Establishment of content determination of 5 saponins in Huoxue zhitong capsules/tablets and confirmation of illegal addition / 中国药房

ABSTRACT

OBJECTIVE To establish a method for simultaneous determination of 5 saponins in Huoxue zhitong capsules/ tablets and to confirm the illegal addition of Panax ginseng ，Panax quiquefolium and stems and leaves of Panax notoginseng . METHODS Ultra-high performance liquid chromatography tandem mass spectrometry was used . The Agilent RRHD Eclipse Plus C18 column was used with mobile phase of water （containing 0.1% formic acid ）-acetonitrile for gradient elution at a flow rate of 0.35 mL/min. The column temperature was 35 ℃andthesamplesizewas 2 μL. Using electrospray ionization source ，negative ion scanning was carried out in multi -reaction monitoring mode . RESULTS The linear ranges of notoginsenoside R 1，ginsenoside Rb 1， ginsenoside Rg 1，ginsenoside Rd ，ginsenoside Re ，ginsenoside Rf （an unique ingredient of P. ginseng），ginsenoside Rb （3 an unique ingredient of stems and leaves of P. notoginseng）and pseudo -ginsenoside F 11（an unique ingredient of P. quiquefolium）were 9.99- 1 499.50，9.99-1 499.50，10.01-1 500.80，9.99-1 499.10，10.00-1 500.20，9.99-1 499.50，10.01-1 500.80，9.99-1 499.00 ng/mL （R2＞0.997）；the detection limits and the quantitative limits were not higher than 2.64 and 8.06 ng/mL，respectively. RSDs of precision，repeatability and stability （24 h）tests were all less than 6%. The average recoveries of saponins in capsules and tablets were 98.72%-102.40% and 95.18%-106.47%，respectively（all RSDs ＜5%，n＝6）. In 18 batches of Huoxue zhitong capsules ，the contents of ginsenoside Re ，ginsenoside Rd ，ginsenoside Rg 1，notoginsenoside R 1 and ginsenoside Rb 1 were 291.79-426.89，427.71- 677.49，2 294.28-3 371.43，571.22-848.19 and 1 841.33-2 959.12 μg/g，respectively；the contents of ginsenoside Rb 3 were no more than 45.02 μg/g. In 22 batches of Huoxue zhitong tablets，the contents of above indicators of P. notoginseng were 44.11-393.83，80.48-549.55，393.36-3 548.57，79.83- 872.60，and 288.64-2 912.66 μg/g，respectively；the contents of ginsenoside Rb 3 were no more than 44.79 μg/g. Ginsenoside Rf and pseudo -ginsenoside F 11 were not detected in the two preparations. CONCLUSIONS The method can be used to determine the contents of saponins in Huoxue zhitong preparations . No illegal addition of P. ginseng and P. quiquefolium are found in 40 batches of preparations ，but the input of P. notoginseng in some batches of tablet samples is less .