An efficient preparative method was successfully developed for isolation and purification of unstable components from medicinal plant extracts, using a combined method of preparative high performance liquid chro-matog...An efficient preparative method was successfully developed for isolation and purification of unstable components from medicinal plant extracts, using a combined method of preparative high performance liquid chro-matography(HPLC) and solid-phase extraction(SPE). The aim of this study was to obtain an effective method with high preparative efficiency and importantly to avoid the transformation of unstable compounds. The preparative HPLC system was based on an LC/MS controlled four-channel autopurification system. The SPE method was performed with a C18 packing material to trap the target compounds and to remove the acidic additive derived from the mobile phase. Using this method, the unstable iridoid glucosides(IGs) as model compounds were successfully isolated and purified from the extract of Hedyotis diffusa Willd. Six IGs(including one new minor IG) and one nucleotide compound were simultaneously obtained, each with a purity of 91% as determined by HPLC. The structures of the isolated compounds were identified by UPLC/Q-TOF MS, UV, 1D and/or 2D NMR. It was demonstrated that the combination of preparative HPLC with SPE is a versatile tool for preparative purification of unstable compounds from complex natural products.展开更多
Centrifugal ultrafiltration after methanol extraction of whole plasma was used as an optimal condition for the preparation of blood plasma before metabonomic studies. The plasma samples from 102 lung cancer patients a...Centrifugal ultrafiltration after methanol extraction of whole plasma was used as an optimal condition for the preparation of blood plasma before metabonomic studies. The plasma samples from 102 lung cancer patients and 34 healthy volunteers were prepared with this approach. With ultra-performance liquid chromatography（UPLC） coupled with quadrupole time-of-flight mass spectrometry（Q-TOF MS） analysis, the samples were investigated in order to find potential disease biomarkers. After data acquisition, orthogonal signal correction partial least squares models were built to differentiate the healthy volunteers from lung cancer patients and to identify metabolites that showed significantly different expression between the two groups. Several metabolite ions were identified as potential biomarkers according to the variable importance in the project（VIP） value in both ion modes. Five lysophosphatidylcholines were further identified as specifically lysoPC 16：0, isomer of lysoPC 16：0, lysoPC 18：0, lysoPC 18：1 and lysoPC 18：2. These results suggest that UPLC coupled with Q-TOF MS is an effective technique for the analysis of plasma metabolites in metabonomic studies.展开更多
Qing-Fei-Pai-Du decoction(QFPDD)is a Chinese medicine compound formula recommended for combating corona virus disease 2019(COVID-19)by National Health Commission of the People's Republic of China.The latest clinic...Qing-Fei-Pai-Du decoction(QFPDD)is a Chinese medicine compound formula recommended for combating corona virus disease 2019(COVID-19)by National Health Commission of the People's Republic of China.The latest clinical study showed that early treatment with QFPDD was associated with favorable outcomes for patient recovery,viral shedding,hospital stay,and course of the disease.However,the effective constituents of QFPDD remain unclear.In this study,an UHPLC-Q-Orbitrap HRMS based method was developed to identify the chemical constituents in QFPDD and the absorbed prototypes as well as the metabolites in mice serum and tissues following oral administration of QFPDD.A total of 405 chemicals,including 40 kinds of alkaloids,162 kinds of flavonoids,44 kinds of organic acids,71 kinds of triterpene saponins and 88 kinds of other compounds in the water extract of QFPDD were tentatively identified via comparison with the retention times and MS/MS spectra of the standards or refereed by literature.With the help of the standards and in vitro metabolites,195 chemical components(including 104 prototypes and 91 metabolites)were identified in mice serum after oral administration of QFPDD.In addition,165,177,112,120,44,53 constituents were identified in the lung,liver,heart,kidney,brain,and spleen of QFPDD-treated mice,respectively.These findings provided key information and guidance for further investigation on the pharmacologically active substances and clinical applications of QFPDD.展开更多
基金Supported by the Science and Technology Plan of Liaoning Province, China(No.2006226002)the Project of the Doctor Fund of Hebei University of Science and Technology, China(No.005121)
文摘An efficient preparative method was successfully developed for isolation and purification of unstable components from medicinal plant extracts, using a combined method of preparative high performance liquid chro-matography(HPLC) and solid-phase extraction(SPE). The aim of this study was to obtain an effective method with high preparative efficiency and importantly to avoid the transformation of unstable compounds. The preparative HPLC system was based on an LC/MS controlled four-channel autopurification system. The SPE method was performed with a C18 packing material to trap the target compounds and to remove the acidic additive derived from the mobile phase. Using this method, the unstable iridoid glucosides(IGs) as model compounds were successfully isolated and purified from the extract of Hedyotis diffusa Willd. Six IGs(including one new minor IG) and one nucleotide compound were simultaneously obtained, each with a purity of 91% as determined by HPLC. The structures of the isolated compounds were identified by UPLC/Q-TOF MS, UV, 1D and/or 2D NMR. It was demonstrated that the combination of preparative HPLC with SPE is a versatile tool for preparative purification of unstable compounds from complex natural products.
基金Supported by the National Natural Science Foundation of China(No.30801513)the Knowledge Innovation Program of Chinese Academy of Sciences(No.KSCX2-YW-R-170)
文摘Centrifugal ultrafiltration after methanol extraction of whole plasma was used as an optimal condition for the preparation of blood plasma before metabonomic studies. The plasma samples from 102 lung cancer patients and 34 healthy volunteers were prepared with this approach. With ultra-performance liquid chromatography（UPLC） coupled with quadrupole time-of-flight mass spectrometry（Q-TOF MS） analysis, the samples were investigated in order to find potential disease biomarkers. After data acquisition, orthogonal signal correction partial least squares models were built to differentiate the healthy volunteers from lung cancer patients and to identify metabolites that showed significantly different expression between the two groups. Several metabolite ions were identified as potential biomarkers according to the variable importance in the project（VIP） value in both ion modes. Five lysophosphatidylcholines were further identified as specifically lysoPC 16：0, isomer of lysoPC 16：0, lysoPC 18：0, lysoPC 18：1 and lysoPC 18：2. These results suggest that UPLC coupled with Q-TOF MS is an effective technique for the analysis of plasma metabolites in metabonomic studies.
基金the National Key Research and Development Program of China(2020YFC0845400)the NSF of China(81922070)+3 种基金Shanghai Science and Technology Innovation Action Plans(20S2190150020S21900900)supported by Shanghai Science and Technology Committeethe Three-year Action Plan of Shanghai TCM Development(ZY-(2018-2020)-CCCX-5001)Program of Shanghai Academic/Technology Research Leader(18XD1403600)。
文摘Qing-Fei-Pai-Du decoction(QFPDD)is a Chinese medicine compound formula recommended for combating corona virus disease 2019(COVID-19)by National Health Commission of the People's Republic of China.The latest clinical study showed that early treatment with QFPDD was associated with favorable outcomes for patient recovery,viral shedding,hospital stay,and course of the disease.However,the effective constituents of QFPDD remain unclear.In this study,an UHPLC-Q-Orbitrap HRMS based method was developed to identify the chemical constituents in QFPDD and the absorbed prototypes as well as the metabolites in mice serum and tissues following oral administration of QFPDD.A total of 405 chemicals,including 40 kinds of alkaloids,162 kinds of flavonoids,44 kinds of organic acids,71 kinds of triterpene saponins and 88 kinds of other compounds in the water extract of QFPDD were tentatively identified via comparison with the retention times and MS/MS spectra of the standards or refereed by literature.With the help of the standards and in vitro metabolites,195 chemical components(including 104 prototypes and 91 metabolites)were identified in mice serum after oral administration of QFPDD.In addition,165,177,112,120,44,53 constituents were identified in the lung,liver,heart,kidney,brain,and spleen of QFPDD-treated mice,respectively.These findings provided key information and guidance for further investigation on the pharmacologically active substances and clinical applications of QFPDD.
