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鼻黏膜递送温敏凝胶的组织残留量体外检测方法研究

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  • 1.浙江工业大学药学院生物制剂与材料实验室,杭州 310014;
    2.浙江大学医学院附属第四医院药剂科,金华 322023;
    3.浙江省量子精密测量协同创新中心,杭州 310023
第一作者 修雪亮 Tel:13840152902; E-mail:xueliangxiu@qq.com
傅姝婷 Tel:19858168323; E-mail:610692181@qq.com
*Tel:(0571)88320218; E-mail:merrigen@126.com

收稿日期: 2020-10-22

  网络出版日期: 2024-06-24

Reseach on methods for in vitro determination of the tissue residues of nasal thermosensitive gel

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  • 1. Laboratory of Biologicals and Biomaterials, College of Pharmacy, Zhejiang University of Technology, Hangzhou 310014, China;
    2. Department of Pharmacy, the Fourth Affiliated Hospital, School of Medicine, Zhejiang University, Jinhua 322023, China;
    3. Zhejiang Provincial Key Laboratory & Collaborative Innovation Center for Quantum Precision Measurement, Hangzhou 310023, China

Received date: 2020-10-22

  Online published: 2024-06-24

摘要

目的:改进组织残留量法冲洗装置,建立更适宜测定鼻黏膜递送凝胶制剂组织粘附性的方法。方法:根据鼻黏液的流变学性质,筛选适宜模拟鼻黏液为黏膜冲洗液,以1.0 mL·min-1的流速冲洗模拟鼻黏液进行组织残留量实验。设计自制冲洗装置并与溶出仪进行比较。同时对比称重法和荧光示踪法对凝胶定量的精密度。结果:以新鲜鸡蛋壳膜为模拟鼻黏膜,在与真实鼻黏液固形物相同的情况下,比较多种模拟黏液剪切速率与黏度的变化趋势后发现,羟乙基纤维素(HEC)-溶菌酶溶液具有剪切变稀的性质,但黏度约为20 mPa·s,比真实鼻黏液偏高,且该溶液不稳定,久置后出现大量絮状沉淀。黏蛋白-溶菌酶各1%溶液和HEC-明胶各1%溶液的黏度接近(约13 mPa·s),即与健康的人鼻黏液黏度一致。但只有HEC-明胶溶液久置后稳定无沉淀生成,具有与健康人鼻黏液最相似的流变学性能,且溶液的荧光强度在测定时间段内稳定(21~22 a.u.),符合试验要求。在对组织残留量测定的方法中,以异硫氰酸荧光素标记的牛血清白蛋白(FITC-BSA)为探针的荧光探针法定量测定的RSD在5%~10%,不仅小于称重法结果,也满足《中华人民共和国药典》要求。使用实验自制装置进行冲洗和荧光示踪法时,组内组织残留量的RSD在5%~10%,重复性好。结论:使用自制冲洗装置,以HEC-明胶溶液作为黏膜冲洗液,荧光示踪法定量测定温敏凝胶在用于药物鼻黏膜递送过程中的组织残留量,这一方法简便可靠。

本文引用格式

修雪亮, 傅姝婷, 刘勇, 李志勇, 马凤森 . 鼻黏膜递送温敏凝胶的组织残留量体外检测方法研究[J]. 药物分析杂志, 2021 , 41(11) : 2007 -2016 . DOI: 10.16155/j.0254-1793.2021.11.19

Abstract

Objective: To improve the flushing device for tissue residual method and to establish a more suitable method for the determination of tissue adhesion of nasal mucosal delivery gel preparations. Methods: According to the rheological properties of nasal mucus,a suitable simulated nasal mucus was selected as the mucosal flushing fluid,and the simulated nasal mucus was rinsed at a flow rate of 1.0 mL·min-1 to carry out the tissue residual test. A self-made flushing device was designed and compared with the dissolution apparatus. And the precision of gel quantification was compared between weighing method and fluorescence tracer method. Results: To simulate the nasal mucosa with fresh egg shell membrane, under the same condition as the real nasal mucus solids, comparing the change trend of the shear rate and viscosity of various simulated mucus, it was found that the hydroxyethyl cellulose(HEC)-lysozyme solution had the properties of shear thinning, but the viscosity was about 20 mPa·s higher than that of real nasal mucus, and the solution was unstable, with a large amount of flocculent precipitation appeared after long-term storage. The viscosity of the mucin-lysozyme (1% each) solution and the HEC-gelatin (1% each) solution were close to about 13 mPa·s, which was consistent with the viscosity of healthy human nasal mucus. However, only the HEC-gelatin solution was stable without precipitation after being stored for a long time, and the fluorescence intensity of the solution was stable during the measurement period (21-22 a.u.), and had the rheological properties most similar to healthy human nasal mucus, which met the test requirements. In the method of determining the amount of tissue residue, the fluorescence probe method using fluorescein isothiocyanate-labeled bovine serum albumin (FITC-BSA) as a probe resulted in a quantitative RSD value between 5% and 10%,which was not only smaller than the result of weighing method, but also met the requirements of the Chinese Pharmacopoeia. When using the experimental self-made device for flushing and fluorescence tracing method, the RSD value in the group was between 5% and 10%, and the repeatability was good. Conclusion: Using a self-made flushing device and hydroxyethyl cellulose gelatin solution as a mucosal rinsing fluid,the tissue residue of thermosensitive gel in nasal mucosa delivery process was quantitatively determined by fluorescence tracing. This method is simple and reliable.

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