Investigation On Quality Consistency Of Safflower Injection With Chemical Fingerprint And Bioassay

Herbal injections are important modernized dosage form of traditional herbal medicines. With their powerful therapeutic effects in treating special diseases, herbal injections have received widespread attention all around the world. However, the increasing body of adverse reactions pertaining to herbal injections have raised public doubts of the safety of herbal injection and have even casted shadow on the development of relevant industries. Currently, quality control (QC) of herbal injection still rely mainly on conventional methods, yet those methods have great difficulty to characterize all chemical information and could not provide much biological information related to clinical safety and efficacy. Therefore, there is an urgent need to establish new methods and models to reduce the adverse reactions.Herbal injections, just like all the other medicines, intrinsically possess both chemical information (e.g. chemical constituents) and biological information (e.g. biological activity), and the quality of any medicine should be chemically and biologically consistent. Base on the aforementioned properties of herbal injection, we tentatively address the issue of quality control of herbal injection by combining chemical detection and biological detection. In this paper, we choose safflower injection as the model medicine as it is commonly used in clinic and it has caused a lot of serious adverse drug reactions. To chemically assess the quality of safflower injection, high performance liquid chromatography was adopted to acquire chemical fingerprints of different safflowers. To biologically assess the quality of safflower inections, because constituents from herbal injections might directly affects target cells when injected into body and anti-thrombin activity is regarded as one therapeutic effect of safflower injection in treating cerebrovascular diseases, we monitored cells responding to different safflowers with the help of RTCA xCELLigence system and the interaction of different safflower injections with thrombin-treated fibrinogen. Different from the conventional approach focusing on chemical information, our study aimed at comprehensively control the quality of herbal injection from chemical and biological perspective and relate the quality control with safety and efficacy of herbal injection in clinic. The main content of our study are as follows:1. From chemical perspective, a chemical fingerprint method was established to characterize the quality of safflower injection.52 different batches of safflower injections were collected or prepared, including 10 batches of normal samples,12 batches of adverse reaction samples which had induced adverse drug reactions,10 batches of high temperature treated samples,10 batches of light treated samples,10 batches of samples which had exposed to air. With the developed HPLC method,25 common peaks in normal safflower injection were assigned. Based on the common peaks in different samples, similarity analysis, PCA analysis, HCA analysis were carried to identify the abnormal samples. The results indicated that similarity analysis could only identify 10 batches of abnormal samples, PCA and HCA analysis both could identify 25 abnormal samples. Therefore, chemical detection could only detect 25 batches of abnormal samples whereas 17 batches of abnormal samples could be identified.2. From cell biological effect perspective, a cell-based biological fingerprinting method was established to evaluate the quality of safflower injectionAllergic reaction takes important proportion of adverse reactions induced by herbal injections including safflower injections, therefore, RBL- 2H3 cell, a mast cell derived cell line which has been extensively utilized to study allergic reaction was selected as the model cell. With the help of RTCA xCELLigence system which could continuously monitor cells status, we established a cell-based biological fingerprinting method for detection the quality of different safflower injections. We extracted parameters including ΔT, AUC, I1, I2 and I3 to perform PCA and HCA analysis. The results indicated that boat PCA and HCA analysis could identify 36 batches of abnormal samples. Therefore,36 batches of abnormal samples could be identified by cell-based biological fingerprinting method whereas 6 abnormal samples could not.3. A comprehensive method for detecting the quality of safflower injection was establish based on combination of biological detection and chemical detection.As has been noticed, none of the established 2 detection methods could identify all the abnormal samples, it is necessary to delve the available data gained from 2 detection methods. Base on the detection results of 2 methods, series integration was adopted to integrating the results. We found that series integration could identify almost all the abnormal samples, this result indicate that combining chemical and biological detection could enhance the ability to control the quality of safflower injection. Based the integrative results, a comprehensive quality control model was established for safflower injection.In conclusion, with safflower injection as the model medicine, a comprehensive approach for quality control of safflower injection was established by integrating chemical and biological detection. This approach could reflect the quality of safflower injection from biological and chemical perspective, in addition, it could, to some extent, relate the detection results to the clinical safety and efficacy of safflower injection. The integrative method could provide reference for establishing quality control methods of other herbal injections.