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Short description The third edition of this popular problem-solving guide includes 11 completely new examples and several updated ones. A must-have resource for every lab wanting to use HPLC efficiently and obtain correct results.
From the contents PREFACE
PART I: Fundamentals Chromatography Chromatographic Figures of Merit The Resolution of Two Peaks Reduced Parameters The Van Deemter Curve Peak Capacity and Number of Possible Peaks Statistical Resolution Probability: Simulation Statistical Resolution Probability: Example Precision and Accuracy of an Analytical Result Standard Deviation Variability of the Standard Deviation Uncertainty Propagation Reproducibility in Trace Analysis Ruggedness Calibration Curves The HPLC Instrument The Detector Response Curve Noise The Playground Presented as an Ishikawa Diagram The Possible and the Impossible
PART II: Pitfalls and Sources of Error Mixing of the Mobile Phase Mobile Phase pH Adjustment of Mobile Phase pH Influence of the Acid Type and Concentration in the Eluent Water as an Unintentional Additive in the Mobile Phase Inadequate Purity of Mobile Phase Water Inadequate Purity of a Mobile Phase Solvent Inadequate Purity of a Mobile Phase Reagent Incomplete Degassing System Peaks and Quantitative Analysis Sample Preparation with Solid Phase Extraction Inadequate Stabilization of the Extraction Solvent Poor Choice of Sample Solvent: Peak Distortion Poor Choice of Sample Solvent: Tailing Sample Solvent and Calibration Curve Impurities in the Sample Formation of a By-Product in the Sample Solution Decomposition by the Sample Vial Artifact Peaks from the Vial Septum Formation of an Associate in the Sample Solution Precision and Accuracy with Loop Injection Injection Technique Injection of Air Sample Adsorption in the Loop Extra-Column Volumes Dwell Volume Elution at t0 Classification of C18 Reversed Phases Different Selectivity of C18 Reversed Phases Different Batches of Stationary Phase Chemical Reaction within the Column Tailing of Phosphate Compounds in the Presence of Steel Recovery and Peak Shape Problems with Proteins Double Peaks from Stable Conformers Influence of Temperature on the Separation Thermal Non-Equilibrium within the Column Influence of the Flow Rate on the Separation Influence of Run Time and Flow Rate on Gradient Separations UV Spectra and Quantitative Analysis UV Detection Wavelength Different Detection Properties of Diastereomers Fluorescence Quenching by Air Detector Overload in UV Detector Overload in ELSD Influence of the Retention Factor on Peak Height Influence of the Flow Rate on Peak Area Leaks in the HPLC Instrument Impairment of Precision as a Result of Noise Determination of Peak Area and Height at High Noise Peak Height Ratios Incompletely Resolved Peaks Area Rules for Incompletely Resolved Peaks Areas of a 1 : 10 Peak Pair Heights of a 1 : 10 Peak Pair Quantitative Analysis of a Small Peak Incompletely Resolved Peaks with Tailing Integration Threshold and Number of Detected Peaks Detector Time Constant and Peak Shape Quantitative Analysis in the 99% Range Correlation Coefficient of Calibration Curves
PART III: Useful Strategies Column Tests Apparatus Tests Wavelength Accuracy of the UV Detector Internal Standards A Linearity Test Rules for Accurate Quantitative Peak Size Determination High-Low Chromatography Control Charts Verification of the Analytical Result by Use of a Second Method Description of Ruggedness Rules for Passing On an HPLC Method Quality Assurance in the Laboratory Standard Operating Procedures Method Validation Some Elements of Validation A Validation Example System Suitability Test From Repeatability to Reproducibility Measurement Uncertainty Formal Quality Assurance Systems