Studies On Quality Control And Material Basis Of Antai Pills

Antai pills(ATP) is a classic prescription in ancient times. It was reported originally from Danggui San in the “Synopsis of Golden Chamber”. It composed of five herbs: Angelicae Sinensis Radix, Chuanxiong Rhizoma, Scutellariae Radix, Paeoniae Radix Alba and Atractylodis Macrocephalae Rhizoma. ATP is widely used as anti-abortion drug to treat pregnancy blood deficiency, threatened abortion, sallow complexion, no appetite and fatigued spirit and lack of strength. Now it is embodied in the first volume of Ministry of Health Medicine Standard of Prescription Drug Preparations. The current method reported in standards and literatures was using microscopic identification, TLC and determination of several bioactive components to evaluate the quality of Antai pills.The characteristics of the Traditional Chinese Medicine is multiplex ingredients, multiplex targets and whole regulation, so the current method is difficult to control the quality of Antai pills. The study using fingerprinting analysis and multicomponent determination to control the quality of ATP, while material basis of ATP were studied. The research contents were as follows:1 Chemical constituents in Antai pills based on UPLC-Q-TOF-MSA new method using ultra high performance liquid chromatography quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was developed for the identification of Antai pills. Separation was performed using an Agilent Eclipse Plus-C18 column(50 mm×2.1 mm, 1.8μm) with the mobile phases consisted of 0.1% formic acid and acetonitrile. The data were collected by the positive and negative ion mode using ESI ion source. Fifty-nine compounds including 5 phenolic acids, 6 monoterpene glycosides, 29 flavonoids, 14 phthalides, 2 phenylethanoid glycosides and 3 sesquiterpene lactones were identified in Antai pills, and the fragmentation pathways were analyzed simultaneously. The study provided a foundation for quality control and material basis of Antai pills.2 UPLC fingerprint and chemical pattern recognition method of Antai pillsTo establish a UPLC fingerprint of Antai pills for reference of the effective quality control. The fingerprint of Antai pills was developed with UPLC. The separation was carried out on an Agilent SB-C18 column(100 mm×2.1 mm, 1.8 μm) with the mobile phase consisting of acetonitrile and 0.4% phosphoric acid at 0.3 m L·min-1. The detective wavelength was 280 nm from 0 to 2 min, 230 nm from 2 to 8 min, 316 nm from 8 to 9.4 min and 280 nm from 9.4 to 35 min. The column temperature was 35℃, and the sample volume was 2 μL. Similarity evaluation combined with cluster analysis(CA), principal components analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were used to evaluate the fingerprint of 12 batches of Antai pills. The common mode of the fingerprint was set up with 24 common peaks, and 9 of them were identified. The similar degrees of 12 batches of sample were over 0.9, they are albiflorin, paeoniflorin, ferulic acid, baicalin, wogonoside, baicalein, wogonin, senkyunolide A, atractylenolide Ⅰ. Totally 12 batches of sample could be classified into three clusters. A brief list of five differential compounds was presented, including Ferulic acid and Baicalein. The method was rapid, reliable, effective and it can be used for quality control of Antai pills.3 Simultaneous determination of eight components in Antai pills by UPLCTo develop an UPLC method for simultaneous determination of albiflorin, paeoniflorin, ferulic acid, wogonoside, baicalein, wogonin, senkyunolide A, atractylenolide Ⅰ in Antai pills. The separation was carried out on an Agilent SB-C18 column(100 mm×2.1 mm, 1.8 μm) with the mobile phase consisting of acetonitrile and 0.4% phosphoric acid at 0.3 m L·min-1.The detective wavelength was 280 nm from 0 to 2 min, 230 nm from 2 to 8 min, 316 nm from 8 to 9.4 min and 280 nm from 9.4 to 35 min. The column temperature was 35℃, and the sample volume was 2 μL. The results demonstrated that the calibration curves for all analyses showed good linearity(r2≥0.9980) within test ranges. And the recovery of method was within the range of 98.0%-103.8%. The method developed was rapid, accurate, reliable and highly sensitive for quantitative analyses of Antai pills.4 Constituents absorbed into blood in Antai pills by UPLC-Q-TOF-MSTo determin the direct effective compounds in vivio of Antai pills by analyze the compositions absorded into the blood. Separation was performed using an Agilent Eclipse Plus-C18 column(50 mm×2.1 mm, 1.8μm) with the mobile phases consisted of 0.1% formic acid and acetonitrile. The data were collected by the positive and negative ion mode using ESI ion source. 44 constituents, including 28 prototypes and 16 metabolites were detected in serum samples, and the metabolic pathways were analyzed. The method could comprehensively analyzed the metabolites so as to provide the evidence for the further research of the material basis of Antai pills.