Development of a novel UHPLC-UV combined with UHPLC-QTOF/MS fingerprint method for the comprehensive evaluation of Nao-Luo-Xin-Tong: multi-wavelength setting based on traditional Chinese medicinal prescription composition
A novel UHPLC-QTOF/MS fingerprint method was developed based on the multi-wavelength detection by ultraviolet detector and the analysis of common peaks by mass spectrometry technique, and was validated in the quality evaluation of traditional Chinese medicinal prescription: Nao-Luo-Xin-Tong (NLXT) decoction. The NLXT is a classical traditional Chinese medicine (TCM) prescription that has been widely used to treat stroke disease over fifty years in the first affiliated hospital of Anhui University of Chinese Medicine. In order to obtain the overall chemical material information of NLXT, the reliable UHPLC-UV methods at four characteristic wavelengths (203 nm, 254 nm, 320 nm and 403 nm) were developed and 8 representative constituents were chosen. Good separation was performed on an Agilent - Eclipse Plus C18 column with a gradient elution at a flow rate of 0.2 mL/min within 60 min. The chromatogram data of twelve batches of NLXT decoction were analyzed by "Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine" (2012 Version). Subsequently, the common pattern was established and the similarity was evaluated. The results showed 40, 19, 13 and 9 common peaks were obtained in chromatograms at the wavelengths of 203 nm, 254 nm, 320 nm and 403 nm, respectively, and the similarities of twelve batches of NLXT decoction were all above 0.97. In addition, the novel UHPLC-QTOF/MS technique in both positive and negative ion mode was applied in the identification and detection of common peaks. The results showed 36 common peaks in all of 47 common peaks were identified or tentatively characterized based on their exact molecular weights, fragmentation patterns and literature data. In a word, the novel UHPLC-QTOF/MS fingerprint method not only give essential data for identification of NLXT decoction but also provide a reference to the quality evaluation of traditional Chinese medicinal prescription.