• Where to begin if you need to perform HPLC analysis?

  First search in the literature for existing methods, do not re-invent the wheel.  Go to Google and search for your compound plus the two pharases "mobile phase" and "liquid chromatography":

  • Waters applications for method development and validation
  • •Modes of liquid chromatography
  • Search a monograph in the International Pharmacopoeia of WHO
  • Selectivity Chart of Columns
  • 2-Method Transfer & Development
  • A review on HPLC Method Development and Validation

Understanding and selecting the right HPLC column בחירת הקולונה המתאימה:

Now, you got an application note or a method from the literature.  Chances are (90%) that you got a reversed phase application for your compound.  What is next? There is a huge selection of reversed phase columns, do I need to buy exactly the same column as the literature shows?  Not necessarily, there is a term called: equivalent column.

• USP Column Equivalency Application
• Characterization of Stationary Phases in RP Chromatography by H. A.Claessens
• Waters' Columns Selectivity Chart Application - Download
• ACDLabs Columns Selector

Optimize and validate the HPLC method אופטימיזציה וולידציה של השיטה:

After you set up your HPLC method, you need to test it on known samples, and you need to validate your method to be sure that it will work properly and accurately from now on.  The next links describe the process and the reason for optimization and validation of HPLC methods.

• A review on HPLC Method Development and Validation
• How To Make a Phosphate Buffer
• Method development and validation overview 2
• Analytical Method Development and Validation

Once you have the method, make sure to assure that it is appropriate for your specific needs, this is done by validation.

• FDA reviewer guidance for HPLC method validation
• Quality by Design (QbD) Considerations for analytical methods: FDA perspective
• Quality Assurance Guidelines - ICH
• FDA Guidance to the industry on validation of bioanalytical methods

Instrumentation Qualification תיקוף מכשירים

Topics related to the hardware of HPLC systems

• Committing to Calibration of Instruments
• Wikipedia: Verification and validation
• Operational Qualification of HPLC Instruments: LCGC North America Vol. 20 No 4 Apr 2002 (Grisanti and Zachowski, Ed: Krull&Swartz)
• The procurement, qualification and calibration of lab Instruments: An Overview
• Performance Qualification of HPLC Instrumentation in Regulated Laboratories

• Preparation of Calibration Curves
• Comparison of analytical calibration methods
• Interactive Computer Models for Analytical Chemistry Instruction    
• Quantitation in LC-MS and LC-MS/MS (Good!)
• Article by N.A. Epshtein: Quantitative work in HPLC  
• Thesis on Method Development and Validation and Quantitation of Drugs

General introduction: books, presentations and overviews

• On-line lecture
• Overview
• Detection parameters: Sampling Rate and Response Time

Light Spectroscopy Detectors

• Principles of Fluoresence Detectors
• Principles of UV detectors
• Principles of Photo-Diode Array (PDA) detectors
• Principles of Refractive Index Detector
• UV Spectroscopy lecture (PPT)
• Refractive Index Explained in animation
• Fluorescence Tutorials in Animations

Aerosol Detectors

• Principles and operation of ELSD
• Charged Aerosol Detection in Pharmaceutical Analysis
• Evaporative Light Scattering Detector (ELSD)

Electrochemical & Conductivity Detectors

• Principles of Conductivity Detector
• Principles of Electrochemical Detecror
• Electrochemical Detection in liquid flow systems - IUPAC
• Ion chromatography and conductivity detector

Mass Spectrometry Detection

• QDa MS Detector
• My page on LCMS
• Considerations for Adapting an HPLC Method for MS Detection
• A Primer of Mass Spectrometry

*  (User guide of Waters' detector: Register to website first) to get the following information on each detector:

• Theory of operation
• Installation procedures
• User procedures
• Maintenance and troubleshooting procedur

Now that you have a validated method, you start working daily, now you need to know how to operate the instrument, chemistry and software smoothly and trouble-free.  For instructions see Operational Instructions

The following are topics related to the proper operation of the HPLC system, chemistry, fluidics, software, plumbing, etc.

• A guide for a trouble-free  operation of HPLC - Hebrew ( by me  210K)
• Guidelines in the operation of an HPLC system
• Selecting water for HPLC by Millipore:  MilliQ system
• Sample preparation - filtration products 
• On-line recipes for Buffers - PFG        
• Performance Verification of HPLC  By Dr. Herman Lam
• SOP for handling and working with analytical standards
• Regeneration of Reversed  Phase  Columns  (LC-GC Europe  July  2003)
• Isranalytica 2006- Unexpected Peaks in the Chromatogram (by Shula Levin 2 MB)
• Cleaning and regeneration of HPLC columns   
• The Troubelshooting guide of Waters  
• HPLC Troubleshooting (Proteomics page)

• Selecting water for HPLC
• Type I ultrapure water crucial for HPLC and UHPLC (ELGA) (pdf)
•  Water Purity for UHPLC by Merck (pdf)

• PierceNet Solvents
• Sigma-Aldrich
• JT Baker solvents
• Spectrum-Chemicals
• Biolab

• On-line recipes for Buffers
• HPLC Buffers-Sigma
  

• Solvent Preparation
• Autosampler Vials
• Thomas Scientific - Solvents preparation
• Sample Preparation Products
• Waters Inc. Filtration products (register first)
• SGE Syringes
• Sample preparation - filtration products

(login to website  first:  http://www.waters.com)

• Preparative chromatography Considerations- by Waters
• Scaling up to Preparative HPLC presentation
• My old presentation on Preparative HPLC
• Scale up of Peptide Separations
• Study of Selectivity and Loadability of Pharmaceutical Compounds on RPLC Columns
• Preparative Loadability as a Function of pH   Preparative Chromatography at High pH
• The Role of the Column in Preparative HPLC   
• Practical Aspects of Preparative HPLC in Pharmaceutical and Development Production   

Ultra-Performace Convergence Chromatography (UPC2)

•  State of the Art Separations by Convergence Chromatography

• Finder of Application
  

• General introduction  with many references.

•   Wikipedia

•   Lipids with UPC2

• Introduction

• Pushing the Boundaries GEN

• Presentation on UPC2
  

• A New Separation Tool for a Broad Range of Analytical Challenges

• SFC solutions: applications

Free pdf Chromatography books: LC and GC ספרים אלקטרוניים בחינם
BookRags-on-line free books:   HPLC 
i-Papers by SCRIBD מאמרים וכתבים בחינם

Chromatographic Science Series 
Textbooks
Journals
Special Topics
Abstracts
News in Separation Sciences
 
Text Books
Amazon: BOOKS ON CHROMATOGRAPHY
Chromatographic Science Books
Analytical Chemistry in a GMP Environment

My Favorite Electronic Library  הספריה האלקטרונית שלי 

• ScienceDirect
• ACS publications
• LCGC Magazine
• Elsevier
• The scientist
• ChemPort Journals
• InterScience- Wiley Chemistry Journals
• Springer Science On-line      Springer Chemistry Journals
• Journal of Chromatography A        Journal of Chromatography B
• JASMS-online
• Journal of Chemical Education
• Journal of Chromatographic Sciences (JCS)

Magazines on-line מגזינים מקצועיים

• http://www.spectroscopyeurope.com/   

• http://www.spectroscopymag.com/spectroscopy/   

• http://www.spectroscopynow.com/view/index.html?tzcheck=1  
  

• http://www.scientistlive.coml   

• http://www.chemindustry.com/index.html  
  

• http://www.chemsoc.org/CFCONF/newrscevents.cfm   

• http://www.sciencedaily.com/ 

Chemistry Journals כתבי עת בכימיה

http://www3.interscience.wiley.com/search/allsearch

ScienceDirect
http://www.sciencedirect.com/science/publications/journal/chemistry

Beilstein Abstracts @ ChemWeb.com
http://chemweb.com/

CAS - Chemical Abstracts Service
http://www.cas.org

ACS publications
http://pubs.acs.org/

ISI - Thomson
http://www.isiwebofknowledge.com/

Kluwer Online
http://www.kluweronline.com/

Taylor & Francis Group
http://www.tandfonline.com/

Ingenta
http://www.ingenta.com/

InfoTrieve
http://www4.infotrieve.com/users/logon.asp?component=AF&dk=/search/databases/newsearch.asp

Biomedical and Life Sciences מדעי החיים

PubMed
http://www.ncbi.nlm.nih.gov/PubMed

Medscape (from WebMD)
http://www.medscape.com

Cambridge Scientific Abstracts
http://www.csa.com/

Free Acess Presentations on the Net מצגות פתוחות לקהל

• Lab Tech Guy on HPLC wash solvents
• Planar Chromatography
• Proteins – Basics you need to know for Proteomics
• Design And Purification Of Proteins
• High Performance Liquid Chromatography
• High Performance Liquid Chromatography HPLC
• Mass Spectrometry
• HPLC Columns and Stationary Phases
• The Basics of Chromatography
• HPLC?
• Tips and Tricks: HPLC Troubleshooting
• Column Troubleshooting
  
• HPLC Course - ppt
• LC-REsources: LC-MS extra slides
• Analytical vs Preparative
  
• Stationary Phases