Abstract
Background Quantitative analysis of the active ingredients of Traditional Chinese Medicine is a research tendency. The objective of this study was to build a novel method, namely, Detection-confirmation-standardization-quantification (DCSQ), for the quantitative analysis of active components in traditional Chinese medicines, without individual reference standard. Methods Danshen (the dried root of Radix Salvia miltiorrhiza) was used as the matrix. The “extraction” function in high-performance liquid chromatography-mass (HPLC-MS) instrument was used to find the peaks corresponding to cryptotanshinone, tanshinone I, and tanshinone IIA in the total ion current (TIC) chromatogram of Danshen. The multicomponent reference standard (MCRS) containing the three tanshinones mainly was prepared by preparative HPLC. The contents of them in the resulting MCRS were determined by NMR; moreover, the constituents of the MCRS were confirmed. The MCRS containing known content of the three tanshinones was used as the reference standard for the quantitative analysis of cryptotanshinone, tanshinone I and tanshinone IIA in Danshen Samples by analytical HPLC. Results The optimized HPLC conditions for the quantitative analysis of the active components in Danshen were established, and the assignments of the extracted peaks were confirmed by analyzing the characteristic fragments in their MS/MS product ion spectra and UV spectra. Then, the MCRS containing the three tanshinones were successfully prepared. The results of determination about the contents by NMR showed linearity fitted with high likelihood and calibration curves possessed high linearity. The results of determination on Danshen Samples obtained through DCSQ exhibited minimal deviations, in contrast to those obtained through individual reference standards. Conclusion The establishing DCSQ is independent and convenient for the quantitative analysis of the active components in TCMs by MCRS, without individual reference standard. This method is a great advance in quantitative analysis for complex composition, especially TCMs.
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