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时间:2020-06-06
《脱毒蓖麻籽粕中蓖麻毒素的提取及测定.pdf》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、国际药学研究杂志2013年4月第4O卷第2期JIntPharmRes,V01.40,No.2,April,2013·213·脱毒蓖麻籽粕中蓖麻毒素的提取及测定李倩,董娜。,武军华,王玉霞,贾培媛,张任鹏,睢大贳[摘要]目的优化双夹心ELISA法检测蓖麻籽粕中蓖麻毒素含量检测方法,并对方法学进行必要的验证。方法以蓖麻籽粕中蓖麻毒素的含量为指标,对提取溶剂、蓖麻籽粕与溶剂体积比、提取时间、提取过程是否匀浆进行单因素实验,选取最佳提取条件。在最佳提取条件下对方法的精密度和准确度进行验证。结果优化出双夹心ELISA法检测蓖麻籽粕中蓖麻毒素
2、的最佳提取溶剂为含0.1%Tween20的磷酸盐缓冲液(PBST),提取溶剂体积为1:lO(质量/体积),匀浆后提取浸泡时间为1h。该方法特异性高,蓖麻籽粕中杂质成分对测定无干扰。蓖麻毒素在3.91—250晷/L浓度范围内线性关系良好,13内和日间精密度测定的相对标准偏差(RSD)值分别小于3.2%和11.8%。同一样品重复测定日间RSD值小于5.5%。结论蓖麻籽粕中的残余蓖麻毒素可采用优化后的实验方法进行有效提取,应用双夹心ELISA方法进行毒素含量的定量分析,该方法为蓖麻籽粕脱毒工艺的确定及脱毒样品的质量控制提供了技术支持。[
3、关键词]蓖麻籽粕;蓖麻毒素;酶联免疫吸附分析[中图分类号]R965.2[文献标识码]A[文章编号]1674-0440(2013)02-0213-06ExtractionanddeterminationofricinindetoxifiedmealsofcastorbeansLIQian,DUNGNa,WUJun.hua,WANGYu—xia,JIAPei.yuan,ZHANGRen.pengr.SUIDa.yun(1.DepartmentofMilitaryToxicologyandBiochemicalPharmacology,
4、InstituteofPharmacologyandToxicology,AcademyofMilitaryMedicalSciences,Beng100850,China;2.SchoolofPharmaceuticalScience,JilinUniversity,Changchun130021,China,3.ChinesePEAGeneralHospital,Bering100853,China)[Abstract]ObjeetiveTooptimizeextractionconditionsofricinfromcast
5、orbeanmealsviaasandwichenzyme—linkedimmu—nosorbentassay(sandwichELISA).MethodsSandwichenzyme—linkedimmunosorbentassay(ELISA)wasusedtodetectricininthecastorbeanmeals.Thesignalfactortestwasdesignedtooptimizethemethodofricinextraction,includingtheextractionsolvents,thera
6、tiobetweenbeanmealandliquidvolume,treatmenttimeandwithorwithouthomogenization.ResultsItwasshownthatphos·phatebufercontaining0.1%Tween20(PBST)wasthebestbuferforricinextraction.TheoptimumextractionratioofbeanmealVSliquidwas1:10(weight/volume),andtheidealtimewas1hforrici
7、nreleasing.Theoptimizedmethodhadahighspecificityandgoodprecisionwithoutthedisturbancefromimpuritiesinthemeals.Goodlinearcurverelationshipwaspresentedbetweentheconcentrationofricinfrom3.91to250g/L.Therelativestandarddeviation(RSD)within—daywas3.2%.Whenthesamesamplewasd
8、etectedrepetitivelytheRSDinter—dayswas5.5%.ConclusionThericinincastorbeanmealcouldbewellextractedbytheoptimizedmethodestabli
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