| Fufang Jiubiying Capsule is a traditional Chinese medicine containing Salvia,Xiangfu and Dongfeng orange;It is mainly used for the treatment of gastroenteritis and diarrhea.It is alsoconsisted to be functioning for pain relief,anti-inflammatory,sedative,antipyretic,and even for treatment of abdominal pain and intestinal diseases.The main active ingredients of Xiangfu are α-cyperone,cyperotundone and nootkatone,all of which are isomers.In the present study,liquid chromatography-tandem mass spectrometry(LC-MS/MS)technology was used to establish a method for simultaneous quantitation of α-cyperone,cyperotundone and nootkatone in Fufang Jiubiying Capsules.In addition,a modified method for simultaneous determination of α-cyperone,cyperotundone and nootkatone in rat plasma has been successfully applied to their pharmacokinetic studies in rats.Furthermore,there are gender differences in metabolism of cyperotundone in rats when using the rat liver microsomal system.These observations are consistent with its pharmacokinetic result between male and female rats.The main details are the following:(1)An LC-MS/MS method for simultaneous determination of the contents of α-cyperone,cyperotundone and nootkatone in the extract of Fufang Jiubiying Capsules has been established.According to the requirements,the methodological verification of a series of related projects was carried out,including: instrument precision,linear range,repeatability,stability,sample recovery rate,and the results were all in line with the relevant standards of quantitative analysis.The average contents of α-cyperone,cyperotundone and nootkatone in the three batches of Fufang Jiubiying Capsules were 437.9 μg/g,848.8 μg/g and 43.0 μg/g,respectively.The method is simple in operation,good in selectivity,high in sensitivity,and accurate and reliable in results.The method is simple in operation,good in selectivity,high in sensitivity,and accurate and reliable in results.(2)A method for simultaneous determination of three isomeric compounds(α-cyperone,cyperotundone and nootkatone)in rat plasma by LC-MS / MS has been established.According to the relevant requirements,the methodological verification of the series of projects mainly includes: selectivity,linear range,precision,accuracy,stability,recovery rate and matrix effect..The results showed that the lower limit of quantification of α-cyperone and cyperotundone was 1 ng/mL,the linear range was 1 to 500 ng/mL;the lower limit of quantification of nootkatone was 5 ng/mL,and the linear range was 5 to 2500 ng/mL.Its Inter-/intro precision,accuracy and stability were in line with the relevant requirements of the relevant quantitative analysis of biological samples,and the recovery were between 88% and 115%.The method has high sensitivity,good selectivity and better resolution,which can be used for the pharmacokinetic study of three isomer in Fufang Jiubiying Capsule extracts in rats.(3)The validated LC–MS/MS method was used to determine the plasma concentration of the three isomer and the pharmacokinetic characteristics in rats after the male and female rats were administrated the Fufang Jiubiying Capsule extracts by single dose intravenous(0.3 g/kg)and a single dose oral administration(3.0 g/kg).The results showed that the concentration of the three isomers in plasma began to decrease rapidly after intravenous injection of the Fufang Jiubiying Capsule extracts in female and male rats,and no significant male-female differences were observed.After oral administration of 3.0 g/kg of Fufang Jiubiying Capsule extracts in female and male rats,there were significant differences in the in vivo pharmacokinetics of cyperotundone,which be absorbed too fast to measure the effective parameters in male rats.(4)The substrate elimination method was used to study the metabolic stability and differences of α-cyperone,cyperotundone and nootkatone in the liver microparticle system of male and female rats respectively.The results showed that cyperotundone was metabolized slowly in the female liver microparticle system but metabolized faster in the female liver microparticle system.The metabolic stability of α-cyperone and nootkatone in the liver microparticle system of male and female rats did not show significant difference. |
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