| The ancient prescription named Shasheng Maidong Decoction came from Wenbingtiaobian written by Jutong Wu in Qing dynasty, which consists of coastal glehnia root, dwarf lilyturf, solomonseal rhizome, radix trichosanthis, white hyacinth bean, mulberry leaf and licorice, while the coastal glehnia root and dwarf lilyturf are sovereign drugs, the solomonseal rhizome and radix trichosanthis are subjects, the white hyacinth bean and mulberry leaf are assistants, the licorice is envoy. The prescription can raise the lung and stomach as well as clear away the heart-fire and promote the secretion of body fluid, which mainly cure such symptoms as tepidity and heat injury by dryness and yin injury of hung and stomach.The research has established the reliable, sensitive and simple UPLCMS/MS method to measure the nine effective constituents in the radix ophiopogon soup at the same time, and firstly established the UPLC-MS/MS method which can measure the internal drug concentration of the three effective constituents in the Shasheng Maidong Decoction at the same time, A single dose of pharmacokinetic experiments were carried out and the respective pharmacokinetic parameters were calculated, which settles a basis for the further research of quality control and the later dosage form transformation research of Shasheng Maidong Decoction, and provides a reference for the physiological disposition and pharmacodynamics material basis of active ingredient in traditional Chinese medicine compound preparations as well as the modern clinical application of Shasheng Maidong Decoction. Part 1 Study on the mothod of quantitative analysis of chemical composition of Shasheng Maidong DecoctionPurpose: Establishing the method for the determination of Rutin, Liquiritin, Psoralen, Xanthotoxin, Bergapten, Monoammonium Glycyrrhizinate, Ophiopogonin D, Methylophiopogonanone A and Methylophiopogonanone B in the Shasheng Maidong Decoction.Method: Adopting the SPE-UPLC-MS/MS method. The chromatographic column was Phenomenex Kenetix C18(50 mm×3 mm, 2.6 ?m), the mobile phase was acetonitrile-0.1% formic acid in water(gradient elution), the column temperature was 25?, while the flow rate was 0.3 m L·min-1; The assay determination was carried out by using the MRM scanning mode, which was ESI model.Result: The linearity range of the sample quantity of Rutin, Liquiritin, Psoralen, Xanthotoxin, Bergapten, Monoammonium Glycyrrhizinate, Ophiopogonin D, Methylophiopogonanone A and Methylophiopogonanone B reached 1.75~700,5.00~1997,0.95~380, 1.75~700,2.00~800,7.00~2799,2.38~950, 0.38~151,0.43~171 ng·m L-1,respectively.(r is 0.9992,0.9997,0.9990,0.9991,0.9998, 0.9993,0.9995,0.9997,0.9990, respectively); RSD of the precision, stability, repetitive testing was less than 3.0%. The range of the sample adding's recovery rate was 95.1%~104.6%, while RSD reached 2.3%?2.9%?2.1%?3.0%?1.2%?2.9%?1.9%?2.6%and 2.8%, respectively.Conclusion: The method is rapid and simple as well as has a good specificity and a high sensitivity, which can be used to measure the content of the nine effective constituents in the Shasheng Maidong Decoction and provides a basis to the quality control and studies on material basis of the Shasheng Maidong Decoction. Part 2 Pharmacokinetic study of the 3 effective composition of the Shasheng Maidong DecoctionPurpose: Establishing the method for the UPLC-MS/MS determination of Liquiritin, Xanthoxin and Methylophiopogonanone A in the plasma of rats, and discussing the pharmacokinetic process of the decoction in the rat's body.Method: The rat's blood was collected when the time reached 0.03?0.08, 0.17, 0.3, 0.66, 1, 1.5, 2, 3, 4, 6, 8, 10, 12 and 24 h after a single dose of lavage of extractive from Shasheng Maidong Decoction for 12 m L·kg-1, respectively, UPLC-MS/MS has measured the plasma concentration of subjects in 0-24 h, the mobile phase was acetonitrile-0.1% formic acid in water, gradient elution, while the flow rate was 0.3 m L·min-1; Using ESI source, the plasma concentration of various components was scanned by the simultaneous detection from the positive and negative ion as well as detected by MRM, then the pharmacokinetic parameter was calculated by DAS 3.0 software.Result: The linear relationship of Liquiritin, Xanthotol and Methylophiopogonanone A at 4.92~315.00 ng·m L-1(r=0.9991), 1.44~92.00 ng· m L-1(r=0.9994)and 0.35~22.00 ng·m L-1(r=0.9992)was good, respectively, all average recovery were greater than 76.5%, all RSD in the daytime were less than 15%. The three kinds of plasma sample were repeatedly frozen and thawed for three times, the stability of the samples was good after placing 15 d at-20? and 4 h at room temperature, and the stability of the analysis of processed plasma samples was also good after placing 4 h at room temperature, the RSD was less than 15.0% in the daytime, the precision and accuracy were all in line with the requirements of biological sample analysis. AUC0-t of Liquiritin, Xanthotol and Methylophiopogonanone A was(718.23±185.55),(22.52±7.53), and(13.55±6.03) ng·h·m L-1, respectively, while T1/2 was 3.61± 2.01, 6.93±7.78 and 3.51±1.92 h, respectively after the lavage of extractive from radix ophiopogon soup.Conclusion: The method is rapid and convenient and can be used for internal quantitative analysis of Liquiritin, Xanthotol and Methylophiopogo-nanone A. |
PDF Full Text Request