曲茎石斛及其近缘种的鉴别与质量评价  

Identification and Quality Evaluation of Dendrobium flexicaule and Its Related Species

作　　者：孙廷 杨玉珍 胡述晓 路遥[3] 张村[4] 胡如善 SUN Ting;YANG Yuzhen;HU Shuxiao;LU Yao;ZHANG Cun;HU Rushan(Nanyang Vocational College of Agriculture,Nanyang 473000,China;Nanyang Landscaping Administration,Nanyang 473002,China;Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing100700,China)

机构地区：[1]南阳农业职业学院,河南南阳473000 [2]南阳市园林绿化管理局,河南南阳473002 [3]北京中医药大学东方医院,北京100078 [4]中国中医科学院中药研究所,北京100700

出　　处：《中国实验方剂学杂志》2022年第20期128-134,共7页Chinese Journal of Experimental Traditional Medical Formulae

摘　　要：目的:对曲茎石斛及其近缘种的药材进行鉴别,并分析其多糖成分、D-甘露糖含量的差异,为石斛药材的准确鉴定及其质量控制提供理论依据。方法:采用DNA条形码和红外光谱技术对9份石斛药材(S1~S9)进行鉴定,并分别采用紫外分光光度法(UV)和高效液相色谱法(HPLC)对石斛多糖和D-甘露糖的含量进行测定,UV检测条件为488 nm;HPLC检测条件为流动相20 mmol·L^(-1)乙酸铵溶液-乙腈(81.5∶18.5),检测波长250 nm。结果:DNA条形码结果显示,样品S1~S3鉴定为金钗石斛;样品S4~S5鉴定为铁皮石斛;样品S6鉴定为霍山石斛;样品S7~S9鉴定为曲茎石斛。一维红外光谱显示,仅3份金钗石斛在波数1570~1467 cm^(-1)处有稳定的特征峰,呈现“W”形,而在842~740 cm^(-1)处无吸收峰;其余石斛则在842~740 cm^(-1)处有稳定吸收峰。一阶导数光谱上在785 cm^(-1)处,霍山石斛呈现“V”形,而其余石斛呈现“W”形;在1110 cm^(-1)处曲茎石斛具有稳定的特征峰。二阶导数光谱中在1125 cm^(-1)处,铁皮石斛呈现“M”形,其余石斛近似“W”形。含量测定结果显示,样品S1~S9的多糖质量分数依次为9.35%、9.12%、32.78%、49.38%、48.97%、32.48%、32.95%、39.41%和25.32%,D-甘露糖质量分数依次为1.39%、0.47%、13.57%、3.04%、33.85%、23.57%、16.64%、17.47%和19.49%。其中曲茎石斛具有着较高的多糖和D-甘露糖含量。结论:DNA条形码和一阶、二阶导数红外光谱均可实现曲茎石斛及其近缘种的鉴别,且红外光谱鉴别方法价格低、操作简便;同时,曲茎石斛具有较高含量的多糖和D-甘露糖,可为快速鉴别曲茎石斛及其近缘种药材提供科学依据,并为曲茎石斛的质量控制和资源利用提供参考。Objective:To identify Dendrobium flexicaule and its related species,and analyze the differences in polysaccharide composition and D-mannose content,so as to provide theoretical basis for the accurate identification and quality control of Dendrobium medicinal materials.Method:Nine samples of Dendrobium(S1-S9)were identified by DNA barcoding and infrared spectroscopy,and the contents of polysaccharides and D-mannose were determined by ultraviolet spectrophotometry(UV)and high performance liquid chromatography(HPLC),respectively.UV detection condition was 488 nm,HPLC detection conditions were the mobile phase of 20 mmol·L^(-1)ammonium acetate solution-acetonitrile(81.5∶18.5)and the detection wavelength at 250 nm.Result:DNA barcoding results showed that samples S1-S3 were D.nobile,samples S4-S5 were D.officinale,sample S6 was D.huoshanense,and S7-S9 were D.flexicaule.One-dimensional infrared spectroscopy showed that only D.nobile had stable characteristics at the wavenumber of 1570-1467 cm^(-1),showing a"W"shape,while no absorption peak was found at the wavenumber of 842-740 cm^(-1),but the other Dendrobium samples had stable absorption peaks at the wavenumber of 842-740 cm^(-1).In the first derivative spectrum,at the wavenumber of 785 cm^(-1),D.huoshanense presented a"V"shape,while the rest of Dendrobium presented a"W"shape.At the wavenumber of 1110 cm^(-1),D.flexicaule had a stable characteristic peak.In the second derivative spectrum,at the wavenumber of 1125 cm^(-1),D.officinale presented an"M"shape,and the rest of Dendrobium was approximately"W"shape.The results of determination showed that the contents of polysaccharides in samples S1-S9 were 9.35%,9.12%,32.78%,49.38%,48.97%,32.48%,32.95%,39.41%and 25.32%,and their contents of D-mannose were 1.39%,0.47%,13.57%,3.04%,33.85%,23.57%,16.64%,17.47%and 19.49%,respectively.Among them,D.flexicaule had high polysaccharide and D-mannose contents.Conclusion:Both DNA barcoding and infrared spectroscopy can be used to identify D.flexicaule and its related species,and inf

关 键 词：曲茎石斛 铁皮石斛 DNA条形码 多糖 红外光谱 质量控制 高效液相色谱法(HPLC) 

分 类 号：R22[医药卫生—中医基础理论] R931[医药卫生—中医学] R28[机械工程—光学工程] O433[理学—光学]