How to Train in HPLC
High Pressure Liquid Chromatography (HPLC) is an analytical instrument used in chemistry to determine the identity of organic compounds. HPLC is widely used in research and in the pharmaceutical industry for precise analysis and complex molecular identification. HPLC equipment must be thoroughly cleaned via the degassage pump cleaning unit prior to every use to ensure the system is clean. HPLC compound analysis is conducted by liquid (the mobile phase) transported throughout the system and separated via the column. HPLC can be a very complicated instrument to use, without fundamental training gained by studying chemistry at university or college. Training a new employee in HPLC can be challenging as many questions will arise. Initially, you will need to ensure that all machine manuals are available, and to assess the level of knowledge, background and qualifications of the trainee.
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Things You'll Need
- HPLC instrument
- Standard Operating Procedures (SOP) manual
- Sample to be analysed
- Standard manufacturer supplied solution, as reference material to measure against the sample
- Computer
- Software
- Printer
Instructions
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Basic Steps to Train in HPLC
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1
Give a brief tour of the HPLC system. Describe the machine used in the laboratory and explain why the company bought it. Provide a short explanation of HPLC and its historic use on the premises and in the business. Give specifics of HPLC by showing where the main components are. Show the trainee where the reservoir is, for the mobile phase. Describe how the entire system can be cleaned by degassage. Show the injector for sample injection and the column where the separation occurs. Describe what type of detector will be used. Explain the computer and software system. Ideally follow this brief training session up by allowing the trainee to observe the HPLC in use.
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2
Make notes of the Standard Operating Procedure for HPLC to aid understanding. Provide the trainee with a copy of the Standard Operating Procedures (SOP). All steps regarding use of HPLC are in the SOP. The SOP provides company guidelines for running HPLC and all employees should use it as reference material when using the equipment. Ensure that the trainee reads the manual with care and understands all aspects of the written explanation. Encourage the trainee to make notes.
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3
Allow the trainee to practice running HPLC. Observe that the trainee prepares the mobile phase, sample and standard solution according to the method specification. Assist the trainee with choice of column and the correct method of connecting to HPLC instrument (detector and injectors), by following the arrow. Observe and assist the trainee to program the method of test designed to run the HPLC. Ensure that they wash the system using the degassage for 15 minutes and that it has been shut down correctly to allow the mobile phases to run through the column. Help the trainee to write the test sequence list that allows the software to organise the HPLC run according to the specification and ensure this is saved.
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4
When the run has finished, observe the trainee in the re-processing of results by writing different variables like weights, volume used to dilute sample and standard solution. Show the trainee how to print the results and to check if they are within the specification. As trainer you will need to sign off and accept responsibility for these results, for traceability. Record the results in the trainee's record of training. Advise the trainee to seek assistance from experienced chemists on future HPLC runs.
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References
Resources
- Photo Credit laboratory image by Alhazm Salemi from Fotolia.com large reading glasses resting on a lined notepad image by Steve Johnson from Fotolia.com
