色譜實驗exp 3

1、Exp 3 The assay of Chloramphenicol by HPLC,1. Purpose1) To study the principle and procedure of HPLC2) To learn the assay method for Chloramphenicol by HPLC3) Grasp the method of establishing and optimization for the

2、condition of HPLC,2. PrincipleChromatography is used both for separation and quantitative analysis of complex sample. The methods for quantitative analysis include methods of normalization, external standard, internal

3、standard (IS) and internal calculation.,3. Instrument and conditionAgilent 1200; Schimadzu LC 20AT/FL; JascoColumn: C18(ODS); 4.6×150mm (4.6×250mm), dp5µmMobile phase: MeOH : H

4、2O : HAc = 55 : 45 : 0.1Flow rate: 1.0ml/min Wavelength: 280nmInjection volume: 20µl （60 ~ 80µl）,4. Chemicals,Standard solutionSample solution:,,,,,5. ExperimentTo inject 20µl of standard solutio

5、n and sample solution respectively into the HPLC and record the chromatogram,6. Result,7. Calculation 1) To construct the calibration curve by plotting the peak areas of analyte versus the concentration

6、 A = a×C + b 2) To calculate the concentration of chloramphenicol according to the calibration curve 3) CX: concentration of sample solution M: labeled amo