HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC): PRINCIPLES, PRACTICES AND PROCEDURES

HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC): PRINCIPLES, PRACTICES AND PROCEDURES

Library of Congress Cataloging-in-Publication Data

Contents

Preface

Chapter 1: Hydrophilic Interaction Liquid Chromatography: Fundamentals and Applications

Abstract

Introduction and Background

Principle and Retention Mechanisms of Hydrophilic Interaction Liquid Chromatography

Stationary Phases in HILIC

Bare Silica

Neutral Cyano, Diol and Amide Derivatized Columns

Amino and Sulfonated S-DVB Ion Exchange Stationary Phases

Sulfoalkylbetaine Zwitterion HILIC Phases

Mobile Phases

Applications of HILIC

Determination of Renal Function Biomarkers, Creatinine and Uric Acid, in Human Fluids

HILIC Analysis of Ascorbic Acid in Fresh Squeezed Orange Juice

Separation and Quantification of Cocaine and Its Decomposed Product, Benzoylecgonine

Summary and Conclusion

Acknowledgments

References

Chapter 2: The New Generation of HPLC Columns: Evolution of Packing Materials

Abstract

Introduction

Theory

Van Deemter equation

Knox Equation

Poppe Plots

Novel Adsorbents and Stationary Phases

Sub-2μm Particles

Core-Shell Particles

Monolithic Columns

Silica-Based Monolithic Columns

Polymer-Based Monolithic Columns

Hydrophilic Interaction Chromatography (HILIC)

Comparison Between Sub-2 µm and Core-Shell Particles

Comparison Between Monolithic and Particle-Packed Columns

Comparative Analysis of Antibiotics Using Different Analytical Columns

Conclusion

References

Chapter 3: High-Performance Liquid Chromatography for Carbohydrate Analysis

Abstract

Introduction

Carbohydrates and Their Conjugates

Carbohydrates

Glycoproteins and Glycopeptides

Proteoglycans and Peptidoglycans

Glycolipids

Lipopolysaccharides

Preparation of Carbohydrate Samples for Analysis

Extraction and Purification

Release of Carbohydrate from Its Conjugates

Hydrolysis of Polysaccharides

Carbohydrate Derivatization

HPLC Separation of Carbohydrates

Reversed Phase (RP) Chromatography

Hydrophobic Interaction Chromatography (HIC)

Normal Phase (NP) Chromatography

Hydrophilic Interaction Liquid Chromatography (HILIC)

Anion Exchange Chromatography (AEC)

Size Exclusive Chromatography (SEC)

Detection Methods for Carbohydrate Analysis

Refractive Index Detection (RID)

Ultraviolet-Visible Detection (UV-Vis)

Fluorescence Detection

Evaporative Light Scattering (ELS) Detection

Laser Light Scattering (LLS) Detection

Electrochemical (EC) Detection

HPLC Application to Chitosan Analysis

Experimental Conditions

Materials

HPLC Analysis

Results and Discussion

Glucosamine Separation And Quantitative Determination

Chitosan Hydrolysis

Effects of Acid Concentration and Temperature on Glucosamine Decomposition

Method Validation

Conclusion

References

Chapter 4: High-Performance Liquid Chromatography for the Analysis of Non-Nucleoside Reverse Transcriptase Inhibitors in Biological Fluids and Pharmaceutical Products

Abstract

Nomenclature

Introduction

Physico-Chemical Properties of Non-Nucleoside Reverse Transcriptase Inhibitors

HPLC Analysis of Non-Nucleoside Reverse

Transcriptase Inhibitors

Sample Pretreatment

a) Protein Precipitation

b) Liquid-Liquid Extraction

c) Solid-Phase Extraction

d) Direct Dissolution

Separation Conditions

Detection

HPLC Methods for Simultaneously Measuring of Non-Nucleoside Reverse Transcriptase Inhibitors

Conclusion

Acknowledments

References

Chapter 5: High-Performance Liquid Chromatography – An Effective Tool for Quality Control of Natural Cholesterol-Lowering Agents

Abstract

Introduction

High-performance liquid Chromatography

Red Yeast Rice

Artichoke

Phytosterols

Omega-3 Fatty Acids

Green Tea

Soybean

Guggulipid

Coenzyme Q10

Taurine

Simultaneous Analysis of Natural

Cholesterol-Lowering Agents

Bioanalytical Methods

Related Techniques

Thin-Layer Chromatography

Gas Chromatography

Capillary Electrophoresis

Conclusion

References

Chapter 6: Measurement of Cholesterol Levels of Lipoprotein Classes by Using Anion-Exchange Chromatography

Abstract

Introduction

Reagent and Instrument Used for AEX-HPLC Method, and the Analysis Condition

Measurement of Cholesterol Levels in 4 Major Lipoprotein Classes

Methods Measurement of Cholesterol Levels in Lipoprotein(A)

Measurement of Cholesterol Levels in HDL and LDL Subfractions

Measurement of Cholesterol Levels in Rabbit Lipoprotein Classes

Monitoring for A Diabetic Patient with Rheumatoid Arthrits by Using AEX-HPLC Method

Comparison between AEX-HPLC Method and Electrophoresis

Conclusion

References

Chapter 7: Recent Trends in LC Analysis of Biological Materials for Determination of Benzodiazepine Drugs

Abstract

Introduction

The Chemical Structure, Physicochemical and Pharmacological Properties of Benzodiazepine-Type Drugs

Characterization and Preparation of Biological Materials for Determination of Benzodiazepines

Sample Preparation of Body Fluids (Urine, Plasma/Serum, Whole Blood)

Sample Preparation of Alternative Matrices (Hair, Nails, Saliva, Vitreous Humor, Finger Prints)

Sample Preparation of Hair and Nails

Sample Preparation of Saliva

Sample Preparation of Vitreous Humor

Sample Preparation of Other Unconventional Materials

LC Methods for Detection and Determination of Benzodiazepine Drugs

Applications in Pharmacological Study

Applications in Toxicological Clinical and Forensic Analysis

Conclusion

References

Chapter 8: HPLC Analytical Study of Methacrylic Monomers Released from Dental Composite Materials

Abstract

Introduction

Sample and Analysis Background Information

Cell Cultures

Treatment of 3T3-Fibroblasts and HGFs with HEMA or TEGDMA, and Preparation of the Samples for HPLC Analysis

Treatment of 3T3-Fibroblasts with HEMA or TEGDMA in Presence of NAC, and Preparation of the Samples for HPLC Analysis

Treatment of HGFs with TEGDMA in Presence of NAC, and Preparation of the Samples for HPLC Analysis

Qualitative and Quantitative Analysis of HEMA and TEGDMA in Intracellular and Extracellular Samples

Statistical Analysis

Monomers Concentrations in Intracellular and Extracellular Samples

Conclusion

References

Chapter 9: HPLC Analysis of Vitamin B1, B2, B3, B6, B9, B12 and Vitamin C in Various Food Matrices

Abstract

Introduction

Vitamin B1 (Thiamine)

Generic Description and Structures – Vitamin B1

Nutritional/Physiological Importance and Dietary Sources – Vitamin B1

Assay Methodology for Vitamin B1 (Vitamins B2, B3 and B6) – General

HPLC Assay Methodology for Vitamin B1

Vitamin B2 (Riboflavin)

Generic Description and Structures – Vitamin B2

Nutritional/Physiological Importance and Dietary Sources – Vitamin B2

Assay Methodology for Vitamin B2 – General

HPLC Assay Methodology for Vitamin B2

Vitamin B3 (Niacin)

Generic Description and Structures – Vitamin B3

Nutritional/Physiological Importance and Dietary Sources – Vitamin B3

HPLC Assay Methodology for Vitamin B3

Vitamin B6 (Pyridoxine)

Generic Description and Structures – Vitamin B6

Nutritional/Physiological Importance and Dietary Sources – Vitamin B6

HPLC Assay Methodology for Vitamin B6

Improvements and Recent Developments: Vitamin B6 Methods

Vitamin B9 (Folate)

Generic Description and Structures – Vitamin B9

Nutritional/Physiological Importance – Vitamin B9

Dietary Sources – Vitamin B9

Assay Methodology for Vitamin B9 – General

HPLC Assay Methodology for Vitamin B9

Improvements and Recent Developments – Vitamin B9 Methods

Vitamin B12 (Cyanocobalamin)

Generic Description and Structures – Vitamin B12

Nutritional/Physiological Importance – Vitamin B12

Dietary Sources – Vitamin B12

Assay Methodology for Vitamin B12 – General

HPLC Assay Methodology for Vitamin B12

Improvements and Recent Developments – Vitamin B12 Methods

Vitamin C (Ascorbic Acid)

Generic Description and Structures – Vitamin C

Nutritional/Physiological Importance and Dietary Sources

Assay Methodology for Vitamin C – General

HPLC Assay Methodology for Vitamin C

References

Chapter 10: Ultra Performance Liquid Chromatography Coupled to Ultraviolet-Vis and Mass Spectrometry Detector for Screening of Organic Acids and Polyphenols in Red Wine

Abstract

Introduction

Materials and Methods

Chemicals and Reagents

Standard Solutions

Wine Samples

Sample Preparation

UPLC Analysis

Equipment

Operating conditions

Assessment of the Trifluroacetic Acid as Matrix Effect

Method Validation

Linearity

Precision

Limits of quantification and detection

Recovery

Results and Discussion

Optimization of the UPLC Methods

Organic acids

Non-pigmented polyphenols

Matrix effect

Anthocyanin pigments

Validation Procedure

Application of the Method to Wine Samples

Conclusion

Acknowledgments

References

Chapter 11: HPLC–UV-Vis–ESI MS Examination of Archeological Fibers: Red Natural Dyes in Italian Textiles from 15th and 16th Centuries

Abstract

Introduction

Experimental

Instrumentation

Chemicals

Examined Fibers

Preparation of Solutions of Standards and Natural Dyestuffs

Preparation of Textile Fibers Prior to Analysis

Results

Discussion

Conclusion

Acknowledgments

References

Index