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אתר הבית של שולה לוין בנושאי כרומטוגרפיה נוזלית

Handouts for HPLC Course

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On-Line HPLC Course קורס מקוון

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Topics of Lectures

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Fundamentals of liquid chromatography יסודות הכרומטוגרפיה הנוזלית - Theory* and Instrumentation*
General Introduction to HPLC - Lecture 
הקדמה כללית
Chromatography Process Movie
סרטון וידאו
Guidelines in the operation of an HPLC system
הנחיות לתפעול נכון של המערכת
Various modes of liquid chromatography
  סוגי הכרומטוגרפיה
Reversed phase chromatography - Mobile Phase Considerations and chemistry and geometry of the columns
כרומטוגרפית היפוך פאזות
Sample Preparation: Solid Phase Extraction (SPE)
הכנת דוגמאות
Detection in liquid chromatography 
Lecture Handouts
Detection in HPLC & Qualification of detectors  
גלאים וכיולם
Photo-diode-array (PDA) detectors: Principle of operation, Peak purity, Spectral matching
גלאי מערך דיודות: עקרון הפעולה, נקיון פיקים והשוואה ספקטרלית

Hebrew Article
מאמר בעברית
Liquid chromatography - mass spectrometry (LC-MS) (ask for the password)
כרומטוגרפיה נוזלית-מס ספקטרוסקופיה
Gradient conditions - Principles and guidelines  
עבודה בגרדיינטים
Quantitation and system suitability
עבודה כמותית
Integration events and calibration methods 
Validation of chromatographic methods - System suitability, precision, accuracy, limit of detection, limit of quantification etc ולידציה של שיטות

Method validation guidelines
Analysis of impurities and degradation products - FDA Guidance
אנליזה של אי נקיונות
Preparative chromatography - considerations in scaling up a separation
כרומטוגרפיה פרפרטיבית
System peaks in liquid chromatography - Their origin and formation, how to live with/without them
שיאי מערכת 
Chiral chromatography for the analysis of enantiomers
כרומטוגרפיה כיראלית
Chiral chromatography article
Analysis of ions - Ion Chromatography (IC)
אנליזה של יונים
Capillary Electrophoresis (CE)
אלקטרופורזה קפילרית 
Ion chromatography course handouts
כרומטוגרפית יונים
Analysis of polymers
- אנליזה של פולימרים -
Gel Permeation Chromatography (GPC) for the determination of molecular weight distribution

GPC course handouts
שיקופיות קורס
Troubleshooting in HPLC
איתור תקלות
High Performance Liquid Chromatography (HPLC) in the pharmaceutical analysis

Please note:   Some of the following documents are Adobe-Acrobat Reader files for viewing and downloading.  You might need to save the PDF file to your computer first by pointing the link with the right mouse and selecting "Save link/target as", then open it with Adobe-Acrobat Reader.


* The type of material and information that is presented in parts of this lecture are similar to the material in the links by  Dr. Yuri Kazakevich and Prof. H.M. McNair's on-line HPLC book.


** Some of the material and information that is presented in parts of this lecture were taken from Waters  seminars around the world


Successful Operation of HPLC System תיפעול נכון של  מערכת


Before beginning to work with any HPLC instrument one must understand the basic principles of liquid chromatography at its various modes. 

The liquid chromatograph is very complex and it consists of sophisticated technology and sometimes delicate and total ignorance in the manner it works might sabotage its parts which are very expensive.  In addition, there are chemical processes of distribution between the two phases, the mobile and stationary, while the analysis goes on, and there is a need for a minimum understanding of their theoretical basis to operate the system correctly.  Also, a minimum understanding of the method, in which the system performs data analysis, is needed so that no false data or results will be obtained in the analysis.


LISTS OF STEPS NEEDED BEFORE ANY RUN BY HPLC  רשימת צעדים מקדימים לפני הרצה :


  • Filter the solvents with membranes with cutoff of 0.22-0.45 mM
  • Use clean and transparent reservoirs through which precipitates and colloids can be distinguished.
  • Make sure that the solvents will be easily mixed with the previous solvents in the same inlets (A, B, C, D).  For example methanol or water should not be placed instead of hexane directly, or any organic solvent should not be placed directly instead of a buffer reservoir.
  • Degass the solvents and purge all the tubing that lead to the pump.
  • Connect the column carefully according to the flow direction marked on it (do not connect directly to the detector).
  • Flow the appropriate solvents through the column at a low flow-rate (0.1-0.5 ml/min) or reach the composition gradually using the appropriate gradient.
  •  Select the appropriate wavelength (or other type of setting) in the detector and wait for stable baseline
  • Prepare the set of methods in the workstation: Instrument method for the control on the system, Processing method for the data processing and the Report method for the report of final results.
  • When the system and the methods are ready a blank run should be performed to test the system and verify that it is clean from interferences.

 The rest of the article  Click   להמשך המאמר 

Layout of the rest of the article:


Solvent delivery system המשאבה
The column הקולונה
Detector  הגלאי
Injector   המזרק
Gradient and isocratic  גרדיינט או איזוקרטי
Data Processing  עיבוד נתונים


This website is intended for academic support of HPLC/LC-MS users in Israel first of all. The rest of the world is welcome too. 

האתר הזה מוקדש קודם כל למשתמשי כרומטוגרפיה נוזלית בישראל - משתמשים מכל רחבי העולם (פירוט למטה) מתכבדים גם

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Copyright (C)1997 Dr. Shulamit Levin
Waters (TC) Israel

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