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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 column הקולונה
• Detector  הגלאי
• Injector   המזרק
• Gradient and isocratic  גרדיינט או איזוקרטי
• Data Processing 
• עיבוד נתונים

• Column Selection -Advisor
• Columns Selectivity Chart
• Vials-Selector
• Plates -Advisor
• Sample-Preparation-Product-Advisor

Chemistry Products for Bio-Separations מוצרי כימיה להפרדות ביולוגיות

איתור חלקי חילוף

  MORE:

• מדריך להפעלת HPLC בעברית  Hebrew guide for HPLC operation (pdf 210K)
• How To Make a Phosphate Buffer
• Preparation of the Waters' Alliance  2690/5 (English) in details פירוט הוראות להכנת מכשיר
• Step by Step Procedure for the Preparation of 2695 unit צעד צעד: הכנת מכשיר לעבודה.
• Preparation of the Waters' 2790/5 Separation unit -Alliance HT הכנת מערכת לעבודה
• Attacking Carry-over Effects מאמר על התמודדות עם
• Maintenance of HPLC with concentrated rough samples המלצות על התמודדות עם דוגמאות קשות
• Switching between Normal phase and Reversed phase הדרכה על מעבר מהיפוך פאזות לפאזה נורמלית
• Hebrew Overview on HPLC (pdf 350k)
• Selecting water for HPLC איך לבחור מים  (Millipore's)
• Solvent Preparation
• The importance of Temperature Control החשיבות של שליטה בטמפרטורה
• Final UPLC Method description: Example   (pdf) טופס דוגמא לכתיבה של שיטה סופית
• Column Archives: LC Troubleshooting LC-GC Europe -  By: John W. Dolan
• Waters applications for method development and validation פיתוח שיטות וולידציה באתר של ווטרס
• "Ghost" peaks: Background contamination

הסבר על הפרופילים של הגרדיינטים במשאבות

Overview (can be used for SOP):

• Peak Purity and Spectral Matching procedures - Experimental and software considerations (English)

• Using Photo-Diode Array Detectors (PDA) for Spectral Analysis of chromatograms
  

• •Principles of Peak Purity and Spectral Matching

• Peak Purity I  ; Peak Purity II;  Peak Purity III;  Library Matching. 
• Instrumental Deviation from Beer's Law  

• Empower - Getting Started with PDA Software Guide

Another Tutorial for self - Training

• Some basic tools for working with Waters' PDA

Overview (can be used for SOP):

• Peak Purity and Spectral Matching procedures - Experimental and software considerations (English)

• Using Photo-Diode Array Detectors (PDA) for Spectral Analysis of chromatograms (Hebrew-
  

• Principles of Peak Purity and Spectral Matching - 

• Peak Purity I  ; Peak Purity II;  Peak Purity III;  Library Matching. 
  
• Instrumental Deviation from Beer\'s Law  

•  PDA Default - Millennium Project (download, save, unzip and restore to your Millennium station) to use with:

Register to Waters\' website and download the pdf guide:

• Empower - Getting Started with PDA Software Guide

Another Tutorial for self - Training

• Some basic tools for working with Waters\' PDA

• JT Baker solvents
• Spectrum-Chemicals
• Merck HPLC/TLC solvents
• PierceNet Solvents

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

• Solvent Preparation
• Autosampler Vials
• Vial Selector  - Plate Selector
• Filter Selector
• Waters Inc. Filtration products (register first)
• SGE Syringes