发酵对胆南星中胆汁酸类成分的影响及胆南星中3种游离胆汁酸含量测定研究
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摘要
考察以猪胆汁为原料的胆南星发酵炮制品及混合蒸制炮制品中主要胆汁酸类成分的组成,建立发酵制胆南星中胆汁酸类成分的含量测定方法。以猪胆汁及天南星、制天南星为原料分别制备发酵法胆南星及混合蒸制法胆南星,采用HPLC-ELSD检测比较2种胆南星中胆汁酸类成分的差异;以3种游离胆汁酸类成分为指标,采用HPLC-ELSD检测发酵制胆南星炮制品中的胆汁酸类成分的含量。采用Agilent Eclipse XDB C_(18)(4.6 mm×250 mm,5μm)色谱柱,以乙腈-0.1%冰醋酸水(55∶45)为流动相,流速为1 mL·min~(-1),柱温为30℃,ELSD:漂移管温度为90℃,氮气流量为2.2 mL·min~(-1)。结果显示混合蒸制法胆南星中主要含有结合型胆汁酸类成分,发酵制胆南星中未见有结合型胆汁酸类成分,主要含游离型胆汁酸类成分。发酵制品中的3种游离型胆汁酸类成分猪胆酸(HCA)、猪去氧胆酸(HDCA)和鹅去氧胆酸(CDCA)均在定量范围内呈良好的线性关系,平均加样回收率在95.99%~104.3%,加样回收符合要求。研究结果表明,胆南星发酵过程可使得胆汁中的结合型胆汁酸类成分转化为游离型胆汁酸。所建方法可以有效控制发酵制胆南星中胆汁酸类成分的含量。
The aim of this study is to analyze the compositions of main bile acids in fermented and mixed processing products of arisame cum bile from pig bile,and to establish a method for content determination of bile acids in fermented Arisaema Cum Bile.Fermented and mixed processing products were prepared from arisaematis rhizome and arisaematis rhizoma preparatum with pig bile respectively.Then the differences in bile acids compositions between such two kinds of products were compared by high performance liquid chromatography and evaporative light-scattering detector(HPLC-ELSD).With three kinds of free bile acid compositions as the indicators,HPLC-ELSD method was adopted to determine the content of bile acid compositions in fermented product,on Agilent Eclipse XDB C18(4.6 mm×250 mm,5μm)chromatographic column,with acetonitrile and 0.1%glacial acetic acid solution(55∶45)as mobile phase,at a flow rate of 1 mL·min~(-1),column temperature of 30℃,drift tube temperature of 90℃,and a nitrogen flow rate of 2.2 mL·min~(-1).The results showed that the bile acids in fermented bile Arisaema were mainly in a free form,while in mixed processing product,the compositions were mainly in a conjugated form.Three kinds of free bile acids,namely porcine cholic acid(HCA),porcine deoxycholic acid(HDCA)and chenodeoxycholic acid(CDCA)in fermented product,showed a good linear relationship in the range of quantification.The average recovery rate was 95.99%-104.3%,complying with the requirements.The results showed that the conjugated bile acids could be transformed into free bile acids during the fermentation of arisaema cum bile.This established method can effectively control the content of bile acids compositions in fermenting arisaema cum bile.
The aim of this study is to analyze the compositions of main bile acids in fermented and mixed processing products of arisame cum bile from pig bile,and to establish a method for content determination of bile acids in fermented Arisaema Cum Bile.Fermented and mixed processing products were prepared from arisaematis rhizome and arisaematis rhizoma preparatum with pig bile respectively.Then the differences in bile acids compositions between such two kinds of products were compared by high performance liquid chromatography and evaporative light-scattering detector(HPLC-ELSD).With three kinds of free bile acid compositions as the indicators,HPLC-ELSD method was adopted to determine the content of bile acid compositions in fermented product,on Agilent Eclipse XDB C18(4.6 mm×250 mm,5μm)chromatographic column,with acetonitrile and 0.1%glacial acetic acid solution(55∶45)as mobile phase,at a flow rate of 1 mL·min~(-1),column temperature of 30℃,drift tube temperature of 90℃,and a nitrogen flow rate of 2.2 mL·min~(-1).The results showed that the bile acids in fermented bile Arisaema were mainly in a free form,while in mixed processing product,the compositions were mainly in a conjugated form.Three kinds of free bile acids,namely porcine cholic acid(HCA),porcine deoxycholic acid(HDCA)and chenodeoxycholic acid(CDCA)in fermented product,showed a good linear relationship in the range of quantification.The average recovery rate was 95.99%-104.3%,complying with the requirements.The results showed that the conjugated bile acids could be transformed into free bile acids during the fermentation of arisaema cum bile.This established method can effectively control the content of bile acids compositions in fermenting arisaema cum bile.
引文
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[3]太平惠民和剂局.太平惠民和剂局方[M].北京:人民卫生出版社,1985:18.
[4]朱震亨.丹溪心法[M].上海:上海科学技术出版社,1959:356.
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[7]李瑶,郁红礼,王卫,等.猪牛羊胆汁HPLC指纹图谱研究[J].中国中药杂志,2018,43(11):56.
[8]游元元,万德光.动物胆汁药用研究进展[J].贵阳中医学院学报,2007,29(4):58.
[9]石岩,耿颖,郑天骄,等.猪胆粉HPLC-ELSD全轮廓谱图主要胆汁酸测定及化学计量学分析[J].药物分析杂志,2017,37(2):283.
[10]鄢海燕,邹纯才.四种动物胆粉中胆酸类成分分析及药理作用比较[J].皖南医学院学报,2012,31(4):274.
[11]陈江宁,单国顺,赵启苗,等.不同胆汁制胆南星中胆酸类成分及其解热作用比较[J].现代药物与临床,2017,32(4):567.
[12]张飞.猪去氧胆酸提取与熊去氧胆酸及其衍生物的合成[D].武汉:武汉轻工大学,2014.
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[19]胡霞敏,石朝周.牛磺结合及其游离胆汁酸在小鼠镇咳祛痰抗炎模型上的作用比较[J].中国临床药学杂志,2001(2):85.
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