《Pharmaceutical Analysis I》课程教学大纲

一、课程基本信息

二、课程目标

（一）总体目标：

药物分析是一门综合性、专业性、实践性和法规性均较强的学科。该课程总体目标是承接基础分析化学在药学研究、药物生产和临床使用过程中的应用。初步使学生了解药物分析与分析化学的区别，理解药物分析的意义，掌握基本药物分析原则和原理。该课程为全英文教学，学生的专业英文的掌握也是该课程的目标之一。

（二）课程目标：

课程目标1：掌握基础知识

 1．1 药物分析相关基本概念

 1．2 药物分析相关国籍法规

 1．3 药物分析相关基础

课程目标2：掌握基本分析方法原理和仪器

 2．1 滴定

 2．2 光谱

 2．3 色谱

 2．4 毛细管电泳

 2．5 质谱

 2．6 其他分析方法和技术

课程目标3：掌握样品处理方法原则和方式

 3．1 样品制备

课程目标4：掌握原料药和最终制剂产品的定性定量分析方法、数据处理原则、质量标准制定原则和条目

 4．1 定性定量方法建立和验证

 4．2 原料药质量标准制定原则和条目

 4．3 最终制剂产品质量标准制定原则和条目

课程目标5：掌握生物样品中药物分析流程和方法

（要求参照《普通高等学校本科专业类教学质量国家标准》，对应各类专业认证标准，注意对毕业要求支撑程度强弱的描述，与“课程目标对毕业要求的支撑关系表一致）（

（三）课程目标与毕业要求、课程内容的对应关系

表1：课程目标与课程内容、毕业要求的对应关系表

（大类基础课程、专业教学课程及开放选修课程按照本科教学手册中各专业拟定的毕业要求填写“对应毕业要求”栏。通识教育课程含通识选修课程、新生研讨课程及公共基础课程，面向专业为工科、师范、医学等有专业认证标准的专业，按照专业认证通用标准填写“对应毕业要求”栏；面向其他尚未有专业认证标准的专业，按照本科教学手册中各专业拟定的毕业要求填写“对应毕业要求”栏。）

三、教学内容

（具体描述各章节教学目标、教学内容等。实验课程可按实验模块描述）

Chapter 1 Introduction to Pharmaceutical Analysis

1. 教学目标

This chapter briefly reviews the life of medical products and the manufacture of medical products according to international regulations and guidelines. Based on this review the major areas and usage of pharmaceutical analysis are identified.

2. 教学重难点

The quality control system.

3. 教学内容

Contents
1.1 Applications and Definitions
1.2 The Life of Medicines
1.3 The Quality of Medical Products
1.4 Summary

Problems:

1. The concept of drug?

2. What’s the main function of excipients?

3. What are the means of QA,QC and GMP?

4. How to assure the safty,protection and well being of the consumer or patient?

5. What requirements should be met before certifying a batch the QP?

6. The significance of the control of drugs’ quality ?

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 2 International Pharmacopoeias, Regulations and Guidelines

1. 教学目标

This chapter reviews a number of laws, guidelines and regulations important in the pharmaceutical production and thus also for the subject pharmaceutical analysis. The manufacture of drugs is international and therefore this chapter focuses on international affairs. But also national laws, guidelines, and regulations are important for manufacturers in order to fulfil the requirements of the relevant authorities. All these laws, guidelines and regulations are subject to regular updates and the latest edition should therefore always be consulted.

2. 教学重难点

The concepts of marketing authorization and manufacturing authorization

3. 教学内容

Contents

2.1 Overview of Legislation
2.2 Legislation and Regulations for Industrial Production
2.3 Life Time of Drugs and Drug Substances
2.4 Pharmacopoeias
2.5 International Harmonization
    2.5.1 International Conference on Harmonization
    2.5.2 Pharmacopoeial Discussion Group
2.6 Legislation and Regulations for Pharmacy Production
2.7 Summary

Problems:

1. What’s applicable for Proprietary Medicinal Products?

2. What’s the situation the marketing authorization may be withdrawn before the expiration by the Ministry of Health?

3. What’s the detail information of the raw materials before production should be controlled?

4. What’s the main elements of the GMP/GLP regulations?

5. What are the ICH guidelines of important relevance for the pharmaceutical analysis?

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 3 Fundamental Chemical Properties, Buffers and pH

1. 教学目标 Chemical analysis is an important part of the quality assessment of drugs. A deeper understanding of the analytical method requires knowledge about both the analytical technique and the chemical properties of the analytes. Therefore a basic knowledge of a number of physicochemical properties of molecules is needed to be able to understand and further develop analytical chemical methods. For example, knowledge of spectroscopic principles and techniques is needed when the choice of the detection technique for a given separation method is made; and knowledge of pH and pKa values is important for the design of many sample preparation techniques. Spectroscopic techniques are treated in separate chapters, and this chapter discusses some important physicochemical and chemical properties of drug substances and focuses on how to utilize the information in an analytical chemical context.

2. 教学重难点

The concepts of pH and pKa

3. 教学内容

Contents
3.1 pH and pKa
3.2 Partition
3.3 Stereochemistry
3.4 Stability Testing
3.5 Summary

Problems:

1. What are the means of pH,pKa and pKb?

2. What is called a buffer system and the purpose?

3. What’s the typical pKa values of following functional groups,such as R-COOH,R-NH₂,R-OH and R-SO₂OH?

4. What’s the influence of the ions?

5. How isomers can be divided into several groups?

6. How long the drug substances are fairly stable?

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 4 Fundamentals of Pharmaceutical Analysis

1. 教学目标

This chapter discusses the basics of pharmaceutical analysis, including the different types of calculations related to pharmaceutical analysis. The chapter also includes a review of simple laboratory equipment, how to make solutions and dilutions, how to calibrate analytical methods, and how to use simple statistics on the analytical data. The chapter concludes with a list of important terms and concepts in pharmaceutical analysis. It is important that you read carefully through this chapter before proceeding to the subsequent chapters. Important terms should be learned.

2. 教学重难点

Presentation of analytical data

3. 教学内容

Contents

4.1 What is a Pharmaceutical (Chemical) Analysis?
4.2 How to Specify Quantities and Concentrations?
4.3 Basic Laboratory Equipment
    4.3.1 The Analytical Balance
    4.3.2 Pipettes
    4.3.3 Volumetric Flasks
    4.3.4 Burettes
4.4 How to Make Solutions and Dilutions
4.5 Calibration of Analytical Methods
4.6 Errors, Accuracy, and Precision
    4.6.1 Systematic and Random Errors
    4.6.2 Accuracy and Precision
4.7 Statistics
    4.7.1 Mean Value and Standard Deviation
    4.7.2 Confidence Intervals
    4.7.3 Comparison of Means with a t-Test
    4.7.4 Q-Test to Reject Outliers
    4.7.5 Linear Regression with the Method of Least Squares
    4.7.6 How to Present an Analytical Result
4.8 Some Words and Concepts
    4.8.1 Analysis and Determination
    4.8.2 Sample Replicates and Measuring Replicates
    4.8.3 Interference
    4.8.4 Blind Samples

Problems:

1. What is a Pharmaceutical (Chemical) Analysis?

2. What should be ensured of sample preparation?

3. How to Specify Quantities and Concentrations?

4. How to calculate concentration in terms of ppm?

5. What are the considerations for the use of the analytical balance?

6. What are the considerations for the use of the pipettes?

7. What are the means of the temperature on pipettes,volumetric flasks and burettes?

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 5 Titrimetric Methods

1. 教学目标

This chapter discusses the principles of quantitative analysis based on titrimetric methods. Titrimetric methods are official methods in the European Pharmacopeia and are used for quality control of active pharmaceutical ingredients (APIs) and excipients. The chapter focuses on titrations based on acid–base reactions and redox reactions and also gives an overview of various principles of endpoint detection. Examples of titrimetric methods for the quantitative determination of APIs and excipients are discussed in Chapter 21.

2. 教学重难点

Non-aqueous Titration

3. 教学内容

Contents

5.1 Introduction
5.2 Acid–Base Titrations
5.3 Acid–Base Titrations in Non-Aqueous Media
5.4 Redox Titrations
5.5 Other Principles of Titration
5.6 Summary

Problems:

1. What are the requirements should be fulfilled to make a titration?

2. What are the characteristics of the reagent in titration reaction?

3. What’s the principle of pH measurements?

4. What is the range of the color change and pH value about the methyl orange and Phenolphthalein?

5. In what situations,can we use redox reactions?

6. How to calculate the pH value of the end point of the acid base titration reaction?

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 6 Introduction to Spectroscopic Methods

1. 教学目标

This chapter presents a brief review of electromagnetic radiation and discusses how molecules and elements absorb and emit electromagnetic radiation. Absorption and emission of electromagnetic radiation are the basis for identification and quantitative determinations in spectroscopic methods such as UV spectrophotometry, IR spectrophotometry, NIR spectrophotometry, atomic absorption spectrometry and atomic emission spectrometry. These methods are presented in subsequent chapters.

2. 教学重难点

Electromagnetic Radiation

3. 教学内容

Contents

6.1 Electromagnetic Radiation
6.2 Molecules and Electromagnetic Radiation
6.3 Atoms and Electromagnetic Radiation
6.4 Summary

Problems:

1. What are categories of spectroscopic methods according to their principle of separation?

2. What’s the electromagnetic radiation in the wavelength region of UV radiation?

3. What is the principle of atomic absorption spectroscopy?

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 7 UV Spectrophotometry

1. 教学目标

This chapter reviews the principles of quantitative determination and identification based on UV spectrophotometry. UV spectrophotometry is an official method in Ph.Eur. It is used for quality control of active ingredients, excipients and pharmaceutical products. Instrumentation is discussed briefly and some practical tips in conjunction with UV spectrophotometry are also discussed in this chapter. Examples of how the technique is used for identification and quantitative determinations with regard to ingredients and pharmaceutical products are discussed in Chapters 21 and 22, respectively.

2. 教学重难点

Instrumentation of UV spectrometry

3. 教学内容

Contents

7.1 Principle of Quantitative Determination
7.2 Principle of Identification
7.3 Which Substances Have Strong UV Absorbance?
7.4 Instrumentation
7.5 Practical Work and Method Development
7.6 Areas of Usage and Performance
7.7 System Testing
7.8 Summary

Problems:

1. What is the principle of UV Spectrophotometry?

2. What does each parameter refer to in Beer’s law?

3. Which substances have strong UV absorbance?

4. What points are essential of the use of methods based on UV spectrophotometry?

5. Which area is particularly suitable for UV spectrophotometry?

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 8 IR Spectrophotometry

1. 教学目标

This chapter reviews infrared (IR) spectrophotometry and the closely related techniquenear infrared (NIR) spectrophotometry. Both techniques are official methods in Ph.Eur. IR spectrophotometry is primarily used for identification of pharmaceutical ingredients and pharmaceutical products. The chapter discussed the information that can be obtained in the different parts of the IR spectra and the instrumentation used. Examples of how the technique is employed for identification of raw materials and products are discussed in Chapters 21 and 22. NIR spectrophotometry is used both for quality control of pharmaceutical ingredient, for process control and for the control of finished products. The information that can be obtained from NIR spectra and the instrumentation used are discussed briefly.

2. 教学重难点

The assignments of functional groups in IR

3. 教学内容

Contents

8.1 IR Spectrophotometry
8.2 Instrumentation
8.3 Scope
8.4 Instrument Calibration
8.5 NIR Spectrophotometry
8.6 Applications
8.7 Summary

Problems:

1. What is the principle of IR spectrophotometry?

2. What wavelength region has been used in pharmaceutical analysis of IR spectrophotometry normally?

3. What differences are between UV spectrometers the IR spectrometers?

4. What does NIR spectrometry applicate for chemical analysis?

5. Which state is the sample of IR spectra

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 9 Atomic Spectrometry

1. 教学目标

This chapter provides general information and defines the procedures used in element determinations by atomic absorption spectrometry (AAS) and atomic emission spectrometry (AES). AAS and AES are official methods of Ph.Eur. and both are used for the quality control of raw materials and products. The instrumentation used and some practical tips in connection with quantification of elements by atomic spectroscopy are also briefly discussed in this chapter.

2. 教学重难点

The differences between AAS and AES

3. 教学内容

Contents

9 Atomic Spectrometry

9.1 Atomic Absorption Spectrometry

9.2 Instrumentation
9.3 Applications and Performance

9.4 Practical Work and Method Development
9.5 Atomic Emission Spectrometry
9.6 Instrumentation
9.7 Summary

Problems:

1. What is the main difference between AAS and AES?

2. Briefly describe the steps of atomization in a flame after the introduction of the sample solution.

3. What are the main components of the atomic absorptionspectrometer?

4. What we need to pay attention to the test solution and atomic absorption?

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 10 Chromatography

1. 教学目标

Chromatography is the name give to a particular family of separation methods where the separation is based on differences in rates of migration when the sample components are transported by a mobile phase through a stationary phase. The mobile phase can be a gas, a liquid or a supercritical fluid and the stationary phase may be a solid, a liquid or a gel. Chromatography is considered the most important analytical technique in pharmaceutical analysis. This chapter looks at the theory of chromatographic separation and at important parameters used to characterize a separation.

2. 教学重难点

Principles of chromatography

3. 教学内容

Contents

10.1 General Principles

10.2 Retention

10.3 Column Efficiency

10.4 Selectivity

10.5 Peak Symmetry

10.6 Resolution

10.7 Chromatographic Techniques

10.8 Summary

Problems:

1. What is the mean of chromatogram? What is an idealized chromatogram?

2. What are the means of t_R, t_M, k, N, H, α, and Rs?

3. How to determine the column efficiency from the peaks in the chromatogram?

4. How to improve separation by altering α value?

5. How to evaluate the resolution?

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 11 Chromatographic Separation Principles

1. 教学目标

Chromatographic techniques involve two phases of which at least one is mobile and the other most often is stationary. The two phases move relative to each other and in most cases the mobile phase moves through a bed of a stationary phase. The stationary phase may be a piece of paper (paper chromatography), or particles of a suitable material placed as a thin layer on an inert support (thin-layer chromatography) or packed into a tube or column (column chromatography). Column chromatography can be executed using a liquid (liquid chromatography) or a gas (gas chromatography) as the mobile phase. When a mixture of analytes is introduced into these systems a number of separation mechanisms influence the partition of analytes between the two phases. This chapter gives an introduction to important separation mechanisms and how different types of analytes can be separated.

2. 教学重难点

Differences between different chromatography

3. 教学内容

Contents

11.1 General Introduction

11.2 Normal Phase Chromatography

11.2.1 Silica

11.2.2 Interactions

11.2.3 Order of Elution

11.2.4 Other Stationary Phases

11.2.5 Mobile Phases

11.2.6 Summary of Normal Phase Chromatography

11.3 Reversed Phase Chromatography

11.3.1 Stationary Phases

11.3.2 Retention Mechanisms

11.3.3 Mobile Phases

11.3.4 Ion-Pair Chromatography

11.3.5 Summary of Reversed Phase Chromatography

11.4 Hydrophilic Interaction Chromatography

11.5 Chiral Separations

11.6 Size Exclusion Chromatography

11.6.1 Principle

11.6.2 Summary of SEC

11.7 Ion Exchange Chromatography

Problems:

1. Why and how the ionic interactions cause strong adsorption and tailing peak?

2. What does the retention depend on? How and Why?

3. Why to say the liquid chromatography is the most important separation principle?

4. How to use organic modifier to change separation selectivity?

5. By using reverse phase chromatography, acids are separated by a pH that suppresses ionization and bases at a pH where they are ionized. Retention of neutral substances is not affected by pH. Why?

6. How to choose chromatography?

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 12 Thin-Layer Chromatography

1. 教学目标

Thin-layer chromatography (TLC) is a liquid chromatography separation technique where the stationary phase is dispersed as a thin layer placed on the surface of a plate typically made of glass or alumina foil. Typical applications are the identification of active ingredients and contaminants in pharmaceutical raw materials and preparations and the analysis of herbals used as drugs or food supplements. The samples, prepared in a volatile solvent, are placed as spots or bands on the plate parallel to one of its edges. The sample solvents are then evaporated and the plate inserted in a tank with a mobile phase at the bottom of the tank. The development tank is closed and the mobile phase migrates up the plate by capillary forces. The analytes in a spot migrate up the plate depending on their distribution between the mobile phase and stationary phase. When the mobile phase has moved a certain distance towards the top of the plate, the TLC plate is removed from the tank and the mobile phase is evaporated from the plate in a fume hood. The position, color and size of the spots in the sample seen on the plate are now compared with the position, color and size of spots in reference substances. More details are given in this chapter.

2. 教学重难点

Selection of method for samples with different properties.

3. 教学内容

Contents

12.1 Introduction

12.2 Apparatus

12.3 TLC Plates

12.4 Stationary Phases

12.5 Mobile Phases

12.6 Chromatographic Development

12.7 Detection

12.8 Applications of TLC

12.9 Quantitative Analysis and Instrumentation

12.10 Summary

Problems:

1. What the mean of Rf?

2. How to choose the stationary phases?

3. How to choose the mobile phases?

4. How to detect the analytes?

5. Describe the application of TLC in pharmaceutical analysis.

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 13 High Performance Liquid Chromatography

1. 教学目标

High performance liquid chromatography (HPLC, or for short LC) is the most commonly used chromatographic technique to determine drugs in pharmaceutical preparations and in biological material. In LCthe mobile phase is a liquid, forced through a column packed with a material that retards the analytes introduced into the system. The analytes are injected into the flow of mobile phase just in front of the separation column. The outlet of the column is connected to a detector where the eluted substances are detected. The separation principle can be any of the principles described in chapter 11. This chapter reviews how the apparatus is constructed and how it works.

2. 教学重难点

The instrumentation and application of HPLC

3. 教学内容

Contents

13.1 Introduction

13.2 The Chromatographic Separation Process

13.3 The Column

13.4 Pumps

13.5 Detectors

13.5.1 UV detector

13.5.2 Fluorescence Detector

13.5.3 Electrochemical Detector

13.5.4 Refractive Index, Evaporative Light Scattering and Corona Discharge Detectors

13.5.5 Combination of Detectors

13.6 Injectors

13.7 Mobile Phases

13.8 Solvents for Sample Preparation

13.9 Reporting the Results

13.10 Summary

Problems:

1. HPLC is the most commonly used chromatographic technique to determine drugs in pharmaceutical preparations and in biological material, why?

2. The column is the heart of the separation process, why?

3. How to choose the detector?

4. Mobile phase is the first choice as a solvent for the sample, why?

5. Describe the application of HPLC in pharmaceutical analysis.

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 14 Gas Chromatography

1. 教学目标

Gas chromatography is a separation technique used for the analysis of volatile substances or substances that can be made volatile. Analytes are separated as a function of different distribution ratios between the mobile and the stationary phase. The mobile phase is a gas under pressure flowing through the heated column containing the stationary phase. In gas chromatography the mobile phase is called the carrier gas. The stationary phase is placed on the inner surface of the column or coated onto particles packed into the column. The column is a long tube made of fused silica or metal. The sample is injected into a heated injector at the beginning of the column where the constituents evaporate. The carrier gas carries the analytes through the column where the substances are distributed between the stationary phase and the mobile phase. Analytes with different distribution ratios between the phases are carried through the column at different speeds towards the detector. The separated substances are detected by a detector placed at the end of the column. Gas chromatography is suitable for the separation of substances which are volatile and thermally stable or can be converted into volatile and thermally stable derivatives.

2. 教学重难点

The principle, instrumentation and application of GC

3. 教学内容

Contents

14.1 Introduction

14.2 Apparatus

14.3 Temperature

14.4 Carrier Gas

14.5 Stationary Phases

14.6 Selectivity in GC

14.7 Columns

14.7.1 Capillary Columns

14.7.2 Packed Columns

14.8 Injection Systems

14.8.1 Injection Systems for Capillary Columns

14.8.2 Injection Systems for Packed Columns

14.9 Detectors

14.9.1 Flame Ionization Detector

14.9.2 Nitrogen–Phosphorus Detector

14.9.3 Thermal Conductivity Detector

14.9.4 Electron Capture Detector

14.9.5 Mass Spectrometry Detector

14.10 Derivatization

14.10.1 Silylation

14.10.2 Alkylation

14.10.3 Acylation

14.11 The Uses of GC

14.12 More Advanced GC techniques

14.13 Summary

Problems:

1. By using capillary column in GC, thin film is used to separate heavy volatile substances, and thicker film is suitable for separation of volatile substances, why?

2. What the relationship between the temperature stability of stationary phase and polarity or molecular weight?

3. How to optimize the temperature of the column to control retention in GC analysis?

4. What the retention of the analyte is determined by?

5. How to choose the detector?

6. Describe the application of GC in pharmaceutical analysis.

4. 教学方法

课堂教授结合提问

5. 教学评价

期中考试

Chapter 15 Capillary Electrophoresis

1. 教学目标

In its basic set-up electrophoresis is a separation technique based on the migration of charged species/ions in an electrical field. Thus it is not based on distribution between two immiscible phases and it is therefore not a chromatographic technique. However, the system can be modified to become a combination of electrophoresis and chromatography. The driving force in electrophoresis is a voltage gradient applied between two electrodes in a buffer system. Charged analytes introduced into the system migrate towards the electrode with opposite charge. In the present text only electrophoresis in capillaries and related techniques are presented.

2. 教学重难点

The principle, instrumentation and application of capillary electrophoresis

3. 教学内容

Contents

15.1 Principle and Theory

15.2 Electroosmotic Flow

15.3 Instrumentation

15.4 The Capillary

15.5 Sample Introduction

15.6 Capillary Zone Electrophoresis; an Example

15.7 Micellar Electrokinetic Chromatography

15.8 Chiral Separations

15.9 Coated Capillaries

15.10 Non-Aqueous CE

15.11 Summary

Problems:

1. What is the mean of EOF? How it affect the mobility of analytes?

2. If the buffer concentration in the sample is more than one-tenth of the running buffer, what will happen? Why?

3. What are the effects of temperature on migration?

4. What does the retention depend on?

5. How to choose the detector?

6. Describe the application of CE in pharmaceutical analysis.

4. 教学方法

课堂教授结合提问

4. 教学评价

期中考试

Chapter 16 Mass Spectrometry

1. 教学目标

This chapter discusses the principles of identification and quantitative analysis based on mass spectrometry (MS). MS is an official technique in the European Pharmacopoeia (Ph.Eur.). MS is mainly used for the analysis of drug substances in biological samples. Various MS techniques will be reviewed, and the most widely used instrumentation will be discussed. Applications of MS are presented in Chapter 23 in connection with the bioanalysis of drug substances.

2. 教学重难点

The principle, basic theory, classification, instrumentation and application of MS

3. 教学内容

Contents

16.1 Introduction

Principle of mass spectrometry

Ionization, fragmentation, separation and detection

Mass spectrum

16.2 Basic Theory

Atomic mass and atomic mass unit

Exact masses and nominal masses

Isotopes and average atomic masses

16.3 Electron Ionization

Principles of electron ionization

Applications of electron ionization

Mass spectrum of electron ionization

16.4 Identification using Electron Ionization Spectra

Why reference EI spectra can be used to identify unknown substance

How to use reference EI spectra

16.5 Characterization of Totally Unknowns using Electron Ionization Spectra

General steps to characterize totally unknowns using EI spectra

Typical fragmentations from the molecular ions

Use of the information that the different isotopes provides

16.6 Chemical Ionization

Principles of chemical ionization

Applications of chemical ionization

Comparison between EI and CI

Negative chemical ionization

16.7 Electrospray Ionization

Principles of electrospray ionization

Applications of electrospray Ionization

Comparison between EI and ESI

16.8 Atmospheric Pressure Chemical Ionization

Principles of atmospheric pressure chemical ionization

Applications of atmospheric pressure chemical ionization

16.9 High-Resolution Mass Spectrometry

Advantages of high-resolution mass spectrometry

Resolving power

16.10 Instrumentation

The general components for a mass spectrometer

Different types of mass analyzer

16.11 Chromatography Coupled with Mass Spectrometry

Full scan and recording of spectra

Selected ion monitoring (SIM)

Selected reaction monitoring (SRM)

16.12 Quantitative GC-MS and LC-MS

Principles of quantitative analysis

16.13 Areas of Usage and Performance     

16.14 Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

Principles of MALDI-MS

Applications of MALDI-MS

16.15 Inductively Coupled Plasma Mass Spectrometry

Principles of ICP-MS

Applications of ICP-MS

Problems:

1. Identify the molecular ion from a given mass spectra

2. How to get the equation: I_m+1 = n  0.011 I_m

3. Explain the isotope patterns of chlorine and bromine

4. How to calculate the resolving power

4. 教学方法

课堂教授结合提问

4. 教学评价

期末考试

Chapter 17 Miscellaneous Chemical Techniques

1. 教学目标

The preceding chapters discuss a broad range of chemical techniques, like high performance liquid chromatography (HPLC), titration, and IR spectroscopy. These techniques are very important for the quality control of pharmaceuticals and for bioanalysis, and therefore they are treated very comprehensively. In addition to these principal techniques, several other techniques are in use for pharmaceutical analysis. This chapter discusses some of these other techniques. Priority is given to techniques officially adopted by the European Pharmacopoeia, and the principles are discussed only briefly. For more comprehensive discussions, the reader has to look in other textbooks

2. 教学重难点

Gel electrophoresis, NMR, Raman Spectrometry

3. 教学内容

Contents

17.1 Potentiometric Determination of Ions using Ion-Selective Electrodes

Potentiometric determination of pH

Principles of ion-selective electrode

Quantification using ion-selective electrode

Usage of ion-selective electrode

17.2 Paper Chromatography

    Principles of paper chromatography

17.3 Supercritical Fluid Chromatography

    Properties of supercritical fluid

Principles of supercritical fluid chromatography

Comparison between SFC, GC and HPLC

Instrumentation of SFC

17.4 Gel Electrophoresis

Principles of gel electrophoresis

Nucleic acid migration in gel electrophoresis

Protein migration in gel electrophoresis

Apparatus of gel electrophoresis

17.5 Iso-Electric Focusing

Definition of iso-electric point

Principles of iso-electric focusing (IEF)

How to build the pH gradient in IEF

Application of IEF

17.6 Nuclear Magnetic Resonance Spectrometry

Principles of NMR

¹H NMR spectrum

17.7 Raman Spectrometry

Principles of Raman spectrometry

Comparison between Raman scattering spectrum and infrared absorption spectrum

Problems:

1. Explain how to use standard addition to do quantification.

2. Explain the differences between SFC, GC and HPLC.

3. How to choose the appropriate approach for specific purpose？

4. 教学方法

课堂教授结合提问

4. 教学评价

期末考试

Chapter 18 Sample Preparation

1. 教学目标

Many samples of pharmaceutical interest have a very complex composition, and they cannot be analyzed directly by chromatographic or spectroscopic methods. Sample preparation is required to reduce the complexity of the sample and to make it compatible with the chromatographic or spectroscopic method. This chapter discusses different strategies for sample preparation. Important terms and parameters used in connection with sample preparation will also be defined and discussed. Applications of sample preparation in pharmaceutical analysis are mainly discussed in Chapters 22 and 23.

2. 教学重难点

Principle and selection of different preparation strategies.

3. 教学内容

Contents

18.1 Why is Sample Preparation Required?

Samples of pharmaceutical interest

Challenges for analyzing drugs in complex

18.2 Main Strategies

Protein precipitation (PP)

Liquid–liquid extraction (LLE)

Solid–liquid extraction (SLE)

Solid-phase extraction (SPE)

18.3 Recovery and Enrichment

Determination of extraction recovery

Determination of enrichment

18.4 Protein Precipitation

Principles of PP

Choices of precipitants in PP

18.5 Liquid–Liquid Extraction

Basic theory of LLE

Choices of extraction solvents

Calculation of recovery inLLE

Multiple extractions

Back-extraction

18.6 Solid–Liquid Extraction

Comparison between LLE and SLE

18.7 Solid Phase Extraction

Principles of SPE

The SPE Column

Conditioning in SPE

Equipment of SPE

Reversed-Phase SPE

Secondary Interactions

Ion Exchange SPE

Mixed-Mode SPE

Normal-Phase SPE

Problems:

1. Calculations of recovery and enrichment.

2. What type of aqueous solution should be used if the analyte is acidic or basic in LLE/SPE?

4. 教学方法

课堂教授结合提问

4. 教学评价

期末考试

Chapter 19 Analytical Chemical Characteristics of Selected Drug Substances

1. 教学目标

This chapter selects a number of drug substances in order to demonstrate the significance of chemical characteristics like solubility, log P and pKa values and the impact these basic physicochemical functionalities have on analytical chemical work.

2. 教学重难点

How to characterize different drugs.

3. 教学内容

Contents

19.1 Amitriptyline and Mianserin

Solubility

Sample preparation

UV spectrum

19.2 Morphine and Codeine

Solubility

Sample preparation

UV spectrum

19.3 Ibuprofen and Naproxen

Solubility

Sample preparation

UV spectrum

19.4 Furosemide

Solubility

Sample preparation

UV spectrum

19.5 Paracetamol (Acetaminophen)

Solubility

Sample preparation

UV spectrum

19.6 Neutral Drugs

Solubility

Sample preparation

UV spectrum

Problems:

1. How to choose the appropriate titration method to determine the concentration of a drug?

4. 教学方法

课堂教授结合提问

4. 教学评价

期末考试

Chapter 20  Quatification and Quality of Analytical Data

1. 教学目标

This chapter focuses on the various aspects of quantitative analysis including calibration methods, the principles of analytical validation and system suitability testing. Calibration methods such as the external standard method, the internal standard method, standard addition and normalization are discussed in detail. The performance characteristics used in the validation process are defined along with typical methods by which it may be measured. Test parameters usually employed in system suitability testing are discussed at the end of this chapter.

2. 教学重难点

Method validation, the standard selection.

3. 教学内容

Contents

20.1 Peak Height and Peak Area

Introduce the definition of peak height and peak area, and explain the differences between these two concepts.

20.2 Calibration Methods

20.2.1 External Standard Method

Introduce the principle, procedure, circumstances to use and some examples of external standard method.

20.2.2 Internal Standard Method

Introduce the procedures, circumstances to use and some examples of internal standard method.   

20.2.3 Standard Addition

Introduce the procedures, circumstances to use and some examples of standard addition method.

20.2.4 Normalization

Introduce the procedures, circumstances to use and some examples of normalization method.

20.3 Validation

Explain what is validation and why do validation.

20.3.1 Analytical Procedure

Introduce the analytical characteristics used in validation.    

20.3.2 Accuracy

Introduce the definition, procedure, report rule and tips of accuracy assessment.    

20.3.3 Precision

Introduce the definition, procedure, report rule and tips of precision assessment.  

20.3.4 Specificity

Introduce the definition, procedure, report rule and tips of specificity assessment.

20.3.5 Detection Limit

Introduce the definition, procedure, report rule and tips of detection limit assessment.     

20.3.6 Quantification Limit

Introduce the definition, procedure, report rule and tips of quatification limit assessment.    

20.3.7 Linearity and Range

Introduce the definition, procedure, report rule and tips of linearity and range assessment.  

20.3.8 Robustness

Introduce the definition, procedure, report rule and tips of robustness assessment.

20.3.9 Test Methods in the European Pharmacopeia

The test methods given in monographs and general chapters of the Ph.Eur. have been validated.

Unless otherwise stated in the monograph or general chapter, validation of the test methods by the analyst is not required.

20.4 System Suitability

Introduce the definition, procedure, report rule and tips of system suitability assessment.

Problems:

1. What do peak height and peak area mean?  

2. What are the commonly used calibration methods?

3. Why does the analytical procedure need validate and how to validate an analytical procedure?

4. 教学方法

课堂教授结合提问

4. 教学评价

期末考试

Chapter 21  Chemical Analysis of Drug Substances

1. 教学目标

This chapter focuses on how raw materials, active pharmaceutical ingredients and excipients for pharmaceutical production are controlled by chemical analysis to ensure that they are of sufficiently high quality. The requirements of their identity and purity is given in monographs in the pharmacopoeias (e.g., Ph.Eur., USP) where detailed tests to be performed are described. We briefly review what pharmaceutical products are and how they typically are produced. Then we discuss how the chemical quality control of raw materials is performed, that is: (i) how to identify a test material to verify that it is the correct ingredient for a given production, (ii) how to make sure that the raw material does not contain significant impurities and (iii) how to determine the quantitative content of the raw material.

2. 教学重难点

How to analyze Drug substances

3. 教学内容

Contents

21.1 What is a Pharmaceutical Raw Material, how is it Produced and why must it be Controlled?

Introduce the concept of pharmaceutical raw material, explain its source and the reasons why it needs to be controlled.     

21.2 The Pharmacopoeias – the Basis for Control of Pharmaceutical Raw Materials

Introduce several typical pharmacopoeias from different countries and their coverage. Specifically focus on the European pharmacopoeia, introduce the major contents of this pharmacopoeia.

Dissect the single monograph, explain the meaning of each subsection included.  

 21.3 Which Contaminants are Found in Raw Materials, What are the Requirements in a Maximum Content and Why?

21.3.1 Well Defined Chemical Compounds

Explain the major sources of impurities contained in the type of well defined chemical compounds.

And separately introduce the limits for water, residue solvents, heavy metals, inorganic material and foreign ions.    

21.3.2 Mixtures of Organic Compounds

More diverse range of contaminants are contained in the type of mixtures of organic compounds. The principle for impurities control in this type of pharmaceutical material is "The higher the toxicity is, the more stringent the requirements for the maximum content of a given impurity. The greater the expected daily consumption is of the raw material in the finished product, the more stringent requirements for maximum content".

21.4 How to Check the Identity of Pharmaceutical Raw Materials

21.4.1 Overview of the Identification Procedures

List the main procedures used for the identification of well defined chemical compounds.

21.4.2 Techniques used for the Identification of Well Defined Chemical Compounds    

21.4.2.1 Infrared Absorption Spectrophotometry

Introduce the brief rationale and sample preparation procedures of infrared absorption spectrophotometry. Give an example based on infrared absorption spectrophotometry.

21.4.2.2 Ultraviolet and Visible Absorption Spectrophotometry

Introduce the brief rationale and tips of ultraviolet and visible absorption spectrophotometry. Give an example based on this method.

21.4.2.3 Thin-Layer Chromatography

Introduce the brief rationale and tips of thin-layer chromatography. Give an example based on thin-layer chromatography.

21.4.2.4 Melting Point

Introduce the brief rationale and procedures of melting point measurement.   

21.4.2.5 Polarimetry

Introduce the brief rationale, sample preparation and procedures of polarimetry. Give an example based on polarimetry.

21.4.2.6 High Performance Liquid Chromatography

Introduce the brief rationale and keys of high performance liquid chromatography. Give an example based on HPLC.

21.4.2.7 Chloride and Sulfate Identification

Introduce the principles of chloride and sulfate identification and give an relative example.

21.5 How to Test for Impurities in Pharmaceutical Raw Materials

21.5.1 Main Purity Tests for Well Defined Chemical Compounds

21.5.1.1 Appearance of Solution

Explain the aim of this test, introduce the detailed procedure and give an relative example.

21.5.1.2 Absorbance

Explain the aim of this test, introduce the detailed procedure and give an relative example.

21.5.1.3 Acidity/Alkalinity

Explain the aim of this test, introduce the detailed procedure and give an relative example: test for acidity or alkalinity in dopamine hydrochloride

21.5.1.4 Optical Rotation

Explain the aim of this test, introduce the detailed procedure and give an relative example.

21.5.1.5 Related Substances

Explain the aim of this test, introduce the detailed procedure and give an relative example.

21.5.1.6 Solvent Residues

Solvent Class 1: carcinogenic or highly toxic, should not be used in the production of pharmaceutical raw materials.

Solvent Class 2: relatively toxic, limited by their permitted daily exposure （PDE)

Concentration (ppm) = (1000×PDE)/dose (PDE: mg/day; dose: g/day)

Solvent Class 3: less toxic and thus are allowed in a little higher concentration (<0.5%, w/w)-loss on drying

21.5.1.7 Foreign Anions

Review the principles of foreign anions (chlorides, sulfates) tests and give an example: test for chlorides and sulfates in furosemide.

21.5.1.8 Cationic Impurities

Heavy metals: Relatively specific for heavy metals like lead (Pb), copper (Cu), iron (Fe) and mercury (Hg)

Sulfated ash: Not specific to the cations, but controls the overall level of inorganic impurities in the raw material

ICP-OES and ICP-MS: more sensitive, for heavy metals test in catalysts

21.5.1.9 Loss on Drying

To evaluate volatile impurities in pharmaceutical raw materials (water, residual solvents and so on)

Not specific, but an overview of the overall level of water and solvents

21.5.1.10 Determination of Water

Why evaluate water separately?

Raw materials are particularly sensitive to traces of water

Raw materials are hydrates

More specific, quantitative test for water than the test of loss on drying is needed

21.5.2 Purity Tests for Raw Materials of the Type of Mixtures of Organic Compounds

21.5.2.1 Oxidizing Substances

Explain the principle and procedure of oxidizing substances test.

21.5.2.2 Acid Value

To determine the total amount of free acids in the raw material (fatty oils and synthetic polymers)

Definition: the amount of potassium hydroxide that must be added to neutralize the free acids in 1 g of the raw material.

21.5.2.3 Hydroxyl Value

To determine the number of OH group of synthetic polymers

Definition: the number of milligrams of potassium hydroxide required to neuralize the acid liberated by acylation of 1 g of the substance.

Principle

21.5.2.4 Iodine Value

To express the degree of unsaturation in 100 g of the substance (oils, fats, oleic acid, stearic acid)

Definition: the number of grams of iodine that can be consumed by (substituted into) 100 g of raw material. It’s typically a maximum number not to be exceed.

21.5.2.5 Peroxide Value

To determine the amount of peroxides in the ingredients (synthetic polymers and fatty oils)

Definition: the number of milliequivalents of active oxygen in 1000 g of the substance (1-100).

21.5.2.6 Saponification Value

A test in some raw materials and especially among the synthetic polymers and fatty oils.

Definition: The number of milligrams of potassium hydroxide needed to neutralize free acids and to saponify (hydrolyze) esters present in 1 g of the raw material.

Principle

21.5.2.7 Unsaponifiable Matter

Introduce the principle and procedure of this test.

21.5.2.8 Other Tests

Ask students learn by themselves after class.

21.5.3 Identification of the Raw Materials of the Type of Mixtures of Organic Compounds

For mixtures of organic compounds, no distinction is normally made between first and second identification, and for some compounds no identification procedures is given at all.Color reactions and different kinds of precipitation reactions are often used in mixtures of organic compounds.

21.6 How to Determine the Purity of Pharmaceutical Raw Materials

21.6.1 Acid–Base Titration in Aqueous Environment

Explain the principles of this test and give an example: Quantitative determination of omeprazole by acid-base titration in aqueous environment according to Ph.Eur. 7th edn (2011) (Box 21.25)

21.6.2 Acid–Base Titration in a Non-Aqueous Environment

Explain the principles of this test and give an example: Quatitative determination of zopiclone by non-aqueous acid-base titration according to Ph.Eur. 7th edn (2011) (Box 21.29)    

21.6.3 Redox Titrations

Explain the principles of this test and give an example: Quatitative determination of ferrous fumarate by redox tritration according to Ph.Eur. 7th edn (2011) (Box 21.30)

21.6.4 High Performance Liquid Chromatography

Explain the principles of this test and give an example: Quantitative determination of simvastatin by HPLC according to Ph.Eur. 7th edn (2011) (Box 21.31)  

21.6.5 UV spectrophotometry

Explain the principles of this test and give an example: Quantitative determination of hydrocortisone by UV spectrophotometry according to Ph.Eur. 7th edn (2011) (Box 21.32)

21.7 How to Control Compounds for Which no Pharmacopoeia Monograph Exists

Give general rationales for quality control of compounds not contained in pharmacopoeia.    

21.8 How are Ph.Eur. and USP Updated

Give Useful websites for pharmacopoeias updating:

European Pharmacopoeia’s free online forum

http://pharmeuropa.edqm.eu

http://www.edqm.eu/en/edqm-homepage-628.html

http://www.newdruginfo.com/

Problems:

1. What is a Pharmaceutical Raw Material, how is it Produced and why must it be Controlled？

2. Which Contaminants are Found in Raw Materials?

3. How to Check the Identity of Pharmaceutical Raw Materials?

4. How to Test for Impurities in Pharmaceutical Raw Materials?

5. How to Determine the Purity of Pharmaceutical Raw Materials?

4. 教学方法

课堂教授结合提问

4. 教学评价

期末考试

Chapter 22  Chemical Analysis of FinalPharmaceutical Products

1. 教学目标

Quality control of the final pharmaceutical product is an important part of the entire pharmaceutical production. Quality control of final pharmaceutical products includes both microbiological tests, pharmaceutical tests, and tests based on chemical analysis (chemical tests). This chapter directs our focus towards the latter subject, namely the chemical tests of final pharmaceutical products. The general rules and requirements are discussed, and the different chemical tests that are normally applied to the final pharmaceutical products are discussed in detail. Because a large number of different methods are used in the pharmaceutical industry, this chapter cannot cover them all. However, with some selected examples, which are discussed in detail, this chapter is intended to give a basic understanding.

2. 教学重难点

How to analyze Drug products

3. 教学内容

Contents

22.1 Quality Control of Final Pharmaceutical Products

22.2 Monographs and Chemical Testing

22.3 Identification of the Active Pharmaceutical Ingredient

22.4 Assay of the Active Pharmaceutical Ingredient

22.5 Chemical Tests for Final Pharmaceutical Products

22.5.1 Test for Related Substances

22.5.2 Uniformity of Content

22.5.3 Dissolution

Problems:

1. How many methods are used in the quality control of final pharmaceutical products?

2. What is the definition of capsules?

4. 教学方法

课堂教授结合提问

4. 教学评价

期末考试

Chapter 23  Analysis of Drugs in Biological Fluids

1. 教学目标

This chapter focuses on analytical methods for the determination of drugs in biological fluids and on the application of bioanalytical methods in drug development, therapeutic drug monitoring, toxicological analysis and doping control analysis. Knowledge of the chemical composition of biological matrices such as blood, serum, plasma and urine is crucial for bioanalytical work and is discussed in detail. The various steps of a bioanalytical method include sampling, sample preparation, separation, detection, identification, calibration and quantification. These steps are discussed on a general basis and highlighted by examples discussing various sample preparation procedures, quantitative determinations and identification of unknown components.

2. 教学重难点

How to analyze Drug drugs in biological fluids

3. 教学内容

Contents

23.1 Introduction

23.1.1 Drug Development

23.1.2 Therapeutic Drug Monitoring

23.1.3 Forensic and Toxicological Analysis

23.1.4 Doping Control Analysis

23.2 The Biological Matrix

23.3 Bioanalytical Methods

23.3.1 Sampling

23.3.2 Sample Preparation

23.3.3 Protein Precipitation

23.3.4 Liquid–Liquid Extraction

23.3.5 Solid-Phase Extraction

23.3.6 Separation

23.3.7 Detection

23.3.8 Calibration and Quantification

23.4 Examples

23.4.1 Sample Preparation

23.4.1.1 Sample Preparation Procedure by LLE

23.4.1.2 Comments to the Procedure

23.4.1.3 Sample Preparation Procedure by LLE and Back Extraction

23.4.1.4 Comments to the Procedure

23.4.1.5 Sample Preparation Procedure by SPE

23.4.1.6 Comments to the Procedure

23.4.1.7 Sample Preparation Procedure by Protein Precipitation

23.4.1.8 Comments to the Procedure

23.4.2 Quantitative Determination

23.4.2.1 Quantitative Determination of Amitriptyline in Serum by LC-MS

23.4.2.2 Comments to the Procedure

23.4.2.3 Determination of Valproic Acid in Serum by GC-MS

23.4.2.4 Comments to the Procedure

23.4.3 Identification

23.4.3.1 Sample Preparation Procedure for Unknown Screening by Mixed Mode Cation
Exchange

23.4.3.2 Comments to the Procedure

23.4.3.3 GC-MS Procedure for Unknown Screening

23.4.3.4 Comments to the Procedure

23.4.3.5 LC-MS-MS Procedure for Unknown Screening

23.4.3.6 Comments to the Procedure

Problems:

1. what is the meaning of HTS and TDM?

2. What is the most importment methods in bioanalylied work?

3. What is the major goals of sample preparation?

4. What is UHPLC and reversed C₁₈?

4. 教学方法

课堂教授结合提问

4. 教学评价

期末考试

四、学时分配

表2：各章节的具体内容和学时分配表

五、教学进度

表3：教学进度表

六、教材及参考书目

（电子学术资源、纸质学术资源等，按规范方式列举）

1. Introduction to Pharmaceutical Chemical analysis

2. Pharmaceutical Analysis for SmallMolecules

七、教学方法

（讲授法、讨论法、案例教学法等，按规范方式列举，并进行简要说明）

1. 讲授法

2. 讨论法

3. 案例教学法

八、考核方式及评定方法

（一）课程考核与课程目标的对应关系

表4：课程考核与课程目标的对应关系表

（二）评定方法

1．评定方法

（例：平时成绩：10%，期中考试：30%，期末考试60%，按课程考核实际情况描述）

2．课程目标的考核占比与达成度分析

表5：课程目标的考核占比与达成度分析表

（三）评分标准