| Since the therapeutic effects of traditional Chinese medicines (TCM) are often the results of the complex interactions of multiply components, in vivo quantitative analysis of multiple bioactive components from TCM in biological samples is essential for understanding and confirming their efficacies and action mechanisms. Ultra high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) is a powerful tool for analysis of complex biological samples since its high sensitivity, high selectivity and high throughput. The methods for simultaneous determination of multiple bioactive components in rat plasma by UHPLC-MS/MS and pharmacokinetic study after oral administration of traditional Chinese medicines were established in this thesis.1. A UHPLC-MS/MS method was developed for simultaneous determination and pharmacokinetic study of phellodendrine, coptisine, jatrorrhizine, berberine, palmatine, baicalin, wogonoside, baicalein, wogonin and emodin in rat plasma after oral administration of Yankening Capsule. After mixing with two internal standards tetrahydropalmatine and rutin, plasma samples were pretreated by protein precipitation with 700μL anhydrous ethanol-acetonitrile (9:1, v/v). The UHPLC separation was carried on a C18 column with gradient elution using a mobile phase composed of methanol and water which containing 0.3% formic acid. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source. The limits of quantification (LOQ) of phellodendrine, coptisine, jatrorrhizine, berberine, palmatine, baicalin, wogonoside, baicalein, wogonin and emodin were 0.50,0.50,0.30,0.30,0.30, 10,3.0,8.0,1.0,8.0 μg·L-1, respectively. The method recoveries of all analytes were found to be within the acceptable range with a range of 90.0%-109%.2. A UHPLC-MS/MS method has been developed and validated for sensitive and rapid determination of 3 phenolic acids,8 flavonoids,2 chromones and 1 lignan from Xiao’er Jiebiao Granules and its pharmacokinetic study in rat plasma using epicatechin and icariin as internal standards. Plasma samples were pretreated by protein precipitation with 350 μL methanol-anhydrous ethanol (9:1, v/v). The limits of quantification (LOQ) of protocatechuic acid, chlorogenic acid, caffeic acid, puerarin, daidzin, prim-o-glucosylcimifugin, cimifugin, rutin, scutellarin, hesperidin, baicalin, arctiin, wogonoside, wogonin were 20,10,80,0.1,0.02,0.1,0.1,1.0,20,6.0,20,50,3.0,2.0 μg·L-1 respectively. The method recoveries of all analytes were found to be within the acceptable range with a range of 88.9%-114%. In conclusion, this method is beneficial for quantification and confirmation assays of multiply components in complex biological samples.3. A method was established for simultaneous determination of protocatechuic acid, protocatechuic aldehyde, chlorogenic acid, caffeic acid, epicatechin, rutin, isochlorogenic acid C, ginsenoside Re, trans-cinnamic acid, icariin, ginsenoside Rb1, tenuifolin, schisandrin, schisantherin, deoxyschizandrin and y-schisandrin in rat plasma after oral administration of Jiannao Anshen pill by UHPLC-MS/MS. After mixing with two internal standards rosmarinic acid and bifendate, plasma samples were pretreated by protein precipitation using 350 μL methanol-anhydrous ethanol (9:1, v/v). All analytes were eluted rapidly within 19 min. The limits of quantification (LOQ) of protocatechuic acid, protocatechuic aldehyde, chlorogenic acid, caffeic acid, epicatechin, rutin, isochlorogenic acid C, ginsenoside Re, trans-cinnamic acid, icariin, ginsenoside Rb1, tenuifolin, schisandrin, schisantherin, deoxyschizandrin and y-schisandrin were 10,5.0,8.0,45,0.40,0.80,8.0,2.0,300,3.0,2.0,20,0.60,1.2,0.80,0.40 ug·L-1, respectively. The extraction recovery and method recovery were found to be within the acceptable range with a range of 78.4%-102% and 91.8%-108%, respectively. The developed method was rapid, simple, reproducible, and successfully applied to simultaneous determination of sixteen active constituents in rat plasma after oral administration of Jiannao Anshen pill. |
