该实验采用LC-MS对银杏二萜内酯葡胺注射液原料中所含黄酮类成分的种类进行筛选,通过紫外-可见分光光度法（UV-Vis）和高效液相色谱法（HPLC-DAD）对银杏二萜内酯葡胺注射液原料中黄酮类成分限量检查灵敏度比较,发现HPLCDAD的检测限约为...该实验采用LC-MS对银杏二萜内酯葡胺注射液原料中所含黄酮类成分的种类进行筛选,通过紫外-可见分光光度法（UV-Vis）和高效液相色谱法（HPLC-DAD）对银杏二萜内酯葡胺注射液原料中黄酮类成分限量检查灵敏度比较,发现HPLCDAD的检测限约为UV-Vis的1/100,即HPLC-DAD检测灵敏度更高。基于LC-MS筛选结果选用槲皮素、山柰素和异鼠李素为对照品,采用Agilent Eclipse plus C18色谱柱（4.6 mm×250 mm,5μm）;0.4%磷酸-甲醇（50∶50）为流动相,等度洗脱;流速1.0 m L·min-1;检测波长360 nm。结果显示,此法专属性、精密度、重复性良好;槲皮素、山柰素和异鼠李素检测限分别为27.6,22.3,29.5μg·L-1;平均回收率在88.3%~91.7%;基于10批次样品检测结果和仪器灵敏度,最终确定银杏二萜内酯葡胺注射液原料中总黄酮醇苷的含量不得过2×10-5。该方法简便、快捷,可操作性好,可用于银杏二萜内酯葡胺注射液原料中黄酮类成分的限度检测。展开更多
Method development for determination of isoflavones in kudzu was achieved by HPLC/UV/ESI-MSD. Us- ing three kudzu species of Pueraria lobata, P. thomsonii and P. edulis, and analyzing the isoflavones sepa- rately by s...Method development for determination of isoflavones in kudzu was achieved by HPLC/UV/ESI-MSD. Us- ing three kudzu species of Pueraria lobata, P. thomsonii and P. edulis, and analyzing the isoflavones sepa- rately by species and from different plant tissues (roots, stems, leaves, flowers and fruits) in each species, a total of 25 isoflavones were identified by their molecular ions and characteristic fragment ion peaks using LC/MSD under MS and MS/MS mode, and in comparison with standard isoflavones. Two main chemical groups were identified: 1) 8-C-glycosyl isoflavone of puerarin and the analogues of 5-OH puerarin, 3’-OH puerarin, 3’-OMe puerarin, and their glycosides;and 2) daidzein, genistein, glycitein and their glycosyl and malonyl derivatives, which are similar to those known in soy. To accurately quantitate total isoflavones, acidic hydrolysis during extraction of kudzu samples was applied to convert the oxygen glycosides into their respective isoflavone aglycones of daidzein, genistein and glycitein, or non-hydrolyzed carbon glycosides of puerarin, 5-OH puerarin, 3’-OH puerarin and 3’-OMe puerarin. Under the multiple optimized conditions, all seven isoflavones in acidic hydrolyzed kudzu extracts were successfully separated within 30 min and quanti- fied individually with calycosin used as internal standard by both UV and MS detectors. For the quantitative study, several standards e.g. 5-OH puerarin, 3’-OH puerarin and 3’-OMe puerarin are not commercially available. Using polyamide, sephdex-LH20 chromatography and Prep-HPLC, we purified these three stan- dards from kudzu extracts and then elucidated their structures by UV, MS and NMR spectrometric methods. This is the first method to simultaneously quantitate all the isoflavones in kudzu.展开更多
目的研究利可君的4个非对映异构体及其有关物质的结构。方法利用液相色谱-二极管阵列检测、液相色谱-串联质谱和液相色谱-核磁共振等联用技术研究利可君及有关物质的结构。采用Phenomnex Luna C18色谱柱(250 mm×4.60 mm ID,5μm),...目的研究利可君的4个非对映异构体及其有关物质的结构。方法利用液相色谱-二极管阵列检测、液相色谱-串联质谱和液相色谱-核磁共振等联用技术研究利可君及有关物质的结构。采用Phenomnex Luna C18色谱柱(250 mm×4.60 mm ID,5μm),水-乙腈-冰醋酸(58∶42∶0.3)为流动相。结果确定了在溶液状态下利可君及其有关物质的结构,发现其均有4个非对映异构体;还对利可君的4个非对映异构体在溶液中互变与稳定性进行比较与量子化学计算,证明其稳定构型为3R4S5R。结论利可君在溶液中非对映异构体相互转化,有一种优势构型。展开更多
文摘该实验采用LC-MS对银杏二萜内酯葡胺注射液原料中所含黄酮类成分的种类进行筛选,通过紫外-可见分光光度法（UV-Vis）和高效液相色谱法（HPLC-DAD）对银杏二萜内酯葡胺注射液原料中黄酮类成分限量检查灵敏度比较,发现HPLCDAD的检测限约为UV-Vis的1/100,即HPLC-DAD检测灵敏度更高。基于LC-MS筛选结果选用槲皮素、山柰素和异鼠李素为对照品,采用Agilent Eclipse plus C18色谱柱（4.6 mm×250 mm,5μm）;0.4%磷酸-甲醇（50∶50）为流动相,等度洗脱;流速1.0 m L·min-1;检测波长360 nm。结果显示,此法专属性、精密度、重复性良好;槲皮素、山柰素和异鼠李素检测限分别为27.6,22.3,29.5μg·L-1;平均回收率在88.3%~91.7%;基于10批次样品检测结果和仪器灵敏度,最终确定银杏二萜内酯葡胺注射液原料中总黄酮醇苷的含量不得过2×10-5。该方法简便、快捷,可操作性好,可用于银杏二萜内酯葡胺注射液原料中黄酮类成分的限度检测。
文摘Method development for determination of isoflavones in kudzu was achieved by HPLC/UV/ESI-MSD. Us- ing three kudzu species of Pueraria lobata, P. thomsonii and P. edulis, and analyzing the isoflavones sepa- rately by species and from different plant tissues (roots, stems, leaves, flowers and fruits) in each species, a total of 25 isoflavones were identified by their molecular ions and characteristic fragment ion peaks using LC/MSD under MS and MS/MS mode, and in comparison with standard isoflavones. Two main chemical groups were identified: 1) 8-C-glycosyl isoflavone of puerarin and the analogues of 5-OH puerarin, 3’-OH puerarin, 3’-OMe puerarin, and their glycosides;and 2) daidzein, genistein, glycitein and their glycosyl and malonyl derivatives, which are similar to those known in soy. To accurately quantitate total isoflavones, acidic hydrolysis during extraction of kudzu samples was applied to convert the oxygen glycosides into their respective isoflavone aglycones of daidzein, genistein and glycitein, or non-hydrolyzed carbon glycosides of puerarin, 5-OH puerarin, 3’-OH puerarin and 3’-OMe puerarin. Under the multiple optimized conditions, all seven isoflavones in acidic hydrolyzed kudzu extracts were successfully separated within 30 min and quanti- fied individually with calycosin used as internal standard by both UV and MS detectors. For the quantitative study, several standards e.g. 5-OH puerarin, 3’-OH puerarin and 3’-OMe puerarin are not commercially available. Using polyamide, sephdex-LH20 chromatography and Prep-HPLC, we purified these three stan- dards from kudzu extracts and then elucidated their structures by UV, MS and NMR spectrometric methods. This is the first method to simultaneously quantitate all the isoflavones in kudzu.
文摘目的研究利可君的4个非对映异构体及其有关物质的结构。方法利用液相色谱-二极管阵列检测、液相色谱-串联质谱和液相色谱-核磁共振等联用技术研究利可君及有关物质的结构。采用Phenomnex Luna C18色谱柱(250 mm×4.60 mm ID,5μm),水-乙腈-冰醋酸(58∶42∶0.3)为流动相。结果确定了在溶液状态下利可君及其有关物质的结构,发现其均有4个非对映异构体;还对利可君的4个非对映异构体在溶液中互变与稳定性进行比较与量子化学计算,证明其稳定构型为3R4S5R。结论利可君在溶液中非对映异构体相互转化,有一种优势构型。
