ABSTRACT
Objective: To establish the fingerprint of Jinzhen Oral Liquid (JOL) by HPLC-UVD-ELSD and determine the main 13 representative components (gallic acid, liquiritin, aloe-emodin-8-O-β-D-glucopyranoside, liquiritigenin, baicalin, chrysin-7-O-β-D- glucoronide, oroxyloside, wogonoside, chrysophal-1-O-β-D-glucopyranoside, chrysophal-8-O-β-D-glucopyranoside, rhein, glycyrrhizic acid, hyodeoxycholic acid and cholic acid) simultaneously, in order to provide reference for the overall quality control of JOL. Methods: The separation was developed on Cosmosil-C18 column (250 mm × 4.6 mm, 5 μm) by gradient elution with methanol-water [containing 0.1% formic acid] at 254 nm, the temperature of drift tube was maintained at 115 ℃ and the carrier gas flow rate was 2.0 L/min. An HPLC-UVD-ELSD fingerprint of JOL was set up, and 15 batches of JOL were evaluated by similarity assay. Furthermore, the contents of the main 13 representative components were determined on the premise of small disparities among batches. Results: The HPLC-UVD-ELSD fingerprint of JOL was established with good separation, and 13 chemical components were determined simultaneously. Fifteen main characteristic peaks [gallic acid (peak 1), liquiritin (peak 5), aloe-emodin-8-O-β-D-glucopyranoside (peak 7), liquiritigenin (peak 11), baicalin (peak 13), chrysin-7-O-β-D-glucoronide (peak 16), oroxyloside (peak 17), wogonoside (peak 18), chrysophal-8-O-β-D-glucopyranoside (peak 19), chrysophal-1-O-β-D-glucopyranoside (peak 20), rhein (peak 24), glycyrrhizic acid (peak 26), (18β,20α)-glycyrrhizic acid (peak 27), hyodeoxycholic acid (peak 28), cholic acid (peak 29)] from four formula of JOL were chemically identified and 29 main characteristic peaks were assigned to individual herbs (peaks 8, 12, 13, 15-18 originate from Scutellariae Radix, peaks 3-5, 10, 11, 25-27 originate from Glycyrrhizae Radix et Rhizoma, peaks 1, 6, 7, 9, 14, 19, 20, 24 originate from Rhei Radix et Rhizoma, peak 2 originates from Fritillariae Ussuriensis Bulbus, peaks 28, 29 originate from Bovis Calculus Artifactus, peaks 21-23 originate from auxiliary materials). The similarity of 15 batches of JOL was about 0.968 to 1.000. Moreover, good linear relationships were found (R2=0.999 0-0.999 9), and the average recovery rates were 96.90%-102.84%. The content range of quantitative components in 15 batches of JOL (gallic acid 51.82-148.27 μg/mL, liquiritin 75.04-130.00 μg/mL, aloe-emodin-8-O-β-D-glucopyranoside 31.72-39.84 μg/mL, liquiritigenin 14.24-43.65 μg/mL, baicalin 610.37-867.40 μg/mL, chrysin-7-O-β-D-glucoronide 12.87-34.09 μg/mL, oroxyloside 62.45-101.48 μg/mL, wogonoside 155.41-205.86 μg/mL, chrysophal-1-O-β-D-glucopyranoside 11.56-23.72 μg/mL, chrysophal-8-O-β-D- glucopyranoside 16.14-36.87 μg/mL, glycyrrhizic acid 222.97-310.32 μg/mL, hyodeoxycholic acid 177.48-239.70 μg/mL, cholic acid 98.54-132.85 μg/mL) was determinated. Conclusion: The qualitative and quantitative methods of HPLC-UVD-ELSD mentioned above provided important evidence for further improving the overall quality standard of JOL.
ABSTRACT
Objective To identify whole ingredients in Niuhuang Qingwei Pills (NQP) and give primary grade evaluation of NQP. Methods Firstly, microscopic characteristics specified by the statutory standard of NQP were summarized. Then new microscopic identification methods were established for Sennae Folium and Gypsum Fibrosum while new thin layer chromatography (TLC) identification methods were established for Bovis Calculus Artifactus, Borneolum Syntheticum, and Rhei Radix et Rhizoma. In addition, microscopic identification method for Menthae Haplocalycis Herba was improved. In this way, whole-ingredient identification of 17 materials were realized. NQP reference drug was developed and applied as accompanying control in whole-ingredient identification and primary grade evaluation of the samples. Results According to the results of 48 samples from 18 manufactures by primary grade evaluation, 10, 37, and 1 samples were classified as superior, qualified, and unqualified, respectively. Conclusion The proposed methods are accurate, simple, and objective, which offers a more comprehensive approach for quality control of NQP. And the methods provide research strategy and experimental basis for further work on establishment of grade standard.