基于体内外关联的六神曲酶活力效价检测方法研究  

Detection Method Study of Enzyme Activity Potency of Massa Medicata Fermentata Based on in Vitro and in Vivo Correlation

作　　者：杨婷[1] 李美延 李佳欣 鲍洋 谭鹏 阳向波 纪奇森 张定堃[1] 许润春[1] YANG Ting;LI Meiyan;LI Jiaxin;BAO Yang;TAN Peng;YANG Xiangbo;JI Qiseng;ZHANG Dingkun;XU Runchun(State Key Laboratory of Southwestern Chinese Medicine Resource,School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137;State Key Laboratory of Quality Evaluation of Traditional Chinese Medicine,National Administration of Traditional Chinese Medicine,Sichuan Academy of Traditional Chinese Medicine,Chengdu6610041;Yaan Xunkang Pharmaceutical Co.,Ltd.,Yaan 625600)

机构地区：[1]成都中医药大学药学院,西南特色中药资源国家重点实验室,成都611137 [2]四川省中医药科学院,国家中医药管理局中药质量生物评价重点研究室,成都610041 [3]雅安迅康药业有限公司,雅安625600

出　　处：《中药药理与临床》2023年第7期20-27,共8页Pharmacology and Clinics of Chinese Materia Medica

基　　金：四川省中医药管理局科学技术研究专项(编号:2021MS530);四川省重点研发计划项目(编号:2022YFS0429)。

摘　　要：目的:建立基于酶活力检测的六神曲生物检定方法,完善六神曲质量控制体系。方法:选择Folin法、pNPP法分别作为消食药六神曲胃蛋白酶和脂肪酶酶活力生物效价测定方法,以消化酶片为阳性药,药物剂量间距为1∶0.6,根据简化概率单位法计算35个批次六神曲的生物效价和可信限度,对精密度、重复性、中间精密度和可靠性进行考察,优化并确定检测条件;采用营养性半固体糊粘度测定方法验证不同酶活力六神曲降低粘度的活力差异;对比研究小鼠胃肠推进与酶活力的相关性差异,评价酶活力效价与体内活力的相关性。结果:建立了基于酶活力检测的六神曲生物检定方法,方法学考察均符合要求,测定了35个批次六神曲样品胃蛋白酶活力效价范围为151.549 U/mg~337.840 U/mg,平均效价为230.622 U/mg,RSD为22.61%;脂肪酶的活力效价范围为31.944 U/mg~616.165 U/mg,平均效价为278.936 U/mg,RSD值为63.67%。营养糊粘度随着六神曲酶活力效价的升高而降低;与正常对照组比较,除S7组外,六神曲各给药组小鼠胃残留率降低,小肠推进率显著提高(P<0.01);小鼠胃残留率、小肠推进率与胃蛋白酶、脂肪酶活力效价的相关性系数分别为0.792、0.835和0.773、0.418,表明消化酶活力效价与体内消食活力高度相关。结论:本文建立的基于消化酶活力的六神曲生物效价测定方法可靠,重复性良好,可为六神曲的质量控制提供科学依据。Objective:To establish a bioassay method for Massa Medicata Fermentata(MMF)based on enzyme activity detection and improve the quality control system of MMF.Methods:The Folin and pNPP methods were used to determine the biological potency of the enzyme activity of MMF pepsase and lipases as the digestant drug.The digestive enzyme tablets were used as the positive drugs,and the interval between the drugs was 1:0.6.The biological potency and confidence range of 35 batches of MMF were calculated by the simplified probit method,and then the precision,repeatability,intermediate precision,and reliability were investigated.The detection conditions were optimized and determined.The nutrient semi-solid paste viscosity method was used to detect the activity difference of MMF samples with different enzyme activity in reducing viscosity.The correlation difference of the gastrointestinal propulsion of mice and enzyme activity was compared to evaluate the correlation between enzyme activity potency and in vivo activity.Results:The bioassay method for MMF based on enzyme activity detection was established,and the methodological investigations met all the requirements.The activity potency of 35 batches of MMF pepsin samples ranged from 151.549 U/mg to 337.840 U/mg.The average activity potency was 230.622 U/mg,and the RSD was 22.61%.The activity potency of lipase samples ranged from 31.944 U/mg to 616.165 U/mg.The average activity potency was 278.936 U/mg,and the RSD was 63.67%.The viscosity of the nutrient paste decreased with the increase in the activity potency of the MMF enzyme.Compared with that in the normal control group,except for the S7 group,the gastric residual rate of mice was reduced in MMF-administered groups,and the intestinal propulsion rate was increased significantly(P0.01).The correlation coefficients of the gastric residual rate and intestinal propulsion rate of mice with pepsin and lipase activity potency were 0.792,0.835,0.773,and 0.418,respectively,indicating that the activity potency of digestive enzymes was

关 键 词：六神曲 消食类中药 消化酶 生物效价 质量评价 

分 类 号：R285[医药卫生—中药学]