Simultaneous Detection Of The Assay Of The Major Constituents And Its Blood Medicinal Concentration In Traditional Chinese Medicine By UPLC-MS/MS

As international exchange and cooperation of medicine increasing extensive and indepth, it has brought new opportunities to the development of traditional Chinese medicine (TCM). And it makes TCM to modernization and globalization. Along with the rapid development of TCM’s modernization and globalization, more and more novel technologies have been widely used in TMC research, development and production. Now, the quality standard of TCM is the composition quality control, but not the quality control of single componen. And its action mechanism and the material base of research is one of the hot topics in the study of modern TCM. In the thesis, using UPLC-MS/MS to simultaneous determination of the major components and its blood concentration in TMCS.The main research contents are as follows:1. Using caffeic acid and icariin as internal standards, a method for simultaneous determination of the assay of protocatechuic acid, protocatechuic aldehyde, chlorogenic acid, scutellarin, isochlorogenic acid C, baicalin, luteolin, apigenin, atractylenolide Ⅲ and atractylenolide Ⅰ and its blood concentration in Fufangxingxiangtu’erfeng capsules were established by UPLC-MS/MS. The separations were performed on a ZORBAX RRHD Eclipse Plus C18column (2.1×50mm,1.8μm) by using water in which containing0.3%formic acid and methanol as mobile phases with the gradient elution at a flow rate of0.3mL/min. The analytes were detected by tandem mass spectrometer in the multiple reaction monitoring (MRM) mode via the switching of positive electrospray ionization (ESI+) and negative electrospray ionization (ESI"). Under the determination results, the ten compounds’calibration curves were range of0.00300-24.0mg/L for protocatechuic acid,0.0170～2.00mg/L for protocatechuic aldehyde,0.0150～30.0mg/L for chlorogenic acid,0.00400～30.0mg/L for scutellarin,0.0105～24.0mg/L for isochlorogenic acid C,0.00300～30.0mg/L for baicalin,0.00300～5.00mg/L for luteolin,0.00600～1.50mg/L for apigenin,0.00150～4.00mg/L for atractylenolide Ⅲ, and0.000600～0.900mg/L for atractylenolide I with detection limits of1.0,11,5.0,1.5,3.5,1.0,1.0,2.0,0.50,0.20μg/L, respectively. The standard recovery was the range of92.5%to106%. And the relative standard deviations were not more than3.1%. In rat plasma, the calibration curves were linear in the range of0.0100～20.0mg/L for protocatechuic acid,0.0500～4.00mg/L for protocatechuic aldehyde,0.0100～2.00mg/L for chlorogenic acid,0.0120～0.900mg/L for scutellarin,0.0300～2.70mg/L for isochlorogenic acid C,0.0150～6.00mg/L for baicalin,0.0100～5.00mg/L for luteolin,0.0100～5.00mg/L for apigenin,0.00300～0.0900 mg/L for atractylenolide Ⅲ, and0.00150-0.0500mg/L for atractylenolide Ⅰ with detection limits of3.0,20,5.0,4.0,11,5.0,3.0,3.0,1.5,0.50μg/L, respectively. The intraday precisions (relative standard deviation, RSD%) were less than2.5%for the ten analytes respectively. The relative recovery is80.5%-109%. The developed method was rapid, simple, accurate and reproducible, and suitable for the quality control and its blood concentration of the Fufangxingxiangtu’erfeng capsules.2. A rapid and sensitive method for simultaneous determination of matrine, protocatechuic acid, chlorogenic acid, scutellarin, quercitrin, baicalin, isoflavoues aglycone, ginsenoside Rgl, ginsenoside Re, luteolin, icariin, ginsenoside Rbl, astragaloside IV, astragaloside Ⅱ, atractylenolide Ⅲ, atractylenolide Ⅰ, cryptotanshinone, tanshinone Ⅰ, and tanshinone ⅡA and its blood concentration in An-Kang-Xin capsule was established by UPLC-MS/MS using internal standards.The mobile phase of gradient elution was acetonitrile-water with0.3%formic acid. The mass spectrometer was operated under the switching of positive ion and negative ion mode with the ESI source and carried out in the MRM mode. Under the optimum conditions, nineteen effective components and four internal standards has a good baselined separation in38minutes, and in a certain concentration range, the calibration curves have good linear relationship (r>0.9900). The developed method was rapid, simple, accurate and reproducible, and can be used in the quality control of An-Kang-Xin capsule which contains nineteen effective components and the determination of blood drug concentration in rat plasma.3. Using codeine phosphate and icariin as internal standards, a method for simultaneous determination of the adenosine, gallic acid, chlorogenic acid, loganin, paeoniflorin, liquiritin, scutellarin, peimisine, peimine, aurantiamarin, peiminine, baicalin, cinnamic acid, linarin, glycyrrhizinic acid, ophiopogonin D, glycyrrhetinic acid, oleanolic acid and its blood concentration in yangyinqingfei capsules were established by UPLC-MS/MS. In30minutes, eighteen effective components were baseline separated in yangyinqingfei capsules. Under the determination results, the detection limits were0.060,27,2.5,13,8.0,2.5,0.50,0.030,0.60,10,0.0060,0.50,100,0.60,2.5,1.5,2.0,2.5μg·L-1for adenosine, gallic acid, chlorogenic acid, loganin, paeoniflorin, liquiritin, scutellarin, peimisine, peimine, aurantiamarin, peiminine, baicalin, cinnamic acid, linarin, glycyrrhizinic acid, ophiopogonin D, glycyrrhetinic acid, oleanolic acid, respectively. The adding standard recovery was the range of95.8%to103%. In rat plasma, the detection limits of eighteen effective components is0.10,150,5.0,13,10,2.0,0.50,0.10,0.60,100,0.010,20,105,30,100,2.0,2.0,12μg/L, respectively. The intraday precisions (relative standard deviation, RSD%) were less than2.9%for the eighten analytes respectively. The relative recovery is80.3%～105%.The method is high selectivity, easy to use, high sensitivity, and has measured yangyinqingfei capsules’eighteen effective components and its blood concentration in.