| Summary: | 碩士 === 國立臺灣大學 === 藥學研究所 === 93 === Sennoside A and sennoside B are the most significant and abundant active components in senna tablets. Clinically, they are used for constipation treatment. However, overdose will cause tormina and severe diarrhea. In order to make sure and determine the doses of sennoside A and sennoside B effectively, this study employed one of the monolithic columns, ChromolithTM performance RP-18e. Compared to the traditional particle-packed column, this one is with lower column pressure and thus it can be analyzed under high flow rate to reduce separation time.
In the course of method development, parameters including concentration and pH value of buffer solution, concentration of ion-pair reagents, volume of organic solvent modifier, column temperature and flow rate of the mobile phase were varied to find out the optimum conditions of separation. The most optimum conditions were : ChromolithTM performance RP-18e (100 �e 4.6 mm) as column, 0.1 M acetate buffer at pH 5.0 and ACN (70:30, v/v) containing 7 mM tetra-n-octylammonium bromide as mobile phase. Flow rate, temperature, and detection wavelength were set at 5 mL/min, 25℃, and 270 nm, respectively. Sennoside A and sennoside B could be separated from other components in 5 minutes.
The optimum condition was validated with the tests of specificity, precision, linearity, limit of detection and accuracy. Plackett-Burman experimental design was adopted for the robustness test. The effects of five experimental factors, including concentration of ion-pair reagent, percentage of organic modifier (acetonitrile), pH and concentration of buffer and flow rate on four response factors, such as resolution, retention time, plate number and tailing factor were investigated. Five experimental factors mentioned above had significant effects on plate number of sennoside A. Concentration of ion-pair reagent, percentage of organic modifier and flow rate especially had greater effect on resolution.
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