Qing Ling Injection In This Article.the Method Research

Traditional Chinese medicine injection is a kind of original formulations in China, but many adverse reactions limit its application and development, tannins is considered as one of the ingredients which cause adverse reactions. At present, the methods for determination of tannins in traditional Chinese medicine injection are restricted to only egg white method and sodium chloride gelatin method. Not only the detection limit and sensitivity of the two methods fail to achieve the requirements, but also it may appear false positive or negative. Therefore, it is highly urgent to set up a method which have better detection limit and sensiti-vity as well as less false results about tannins detection in traditional Chinese medicine injection.This subject takes Qingkailing injection as the object to detect-the tannins in Lonicera japonica, gardenia, and Radix Isatidis by using physical and chemical reaction, thin layer chromatography (TLC), instrumental analysis including high performance liquid chromato-graphy (HPLC) and liquid chromatography-mass spectrometry (LC-MS) method,etc. with Lonicera japonica being most important research object, a better detection method for tannins will be established and applied to Qingkailing injection. From the perspective of tannins, using the established method to make a evaluation on the quality of Qingkailing injection and even other traditional Chinese medicine injections, so that improving the quality evaluation system of TCM injections.Thesis research and the results are as follows:(1) Physical and chemical reaction, thin layer chromatography preliminary detect tannins in Qingkailing injection. The results showed that tannins exist in Lonicera japonica and gardenia, but might not in Radix Isatidis.(2) Instrument analysis results:firstly, the hydrolysable tannins in Lonicera japonica were converted by acid hydrolysis to gallic acid and were detected by HPLC, the result showed that Lonicera japonica contained a certain amount of hydrolysis tannins. secondly, in order to detected the different types of tannins in Lonicera japonica, a rapid, sensitive and selective method, high performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) was used to detect the gallic acid after acid hydrolysis, the catechin, epicatechin and benzylthioether of (epi)catechin in Lonicera japonica after thiolysis, the result showed that Lonicera japonica contained a certain amount of hydrolysis tannins and condensed tannins. The established method could detect hydrolysable tannins and condensed tannins in Lonicera japonica rapidly. Thirdly, the identification of the dimer of catechin in50%methanol aqueous solution of Lonicera japonica was carried out by HPLC-ESI-MS/MS.Fourthly, consider the method of column chromatography pre-processing could achieve enrich-ment and purification of trace component, in the study, we investigated the AB-8macroporous adsorption resin’s effects on enriching and purifying the Lonicera japonica sample, but the result was not perfect. Then the solid phase extraction column (SPE) was applied to enrich and purify the sample and the result showed that Waters Oasis HLB SPE was better to purify gallic acid, the microconstituent than AB-8in Lonicera japonica hydrolysate, the content of gallic acid detected in hydrolysate was1.667ug·g-1.However,it was rather difficult to develop determination method systematically for gallic acid because of complicated hydrolytic process. Whereas, the established SPE-HPLC method could provide reference for quantity control of microconstituents in a complex system of traditional Chinese medicine.(3)To establish a method for determining hydrolysable tanninsThrough selected Geranium carolinianum L. which contained hydrolysable tannins as positive medicinal materials, and investigated extraction method and acid hydrolysis method, to establish a complete determination method of hydrolysable tannins, which had a low limit of detection, high sensitivity and good specificity, it could be used to determine whether there are hydrolyzed tannins in a complex system of traditional Chinese medicine. The established detection method was applied to Lonicera japonica and Qingkailing injection, but didn’t detect gallic acid and ellagic acid after acid hydrolysis. It was shown that there were utmost low levels of hydrolysable tannins in Lonicera japonica and Qingkailing injection or no hydroly-sable tannins in Qingkailing injection.