing pump.

9 Vendor’s Report – SCIEX

Figure 9.1 QTRAP^® 6500 System ion path: curved collision cell with orifice (OR), QJet (Q0) for ion transfer, RF quadrupoles, and detector. Reproduced with kind permission of SCIEX.
Figure 9.2 Comparison of “traditional” (a), fast (b), and UHPLC gradients (c). Reproduced with kind permission of SCIEX.
Figure 9.3 MRM transition 331/127 can be taken for quantitation, while the MRM transition 331/99 shows matrix interference. MRM³ transition 331∕99∕71 has almost no matrix interference. Reproduced with kind permission of SCIEX.
Figure 9.4 Extracted ion chromatogram (XIC) from a sample (a) compared with an XIC from a control standard (b). Identification with exact mass, retention time, isotopic pattern and the calculated sum formula (c). MS spectrum (d) and MS/MS spectrum (e) in comparison to a MS/MS spectrum from the spectral library. Reproduced with kind permission of SCIEX.

Preface

LC/MS coupling has developed from a method for experts in research to a wellproven technique for users in their daily routine. Hence, this book is dedicated exclusively to LC/MS coupling.

It is our goal to give LC/MS users detailed information in order to use their LS/MS application in an optimal manner. Colleagues who have authored articles in my previous books have therefore revised and updated their articles. Furthermore, new articles from LC/MS practitioners were added. When writing those articles, it was most important to us to have an eye on practice, but compact background knowledge is also given. I hope that the analyst in development as well as the user in daily routine will find inspiration and tips for optimal usage of LC/MS coupling.

My special thanks goes to Wolfgang Dreher for his critical comments to this manuscript, furthermore to my author colleagues, who put down their experience and knowledge in writing despite their limited time resources. I would like to thank WILEY-VCH and in particular Reinhold Weber and Martin Preuss for the good and close cooperation.

Blieskastel, March 2017

Stavros Kromidas

The Structure of HPLC-MS for Practitioners

The book contains ten chapters that are divided into four parts:

1–3: Part I Overview, Pitfalls, Hardware Requirements
4: Part II Tips, Examples
5–7: Part III User Reports
8–10: Part IV Vendor’s reports, Trends

Part I

In Chapter 1 Oliver Schmitz overviews the State of the art of LC/MS coupling and opposes different modes. In Chapter 2 Markus Martin shows Technical aspects and pitfalls of LC/MS hyphenation and provides precise and specific hints how LC/MS coupling can successfully be established in daily routine. Other topics of Chapter 2 are method development as well as method transfer. Thorsten Teutenberg and co-authors provide a great many of suggestions in Chapter 3 (Requirements of LC hardware for the coupling of different mass spectrometers), as to arrange LC/MS coupling as optimal as possible. Among other things complex samples and miniaturization play an important role.

Part II

In Chapter 4 Friedrich Mandel offers a numerous of LC/MS Tips addressing different topics of LC/MS coupling.

Part III

Chapter 5 contains examples and experience reports from users and service engineers: LC/MS coupling is often linked to life science and environmental analysis. Alban Muller and Andreas Hofmann show in Chapter 5 a concrete example of LC/MS coupling in ion chromatography as an unfamiliar application. In Chapter 6 Edmond Fleischer shows on the basis of 4 examples coming from the field of synthesis how to proceed if characterization of impurities are on focus (Problem solving with HPLC-MS – a practical view from practitioners). Oliver Müller (LC/MS from the perspective of a maintenance engineer) undertakes a virtual walk across a MS and gives hints how to handle the problem “impurities in LC/MS”.

Part IV

Finally, in Part IV (Chapter 8–10, Report of device manufacturers – article by Agilent, SCIEX, and ThermoScientific) three manufacturers introduce briefly their newest products and evaluate the future of HPLC-MS coupling.

We think the style and structure of The HPLC Expert has proven itself, so those were kept the same in the subsequent book: the book need not to be read linearly. All chapters present self-contained modules – “jumping” between chapters is always possible. That way we try to keep the character of the book as a reference book for LC/MS users. The reader may benefit therefrom.

List of Contributors

Dr.Claudia vom Eyser

Institut für Energie- und
Umwelttechnik e. V.
Bliersheimer Straße 58–60
47229 Duisburg
Germany

Dr. EdmondFleischer

MicroCombiChem e. K.
Rheingaustraße 190–196
Building E512
65203 Wiesbaden
Germany

Terence Hetzel

Institut für Energie- und
Umwelttechnik e. V.
Bliersheimer Straße 58–60
47229 Duisburg
Germany

Dr.AndreasHofmann

Novartis
Institutes for BioMedical Research
Novartis Campus
4056 Basel
Switzerland

Dr.FriedrichMandel

Friedrich-Speidel-Straße 43
76307 Karlsbad
Germany

Dr.Markus M.Martin

Thermo Fischer Scientific
Dornierstraße 4
82110 Germering
Germany

Oliver Müller

Fischer Analytics GmbH
Duhlwiesen 32
55413 Weiler bei Bingen
Germany

AlbanMuller

Novartis Institutes for BioMedical
Research
Novartis Campus
4056 Basel
Switzerland

Dr.Christoph Portner

Tauw GmbH
Richard-Löchel-Straße 9
47441 Moers
Germany

Dr. Detlev Schleuder

AB SCIEX Germany GmbH
Landwehrstraße 54
64293 Darmstadt
Germany

Prof. Dr. Oliver J. Schmitz

University of Duisburg–Essen
Faculty of Chemistry
Applied Analytical Chemistry
Campus Essen, S05 T01 B35
Universitätsstr. 5
45141 Essen
Germany

Dr.TerrySheehan

Director MS Business Development
Agilent Technologiesy
5301 Stevens Creek Blvd, 3U-WI
Santa Clara, CA 95051
USA

Dr.ThorstenTeutenberg

Institut für Energie- und
Umwelttechnik e. V.
Bliersheimer Straße 58–60
47229 Duisburg
Germany

Dr.Jochen Türk

Institut für Energie- und
Umwelttechnik e. V.
Bliersheimer Straße 58–60
47229 Duisburg
Germany

Dr.Steffen Wiese

Institut für Energie- und
Umwelttechnik e. V.
Bliersheimer Straße 58–60
47229 Duisburg
Germany

The HPLC-MS Handbook for Practitioners

Editor

Preface

The Structure of HPLC-MS for Practitioners

Part I

Part II

Part III

Part IV

List of Contributors

Part I
Overview, Pitfalls, Hardware-Requirements