目的建立18个品种苦荞麦Fagopyrum tataricum指纹图谱,并以芦丁为内标物,采用一测多评法(quantitative analysis of multi-components by single-marker,QAMS)同时检测苦荞麦中葫芦巴碱、表儿茶素、山柰酚-3-O-芸香糖苷、槲皮素、山柰...目的建立18个品种苦荞麦Fagopyrum tataricum指纹图谱,并以芦丁为内标物,采用一测多评法(quantitative analysis of multi-components by single-marker,QAMS)同时检测苦荞麦中葫芦巴碱、表儿茶素、山柰酚-3-O-芸香糖苷、槲皮素、山柰酚和大黄素含量,以期对不同产地苦荞麦进行质量控制。方法采用超高效液相-三重四级杆液质联用技术(UPLC-QQQ-MS/MS),首次以一级全扫(Full-MS)和多反应监测(multiple reaction monitoring,MRM)2种采集模式相结合的方式建立UPLC-QQQ-MS/MS指纹图谱共有模式,采用夹角余弦法进行相似度评价。以ThermoFisher Accucore^(TM)C_(18)色谱柱(100 mm×3.0 mm,2.6μm);流动相为乙腈-0.1%甲酸水溶液,体积流量0.3 mL/min,柱温30℃,梯度洗脱。以芦丁作为内标物,建立苦荞中其他6种指标成分的相对校正因子(fs/x),从而计算各待测成分的量,并将QAMS计算值与外标法(external standard method,ESM)实测值进行比较,以验证QAMS的准确性和方法适宜性。运用聚类分析、主成分分析、加权分析等化学计量学分析手段建立苦荞麦综合质量优劣评价方法。结果建立了苦荞麦的UPLC-QQQ-MS/MS指纹图谱,在Full-MS扫描模式下标定了15个共有峰,各品种间相似度在0.399~0.973;同时以MRM扫描模式结合对照品对其中芦丁等7种指标成分进行了峰指认。以芦丁为内标物,与葫芦巴碱、表儿茶素、山柰酚-3-O-芸香糖苷、槲皮素、山柰酚和大黄素6种有效成分的建立的fs/x重现性良好,分别为0.283、1.338、0.654、1.162、8.509、0.709。且采用QAMS测定的18个品种苦荞麦中7种指标成分含量与ESM的实测值之间无显著性差异(P>0.05)。化学计量学分析结果表明,芦丁、山柰酚-3-O-芸香糖苷和山柰酚是影响苦荞麦质量的主要潜在标志物,质量排序相对靠前的苦荞麦产区主要集中在海拔高、光照强、昼夜温差大的中国西南地区,而指纹图谱相似度差异较明显的荞杂2号、综甜2号和通荞1号排在后3位。结论所建立的指纹图谱结合QAMS简便可行,结果准确,化学计量学分析方法全面客观,可为苦荞麦品质评价和质量控制提供参考和依据。展开更多
Objective To investigate excretion profiles of the four major anti-oxidant active catechins, (-) epigallo-catechin-3-gallate (EGCG), (-) epicatechin-3-gallate (ECG), (-) epigallocatechin (EGC), and epicatechin (EC) in...Objective To investigate excretion profiles of the four major anti-oxidant active catechins, (-) epigallo-catechin-3-gallate (EGCG), (-) epicatechin-3-gallate (ECG), (-) epigallocatechin (EGC), and epicatechin (EC) in tea polyphenols (TP) in rats in order to provide experimental data for clinical uses and development of TP as a novel drug. Methods The above four catechins in urine, bile, and feces were simultaneously determined by high performance liquid chromatography coupled with ultraviolet absorption detector (HPLC-UV) assay with a binary gradient elution. The samples were extracted by ethyl acetate prior to HPLC. The quantification was carried out by peak area internal standard method. Following iv dosing TP 100 mg/kg to rats, the samples were collected at different time intervals up to 8 h (urine and bile) and 24 h (feces). Results The urinary Ae, 0-8 h (cumulative excretion amount over 8 h) of EGCG, ECG, EGC, and EC were, on the average, 150.83, 30.75, 116.69, and 254.56 μg, corresponding to fe, 0-8 h (cumulative excretion fraction of dose over 8 h) of 1.45%, 0.84%, 7.88%, and 10.73%, respectively; the biliary Ae, 0-8 h were 12.61, 42.64, 6.61, and 1.24 μg, corresponding to the fe, 0-8 h of 0.12%, 1.16%, 0.45%, and 0.053%,respectively. For fecal excretion, only EGCG and EGC were detected with Ae, 0-24 h of 7.38 μg (fe, 0-24 h of 0.07%) and 157 μg (fe, 0-24 h of 9.99 %), respectively. The fe, total (the total fe of 3 excretory routes) were 18.32%, 10.78%, 2.00%, and 1.64% for EGC, EC, ECG, and EGCG, respectively. Conclusion EGCG and EC are mainly excreted in urine, ECG in bile, and EGC in feces by reference to their Ae and fe. The excretion of the four catechins based on fe, total is ranked in order of EGC > EC > ECG > EGCG. Only small amount of four catechins are recovered in urine, bile, and feces, indicating an extensive metabolic conversion of catechins in the rat body.展开更多
文摘目的建立18个品种苦荞麦Fagopyrum tataricum指纹图谱,并以芦丁为内标物,采用一测多评法(quantitative analysis of multi-components by single-marker,QAMS)同时检测苦荞麦中葫芦巴碱、表儿茶素、山柰酚-3-O-芸香糖苷、槲皮素、山柰酚和大黄素含量,以期对不同产地苦荞麦进行质量控制。方法采用超高效液相-三重四级杆液质联用技术(UPLC-QQQ-MS/MS),首次以一级全扫(Full-MS)和多反应监测(multiple reaction monitoring,MRM)2种采集模式相结合的方式建立UPLC-QQQ-MS/MS指纹图谱共有模式,采用夹角余弦法进行相似度评价。以ThermoFisher Accucore^(TM)C_(18)色谱柱(100 mm×3.0 mm,2.6μm);流动相为乙腈-0.1%甲酸水溶液,体积流量0.3 mL/min,柱温30℃,梯度洗脱。以芦丁作为内标物,建立苦荞中其他6种指标成分的相对校正因子(fs/x),从而计算各待测成分的量,并将QAMS计算值与外标法(external standard method,ESM)实测值进行比较,以验证QAMS的准确性和方法适宜性。运用聚类分析、主成分分析、加权分析等化学计量学分析手段建立苦荞麦综合质量优劣评价方法。结果建立了苦荞麦的UPLC-QQQ-MS/MS指纹图谱,在Full-MS扫描模式下标定了15个共有峰,各品种间相似度在0.399~0.973;同时以MRM扫描模式结合对照品对其中芦丁等7种指标成分进行了峰指认。以芦丁为内标物,与葫芦巴碱、表儿茶素、山柰酚-3-O-芸香糖苷、槲皮素、山柰酚和大黄素6种有效成分的建立的fs/x重现性良好,分别为0.283、1.338、0.654、1.162、8.509、0.709。且采用QAMS测定的18个品种苦荞麦中7种指标成分含量与ESM的实测值之间无显著性差异(P>0.05)。化学计量学分析结果表明,芦丁、山柰酚-3-O-芸香糖苷和山柰酚是影响苦荞麦质量的主要潜在标志物,质量排序相对靠前的苦荞麦产区主要集中在海拔高、光照强、昼夜温差大的中国西南地区,而指纹图谱相似度差异较明显的荞杂2号、综甜2号和通荞1号排在后3位。结论所建立的指纹图谱结合QAMS简便可行,结果准确,化学计量学分析方法全面客观,可为苦荞麦品质评价和质量控制提供参考和依据。
基金support was provided by Dalian Municipal Fund of Science and Technology (2002B4NS044)Kangbosi Pharmaceutical Co. Ltd, China
文摘Objective To investigate excretion profiles of the four major anti-oxidant active catechins, (-) epigallo-catechin-3-gallate (EGCG), (-) epicatechin-3-gallate (ECG), (-) epigallocatechin (EGC), and epicatechin (EC) in tea polyphenols (TP) in rats in order to provide experimental data for clinical uses and development of TP as a novel drug. Methods The above four catechins in urine, bile, and feces were simultaneously determined by high performance liquid chromatography coupled with ultraviolet absorption detector (HPLC-UV) assay with a binary gradient elution. The samples were extracted by ethyl acetate prior to HPLC. The quantification was carried out by peak area internal standard method. Following iv dosing TP 100 mg/kg to rats, the samples were collected at different time intervals up to 8 h (urine and bile) and 24 h (feces). Results The urinary Ae, 0-8 h (cumulative excretion amount over 8 h) of EGCG, ECG, EGC, and EC were, on the average, 150.83, 30.75, 116.69, and 254.56 μg, corresponding to fe, 0-8 h (cumulative excretion fraction of dose over 8 h) of 1.45%, 0.84%, 7.88%, and 10.73%, respectively; the biliary Ae, 0-8 h were 12.61, 42.64, 6.61, and 1.24 μg, corresponding to the fe, 0-8 h of 0.12%, 1.16%, 0.45%, and 0.053%,respectively. For fecal excretion, only EGCG and EGC were detected with Ae, 0-24 h of 7.38 μg (fe, 0-24 h of 0.07%) and 157 μg (fe, 0-24 h of 9.99 %), respectively. The fe, total (the total fe of 3 excretory routes) were 18.32%, 10.78%, 2.00%, and 1.64% for EGC, EC, ECG, and EGCG, respectively. Conclusion EGCG and EC are mainly excreted in urine, ECG in bile, and EGC in feces by reference to their Ae and fe. The excretion of the four catechins based on fe, total is ranked in order of EGC > EC > ECG > EGCG. Only small amount of four catechins are recovered in urine, bile, and feces, indicating an extensive metabolic conversion of catechins in the rat body.
