ABSTRACT
Objective: To establish a method for the simultaneous content determination of 14 chemical components such as D- amygdalin, puerarin, hesperidin in Fenghan Ganmao Granules (FGG) by UHPLC-UV wavelength switching method, and chemometric analysis was used to analyze the quality differences. Methods: Separation was performed on an Agilent Poroshell 120 EC-C18 (150 mm × 2.1 mm, 2.7 μm) with a gradient elution of acetonitrile and 0.1% phosphoric acid. The content of 14 chemical components in 68 batches of samples from five manufacturers was determined by switching wavelength (210, 254, 310 nm). The radar chart, similarity evaluation, heat map and hierarchical clustering analysis, and principal component analysis (PCA) were used for data analysis. Results: The content of each component was as follows, D-amygdalin 0.063-3.885 mg/g, 3'-hydroxy puerarin 0.012-1.540 mg/g, puerarin 0.036-4.017 mg/g, 3'-methoxy puerarin 0.016-1.837 mg/g, puerarin-6″-O-xyloside 0.004-0.449 mg/g, mirificin 0.021-2.076 mg/g, daidzin 0.010-1.527 mg/g, prim-O-glucosylcimifugin 0.007-0.471 mg/g, 5-O-methylvisammioside 0.062-1.029 mg/g, hesperidin 0.210-8.453 mg/g, rosmarinic acid 0.001-0.237 mg/g, oxypeucedanin hydrate 0.007-0.204 mg/g, glycyrrhizic acid 0.056-1.311 mg/g, oxypeucedanin 0.002-0.042 mg/g, respectively. Chemometric analysis showed that there were some differences among the samples from different manufacturers, and the samples from the same manufacturer were more consistent. Conclusion: The method is simple, reproducible, and specific, which provides a reference method for the overall quality evaluation of FGG.
ABSTRACT
This study aimed to establish characteristic chromatogram and content determination method for Chan Taoren formula granules,evaluate the production processes of Chan Taoren formula granules based on the correlation of characteristic chromatogram and the transfer rate of D-amygdalin,and clarify the key control points. The optimized analytical method was carried out on a Waters CORTECS C18 column(4. 6 mm×150 mm,2. 7 μm) with acetonitrile-0. 1% phosphoric acid aqueous solution as the mobile phase at a flow rate of 0. 6 m L·min-1. The detection wavelength was 207 nm,and the column temperature was 20 ℃ ． As compared with the standard decoction of Chan Taoren,there were five characteristic peaks in the decoction pieces,extracts,concentrates,spray-dried powders and formula granules,basically consistent in relative retention time and peak pattern; in addition,the transfer rate of D-amygdalin from Chan Taoren pieces to the formula granules was within the transfer rate range of standard decoction. The average transfer rate of D-amygdalin was 56.65%,72.85%,94.58% and 99.29% respectively in the extraction,concentration,spray drying and granulation processes. Therefore,the factors affecting D-amygdalin in the extraction process were further studied. The results showed that D-amygdalin was easily converted to L-amygdalin in the water extraction process,leading to a low transfer rate of D-amygdalin in this process.D-amygdalin was unstable under alkaline conditions and prone to isomerization. Both liquid to solid ratio and extraction time had significant effects on the extraction rate of D-amygdalin. In this study,the key links in the production process of Chan Taoren formula granules was clarified based on the characteristic chromatogram and the quantity transmission of D-amygdalin,which provided a theoretical basis for production and quality control.
Subject(s)
Amygdalin , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Quality Control , WaterABSTRACT
Objective To evaluate the safety limits of amygdalin in the Chinese patent medicines and Chinese herbal medi-cines,so as to provide reference for establishing the standard in the safety limits of endogenous cyanide glycosides as toxic ingredients. Methods The toxicity differences between D-amygdalin and L-amygdalin were explored by CCK-8 assay. The daily safe dose of amyg-dalin was deduced by the no-observed-adverse-effect level(NOAEL)of cyanide. Based on the deduced results,the safety limits of amygdalin in thirty-six Chinese patent medicines and three different kinds of Chinese herbal medicine were calculated. Results The half maximal inhibitory concentration of D-amygdalin and L-amygdalin was(3.5±2.0)and(6.6±0.4)mmol/L,respectively,and the daily safe dose of amygdalin was 104.30 mg. Conclusion The toxicity of D-amygdalin was approximately two times that of L-amygda-lin. To establish the safety limits of amygdalin,the sum content of the epimer could be regarded as the D-amygdalin itself to enlarge the safety scope. The content of amygdalin of three Chinese patent medicines was determined to exceed the maximum safety limit for all measured batches,indicating some kind of safety risk. Moreover,in the researches on safety limit establishment,the toxicity-alleviat-ed processing effect should be included.