A sensitive and specific HPGPC-FD method for the study of pharmacokinetics and tissue distribution of Radix Ophiopogonis polysaccharide in rats
A sensitive and specific HPGPC-FD method for the study of pharmacokinetics and tissue distribution of Radix Ophiopogonis polysaccharide in rats
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DOI：
10.1002/bmc.1369
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发表时间：
2010-08-01
影响因子：
1.8
通讯作者：
Feng, Yi
中科院分区：
文献类型：
--
作者：
Lin, Xiao;Wang, Zhuojun;Feng, Yi
对从黄芪中提取的重均分子量为4.8kDa的禾本科型果聚糖（ROP）的抗心肌缺血活性的兴趣使其药代动力学和组织分布的研究成为必要。为此，开发了一种简单的HPGPC-FD方法，用于检测血浆和大鼠组织（心、肝、脾、肺、肾、脑和胃）中的FITC-ROP（异硫氰酸荧光素标记的ROP）。在Shodex Sugar KS-802高效凝胶柱上分离分析物，以0.1磷酸盐缓冲液（pH 7.0）为移动的相，流速为0.5 mL/min，在λ（ex）495 nm和λ（em）515 nm处进行荧光检测。在所有研究的生物样品中，FITC-ROP的校准曲线在0.25-20.0或50.0 μ g/mL范围内呈线性，相关系数>0.995。分析的日间和日内精确度不超过10%，血浆的测定准确度范围为93%至105%，组织匀浆的测定准确度范围为89%至108%。该方法适用于ROP的药代动力学和组织分布的研究，所得结果对ROP的进一步药物开发具有重要指导意义，表明该方法在日益重要的糖基药物中具有广阔的应用前景。版权所有（C）2009约翰威利父子有限公司
Interest in antimyocardial ischemic activity of a graminan-type fructan with a weight average molecular weight of 4.8 kDa extracted from Radix Ophiopogonis (ROP) has necessitated the study of its pharmacokinetics and tissue distribution. For that, a simple HPGPC-FD method was developed for the sensitive and specific determination of FITC-ROP (fluorescein-isothiocyanate-labeled ROP) in plasma and rat tissues (heart, liver, spleen, lung, kidney, brain and stomach). The analyte was separated on a Shodex Sugar KS-802 high-performance gel column with 0.1 on phosphate buffer (pH 7.0) as mobile phase at a flow rate of 0.5 mL/min, and fluorescence detection at lambda(ex) 495 nm and lambda(em) 515 nm. The calibration curve for FITC-ROP was linear over the range 0.25-20.0 or 50.0 mu g/mL in all studied biosamples with correlation coefficients >0.995. The inter-day and intra-day precisions of analysis were not more than 10%, and assay accuracy ranged from 93 to 105% for plasma and from 89 to 108% for tissue homogenates. This method has been confirmed here to be suitable for the study of pharmacokinetics and tissue distribution of ROP and the achieved results are highly instructive for the further pharmaceutical development of ROP, suggesting the promising application of the method to the increasingly important carbohydrate-based drugs. Copyright (C) 2009 John Wiley & Sons, Ltd.