SPE-HPLC method for the determination of four flavonols in rat plasma and urine after oral administration of Abelmoschus manihot extract
SPE-HPLC method for the determination of four flavonols in rat plasma and urine after oral administration of Abelmoschus manihot extract
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SPE-HPLC法测定口服秋葵提取物后大鼠血浆和尿液中4种黄酮醇的含量
DOI：
10.1016/j.jchromb.2006.12.043
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发表时间：
2007-06-01
影响因子：
3
通讯作者：
Guo, Dean
中科院分区：
文献类型：
--
作者：
Lai, Xianyin;Zhao, Yuying;Guo, Dean
建立了固相萃取-高效液相色谱法同时测定大鼠口服黄蜀葵总黄酮（TFA）后血浆和尿液中异槲皮苷（1）、冬青叶素（2）、杨梅素（3）、槲皮素-3 '-O-D-葡萄糖苷（4）和槲皮素（5）等黄酮醇类化合物的含量。以黄芩苷（6）和山奈酚（7）为内标物。使用Winchem（TM）C-18反相柱通过固相萃取预处理血浆和尿液样品。血浆和尿液提取物分别在YMC-Pack ODS-A C-18和Thermo ODS-2 HYEPRSIL C-18反相柱上进行分析，采用乙腈-0.1%磷酸的移动的流动相。用不同的洗脱梯度进行HPLC分析。流速为1.0 mL/min，检测波长为370 nm。血浆中黄酮醇2-5的校准范围分别为0.011-2.220、0.014-2.856、0.022-4.320和0.028-5.600 μ g/mL。在尿液中，黄酮醇1、2、4和5的校准范围分别为2.00-16.00、8.56-102.72、2.70-21.60和3.00-24.00 μ g/mL。日内和日间RSD分别小于5.40%和4.89%，小于3.96%和6.85%。初步研究了大鼠口服TFA后血浆和尿液中黄酮醇的浓度，结果表明，本方法适用于大鼠血浆和尿液中黄酮醇的测定。(c)2007 Elsevier B. V.保留所有权利。
A SPE-HPLC method was developed and validated for the simultaneous determination of flavonols, isoquercitrin (1), hibifolin (2), myricetin (3), quercetin-3'-O-D-glucoside (4) and quercetin (5) in rat plasma and urine after oral administration of the total flavonoids from Abelmoschus manihot (TFA). The astragalin (6) and kaempferol (7) were used as internal standards (IS). Plasma and urine samples were pretreated by solid-phase extraction using Winchem (TM) C-18 reversed-phase cartridges. Analysis of the plasma and urinary extract was performed on YMC-Pack ODS-A C-18 and Thermo ODS-2HYEPRSIL C-18 reversed-phase column, respectively and a mobile phase of acetonitrile-0.1% phosphoric acid was employed. HPLC analysis was conducted with different elution gradients. The flow rate was 1.0 mL/min and the detection wavelength was set at 370 nm. Calibration ranges in plasma for flavonols 2-5 were at 0.011-2.220, 0.014-2.856, 0.022-4.320, and 0.028-5.600 mu g/mL, respectively. In urine calibration ranges for flavonols 1, 2, 4 and 5 were at 2.00-16.00, 8.56-102.72, 2.70-21.60, and 3.00-24.00 mu g/mL, respectively. The RSD of intra- and inter-day was less than 5.40% and 4.89% in plasma, and less than 3.96% and 6.85% in urine for all the analyses. A preliminary experiment to investigate the plasma concentration and urinary excretion of the flavonols after oral administration of TFA to rats demonstrated that the present method was suitable for determining the flavonols in rat plasma and urine. (c) 2007 Elsevier B.V. All rights reserved.