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H P L C

H P L C. high pressure liquid chromatography. Analyzing Vitamins B1 and B2 Using Ion Pairing. Travis Crisp and Joanna Magner Mentor: Dr. Christine Pastorek. E q u i p m e n t and chemicals. HPLC (High Pressure Liquid Chromatography): C18 column

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H P L C

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  1. H P L C high pressure liquid chromatography Analyzing Vitamins B1 and B2 Using Ion Pairing Travis Crisp and Joanna Magner Mentor: Dr. Christine Pastorek

  2. E q u i p m e n tand chemicals • HPLC(High Pressure Liquid Chromatography): C18 column Settings: 280nm, 2.5mL/min (4000psi), isocratic conditions (10 MeOH : 90 aq) • Balances • Glassware: beakers, volumetric flasks, graduated cylinders, etc. • Chemicals:Methanol (MeOH), deionized and milli-pore H2O, Thiamine (Vitamin B1), Riboflavin (Vitamin B2), Acetic Acid, NaOSA, Mobile Phase

  3. P r o c e d u r e • Determine best wave length to run lamp • Create mobile phase(s) and sample solution(s) • Run pump and inject sample • Allow solution to flow through system completely • Analyze results

  4. P r o b l e m s E n c o u n t e r e d • Inconsistent labeling of mobile phase • Column problems (polarity, “clogs”) • Air bubbles in tubing • B1 disappearing in later tests • Over pressurization of pump • Estimations and assumptions in sample preparation

  5. Results • B1 was more polar compared to B2 • Mobile phase content changed run time • Air bubbles cause difficulties • Composition of the column changes over time • B2, in real samples, was detected

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