Qualitative and quantitative analysis of traditional Chinese medicine Niu Huang Jie Du Pill using ultra performance liquid chromatography coupled with tunable UV detector and rapid resolution liquid chromatography coupled with time-of-flight tandem mass spectrometry.

An ultra performance liquid chromatography coupled with tunable UV detector (UPLC-TUV) and rapid resolution liquid chromatography coupled with time-of-flight tandem mass spectrometry (RRLC-Q-TOF) method was developed for the quality assessment of Niu Huang Jie Du Pill (NHJDP), a commonly used traditional Chinese medicine (TCM). Ten compounds were simultaneously identified by electrospray ion mass spectrometry (ESI/MS) and comparison with reference standards and literature data. All of them were quantified by UPLC method. Baseline separation was achieved on an ODS-140HTP C(18) column (2.3mum, 100mmx2.1mm I.D.) with linear gradient elution of acetonitrile-0.1% formic acid. This developed method provides good linearity (r(2)>0.9996), repeatability (RSD<3.63%), intra- and inter-day precisions (RSD<0.86%) with accuracies (97.88-101.56%) and recovery (98.88-101.92%) of 10 major constituents, namely baicalin, baicalein, wogonoside, wogonin, glycyrrhizic acid, liquiritin, rhein, emodin, chrysophanol and physcion. In addition, the principal component analysis (PCA) coupled with the UPLC fingerprint was applied to classify the NHJDP samples according to their manufacture corporation. This proposed method with high sensitivity and selectivity was successfully utilized to analyze 10 major bioactive compounds in 30 batches of NHJDPs, and the results demonstrate that this analytical method is simple and suitable for the original discrimination and quality control of this TCM.