| Qingfei mixture, one of the classical Zhejiang cancer hospital preparations, which consists of ten Chinese herbs including herba hedyotidis diffusae, bulbus fritillariae thunbergii, spica prunellae, herba scutellariae barbatae, rhizoma imperatae, herba agrimoniae, herba scutellariae barbatae, rhizoma paridis(paris polyphlla smith var.chinensis(franch.)hara), radix stephaniae tetrandrae and gekko chinensis, is widely used in treating lung cancer in different phases, pneumonia and radiation pneumonitis. Though its effectiveness has been well documented during long-term clinical practice and its mechanism is revealing, its quality control methods is simple and crude. In the present study, HPLC-fingerprint analytical methods and quantitative methods of bioactive compounds were developed for the quality control of Qingfei mixture and spica prunellae respectively.In this study, an HPLC method for simultaneous determination of caffeic acid, rosmarinic acid, oleanolic acid and ursolic acid in spica prunellae was established. After ultrasonic extraction with 75 % ethanol solution (containing 1 % formic acid, v/v), the ethanol-extract of spica prunellae was analyzed by RP-HPLC on an Elite SinoChrom ODS-AP column using gradient elution of 0.01 % phosphoric acid (A) and acetonitrile (B) at a flow rate of 0.9-1.0 mL·min-1. A wavelength switch program was used for detection at 330 nm (0-33min) and 203 nm (33-40min). The column temperature was set at 20℃and injection volume was 50μL.The calibration curves of all analytes were linear. The average recoveries were 93.7 %-105.2 % with RSDs not more than 4.5 %. The contents of caffeic acid, rosmarinic acid, ursolic acid and oleanolic acid in spica prunellae were 0.0401-0.0968 mg·g-1, 0.99-2.57 m·g-1, 0.243-0.556 m·g-1 and 4.06-8.13 m·g-1, respectively. The described method is sensitive, convenient and accurate, and is suitable for the simultaneous determination of caffeic acid, rosmarinic acid, oleanolic acid and ursolic acid in spica pnmellae.A quality assessment method of spica pnmellae from different habitats was developed by HPLC chemical fingerprint analysis, principle component analysis and cluster analysis. The fingerprint analysis method of spica pnmellae was developed by HPLC with an Elit SinoChrom ODS-AP column at 35℃and detected at 203nm, using 0.01% phosphoric acid (A) together with acetonitrile (B) as gradient elution at a flow rate of 1.0 mL·min-1.Eleven batches of spica pnmellae from different sources were determined and the data achieved were evaluated based on similarity analysis, principal component analysis and cluster analysis. The fingerprint of 11 batches of spica pnmellae showed 17 common peaks. As a result of similarity analysis, it showed a high similarity between all samples.Samples were classified as 4 clusters by principal component analysis and cluster analysis, consistented with their habitats. The results indicated that the quality of determined spica pnmellae is stable. The method is sensitive, convenient and accurate, and is suitable for identification and evaluation the quality of spica pnmellae.Moreover, a simple, reliable and accurate method for the simultaneous separation and determination of 10 active components (protocatochic acid , chlorogenic acid , caffeic acid , p-coumaric acid , rosmarinic acid , scutellarin and apigenin ) in qingfei mixture was developed using HPLC coupled with diode array detection. The chromatographic separation was performed on a Elite SinoChrom ODS-AP column with gradient elution of 0.01 % phosphoric acid (A) and acetonitrile (B) at a flow rate of 1.0 mL·min-1. A wavelength switch program was used for detection. The column temperature was set at 50℃and injection volume was 20μL. Good linear behaviors over the investigated concentration ranges were observed for all the analytes. The recoveries, measured at three concentration levels, varied from 94.4 to 98.5%. The validated method was successfully applied to the simultaneous determination of these active components in qingfei mixture from different production batches.At last, the fingerprint analysis method was established by RP-HPLC with a Elit SinoChrom ODS-AP column at 50℃and detected at 330nm,using 0.01% phosphoric acid(A) together with acetonitrile(B) as gradient elution at a flow rate of 1.0 mL·min-1.Sixteen batches of qingfei mixture were determined and the data achieved were evaluated based on similarity analysis. The fingerprint of all samples showed 16 common peaks. As a result of similarity analysis, it showed a high similarity between all samples. The results indicated that the quality of qingfei mixture is stable. The method is sensitive, convenient and accurate, and is suitable for identification and evaluation the quality of qingfei mixture.
|
PDF Full Text Request