Study On The Quality Control Of The Correlation Between The Establishment Of HPLC-UPLC-MS Fingerprint Of Total Alkaloids Of Stemona Tuberosa Lour And Its Origins

Objective: To carry out field visits and sample gathering of Stemona Tuberosa Lour in 31 counties and cities of 11 provinces across the country.The HPLC fingerprints of total alkaloids of Stemona Tuberosa Lour samples of 31 origins are studied,and the HPLC fingerprints of total alkaloids of Stemona Tuberosa Lour from the country are subjected to reference peak identification through the ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS/MS).The correlation between the HPLC fingerprint and the origin is established.Experimental data is provided for the quality control of medicinal materials of Stemona Tuberosa Lour in order to provide a scientific basis for better development and utilization of this traditional Chinese herbal medicine.Methods: 1.By means of hand-held GPS,field visits and sample collection of Stemona Tuberosa Lour were made in 31 counties and cities of 11 provinces across the country.Gathered medicinal herbs were processed in the origins in accordance with the methods under Chinese Pharmacopoeia 2015 Edition.2.Through the single factor investigation method,the classical bio-alkali-acid precipitation was optimized to prepare the test solution of total alkaloids of Stemona Tuberosa Lour and to determine the liquid chromatographic conditions,the gathered data is given the clustering analysis,its fingerprints are established,and the Chinese medicine chromatography fingerprint similarity evaluation system 2012.1 version software is adopted to conduct the common peak confirmation and similarity evaluation of 31 batches of sample categories.The major principal component factors of the reference peak in HLPC fingerprints are analyzed by SPSS 22.0 statistical software.3.By using the ultra-high performance liquid chromatography-tandem mass spectrometry,the reference peaks in HLPC fingerprints are analyzed and identified.4.Through the method of combining the HPLC fingerprint and the origin distribution,the HPLC fingerprint evaluation system of HPLC fingerprints of Stemona Tuberosa Lour is established,in which the origins are combined with the HPLC fingerprints.Results: 1.Detailed habitat study and record of medicinal materials of Stemona Tuberosa Lour from 31 producing areas in the country were made,and the samples were processed.The data analysis software SPSS 22.0 and the chromatographic evaluation system software were used to have calibrated eight chromatographic peaks with good stability,strong absorption peaks and obvious characteristics as the reference peaks of HPLC fingerprints of the total alkaloids of Stemona Tuberosa Lour,31 batches of Stemona Tuberosa Lour were discussed by classification and divided into 3 categories,with the similarity of the fingerprints of total alkaloids of each category greater than 0.90.After analysis of principal component factors,two principal component factors were obtained,standing for 78.30%.3.The ultra-high performance liquid chromatographytandem mass spectrometry was utilized to identify the reference peaks of HPLC fingerprints of the total alkaloids of Stemona Tuberosa Lour,and six components including Croomine,isotuberostemonine,neotuberostemonine,tuberostemonine,tuberostemonin e H and oxytuberstemonine were determined preliminarily.4.The classification analysis of the origins and HPLC fingerprints shows the consistent congruity of fingerprints and origins.Conclusion: This experiment has established the HPLC fingerprints of total alkaloids of Stemona Tuberosa Lour,employed a system of combining the fingerprints with the quantification of the index components,and exhibited the correlation of integral characteristics to individual characteristics of the active ingredients of Chinese medicinal materials and correlation of origins to difference between components to control the quality of medicinal materials,providing a data support for further optimization and improvement of the quality standard of medicinal materials of Stemona Tuberosa Lour.,and furnishing a theoretical basis for assessment of its quality,and recyclable development and utilization.