Qualitative and Quantitative Study of Tibetan Medicine Thlaspi semen / 中国药房

ABSTRACT

OBJECTIVE： To establish the qualitative and quantitative control method of Tibetan medicine Thlaspi semen. METHODS： TLC and HPLC method were used to identify and determine flavonoids isovitexin， swertisin and glucosinolates sinigrin from 15 batches of T. semen. The stationary phases identified by TLC of flavonoids and glucosinolates were polyamide film and high performance silica gel GF254. The developing agents were trichloromethane-methanol-glacial acetic acid （11 ∶ 1 ∶ 1，V/V/V） and ethyl acetate-methanol- triethylamine （4 ∶ 5 ∶ 0.5，V/V/V）. In chromatogram condition of content determination of isovitexin and swertisin， the separation was performed on CAPCELL PAK MGⅡ C18 column with mobile phase composed of acetonitrile-0.4％ glacial acetic acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 336 nm. In chromatogram condition of content determination of sinigrin， the separation was performed on CAPCELL PAK MGⅡ C18 column with mobile phase composed of acetonitrile-0.02 mol/L tetrabutylammonium hydrogen sulfate （15 ∶ 85，V/V，pH 6） at the flow rate of 1.0 mL/min. The detection wavelength was set at 227 nm. RESULTS： In TLC identification chromatogram， spots corresponding to isovitexin， swertisin and sinigrin control were detected in test samples. The linear ranges of isovitexin， swertisin and sinigrin were 1.26-79.00， 1.21-75.38， 12.80-640.00 μg/mL， respectively （all r≥0.999 5）. The limits of detection （LODs） were 0.09， 0.12， 0.15 μg/mL， and limits of quantitation （LOQs） were 0.39， 0.43， 0.54 μg/mL， respectively. RSDs of precision， stability （24 h） and reproducibility tests were all lower than 2.0％（n＝6）. The recoveries were 99.1％， 97.0％ and 98.1％， and RSDs were 1.9％， 1.8％， 1.8％（n＝6），respectively. The contents of isovitexin， swertisin and sinigrin in 15 batches of T. semen were 0.013-0.090， 0.020-0.130 and 18.92-40.75 mg/g， respectively. CONCLUSIONS： Established quality control method is simple， reproducible and stable， and can be used for the quality control of Tibetan medicine T. semen.