ABSTRACT
Objective:Based on pharmacokinetics， the antitussive and expectorant related quality markers （Q-marker） of Trichosanthis Fructus were screened from diosmetin-7-<italic>O</italic>-glucopyranoside， diosmetin， apigenin， vanillic acid and cinnamic acid， and the candidate Q-marker was evaluated by multivariate statistical method. Method:Six healthy rats were randomly selected and the 70% ethanol extract of Trichosanthis Fructus （dose of 20 g·kg<sup>-1</sup>） was given by intragastric administration. Blood was collected from the orbital vein at different time points， and the plasma concentrations of 5 components （diosmetin-7-<italic>O</italic>-glucopyranoside， diosmetin， apigenin， vanillic acid and cinnamic acid） from Trichosanthis Fructus were detected simultaneously by high performance liquid chromatography-triple quadrupole tandem mass spectrometry （HPLC-QqQ-MS/MS）. The main detection conditions were as following：mobile phase of 0.2% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-4 min， 6%-23%B； 4-5 min， 23%-59.5%B； 5-10 min， 59.5%-60%B）， flow rate of 0.5 mL·min<sup>-1</sup>， the detection wavelength at 254 nm， electrospray ionization （ESI）， positive ion mode detection， multiple reaction monitoring （MRM） mode scanning， scanning range of <italic>m</italic>/<italic>z</italic> 50-1 500. Diosmetin-7-<italic>O</italic>-glucopyranoside， diosmetin， apigenin and vanillic acid with clear pharmacokinetic behaviors were selected as candidate Q-marker about antitussive and expectorant of Trichosanthis Fructus. The contents of these components in 9 batches of medicinal materials were determined and the main detection conditions were the same as the pharmacokinetic study. SPSS 21.0 was used for cluster analysis and principal component analysis （PAC） based on the results of determination. Result:The pharmacokinetic results showed that the area under concentration-time curve （AUC<sub>0-</sub><italic><sub>t</sub></italic>） of 4 components （diosmetin-7-<italic>O</italic>-glucopyranoside， diosmetin， apigenin and vanillic acid） were （111.28±9.94）， （27.08±2.76）， （1 376.12±101.86）， （631.32±64.72） μg·h·L<sup>-1</sup>， respectively. The 9 batches of Trichosanthis Fructus samples were clustered into 3 groups by systematic cluster analysis. The clustering results were related to the variety of Trichosanthis Fructus and also affected by the origin. The PCA results showed that the comprehensive scores of Gaotang Trichosanthis Fructus， Shanxi Trichosanthis Fructus， Hebei Ben Trichosanthis Fructus were 1.919， 1.356 and 0.299， respectively， ranking in the top 3 among all samples. The comprehensive scores of Nongkeyuan No. 1， Hebei Trichosanthis Fructus and Nongkeyuan No. 2 were -0.804， -1.085， -1.120， respectively， which were in the last 3 positions among all samples. Conclusion:The pharmacokinetic characteristics and quality evaluation of diosmetin-7-<italic>O</italic>-glucopyranoside， diosmetin， apigenin and vanillic acid meet the requirements about antitussive and expectorant related Q-marker of Trichosanthis Fructus.
ABSTRACT
Objective: To develop an analysis method based on N-propylethylenediamine (PSA)-HPLC-QQQ-MS/MS for the determination of 12 kinds pesticide residues in Fritiliariae Thunbergii Bulbus and to quantify them in 20 batches of Fritiliariae Thunbergii Bulbus produced in various places in Zhejiang province. Methods: Fritiliariae Thunbergii Bulbus samples were extracted with acetonitrile and purified by the small column of PSA. The prepared samples were analyzed by HPLC-QQQ-MS/MS in multiple reaction monitoring (MRM) mode, and the pesticides were qualitified by the internal standard method. Results: All the 12 pesticides showed good linearities in their reasonable range (r = 0.9986-0.9998), and the average recoveries of all the pesticides were in the range of 68.1%-108.0% at three spiked levels of 90, 300, and 900 ng/mL. The RSD values were in the range of 1.1%-6.1%, and the LODs of each pesticide were all in the range of 0.08-1.0 μg/kg. Conclusion: The method is suitable for the multiresidues analysis of pesticides in Fritiliariae Thunbergii Bulbus simultaneously and the quality of Fritiliariae Thunbergii Bulbus is basically good for the safty although it contains a trace of several pesticide residues.