不同来源、不同等级猪苓饮片中总多糖的含量测定
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摘要
目的:建立猪苓饮片总多糖的含量测定方法,分析不同来源、不同等级猪苓饮片中的总多糖含量,为制定猪苓饮片含量限度及饮片商品等级划分标准提供参考依据。方法:采用高效凝胶色谱法-示差折光-多角度激光检测器联用技术测定猪苓多糖的相对分子质量及其分布情况,选择与总多糖相对分子质量相近的葡聚糖为对照品,采用正交试验和单因素试验优化猪苓总多糖含量测定的前处理条件,采用蒽酮-硫酸显色法,在630 nm处测定13批不同产地猪苓饮片和39批不同等级猪苓饮片中总多糖的含量。结果:含量测定方法学考察的线性关系、稳定性、精密度、重复性及加样回收率结果均符合要求。不同产地猪苓饮片中总多糖的质量分数在0. 87%～1. 39%。不同等级猪苓饮片中总多糖质量分数为一等饮片1. 40%,二等饮片1. 21%,三等饮片1. 03%。结论:建立的含量测定方法灵敏度高、重复性好,可用于猪苓饮片的总多糖含量测定。不同来源的猪苓饮片总多糖含量有一定差异。不同等级猪苓饮片总多糖含量呈现一定的规律性,一等饮片中总多糖含量最高,二等饮片总多糖含量次之,三等饮片总多糖含量最低,对于制定猪苓饮片含量限度及饮片商品等级划分标准具有重要参考价值。
Objective: To establish a method for determining the content of total polysaccharides in decoction pieces of Polyporus,analyze the content of total polysaccharides in samples with different sources and grades. Method: The relative molecular weight and the polydispersity index of polysaccharides in decoction pieces of Polyporus were measured by a high performance gel chromatography coupled with a multi-angle laser light scattering and refractive index system. Dextran with similar molecular weight as polysaccharides was selected as the reference substance. Orthogonal experiment and single factor tests were used to optimize the pretreatment conditions for the determination of total polysaccharides in Polyporus. Polysaccharides in Polyporus with different areas and grades were determined by anthrone-sulfuric acid colorimetric method at 630 nm. Result: The linearity,stability,precision, repeatability and recovery rate of the established method all reached the standards,respectively. The content of total polysaccharides in samples from different areas ranged from 0. 87% to 1. 39%.The content of total polysaccharides in samples with different grades was 1. 40% for first-grade pieces,1. 21% for second-grade pieces,and 1. 03% for third-grade pieces. Conclusion: The established method is simple,accurate and reproducible,and it can be used for the determination of polysaccharides in decoction pieces of Polyporus.The content of polysaccharides in samples from different origins varies greatly. The content of polysaccharides in samples with different grades shows a certain regularity. The content of polysaccharides is the highest in the firstgrade pieces,followed by the content in the second-grade,and the lowest in the third-grade. The results can provide a reference for formulating limits for the content of total polysaccharides and the grade standard of decoction pieces of Polyporus.
Objective: To establish a method for determining the content of total polysaccharides in decoction pieces of Polyporus,analyze the content of total polysaccharides in samples with different sources and grades. Method: The relative molecular weight and the polydispersity index of polysaccharides in decoction pieces of Polyporus were measured by a high performance gel chromatography coupled with a multi-angle laser light scattering and refractive index system. Dextran with similar molecular weight as polysaccharides was selected as the reference substance. Orthogonal experiment and single factor tests were used to optimize the pretreatment conditions for the determination of total polysaccharides in Polyporus. Polysaccharides in Polyporus with different areas and grades were determined by anthrone-sulfuric acid colorimetric method at 630 nm. Result: The linearity,stability,precision, repeatability and recovery rate of the established method all reached the standards,respectively. The content of total polysaccharides in samples from different areas ranged from 0. 87% to 1. 39%.The content of total polysaccharides in samples with different grades was 1. 40% for first-grade pieces,1. 21% for second-grade pieces,and 1. 03% for third-grade pieces. Conclusion: The established method is simple,accurate and reproducible,and it can be used for the determination of polysaccharides in decoction pieces of Polyporus.The content of polysaccharides in samples from different origins varies greatly. The content of polysaccharides in samples with different grades shows a certain regularity. The content of polysaccharides is the highest in the firstgrade pieces,followed by the content in the second-grade,and the lowest in the third-grade. The results can provide a reference for formulating limits for the content of total polysaccharides and the grade standard of decoction pieces of Polyporus.
引文
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[13]苏靖.猪苓药用多糖的含量测定[J].内蒙古中医药,2012,31(16):43.
[14]匡扶,朱照静,马俐丽,等.葡聚糖T20作为白芨多糖含量测定标准品可行性研究[J].重庆医科大学学报,2008,33(5):570-574.
[15]杨莉,王志华,陶健生,等.黄芪中黄芪多糖含量测定方法的比较[J].中国医药工业杂志,2005,36(9):562-563.
[16]罗爱勤,陈亮,李菁.灵芝孢子多糖含量不同方法测定的比较[J].中国中医药现代远程教育,2016,14(19):132-134.
[17]李志洲.回归正交试验设计优化猪苓多糖的提取工艺[J].光谱实验室,2012,29(5):3047-3050.
[18]苏德龙,史红波,裴福成,等.正交试验法研究猪苓多糖提取工艺[J].现代中药研究与实践,2002,16(1):27-28.
[19]冯航,王虹.猪苓多糖的提取及含量测定[J].陕西农业科学,2014,60(8):8-10.
[20]鲁文静,周密,梁宗锁.猪苓药材质量影响因素及质量评价的研究进展[J].中国实验方剂学杂志,2013,19(17):366-370.
[2]聂红,马安伦,沈佰华,等.复方猪苓多糖对小鼠免疫功能的调节[J].细胞与分子免疫学杂志,2000,16(5):384-386.
[3]江泽波,李思明,赵晋,等.猪苓多糖对LPS诱导的J774炎症模型的抗炎作用及其机制[J].中国实验方剂学杂志,2015,21(3):156-159.
[4]邵玉英,刘培民.猪苓汤含药血清对体外培养k562及PG细胞nm23基因表达的影响[J].世界中西医结合杂志,2009,4(9):627-629.
[5] BI Y P,MIAO Y,HAN Y,et al. Biological and physicochemical properties of two polysaccharides from the mycelia of Grifola umbellate[J]. Carbohydr Polym,2013,95(2):740-745.
[6]王林丽,吴寒寅,罗桂芳.猪苓的药理作用及临床应用[J].中国药业,2000,9(10):58-59.
[7]李佩文,刘韶辉,罗劲华,等.口服猪苓多糖联合或序贯拉米夫定治疗慢性乙型肝炎的疗效[J].广东医学,2005,26(9):1278-1280.
[8]吕宝璋,吴忠忱,单京瑞,等.柴胡和猪苓多糖的生化作用及抗辐射损伤原理的研究[J].解放军医学杂志,1984,9(1):9-12.
[9]张辉,王统康,陈红.猪苓多糖抗诱变性的研究[J].中成药,1993,15(3):29-29.
[10]张国伟,李彩霞,王艳峰,等.高效液相法与硫酸-蒽酮法测定猪苓多糖含量比较[J].天然产物研究与开发,2011,23(6):1099-1102.
[11]王瑞海,叶迎,许京,等.甘肃红芪和黄芪总多糖含量测定对比[J].中国实验方剂学杂志,2017,23(22):77-83.
[12]夏琴,李敏,周进,等.不同产地、商品规格及生长年限猪苓麦角甾醇及多糖的含量分析[J].中草药,2015,38(1):45-48.
[13]苏靖.猪苓药用多糖的含量测定[J].内蒙古中医药,2012,31(16):43.
[14]匡扶,朱照静,马俐丽,等.葡聚糖T20作为白芨多糖含量测定标准品可行性研究[J].重庆医科大学学报,2008,33(5):570-574.
[15]杨莉,王志华,陶健生,等.黄芪中黄芪多糖含量测定方法的比较[J].中国医药工业杂志,2005,36(9):562-563.
[16]罗爱勤,陈亮,李菁.灵芝孢子多糖含量不同方法测定的比较[J].中国中医药现代远程教育,2016,14(19):132-134.
[17]李志洲.回归正交试验设计优化猪苓多糖的提取工艺[J].光谱实验室,2012,29(5):3047-3050.
[18]苏德龙,史红波,裴福成,等.正交试验法研究猪苓多糖提取工艺[J].现代中药研究与实践,2002,16(1):27-28.
[19]冯航,王虹.猪苓多糖的提取及含量测定[J].陕西农业科学,2014,60(8):8-10.
[20]鲁文静,周密,梁宗锁.猪苓药材质量影响因素及质量评价的研究进展[J].中国实验方剂学杂志,2013,19(17):366-370.