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<p>List of Contributors xiii</p> <p>Foreword xv</p> <p>The Structure of “The HPLC-Expert 2” xvii</p> <p><b>1 When Should I UseMy UHPLC as a UHPLC? 1</b><br /><i>Stavros Kromidas</i></p> <p>1.1 Introduction 1</p> <p>1.2 What Do IWant to Achieve and What Is a UHPLC Capable of? 2</p> <p>1.3 What Is Required from an HPLC Method? 3</p> <p>1.4 The UHPLC in Routine Use – A Brief Report 17</p> <p>1.5 How Can the Potential of UHPLC Effectively Be Fully Exploited? (See Also Chapters 2, 3, and 9) 20</p> <p>1.6 Summary and Outlook 22</p> <p>References 25</p> <p><b>Part I Hardware and Software, Separation Modes, Materials 27</b></p> <p><b>2 The Modern HPLC/UHPLC Device 29</b></p> <p>2.1 The Modern HPLC/UHPLC System 29<br /><i>Steffen Wiese and Terence Hetzel</i></p> <p>Acknowledgment 50</p> <p>References 50</p> <p>2.2 TheThermostate of Columns – A Minor Matter 52<br /><i>Michael Heidorn and Frank Steiner</i></p> <p><b>3 The Issue of External Band Broadening in HPLC/UHPLC Devices 73</b><br /><i>Monika Dittmann</i></p> <p>3.1 Introduction 73</p> <p>3.2 Theoretical Background 74</p> <p>3.3 Extracolumn Dispersion in (U)HPLC Systems 78</p> <p>3.4 Impact of External Contributions in Different Application Areas 90</p> <p>3.5 Optimization of HPLC/UHPLC Systems 94</p> <p>3.6 Conclusions 97</p> <p>References 98</p> <p><b>4 The Gradient; Requirements, Optimal Use, Hints, and Pitfalls 101</b><br /><i>Frank Steiner</i></p> <p>4.1 Instrumental Influences in Gradient Elution – An Overview 101</p> <p>4.2 Gradient Elution Technology and How to Systematically Characterize Gradient Instrumentation 117</p> <p>References 169</p> <p><b>5 Requirements of LC-Hardware for the Coupling of Different Mass Spectrometers 171</b><br /><i>Terence Hetzel, Thorsten Teutenberg, Christoph Portner, and Jochen Tuerk</i></p> <p>5.1 Introduction 171</p> <p>5.2 From Target Analysis to Screening Approaches 171</p> <p>5.3 What Should Be Considered for UHPLC/MS Hyphenation? 173</p> <p>5.4 Target Analysis Using Triple-Quadrupole Mass Spectrometry 178</p> <p>5.5 Screening Approaches Using LC-MS 185</p> <p>5.6 Miniaturization – LC-MS Quo Vadis? 189</p> <p>References 192</p> <p><b>6 2D chromatography – Opportunities and limitations 193</b><br /><i>Thorsten Teutenberg and Juri Leonhardt</i></p> <p>6.1 Introduction 193</p> <p>6.2 Why Two-Dimensional HPLC? 193</p> <p>6.3 Peak Capacity of One- and Two-Dimensional Liquid Chromatography 195</p> <p>6.4 Modulation 200</p> <p>6.5 Practical Problems of Online LC × LC 203</p> <p>6.6 Development of a Miniaturized LC× LC System 204</p> <p>6.7 Real Applications 207</p> <p>6.8 Advantages of the MS/MS Functionality 211</p> <p>6.9 General Comments to Specific Aspects of an LC × LC System 211</p> <p>6.10 Method Development and Gradient Programming 215</p> <p>6.11 Presentations of the Instrument Manufacturers (in Alphabetical Order) 215</p> <p>6.12 2D LC – Quo Vadis? 217</p> <p>References 219</p> <p><b>7 Materials in HPLC and UHPLC – What to Use for Which Purpose 223</b><br /><i>Tobias Fehrenbach and Steffen Wiese</i></p> <p>7.1 Introduction 223</p> <p>7.2 Requirements for Materials in UHPLC 225</p> <p>7.3 Flow Paths in UHPLC Systems 227</p> <p>7.4 Low-Pressure Flow Path 229</p> <p>7.5 High-Pressure Flow Path 231</p> <p>7.6 When and Why Can an Inert UHPLC System Be Required? 248</p> <p>References 261</p> <p><b>Part II Experience Reports, Trends 269</b></p> <p><b>8 What a Software has to Possess in Order to Use the Hardware Optimally 271</b><br /><i>Arno Simon</i></p> <p>8.1 Functionality and Handling 271</p> <p>8.2 Data Exchange 277</p> <p>8.3 From PCs Scalability to Global Installation 278</p> <p><b>9 Aspects of the Modern HPLC Device – Experience Report of an Operator 281</b><br /><i>Steffen Wiese and Terence Hetzel</i></p> <p>9.1 Introduction 281</p> <p>9.2 Determination of the Gradient Delay Volume 281</p> <p>9.3 High-Throughput Separations 285</p> <p>9.4 Method Transfer between UHPLC Systems of Different Manufacturers 287</p> <p>9.5 Application of Elevated Temperatures 290</p> <p>9.6 Large-Volume Injection (LVI) 293</p> <p>9.7 UHPLC Separation with 1mm ID Columns 296</p> <p>Acknowledgment 299</p> <p>References 299</p> <p><b>10 Experiences of an Independent Service Engineer – Hints and Recommendations for an Optimal Operation of Agilent and Waters-Devices 301</b><br /><i>Siegfried Chroustovsky</i></p> <p>10.1 Introduction 301</p> <p>10.2 The Degasser, Principles 301</p> <p>10.3 The Pump, Principles 303</p> <p>10.4 The Autosampler, Principles 306</p> <p>10.5 The UV Detector, Principles 308</p> <p><b>11 The Analyte, the Question, and the UHPLC – The Use of UHPLC in Practice 311</b><br /><i>Stefan Lamotte</i></p> <p>11.1 Introduction 311</p> <p>11.2 When Does It Make Sense to Use UHPLC and When Should I Better Use Conventional HPLC? 311</p> <p>11.3 Dissolution Tests in Pharmaceutical Industry 313</p> <p>11.4 Method Development and Optimization 314</p> <p>11.5 Typical “Classical” Liquid Chromatographic Analysis 314</p> <p>11.6 Fast (Most Second) Dimension of Multidimensional Chromatography 315</p> <p>11.7 Separation of (Bio)polymers 316</p> <p>11.8 Process Analysis (PAT) 316</p> <p>11.9 Conclusion 316</p> <p>References 316</p> <p><b>12 Report of Device Manufacturers – Article by Agilent, Shimadzu, and ThermoScientific 319</b></p> <p>12.1 Agilent Technologies 319<br /><i>Jens Trafkowski</i></p> <p>References 328</p> <p>12.2 HPLC Current Status and Future Development 328<br /><i>Björn-Thoralf Erxleben</i></p> <p>12.3 Thermo Fisher Scientific, Germering 334<br /><i>Frank Steiner</i></p> <p>About the Authors 349</p> <p>Index 355</p>

Stavros Kromidas studied biology and chemistry at the University of Saarbrucken, where he obtained his Ph.D. degree on the development of new chiral stationary phases for HPLC in 1983. After working for Waters GmbH for five years, he founded NOVIA GmbH, a provider of professional training and consulting in analytical chemistry, serving as the CEO until 2001. Since 2001 he works as an independent consultant for analytical chemistry, based in Saarbrucken (Germany). For more than 20 years he has regularly held lectures and training courses on HPLC, and has authored numerous articles and several books on various aspects of chromatography.

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