‘To raise new questions, new possibilities, to regard old problems from a new angle, requires creative imagination and marks real advance in science’ – Albert Einstein.
With the same pace and eagerness to learn AAPS student chapter hosted another workshop and this time for learning high performance liquid chromatography (HPLC) and it’s instrumental training. For second consequent time we were fortunate to have Joe with us for the workshop held on on 23rd Oct, 2015. The highly trained and very professional Joe introduced HPLC with the theoretical concept of chromatography. With a simple yet informative video he described its operation. The lecture proceeded discussing different challenges and their ‘quick fix solutions’. Thereafter qualified candidates were selected for instrumental training held on 30th Oct 2015.They had hands on experience with the HPLC under Joe Ocando’s supervision.
Answers from the debrief
How does it work? The components involve a reservoir that holds the solvent, a high-pressure pump, an injector/sample manager and the HPLC column. A detector is needed to see the separated compound bands as they elute from the HPLC column. The mobile phase exits the detector and can be sent to waste, or collected, as desired. When the mobile phase contains a separated compound band, HPLC provides the ability to collect this fraction of the eluate containing that purified compound for further study.He emphasized how important it is to understand a chromatogram and its extrapolation for the compounds desired in separation.
How do I choose? The importance of detector was explained along with how a choice is made among many different types of detectors, depending upon the characteristics and concentrations of the compounds that need to be separated and analyzed. In a conventional UV-Vis detector, manual or a motor governed mechanical movement is required to select a specific wavelength. But a photodiode array UV-Vis acquires data at each wavelength by electrical scanning.The wavelength is therefore reproducible in a PDA instrument and is much better than the conventional mechanical scanning UV-Vis spectrophotometer.
What is important before I start HPLC?
- Precautions like purging and stabilizing column before method run
- System suitability parameters and their weighing during method development and validation.
- Sample preparation
- Column maintenance and efficiency considerations
Yes we are competent in carrying out analytical research. Joe was generous to have helped one of our graduate students by recommending the software version required for specific system from the Waters company. His advice had impacted the student’s research work significantly. He’s in depth knowledge of Analytical Chemistry and commercialized approach for selecting an equipment specific for the use always comes handy for our fellow researchers.