| Lonicerae Japonicae Flos was a commonly used traditional Chinese medicine,contained the version of the calendar year "pharmacopoeia of the People's Republic of China",had the effect of the heat-clear and detoxification and the dispel wind.It was prescribed as the main ingredient in many traditional herb formulas and widely used in the clinical compatibility of traditional Chinese medicine for many diseases,and also for many Chinese patent medicine and health medicine raw materials.In recent years,with their considerably different price,Lonicerae Flos was the main fraudulent herb of Lonicerae Japonicae Flos in the market and these two herbs had the different appearances and chemical constituents,but quite similar properties and indications,resulting in Lonicerae Japonicae Flos adulteration was serious.In addition,the higher content of the saponin group in Lonicerae Flos had many hidden dangers in terms of safety.It was difficult to reflect the quality of Lonicerae Japonicae Flos only by the main components of chlorogenic acid and Luteolin-7-O-glucoside as the quality control index.Therefore,in view of the serious adulteration of Lonicerae Japonicae Flos and the traditional methods for detecting Lonicerae Japonicae Flos aduheration which was not suitable for the rapid analysis of industrial production and required longer analysis time and consumed more organic solvent as mobile phase,it was necessary to develop and validate a simultaneously qualitative and quantitative determination of rapid resolution liquid chromatography analysis method on multiple components in Lonicerae Japonicae Flos,which achieved the overall quality control of Lonicerae Japonicae Flos herbs and products effectively.It was a low cost and rapid methods to address this issue.A UHPLC-DAD/ELSD method was analyzed and studied to simultaneously establish fingerprints determine the concentration of nine constituents,including neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,luteolin-7-O-glucoside,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C,maranthoidin B and dipsaccside B in Lonicerae Japonicae Flos and Lonicerae Flos.Furthermore,the method could easily distinguish Lonicerae Flos from Lonicerae Japonicae Flos by the saponin group within 8 min.Nine constituents of Lonicerae Japonicae Flos were identified and showed good linear relationship in the ranges of the test concentrations.The results of the precision,stability,repeatability and recovery tests were in compliance with the requirements(RSDs < 3%).The average recoveries were in the ranges of 98.71%~101.1%.It indicated that the analytical method was accurate and reliable.Compared with the traditional method,it also had the advantages of short analysis time,good separation effect,less solvent consumption and easy operating.15 batches of Lonicerae Japonicae Flos,9 batches of Lonicerae Flos and 6 batches of Lonicerae Japonicae Flos extracts were detected and analyzed by this method.The results of the analysis could effectively evaluate the quality of the herbs and extracts from different sources.Therefore,a reliable quality analysis method for the medicinal materials,could not only more comprehensively reflect and effectively control the internal quality of Lonicerae Japonicae Flos and their extract,but also improve their quality standards to provide a basis for reference of other medicinal herbs and preparations which contain Lonicerae Japonicae Flos.In addition,it could also be used as reference to achieve similar purposes of other herb materials.Based on the results of the single-factor tests and the Box-Behnken central composite experimental design principles,a response surface methodology which was designed to optimize the extraction process of Lonicerae Japonicae Flos based on the content of total phenolic acid was investigated.It was used to study the concentration of ethanol(45~75%),material ratio(10~30 times)and extraction time(1~3 h).Combining the optimization results of response surface methodology and the actual situation of manufacture,the extract conditons were confirmed as follows: material ratio 1:30,concentration of ethanol 65%,pH 3,extraction temperature 90 ?,extraction t ime 1 hours for two times. |
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