Mingjuan Zhu1, Pinqing Wei1, Qian Peng1, Shengying Qin2, Yuan Zhou1, Ren Zhang3, Chenchen Zhu4, Lei Zhang1. 1. School of Pharmaceutical Sciences, Guangzhou University of Traditional Chinese Medicine, Guangzhou, P. R. China. 2. Clinical Experimental Centre, First Affiliated Hospital of Jinan University, Guangzhou, P. R. China. 3. The College of Fundamental Medical Science, Guangzhou University of Chinese Medicine, Guangzhou, P. R. China. 4. Institute of Clinical Pharmacology, Guangzhou University of Chinese Medicine, Guangzhou, P. R. China.
Abstract
INTRODUCTION: Coumarin and alkaloids are the major bioactive constituents of Toddalia asiatica, playing an important role in various biological activities such as anti-inflammatory, analgesic, anti-bacterial and anti-tumour. OBJECTIVE: To establish a method that will simultaneously determine the coumarins and alkaloids compounds in T. asiatica and identify their characteristic fragmentation patterns, while combining fingerprints and chemical identification with chemometrics for discrimination and quality assessment of T. asiatica samples. METHODOLOGY: Qualitative characterisation of coumarins and alkaloids compounds in the methanol extracts of T. asiatica was determined by ultra-high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS). Quantitative analysis relies on high-performance liquid chromatography with a diode array detector (HPLC-DAD). RESULTS: A total of 59 components were characterised by UPLC-QTOF-MS/MS, including 29 coumarin, 25 alkaloids, one phenolic acid and four flavonoids. While the 19 characteristic components out of 23 common peaks in the chromatographic fingerprints of T. asiatica were confirmed. Quantitative analysis of seven major compounds from 18 samples were simultaneously detected by HPLC-DAD at wavelengths of 280 nm. The samples were classified into three groups by hierarchical clustering analysis (HCA) combined with principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) which screened out the main chemical markers responsible for the samples differences. CONCLUSION: Fingerprints combined with chemometrics and chemical identification are a simple, rapid and effective method for the quality control of T. asiatica.
INTRODUCTION:Coumarin and alkaloids are the major bioactive constituents of Toddalia asiatica, playing an important role in various biological activities such as anti-inflammatory, analgesic, anti-bacterial and anti-tumour. OBJECTIVE: To establish a method that will simultaneously determine the coumarins and alkaloids compounds in T. asiatica and identify their characteristic fragmentation patterns, while combining fingerprints and chemical identification with chemometrics for discrimination and quality assessment of T. asiatica samples. METHODOLOGY: Qualitative characterisation of coumarins and alkaloids compounds in the methanol extracts of T. asiatica was determined by ultra-high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS). Quantitative analysis relies on high-performance liquid chromatography with a diode array detector (HPLC-DAD). RESULTS: A total of 59 components were characterised by UPLC-QTOF-MS/MS, including 29 coumarin, 25 alkaloids, one phenolic acid and four flavonoids. While the 19 characteristic components out of 23 common peaks in the chromatographic fingerprints of T. asiatica were confirmed. Quantitative analysis of seven major compounds from 18 samples were simultaneously detected by HPLC-DAD at wavelengths of 280 nm. The samples were classified into three groups by hierarchical clustering analysis (HCA) combined with principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) which screened out the main chemical markers responsible for the samples differences. CONCLUSION: Fingerprints combined with chemometrics and chemical identification are a simple, rapid and effective method for the quality control of T. asiatica.