Study On Quality Control Method Of Polygonum Bistorta L. And Injectio Folii Isatidis Composita

Injectio Folii Isatidis Composita is prepared from Folium Isatidis, Flos Lonicerae, Radix et Rhizoma Rhei, Rhizoma et radix notopterygii and Rhizoma Bistortae. Rhizoma Bistortae is the dried rhizomes of Polygonum bistorta L. of Polygonaceae. The other four herbs of the prescription have been investigated by other students of our group. In this thesis, quality control method of Polygonum bistorta L and Injectio Folii Isatidis Composita. are presented.Firstly, a study on quality control of Polygonum bistorta L. was carried out. According to the theory of casein method, the test condition was optimized and the activated tannins in Polygonum bistorta L were determined. Two bioactive constituents namely gallic acid and chlorogenic acid in Polygonum bistorta L. samples were determined simultaneously by high performance liquid chromatography. The samples were separated on a Diamonsil C₁₈ column (4.6 mm×250 mm, 5μm) with the mobile phase of methanol-0.4% acetic acid (25:75, v/v). The flowrate was 0.8 mL·min^-1 , the detection was set at 290 nm and the column temperature at roomtemperature. The linear range was 4.24～106μg·mL^-1 with a regression coefficient of 0.9996 for gallic acid and 5.0～125μg·mL^-1 with a regression coefficient of 0.9998 for chlorogenic acid. With chlorogenic acid as reference substance, HPLC-Fingerprints that can indicate substances existing in Polygonum bistorta L. was established with ultraviolet detection. All constituents were separated completely in 60 min. Peaks existing in all sample chromatograms were assigned as "common pesks" for Polygonum bistorta L. There had 12 "common peaks" in the fingerprint. When the fingerprint data were analyzed by using Cluster Analysis with Squared Euclidean distance as the measure, Polygonum bistorta L. can be differentiated definitely from its fakes. The HPLC-FPS similarities of Polygonum bistorta L. samples were evaluated by Similarity Evaluation System for Chromatographic Fingerprint of TCM of the Chinese Pharmacopoeia Committee. Good similarities with cosine coefficient above 0.90 were found in fingerprints of 12 Polygonum bistorta L samples from different sources. Under the same condition, four fake samples were analyzed. The similarities between their HPLC-FPS and the common pattern of Polygonum bistorta L were all lower than 0.90.Secondly, five water-soluble constituents namely cytidine, hypoxanthine, uridine, xanthine and guanosine in the aqueous extract of Isatis indigotica Fort. were identified by HPLC/MS². After separation of the aqueous extract of Isatis indigotica Fort. by HPLC, the eluates of five peaks were collected separatively and analysed by MS². The structures of constituents in the collected eluates were characterized by comparing their retention times, UV spectra and fragment pattern in the MS² spectrum with those of corresponding standards. Then, a method for determining uridine in Injeetio Folii Isatidis Composita was developed. The analytes were separated on Diamonsil C₁₈ (4.6 mm×200 mm, 5μm) column with methanol-water (5:95, v/v) as mobile phase and detected at 254 nm at room temperature. This method was simple, accurate, robust and it was helpful for the quality control of Injectio Folii Isatidis Composita.Finally, the HPLC-FPS of Injectio Folii Isatidis Composita were analyzed and evaluated. Uridine was choosed as the reference substance, 20 "common peaks" were achieved after analyzing the injection samples of different factories and HPLC-FPS common pattern of the injection was developed. Analyzed by Similarity Evaluation System for Chromatographic Fingerprint of TCM of the Chinese Pharmacopoeia Committee, a similarity threshold of 0.85 was set up for Injeetio Folii Isatidis Composita. Furthermore, by comparing the fingerprints of the injection, the intermediate and each herb decoction, the correlation of peaks among their fingerprints were found. The developed method offered guarantee for the stability of the intermediate and final products of Injectio Folii Isatidis Composita from batch to batch and it was beneficial to improving its quality.