Quantitative determination of insulin entrapment efficiency in triblock copolymeric nanoparticles by high-performance liquid chromatography
Quantitative determination of insulin entrapment efficiency in triblock copolymeric nanoparticles by high-performance liquid chromatography
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DOI：
10.1016/j.jpba.2005.10.016
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发表时间：
2006-04-11
影响因子：
3.4
通讯作者：
Zhang, ZR
中科院分区：
文献类型：
--
作者：
Xu, XL;Fu, Y;Zhang, ZR
建立了一种快速、有效的三嵌段共聚物纳米粒胰岛素包封率(EE)的测定方法，采用反相高效液相色谱法，在低流速下检测。方法采用Shimadzu Shim-pack VP-ODS柱(150 mm×4.6 mm，5µm，日本东京千代田区)，流动相为0.2M无水硫酸钠溶液，用磷酸和乙腈(73：27，v/v)调节至pH 2.3，流速为0.8mlmin(-1)，检测波长214 nm。对方法的选择性、线性、精密度、准确度、溶液稳定性、检出限和定量限进行了验证。线性范围为2.0~500.0 mgml(-1)，检出限为8 ng，定量限为20 ng。在8-100微克毫升(-1)浓度范围内，平均回收率为98.96%(RSD=2.51%，n=9)。批内和批间变异系数均小于2.24%。该方法具有前处理简单、分离速度快、专属性强、精密度高等优点，可用于商品原料中胰岛素的直接分析、纳米粒制剂、药物释放及稳定性研究。(C)2005 Elsevier B.V.保留所有权利。
A rapid and effective isocratic chromatographic procedure was described in this paper for the determination of insulin entrapment efficiency (EE) in triblock copolymeric nanoparticles using reversed-phase high-performance liquid chromatography (RP-HPLC) with an ultraviolet/visible detector at low flow rate. The method has been developed on a Shimadzu Shim-pack VP-ODS column (150 mm x 4.6 mm, 5 mu m, Chiyoda-Ku, Tokyo, Japan) using a mixture of 0.2 M sodium sulfate anhydrous solution adjusted to pH 2.3 with phosphoric acid and acetonitrile (73:27, v/v) as mobile phase at the flow rate of 0.8 ml min(-1) and a 214 nm detection. The method was validated in terms of selectivity, linearity, precision, accuracy, solution stability, limit of detection (LOD) and limit of quantification (LOQ). The calibration Curve was linear in the concentration range of 2.0-500.0 mu g ml(-1), and the limits of detection and quantitation were 8 and 20 ng, respectively. The mean recovery of insulin from spiked samples, in a concentration range of 8-100 mu g ml(-1), was 98.96% (R.S.D. = 2.51%, n = 9). The intra- and inter-assay coefficients of variation were less than 2.24%. The proposed method has the advantages of simple pretreatment, rapid isolation, high specificity and precision, which can be used for direct analysis of insulin in commercially available raw materials, formulations of nanoparticles, and drug release as well as stability studies. (c) 2005 Elsevier B.V. All rights reserved.