美花石斛的质量研究及其与近似种石斛的比较
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摘要
目的建立美花石斛生物碱、联苄类、黄酮类成分的TLC鉴别方法,以及石斛酚、甘露糖与葡萄糖含量测定方法,并与霍山石斛、金钗石斛、流苏石斛、铁皮石斛进行比较,为其质量控制提供科学依据。方法采用TLC法鉴别生物碱、联苄类、黄酮类成分;石斛酚含量测定:采用HPLC法,色谱柱为Kromasil 100-5C_(18);流动相为乙腈-水(36∶64);检测波长为210 nm;流速为0.5 m L·min~(-1);柱温为30℃。甘露糖与葡萄糖的含量测定:采用酸水解柱前衍生HPLC方法,色谱柱为Kromasil 100-5 C_(18);流动相为乙腈-0.02 mol·L~(-1)乙酸铵溶液(20∶80);检测波长为250 nm;流速为1.0 m L·min~(-1);柱温为30℃。结果 TLC鉴别中美花石斛分别检出2个生物碱主斑点、包括石斛酚在内的2个联苄类成分主斑点、包括柚皮素在内的5个荧光主斑点,与霍山石斛、金钗石斛、流苏石斛或铁皮石斛有一定区别。石斛酚、甘露糖、葡萄糖线的性关系良好(r=0.999 9);平均加样回收率分别为101.5%(RSD为1.95%)、97.9%(RSD为1.94%)、100.0%(RSD为1.36%);10批美花石斛的石斛酚含量为0.0739~0.171 mg·g~(-1);7批美花石斛的甘露糖与葡萄糖含量分别为5.07%~9.42%、8.43%~13.71%,甘露糖和葡萄糖含量之和为13.50%~22.61%。结论 TLC鉴别方法具有一定专属性,可作为美花石斛的鉴别方法之一;含量测定方法准确可靠,重复性好,可作为美花石斛的定量指标。
Objective To establish TLC methods for alkaloids,bibenzyls and flavonoid constituents from Dendrobium loddigesii;and set up HPLC methods for content determination of gigantol,D-mannose and D-glucose in Dendrobium Loddigesii. Comparision with Dendrobioum huoshanense, Dendrobium nobile, Dendrobium fimbriatum and Dendrobium officinale was conducted to provide the scientific basis for its quality control. Methods TLC methodswere developed to identify alkaloids, bibenzyls and flavonoids. HPLC was performed on Kromasil 100-5 C_(18) column, with acetonitrile-water(36∶64)as mobile phase, and a detection wavelength of 210 nm. The columntemperature was 30 ℃, and flow rate was 0.5 m L · min~(-1). The pre-column derivation HPLC method for thedetermination of D-mannose and D-glucose was established. HPLC was performed on Kromasil 100-5 C_(18) column,with acetonitrile-water(36∶64)as mobile phase,and a detection wavelength at 250 nm. The column temperaturewas 30 ℃,and flow rate was 1.0 m L·min~(-1). Results Under three TLC conditions,two main alkaloids spots,two main bibenzyls that including gigantol spots,and 5 fluorescence main spots that including naringenin,were foundfrom Dendrobium Loddigesii, which showed a certain difference with Dendrobioum huoshanense, Dendrobium nobile,Dendrobium fimbriatum and Dendrobium officinale. Linear relationships were all good with the HPLC methods(r=0.9999). The average recoveries of gigantol,D-mannose and D-glucose were 101.5%(RSD=1.95%),97.9%(RSD=1.94%),100.0%(RSD=1.36%)(n = 6),respectively. Contents in 10 batches of Dendrobium Loddigesii were0.0739-0.171 mg·g~(-1) for gigantol;the contents of D-mannose and D-glucose(Cmand Cg)in the 7 batches were5.07%-9.42%,8.43%-13.71%,respectively. Conclusion TLC identification methods have special properties,which can be used as one of the identification methods of dendrobiums. The methods for content determination wereaccurate,reliable and reproducible,which can be used as quantitative indexes of dendrobiums.
Objective To establish TLC methods for alkaloids,bibenzyls and flavonoid constituents from Dendrobium loddigesii;and set up HPLC methods for content determination of gigantol,D-mannose and D-glucose in Dendrobium Loddigesii. Comparision with Dendrobioum huoshanense, Dendrobium nobile, Dendrobium fimbriatum and Dendrobium officinale was conducted to provide the scientific basis for its quality control. Methods TLC methodswere developed to identify alkaloids, bibenzyls and flavonoids. HPLC was performed on Kromasil 100-5 C_(18) column, with acetonitrile-water(36∶64)as mobile phase, and a detection wavelength of 210 nm. The columntemperature was 30 ℃, and flow rate was 0.5 m L · min~(-1). The pre-column derivation HPLC method for thedetermination of D-mannose and D-glucose was established. HPLC was performed on Kromasil 100-5 C_(18) column,with acetonitrile-water(36∶64)as mobile phase,and a detection wavelength at 250 nm. The column temperaturewas 30 ℃,and flow rate was 1.0 m L·min~(-1). Results Under three TLC conditions,two main alkaloids spots,two main bibenzyls that including gigantol spots,and 5 fluorescence main spots that including naringenin,were foundfrom Dendrobium Loddigesii, which showed a certain difference with Dendrobioum huoshanense, Dendrobium nobile,Dendrobium fimbriatum and Dendrobium officinale. Linear relationships were all good with the HPLC methods(r=0.9999). The average recoveries of gigantol,D-mannose and D-glucose were 101.5%(RSD=1.95%),97.9%(RSD=1.94%),100.0%(RSD=1.36%)(n = 6),respectively. Contents in 10 batches of Dendrobium Loddigesii were0.0739-0.171 mg·g~(-1) for gigantol;the contents of D-mannose and D-glucose(Cmand Cg)in the 7 batches were5.07%-9.42%,8.43%-13.71%,respectively. Conclusion TLC identification methods have special properties,which can be used as one of the identification methods of dendrobiums. The methods for content determination wereaccurate,reliable and reproducible,which can be used as quantitative indexes of dendrobiums.
引文
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[14]CHEN H H,HUANG Y C,Huang D D,et al.Protective effect of gigantol against hydrogen peroxideinduced apoptosis in rat bone marrow mesenchymal stem cells through the PI3K/Akt pathway[J].Molecular Medicine Reports,2018,17(2):3267-3273.
[15]李玲,邓晓兰,赵兴兵,等.铁皮石斛化学成分及药理作用研究进展[J].肿瘤药学,2011,1(2):90-94.
[16]张红玉,戴关海,马翠,等.铁皮石斛多糖对S180肉瘤小鼠免疫功能的影响[J].浙江中医杂志,2009,45(05):380-381.
[2]冯耀南,刘明,刘俭,等.中药材商品规格质量鉴别[M].广州:暨南大学出版社,1995:193-196.
[3]陈康,劳燕霞.五种石斛药材的导数光谱及薄层鉴别[J].广东药学,1996,6(4):29-30.
[4]陈化,陈竹,梁斌,等.紫外-可见分光光度法测定环草石斛中石斛多糖的含量[J].中国药房,2011,22(39):3690-3692.
[5]白音,王文全,包英华,等.不同产地美花石斛形态及多糖含量比较研究[J].中药材,2007,30(2):130-132.
[6]黄月纯,叶家宏,戴亚峰,等.霍山石斛与美花石斛多糖柱前衍生HPLC特征图谱比较[J].中药新药与临床药理,2014,25(6):725-730.
[7]李春燕,陆羽,陈烨,等.环草石斛化学成分研究[J].中山大学学报(自然科学版),2013,52(3):73-76.
[8]周楚娟,任晋,张俊仪,等.美花石斛HPLC特征图谱研究及其与水煎液相关性分析[J].中药新药与临床药理,2017,28(5):663-667.
[9]中华人民共和国药典委员会.中国药典(一部)[M].北京:中国医药科技出版社,2015:92,282.
[10]卢文芸,张宇斌,洪化鹏,等.环草石斛野生植株与组培苗总生物碱和多糖含量的比较[J].安徽农业科学,2006,34(8):1606-1607.
[11]徐蓓,杨莉,陈崇崇,等.黄草类石斛的薄层色谱鉴别研究[J].中国药品标准,2010,11(2):99-103.
[12]孟海涛.安徽霍山产三种石斛HPLC化学指纹研究[D].合肥:合肥工业大学,2015:51-54.
[13]CHEN H H,HUANG Y C,HUANG J B,et al.Gigantol attenuates the proliferation of human liver cancer Hep G2 cells through the PI3K/Akt/NF-κB signaling pathway[J].Oncology Reports,2017,37(2):865-870.
[14]CHEN H H,HUANG Y C,Huang D D,et al.Protective effect of gigantol against hydrogen peroxideinduced apoptosis in rat bone marrow mesenchymal stem cells through the PI3K/Akt pathway[J].Molecular Medicine Reports,2018,17(2):3267-3273.
[15]李玲,邓晓兰,赵兴兵,等.铁皮石斛化学成分及药理作用研究进展[J].肿瘤药学,2011,1(2):90-94.
[16]张红玉,戴关海,马翠,等.铁皮石斛多糖对S180肉瘤小鼠免疫功能的影响[J].浙江中医杂志,2009,45(05):380-381.