A simple, sensitive and reproducible high performance liquid chromatography-mass spectrometry coupled with electrospray ionization method for simultaneous separation and determination of adenine, adenosine and （urid...A simple, sensitive and reproducible high performance liquid chromatography-mass spectrometry coupled with electrospray ionization method for simultaneous separation and determination of adenine, adenosine and （uridine） was developed. The analytical column is a 2.0 mm×150 mm Shimadzu VP-ODS column and volume fraction of the mobile phase is （86.5%）water, 12.0%methanol and 1.5%formic acid. 2-chloroadenosine was used as internal standard. Selective ion monitoring mode and selective ion monitoring ions at ratio of mass to electric charge of 136 for adenine, 268 for adenosine and 267 for uridine were chosen for quantitative analysis of the three active components. The results show that the regression equations and linear range are Y=0.062X+（0.005） and 2.0140.0 (μg·mL-1)for adenine, Y=0.049X+0.004 and 4.0115.0 μg·mL-1 for uridine, （Y=0.154X）+0.014 and （1.0125.0） μg·mL-1 for adenosine. The limits of detection are 0.6 μg·mL-1 for adenine, 1.0 μg·mL-1for uridine and (0.2 μg·mL-1) for adenosine. The recoveries of the three constituents are from 96.6% to 103.2%.展开更多
AIM To develop a reliable and simple method to identify important biological metabolites and relevant pathways for taurine in hepatic stellate cells(HSCs), in order to provide more data for taurine therapy.METHODS All...AIM To develop a reliable and simple method to identify important biological metabolites and relevant pathways for taurine in hepatic stellate cells(HSCs), in order to provide more data for taurine therapy.METHODS All the biological samples were analyzed by using highperformance liquid chromatography-time electrospray ionization/quadrupole-time of flight mass spectrometry. Principal component analysis and partial least squares discriminant analysis were used to identify statistically different metabolites for taurine in HSCs, and metabolomic pathway analysis was used to do pathway analysis for taurine in HSCs. The chemical structure of the related metabolites and pathways was identified by comparing the m/z ratio and ion mode with the data obtained from free online databases.RESULTS A total of 32 significant differential endogenous metabolites were identified, which may be related to the mechanism of action of taurine in HSCs. Among the seven relevant pathways identified, sphingolipid metabolism pathway, glutathione metabolism pathway and thiamine metabolism pathway were found to be the most important metabolic pathways for taurine in HSCs.CONCLUSION This study showed that there were distinct changes in biological metabolites of taurine in HSCs and three differential metabolic pathways including sphingolipid pathway, glutathione pathway and thiamine metabolism pathway might be of key importance in mediating the mechanism of action of taurine in HSCs.展开更多
A support vector regression based on the mean impact value (MIV) model was constructed to identify the bioactive compounds inhibiting proliferation of He La cells in a combination of turmeric (Curcuma longa L.)and liq...A support vector regression based on the mean impact value (MIV) model was constructed to identify the bioactive compounds inhibiting proliferation of He La cells in a combination of turmeric (Curcuma longa L.)and liquorice (Glycyrrhiza) extracts.The quantitative chemical fingerprint from 50 batches of turmeric and liquorice extracts was established using high performance liquid chromatography hyphenated to an ultraviolet visible detector.Qualitative results were obtained using ultra performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry.A total of 46 peaks (peaks 1–15 from turmeric and 16–46 from liquorice) were selected as "common peaks" for analysis.The inhibitory effect of the combined extracts on He La cells was measured by MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.It was found that 15 compounds (peaks:8,12,30,24,46,11,14,9,3,1,44,18,7,45 and 43)possessing high absolute MIV exhibited a significant correlation with the cytotoxicity against He La cells; most of these have already been confirmed with potential cytotoxicity in previous research.The important potential application of the present model can be extended to help discover active compounds from complex herbal medicine prior to traditional bioassay-guided separation.It is considered that this could be a useful tool for redeveloping herbal medicine based on the use of these active compounds.展开更多
To identify acute renal allograft rejection biomarkers in human serum, two-dimensional differential in-gel electrophoresis （2-D DIGE） and reversed phase high-performance liquid chromatography （RP-HPLC） followed by...To identify acute renal allograft rejection biomarkers in human serum, two-dimensional differential in-gel electrophoresis （2-D DIGE） and reversed phase high-performance liquid chromatography （RP-HPLC） followed by electrospray ionization mass spectrometry （ESI-MS） were used. Serum samples from renal allograft patients and normal volunteers were divided into three groups： acute rejec- tion （AR）, stable renal function （SRF） and normal volunteer （N）. Serum samples were firstly processed using Multiple Affinity Removal Column to selectively remove the highest abundance proteins. Differentially expressed proteins were analyzed using 2-D DIGE. These differential protein spots were excised, digested by trypsin, and identified by RP-HPLC-ESI/MS. Twenty-two differentially expressed proteins were identified in serum from AR group. These proteins included complement C9 precursor, apolipoprotein A-IV precursor, vitamin D-binding protein precursor, beta-2-glycoprotein 1 precursor, etc. Vitamin D-binding protein, one of these proteins, was confirmed by ELISA in the independent set of serum samples. In conclusion, the differentially expressed proteins as serum biomarker candidates may provide the basis of acute rejection noninvasive diagnosis. Confirmed vitamin D-binding protein may be one of serum biomarkers of acute rejection. Furthermore, it may provide great insights into understanding the mechanisms and potential treatment strategy of acute rejection.展开更多
建立了鸡肉中3种氯霉素类抗生素残留的高效液相色谱 电喷雾电离三级四极杆质谱(HPLC ESI MS MS)测定法。该方法采用多反应监测(MRM)负离子模式,可一次对鸡肉中的氯霉素、甲砜霉素和氟甲砜霉素进行定性和定量。该方法仅需1g样品,并省去...建立了鸡肉中3种氯霉素类抗生素残留的高效液相色谱 电喷雾电离三级四极杆质谱(HPLC ESI MS MS)测定法。该方法采用多反应监测(MRM)负离子模式,可一次对鸡肉中的氯霉素、甲砜霉素和氟甲砜霉素进行定性和定量。该方法仅需1g样品,并省去固相萃取步骤,具有操作简便、有机试剂消耗量少、测定周期短等优点。方法的检出限为0 010μg/kg,测定低限为0 100μg/kg,线性范围为0 050～1 00μg/L,加标回收率为69 0%～92 8%,相对标准偏差为6 3%～12 9%。展开更多
文摘A simple, sensitive and reproducible high performance liquid chromatography-mass spectrometry coupled with electrospray ionization method for simultaneous separation and determination of adenine, adenosine and （uridine） was developed. The analytical column is a 2.0 mm×150 mm Shimadzu VP-ODS column and volume fraction of the mobile phase is （86.5%）water, 12.0%methanol and 1.5%formic acid. 2-chloroadenosine was used as internal standard. Selective ion monitoring mode and selective ion monitoring ions at ratio of mass to electric charge of 136 for adenine, 268 for adenosine and 267 for uridine were chosen for quantitative analysis of the three active components. The results show that the regression equations and linear range are Y=0.062X+（0.005） and 2.0140.0 (μg·mL-1)for adenine, Y=0.049X+0.004 and 4.0115.0 μg·mL-1 for uridine, （Y=0.154X）+0.014 and （1.0125.0） μg·mL-1 for adenosine. The limits of detection are 0.6 μg·mL-1 for adenine, 1.0 μg·mL-1for uridine and (0.2 μg·mL-1) for adenosine. The recoveries of the three constituents are from 96.6% to 103.2%.
基金Supported by National Natural Science Foundation of China,No.81360595 and No.81360532Guangxi Natural Science Foundation Program,No.2014GXNSFDA118027+1 种基金Bagui Scholars Foundation Program of GuangxiSpecial-term Experts Foundation Program of Guangxi
文摘AIM To develop a reliable and simple method to identify important biological metabolites and relevant pathways for taurine in hepatic stellate cells(HSCs), in order to provide more data for taurine therapy.METHODS All the biological samples were analyzed by using highperformance liquid chromatography-time electrospray ionization/quadrupole-time of flight mass spectrometry. Principal component analysis and partial least squares discriminant analysis were used to identify statistically different metabolites for taurine in HSCs, and metabolomic pathway analysis was used to do pathway analysis for taurine in HSCs. The chemical structure of the related metabolites and pathways was identified by comparing the m/z ratio and ion mode with the data obtained from free online databases.RESULTS A total of 32 significant differential endogenous metabolites were identified, which may be related to the mechanism of action of taurine in HSCs. Among the seven relevant pathways identified, sphingolipid metabolism pathway, glutathione metabolism pathway and thiamine metabolism pathway were found to be the most important metabolic pathways for taurine in HSCs.CONCLUSION This study showed that there were distinct changes in biological metabolites of taurine in HSCs and three differential metabolic pathways including sphingolipid pathway, glutathione pathway and thiamine metabolism pathway might be of key importance in mediating the mechanism of action of taurine in HSCs.
基金financial support provided by the National Natural Science Foundation of China(No.81102900)
文摘A support vector regression based on the mean impact value (MIV) model was constructed to identify the bioactive compounds inhibiting proliferation of He La cells in a combination of turmeric (Curcuma longa L.)and liquorice (Glycyrrhiza) extracts.The quantitative chemical fingerprint from 50 batches of turmeric and liquorice extracts was established using high performance liquid chromatography hyphenated to an ultraviolet visible detector.Qualitative results were obtained using ultra performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry.A total of 46 peaks (peaks 1–15 from turmeric and 16–46 from liquorice) were selected as "common peaks" for analysis.The inhibitory effect of the combined extracts on He La cells was measured by MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.It was found that 15 compounds (peaks:8,12,30,24,46,11,14,9,3,1,44,18,7,45 and 43)possessing high absolute MIV exhibited a significant correlation with the cytotoxicity against He La cells; most of these have already been confirmed with potential cytotoxicity in previous research.The important potential application of the present model can be extended to help discover active compounds from complex herbal medicine prior to traditional bioassay-guided separation.It is considered that this could be a useful tool for redeveloping herbal medicine based on the use of these active compounds.
基金supported by a grant from National Basic Research 973 Program of China (No.2009CB522407)
文摘To identify acute renal allograft rejection biomarkers in human serum, two-dimensional differential in-gel electrophoresis （2-D DIGE） and reversed phase high-performance liquid chromatography （RP-HPLC） followed by electrospray ionization mass spectrometry （ESI-MS） were used. Serum samples from renal allograft patients and normal volunteers were divided into three groups： acute rejec- tion （AR）, stable renal function （SRF） and normal volunteer （N）. Serum samples were firstly processed using Multiple Affinity Removal Column to selectively remove the highest abundance proteins. Differentially expressed proteins were analyzed using 2-D DIGE. These differential protein spots were excised, digested by trypsin, and identified by RP-HPLC-ESI/MS. Twenty-two differentially expressed proteins were identified in serum from AR group. These proteins included complement C9 precursor, apolipoprotein A-IV precursor, vitamin D-binding protein precursor, beta-2-glycoprotein 1 precursor, etc. Vitamin D-binding protein, one of these proteins, was confirmed by ELISA in the independent set of serum samples. In conclusion, the differentially expressed proteins as serum biomarker candidates may provide the basis of acute rejection noninvasive diagnosis. Confirmed vitamin D-binding protein may be one of serum biomarkers of acute rejection. Furthermore, it may provide great insights into understanding the mechanisms and potential treatment strategy of acute rejection.
