阿胶酶解物抗氧化肽的分离与质谱分析

采用阴离子交换纤维及Sephadex G-25柱色谱法对阿胶酶解物进行分离纯化,以DPPH,ABTS法对阿胶酶解物及其不同部位进行体外抗氧化能力测定,从阿胶酶解物中筛选出体外抗氧化活性强的部位,命名为GFC-1.GFC-1质量浓度为2.0g·L-1时对DPPH清除率为47.95％,0.40 g· L-1时对ABTS清除率为97.20％;利用LC-ESI-MS/MS结合TurboSEQUEST检索软件及Swiss-Prot数据库从GFC-1中鉴定出9个小分子肽,并从中识别出高重复核心序列GPAGPP* GPP*(P*为羟脯氨酸).

电喷雾离子阱二级质谱法测定人血浆中利培酮浓度

目的 建立电喷雾离子阱二级质谱法(LC-ESI-MS/MS)测定人血浆中利培酮浓度.方法 色谱柱DiamonsilTM C18(4.6mm×150mm,5μm);流动相:乙腈(1%甲酸):0.02mol·L-1醋酸铵=(60:40,V:V),流速:0.8 mL·min-1;柱温:25℃;进样体积:10μL;质谱条件为电喷雾离子源,检测方式为正离子多离子反应监测(MRM),用于定量分析的离子为利培酮m/z411→191,内标替米沙坦m/z516→497,生物样本采用醋酸乙酯:二氯甲烷(4:1)液液萃取处理.结果 利培酮血药浓度线性范围为0.5～50 μg·L-1,线性方程为C=2.38F-0.24,r=0.999(n=7),低检测浓度为0.1μg·L-1,高中低三个浓度的提取回收率分别为71.70%,64.58%,64.93%,日内、日间RSD均小于15%.结论 本法灵敏、准确、快速,可用于临床常规血药浓度测定和药动学研究.

大鼠饲料对黄酮类成分药代动力学试验的干扰

目的:研究黄酮类成分药代动力学试验中,大鼠饲料对测定结果的干扰.方法:采用LC-ESI-MS/MS法对大鼠饲料和血浆中的黄酮进行分离,并根据其鉴定.结果:在大鼠标准饲料中检测到5个柑橘类黄酮(柚皮苷、柚皮素、橙皮苷、橙皮素和槲皮素)和2个大豆异黄酮(大豆素、大豆苷),同时在采用该饲料喂养的大鼠血浆中也检测到这7个黄酮.结论:在进行黄酮类成分的药代动力学试验时,饲料来源的黄酮会干扰血浆中该物质的测定,影响研究结果的准确性.

A selective, sensitive and high throughput liquid chromatography-tandem mass spectro-metry (LC-ESI-MS/MS) method has been developed for separation and quantification of metoprolol enantiomers on a chiral Lux Amylose-2 (250 mm×4.6 mm, 5 mm) column. Solid phase extraction of (S)-(-)- and (R)-(t)-metoprolol and rac-metoprolol-d6 as an internal standard (IS) was achieved on Lichrosep DVB HL cartridges employing 200 mL human plasma. Both the analytes were chromatographically separated with a resolution factor of 2.24 using 15 mM ammonium acetate in water, pH 5.0 and 0.1% (v/v) diethyl amine in acetonitrile (50:50, v/v) as the mobile phase within 7.0 min. The precursor-product ion transitions for the enantiomers and IS were monitored in the multiple reaction monitoring and positive ionization mode. The method was validated over the concentration range of 0.500-500 ng/mL for both the enantiomers. Matrix effect was assessed by post-column analyte infusion experiment and the mean extraction recovery was greater than 94.0% for both the enantiomers at all quality control levels. The stability of analytes was evaluated in plasma and whole blood under different storage conditions. The method was successfully applied to a clinical study in 14 healthy volunteers after oral administration of 200 mg metoprolol tablet under fasting conditions. The assay reproducibility is shown by reanalysis of 68 incurred samples. The suitability of the developed method was assessed in comparison with different chromatographic methods developed for stereoselective analysis of metoprolol in biological matrices.