目的:通过采用超高效液相色谱—四极杆-静电场轨道阱高分辨质谱联用仪(Q Exactive Plus UHPLC/HRMS),建立对人尿中42种小肽类禁用物质的检测方法。被检测的小肽类禁用物质涵盖了世界反兴奋剂机构(WADA)禁用清单中的生长激素促分泌剂类(G...目的:通过采用超高效液相色谱—四极杆-静电场轨道阱高分辨质谱联用仪(Q Exactive Plus UHPLC/HRMS),建立对人尿中42种小肽类禁用物质的检测方法。被检测的小肽类禁用物质涵盖了世界反兴奋剂机构(WADA)禁用清单中的生长激素促分泌剂类(GHS)、生长激素释放肽类(GHRP)、促性腺激素释放因子(GnRHs)、人类生长激素(h GH)、生长因子以及生长因子调节剂等分类的定性物质、片段及代谢物,还包括了两种利尿剂去氨加压素(Desmopressin)和苯赖加压素(Felypressin)及其代谢物。方法:使用直接进样方法对尿液样本进行预处理,取待测样品加入内标溶液涡旋混匀,以10000 r/min离心10分钟后。取上清液与稀释溶液混匀,在超高效液相色谱-Q Exactive Plus组合型四极杆Orbitrap质谱仪上进行分析。结果:采用全扫描(Full Scan)和平行反应监测(PRM)模式对含有42种小肽类禁用物质及内标的阳性质量控制样品(QCP)进行定性分析。在最低要求检测浓度(MRPL)下,该方法均可以正确检出尿样中的小肽类目标化合物(其信噪比S/N>3),可满足WADA的相关技术文件的要求。结论:该方法通过直接进样的前处理方法,使得样品前处理程序变得简单、高效,同时因为采用了Q Exactive Plus UHPLC/HRMS,尽可能多地保留了样品检测的灵敏度和特异性,从而使得该方法的灵敏度、特异性以及检出限等均可满足WADA对此类药品检测能力的要求。展开更多
Objective:The present study aimed to evaluate the therapeutic effect and explore the underlying mechanisms of Longxue Tongluo Capsule(LTC)on ischemic stroke rats.Methods:Twenty-six rats were randomly divided into four...Objective:The present study aimed to evaluate the therapeutic effect and explore the underlying mechanisms of Longxue Tongluo Capsule(LTC)on ischemic stroke rats.Methods:Twenty-six rats were randomly divided into four groups,including sham group,sham+LTC group,MCAO group,and MCAO+LTC group.Ischemic stroke rats were simulated by middle cerebral artery occlusion(MCAO),and LTC treatment group were orally administrated with 300 mg/kg of LTC once daily for seven consecutive days.LTC therapy was validated in terms of neurobehavioral abnormality evaluation,cerebral infarct area,and histological assessments.The plasma metabolome comparisons amongst different groups were conducted by UHPLC-Q Exactive MS in combination with subsequent multivariate statistical analysis,aiming to finding the molecules in respond to the surgery or LTC treatment.Results:Intragastric administration of LTC significantly decreased not only the neurobehavioral abnormality scores but also the cerebral infarct area of MCAO rats.The interstitial edema,atrophy,and pyknosis of glial and neuronal cells occurred in the infarcted area,core area,and marginal area of cerebral cortex were improved after LTC treatment.A total of 13 potential biomarkers were observed,and Youden index of 11 biomarkers such as LysoPC,SM,and PE were more than 0.7,which were involved in neuroprotective process.The correlation and pathway analysis showed that LTC was beneficial to ischemic stroke rats via regulating glycerophospholipid and sphingolipid metabolism,together with nicotinate and nicotinamide metabolism.Heatmap and ternary analysis indicated the synergistic effect of carbohydrates and lipids may be induced by flavonoid intake from LTC.Conclusion:The present study could provide evidence that metabolomics,as systematic approach,revealed its capacity to evaluate the holistic efficacy of TCM,and investigate the molecular mechanism underlying the clinical treatment of LTC on ischemic stroke.展开更多
Objective:Pseudostellaria heterophylla has been paid more attention in recent years,mainly as a medicine food homology plant.The content determination of P.heterophylla is not specified in the Chinese Pharmacopoeia(ve...Objective:Pseudostellaria heterophylla has been paid more attention in recent years,mainly as a medicine food homology plant.The content determination of P.heterophylla is not specified in the Chinese Pharmacopoeia(version 2020).The environmental conditions in different production areas could exert an influence on the quality of P.heterophylla.The purpose of this study is to discriminate P.heterophylla collected from different geographical origins of China.Methods:In this study,the content of polysaccharide in 28 batches of P.heterophylla was determined using phenol–sulfuric acid.HPLC fingerprints were established under optimised HPLC-PDA methods.Subsequently,the similarity analysis(SA)and the quantification of heterophyllin B were analyzed.The metabolites of P.heterophylla were identified and evaluated using UHPLC-Q Exactive HF orbitrap MS system.Principal component analysis(PCA),partial least squares discriminant analysis(PLS-DA),hierarchical cluster analysis(HCA)and orthogonal PLS-DA(OPLS-DA)were performed based on all peak areas.Results:The polysaccharide content in Guizhou and Jiangsu was higher than that of other production areas,which varied significant from different origins.While the content of heterophyllin B in Anhui and Jiangsu was high.The correlation coefficients of HPLC fingerprints for 28 batches samples ranged from 0.877 to 0.990,and the characteristic map can be used to identify and evaluate the quality of P.heterophylla.The samples from Fujian,Guizhou,Jiangsu provinces can be relatively separated using multivariate statistical analysis including PCA,PLS-DA,HCA,OPLS-DA,indicating that their metabolic compositions were significantly different.Ultimately,a total of 15 metabolites which were filtrated by a VIPvalue>1 and a P-value<0.05 associated with the separation of different origins were identified.Conclusion:HPLC fingerprint was established to evaluate the quality and authenticity of P.heterophylla.The present work showed that the difference of geographic distributions had an influence on th展开更多
基金financially supported by the National Natural Science Foundation of China(81573572,81530097,32101957)the National Key R&D Program of China(2022YFC2010104)Agricultural Science and Technology Innovation Program of Institute of Food Science and Technology,Chinese Academy of Agricultural Sciences(CAAS-ASTIP-Q2022-IFST-08)。
文摘Objective:The present study aimed to evaluate the therapeutic effect and explore the underlying mechanisms of Longxue Tongluo Capsule(LTC)on ischemic stroke rats.Methods:Twenty-six rats were randomly divided into four groups,including sham group,sham+LTC group,MCAO group,and MCAO+LTC group.Ischemic stroke rats were simulated by middle cerebral artery occlusion(MCAO),and LTC treatment group were orally administrated with 300 mg/kg of LTC once daily for seven consecutive days.LTC therapy was validated in terms of neurobehavioral abnormality evaluation,cerebral infarct area,and histological assessments.The plasma metabolome comparisons amongst different groups were conducted by UHPLC-Q Exactive MS in combination with subsequent multivariate statistical analysis,aiming to finding the molecules in respond to the surgery or LTC treatment.Results:Intragastric administration of LTC significantly decreased not only the neurobehavioral abnormality scores but also the cerebral infarct area of MCAO rats.The interstitial edema,atrophy,and pyknosis of glial and neuronal cells occurred in the infarcted area,core area,and marginal area of cerebral cortex were improved after LTC treatment.A total of 13 potential biomarkers were observed,and Youden index of 11 biomarkers such as LysoPC,SM,and PE were more than 0.7,which were involved in neuroprotective process.The correlation and pathway analysis showed that LTC was beneficial to ischemic stroke rats via regulating glycerophospholipid and sphingolipid metabolism,together with nicotinate and nicotinamide metabolism.Heatmap and ternary analysis indicated the synergistic effect of carbohydrates and lipids may be induced by flavonoid intake from LTC.Conclusion:The present study could provide evidence that metabolomics,as systematic approach,revealed its capacity to evaluate the holistic efficacy of TCM,and investigate the molecular mechanism underlying the clinical treatment of LTC on ischemic stroke.
基金supported by the National Key R&D Program of China(No.2019YFC1710604)the National Key Research and Development Program(No.2020YFA0907903)+2 种基金the key Project at Central Government Level(No.2060302)Tianjin Science and Technology Plan Project(No.19YFZCSY00170)the National Natural Science Foundation of China(No.81872956)。
文摘Objective:Pseudostellaria heterophylla has been paid more attention in recent years,mainly as a medicine food homology plant.The content determination of P.heterophylla is not specified in the Chinese Pharmacopoeia(version 2020).The environmental conditions in different production areas could exert an influence on the quality of P.heterophylla.The purpose of this study is to discriminate P.heterophylla collected from different geographical origins of China.Methods:In this study,the content of polysaccharide in 28 batches of P.heterophylla was determined using phenol–sulfuric acid.HPLC fingerprints were established under optimised HPLC-PDA methods.Subsequently,the similarity analysis(SA)and the quantification of heterophyllin B were analyzed.The metabolites of P.heterophylla were identified and evaluated using UHPLC-Q Exactive HF orbitrap MS system.Principal component analysis(PCA),partial least squares discriminant analysis(PLS-DA),hierarchical cluster analysis(HCA)and orthogonal PLS-DA(OPLS-DA)were performed based on all peak areas.Results:The polysaccharide content in Guizhou and Jiangsu was higher than that of other production areas,which varied significant from different origins.While the content of heterophyllin B in Anhui and Jiangsu was high.The correlation coefficients of HPLC fingerprints for 28 batches samples ranged from 0.877 to 0.990,and the characteristic map can be used to identify and evaluate the quality of P.heterophylla.The samples from Fujian,Guizhou,Jiangsu provinces can be relatively separated using multivariate statistical analysis including PCA,PLS-DA,HCA,OPLS-DA,indicating that their metabolic compositions were significantly different.Ultimately,a total of 15 metabolites which were filtrated by a VIPvalue>1 and a P-value<0.05 associated with the separation of different origins were identified.Conclusion:HPLC fingerprint was established to evaluate the quality and authenticity of P.heterophylla.The present work showed that the difference of geographic distributions had an influence on th
