Development and validation of HPLC method for imiquimod determination in skin penetration studies
Development and validation of HPLC method for imiquimod determination in skin penetration studies
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DOI：
10.1002/bmc.1075
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发表时间：
2008-12-01
影响因子：
1.8
通讯作者：
Lopes Badra Bentley, Maria Vitoria
中科院分区：
文献类型：
--
作者：
De Paula, Daniel;Martins, Carolina Azenha;Lopes Badra Bentley, Maria Vitoria
一个简单，快速和灵敏的分析方法，用于测量皮肤样品中的咪喹莫特体外渗透研究后，已开发和验证。在Franz扩散池中用猪皮进行体外渗透研究。采用胶带剥离技术将角质层（SC）与有活力的表皮和真皮分离。使用7：3（v/v）甲醇：乙酸盐缓冲液（100 mm，pH 4.0）溶液和超声处理从皮肤样品中提取咪喹莫特。咪喹莫特通过使用C-8柱的H-PLC和在242 nm处的UV检测进行分析。使用的移动的流动相为乙腈：乙酸盐缓冲液（pH 4.0，100 mM）：二乙胺（30：69.85：0.15，v/v），流速为1 mL/min。咪喹莫特在4.1 min洗脱，运行时间限制为6.0 min。该方法在以下浓度范围内呈线性：SC和胶带剥离皮肤均为100-2500 ng/mL，受体溶液为20-800 ng/mL。在定量限下，日内和日间准确度和精密度值均低于20%。回收率范围为80 - 100%。该方法足以测定来自皮肤样品的咪喹莫特，使得能够通过体外研究确定优喹莫特的皮肤渗透特性。版权所有（C）2008约翰威利父子有限公司
A simple, rapid and sensitive analytical procedure for the measurement of imiquimod in skin samples after in vitro penetration studies has been developed and validated. In vitro penetration studies were carried out in Franz diffusion cells with porcine skin. Tape stripping technique was used to separate the stratum corneum (SC) from the viable epidermis and dermis. Imiquimod was extracted from skin samples using a 7:3 (v/v) methanol:acetate buffer (100 mm, pH 4.0) solution and ultrasonication. Imiquimod was analyzed by H-PLC using C-8 column and UV detection at 242 ran. The mobile phase used was acetonitrile:acetate buffer (pH 4.0, 100 mM):diethylamine (30:69.85:0.15, v/v) with flow rate 1 mL/min. Imiquimod eluted at 4.1 min and the running time was limited to 6.0 min. The procedure was linear across the following concentration ranges: 100-2500 ng/mL for both SC and tape-stripped skin and 20-800 ng/mL for receptor solution. Intra-day and inter-day accuracy and precision values were lower than 20% at the limit of quantitation. The recovery values ranged from 80 to 100%. The method is adequate to assay imiquimod from skin samples, enabling the determination of the cutaneous penetration profile of uniquimod by in vitro studies. Copyright (C) 2008 John Wiley & Sons, Ltd.