不同寡糖体外透皮吸收荧光定量分析方法的建立与应用  

Establishment and application of fluorescence quantitative analysis method for transdermal absorption of different oligosaccharides in vitro

作　　者：杨荣荣[1] 王力 李鑫[1] 杨程[1] 王世欣[1,2] 李春霞 YANG Rongrong;WANG Li;LI Xin;YANG Cheng;WANG Shixin;LI Chunxia(Key Laboratory of Marine Drugs,Ministry of Education,Shandong Key Laboratory of Glycoscience and Glycotechnology,School of Medicine and Pharmacy,Ocean University of China,Qingdao 266003,China;Marine Biomedical Research Institute of Qingdao,Qingdao 266071,China;Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology,Qingdao 266237,China)

机构地区：[1]中国海洋大学海洋药物教育部重点实验室,山东省糖科学与糖工程重点实验室,医药学院,山东青岛266003 [2]青岛海洋生物医药研究院,山东青岛266071 [3]青岛海洋科学与技术国家实验室海洋药物与生物制品功能实验室,山东青岛266237

出　　处：《中国海洋药物》2024年第2期1-10,共10页Chinese Journal of Marine Drugs

基　　金：山东省自然科学基金项目(ZR2021QH144);山东省重点研发计划项目(2021ZDSYS22)资助。

摘　　要：目的建立乳膏中不同寡糖透皮吸收的异硫氰酸荧光素(FITC)荧光定量分析方法,并考察寡糖体外透皮性能。方法采用Franz扩散池,以巴马香猪皮为载体,参照《中国药典》(2020年版四部9012通则)中生物样品定量分析方法验证指导原则,建立不同寡糖的FITC荧光定量分析方法,并测定乳膏中不同寡糖体外透皮含量。结果本文建立了乳膏中壳寡糖(COS)、褐藻寡糖(AOS)、葡寡糖(GOS)的体外透皮吸收的FITC荧光定量分析方法。结果显示,皮中及皮下FITC-寡糖的质量浓度与荧光强度呈良好的线性关系,其中FITC-COS为0.92~45.83μg/mL,FITC-GOS为0.71~42.44μg/mL,FITC-AOS为0.28~27.85μg/mL(皮中)、0.28~16.71μg/mL(皮下),且R^(2)>0.99。该方法准确度、精密度和稳定性考察结果符合指导原则要求。结论建立了乳膏中不同寡糖透皮吸收的FITC荧光定量分析方法,且发现皮中及皮下的寡糖含量均为壳寡糖>褐藻寡糖>葡寡糖,其中壳寡糖的皮中滞留量为(35.81±2.09)μg/cm^(2),皮下透过量为(118.72±6.91)μg/cm^(2)。Objective The quantitative analysis method of fluorescein isothiocyanate(FITC)of different oligosaccharides in cream was established,and the transdermal ability of oligosaccharides was investigated in vitro.Methods Using Franz diffusion cell,the FITC fluorescence quantitative analysis method of different oligosaccharides was established by using Bama pig skin as the carrier and referring to the verification guidelines for quantitative analysis methods of biological samples in Chinese Pharmacopoeia(2020 edition,volumeⅣ,9012 general rules),and the transdermal content of different oligosaccharides in cream was determined.Results A fluorescence quantitative analysis method for the transdermal absorption of chitosan oligosaccharide(COS),alginate oligosaccharide(AOS)and glucan oligosaccharide(GOS)in cream was established.The results showed that the mass concentration of FITC-fluorescence in and under the skin had a good linear relationship with fluorescence intensity,of which FITC-COS was 0.92-45.83μg/mL,FITC-GOS was 0.71-42.44μg/mL,and FITC-AOS was 0.28-27.85μg/mL(in the skin)and 0.28-16.71μg/mL(under the skin),and R^(2)>0.99.The accuracy,precision and stability of the method were in line with the requirements of the guiding principles.Conclusion The quantitative analysis method of FITC-fluorescence of different oligosaccharides in cream was established,and the results showed that the retention amount in the skin and the subcutaneous penetration amount was chitosan oligosaccharide>alginate oligosaccharide>glucan oligosaccharide.The retention amount of chitosan oligosaccharide in the skin was(35.81±2.09)μg/cm^(2),and the subcutaneous penetration amount was(118.72±6.91)μg/cm^(2)。

关 键 词：FITC-寡糖 体外透皮吸收 定量分析 方法学验证 

分 类 号：R917[医药卫生—药物分析学]