张嵩,王维宁,陈芳芳,张琳,袁丹
ZHANG Song1,WANG Wei-ning2,CHEN Fang-fang1,ZHANG Lin1,YUAN Dan1(1.School of Traditional Chinese Materia Medica

• 1:沈阳药科大学中药学院
• 2:辽宁省食品药品检验所

摘要(Abstract)：

目的建立哈蟆油及其类似品黑龙江林蛙油、桓仁林蛙油与伪品蟾蜍油、牛蛙油和青蛙油的定性和定量方法。方法系统地研究不同产地哈蟆油及其类似品与伪品的膨胀度、薄层色谱法等定性鉴别方法,并采用高效液相色谱法、凯氏定氮法、氯胺T比色法和茚三酮柱后衍生化-氨基酸分析法等,分别测定哈蟆油及其类似品与伪品中1-甲基海因、激素、总蛋白质、胶原蛋白和氨基酸的含量。结果哈蟆油、黑龙江林蛙油及桓仁林蛙油膨胀度均大于75.0,青蛙油膨胀度范围为52.5～95.4,而蟾蜍油和牛蛙油膨胀度均低于40.0。蟾蜍油和牛蛙油薄层色谱图中有2个特征性斑点,与哈蟆油、黑龙江林蛙油、桓仁林蛙油及青蛙油的色谱图有明显区别。哈蟆油及其类似品与伪品中均检测出1-甲基海因,其含量质量分数为1.51×10-4%～3.72×10-2%。哈蟆油及其类似品与伪品中雌二醇、雌酮、睾酮和孕酮等4种激素含量为微量。哈蟆油及其类似品总蛋白质含量、氨基酸含量明显高于蟾蜍油、牛蛙油和青蛙油等伪品,但胶原蛋白含量明显低于3种伪品。结论系统地研究了不同产地哈蟆油及其类似品与伪品的定性和定量分析方法,为哈蟆油质量综合评价和控制提供了详实的实验数据,对今后药典哈蟆油质量标准的修订具有重要的参考价值。
Objective To establish identification and quantitation methods for Ranae Oviductus,its analogous and adulterant materials.Methods Several identification methods for the oviducts of Rana sp.collected from different habitats were investigated,inclusive of swelling capacity test and TLC test.Also some quantitative determinations were performed,inclusive of determination of 1-methylhydantoin and hormones using HPLC method,total protein using Kjeldahl method,collagen using chloramine T colorimetric method,and eighteen kinds of amino acids using ninhydrine post-column derivatization-amino acid analysis method or colorimetric method,respectively.Results The swelling capacities of the oviducts of R.temporaria chensinensis,R.amureansis and R.huanrenensis were more than 75.0,while those of the oviduct of R.nigromaculata were 52.5-95.4,and those of the oviducts of B.bufo gargarizans and R.catesbeiana were lower than 40.0.Two characteristic spots with white blue fluorescence displayed in the TLC chromatograms of the oviducts of B.bufo gargarizans and R.catesbeiana,which may be used as markers to distinguish both adulterants from the oviducts of R.temporaria chensinensis,R.amureansis,R.huanrenensis and R.nigromaculata.1-methylhydantoin was detected in all of the true,analogous and adulterant materials,with the contents ranging from 1.51×10-4% to 3.72×10-2%.Estradiol,estrone,testosterone and progesterone were found in trace in all the tested samples.The higher contents of total protein and amino acids and lower contents of collegen were detected in true and analogous materials than those in adulterant materials.Conclusions The identification and quantitation methods of Ranae Oviductus,its analogous and adulterant materials collected from different habitats,may provide detailed experimental data for chemically evaluating and quality control of Ranae Oviductus,and they are valuable in revising the qualitative standard of Ranae Oviductus in Chinese Pharmacopoeia in future.

关键词(KeyWords)： 哈蟆油;伪品;定性鉴别;1-甲基海因;蛋白质类;含量测定
Ranae Oviductus;adulterants;identification;1-methylhydantoin;protein;quantitation

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 张嵩,王维宁,陈芳芳,张琳,袁丹
ZHANG Song1,WANG Wei-ning2,CHEN Fang-fang1,ZHANG Lin1,YUAN Dan1(1.School of Traditional Chinese Materia Medica

DOI: 10.14066/j.cnki.cn21-1349/r.2012.12.008

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