丹霞种、云南广南种铁皮石斛中主要黄酮苷的分离鉴定
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摘要
目的:对2个道地种源(丹霞种、云南广南种)铁皮石斛中的黄酮苷成分进行提取分离、纯化及结构鉴定,为确认铁皮石斛的共性黄酮苷成分提供实验依据。方法:(1)采用70%乙醇提取两个产地铁皮石斛叶的总黄酮成分,依次采用有机溶剂石油醚、乙酸乙酯、水饱和正丁醇萃取粗提物,运用大孔吸附树脂AB-8,LH-20型羟丙基葡聚糖凝胶(Sephadex LH-20)以及ODS柱色谱对水饱和正丁醇萃取部位进行分离纯化,通过化合物的理化性质结合HPLC,ESI-MSn,1H-NMR,13C-NMR等现代波谱学方法鉴定黄酮苷结构。(2)利用HPLC分析比较不同道地种源(丹霞种、云南广南种、广西铁皮兰种、浙江本地种)铁皮石斛的黄酮苷共有成分。结果:从2个产地铁皮石斛叶中共分离得到了5个黄酮苷化合物,经鉴定为芦丁,新西兰牡荆苷Ⅱ,新西兰牡荆苷Ⅰ,佛莱心苷,异佛莱心苷。4个不同道地种源(丹霞种、云南广南种、广西铁皮兰种、浙江本地种)铁皮石斛茎中均能检出新西兰牡荆苷Ⅱ,新西兰牡荆苷Ⅰ特征峰,其中新西兰牡荆苷Ⅱ丰度相对较大,而佛莱心苷、异佛莱心苷在云南广南种、广西铁皮兰种中出峰明显,芦丁则在丹霞种中尤为突出。结论:新西兰牡荆苷Ⅱ为铁皮石斛不同道地种源(丹霞种、云南广南种、广西铁皮兰种、浙江本地种)的共性黄酮苷成分,可作为铁皮石斛特征图谱的参照物。
Objective: To extract,isolate,purify and identify the structures of the flavonoid glycoside in Dendrobium officinale from two different origin places(Danxia species and Yunnan Guangnan species), and provide experimental reference for confirming the common flavonoid glycoside components in D. officinale.Method:(1)70% ethanol was applied to extract the total flavonoids in leaves of D. officinale from two different species. Organic solvents petroleum ether,acetic ether and water saturated n-butyl alcohol were used in turn to extract the crude extraction. Then AB-8 Macroporous resin,Sephadex LH-20 and ODS chromatographic column were applied to isolate and purify the water saturated n-butyl alcohol extraction fraction. The structures of flavonoid glycoside were identified by studying physicochemical property,applying modern spectroscopy method like HPLC,ESI-MSn,1 H-NMR,13 C-NMR,etc.(2)HPLC characteristic spectrum technique was used to analyse and compare the common flavonoid glycoside components in Dendrobium officinale from different origin places(Danxia species,Yunnan Guangnan species, Guangxi Tiepilan species and Zhejiang native species). Result: Five flavonoid glycoside compounds were isolated from the crude extractions of the leaves of D. officinale from two different species,and they were identified as rutin, vicenin Ⅱ, vicenin Ⅰ, violanthin and isoviolanthin. The characteristic spectrum of vicenin Ⅱ and viceninⅠwere detected in stems of D. officinale from four different origin places(Danxia species,Yunnan Guangnan species,Guangxi Tiepilan species and Zhejiang native species),and vicenin Ⅱ had a better separation degree in the characteristic spectrum. However,the characteristic spectrum of violanthin and isoviolanthin were more obvious in Yunnan Guangnan species and Guangxi Tiepilan species,while rutin was obvious in the Danxia species. Conclusion: Vicenin Ⅱis the common flavonoid glycosides component in D. officinale from different origin places(Danxia species,Yunnan Guangnan species,Guangxi Tiepilan species and Zhejiang native species),and can be used as the internal reference material for the characteristic spectrum of D. officinale.
Objective: To extract,isolate,purify and identify the structures of the flavonoid glycoside in Dendrobium officinale from two different origin places(Danxia species and Yunnan Guangnan species), and provide experimental reference for confirming the common flavonoid glycoside components in D. officinale.Method:(1)70% ethanol was applied to extract the total flavonoids in leaves of D. officinale from two different species. Organic solvents petroleum ether,acetic ether and water saturated n-butyl alcohol were used in turn to extract the crude extraction. Then AB-8 Macroporous resin,Sephadex LH-20 and ODS chromatographic column were applied to isolate and purify the water saturated n-butyl alcohol extraction fraction. The structures of flavonoid glycoside were identified by studying physicochemical property,applying modern spectroscopy method like HPLC,ESI-MSn,1 H-NMR,13 C-NMR,etc.(2)HPLC characteristic spectrum technique was used to analyse and compare the common flavonoid glycoside components in Dendrobium officinale from different origin places(Danxia species,Yunnan Guangnan species, Guangxi Tiepilan species and Zhejiang native species). Result: Five flavonoid glycoside compounds were isolated from the crude extractions of the leaves of D. officinale from two different species,and they were identified as rutin, vicenin Ⅱ, vicenin Ⅰ, violanthin and isoviolanthin. The characteristic spectrum of vicenin Ⅱ and viceninⅠwere detected in stems of D. officinale from four different origin places(Danxia species,Yunnan Guangnan species,Guangxi Tiepilan species and Zhejiang native species),and vicenin Ⅱ had a better separation degree in the characteristic spectrum. However,the characteristic spectrum of violanthin and isoviolanthin were more obvious in Yunnan Guangnan species and Guangxi Tiepilan species,while rutin was obvious in the Danxia species. Conclusion: Vicenin Ⅱis the common flavonoid glycosides component in D. officinale from different origin places(Danxia species,Yunnan Guangnan species,Guangxi Tiepilan species and Zhejiang native species),and can be used as the internal reference material for the characteristic spectrum of D. officinale.
引文
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[13] ZHANG Y,LUO J,HAN C et al. Bioassay-guided preparative separation of angiotensin-converting enzyme inhibitory C-flavone glycosides from Desmodium styracifolium by recycling complexation high-speed counter-current chromatography[J]. J Pharm Biomed Anal,2015,102:276-281.
[14]刘晖晖,于永明,张涛,等.半边铁角蕨化学成分研究[J].中国药学杂志,2017,52(2):100-104.
[15] Flamini G. Flavonoids and other compounds from the aerial parts of Viola etrusca[J]. Chem Biodivers,2007,4(2):139-144.
[16]周桂芬,吕圭源.铁皮石斛叶中黄酮碳苷类成分的高效液相指纹图谱及指标成分的含量测定[J].中国药学杂志,2012,47(11):889-893.
[2]高正华,杨兵勋,陈立钻.铁皮石斛的研究进展[J].中国现代应用药学,2008,25(8):692-695.
[3]孙恒,胡强,金航,等.铁皮石斛化学成分及药理活性研究进展[J].中国实验方剂学杂志,2017,23(11):225-234.
[4]管惠娟.铁皮石斛的化学成分及指纹图谱研究[D].沈阳:沈阳药科大学,2009:
[5]李燕,王春兰,王芳菲,等.铁皮石斛中的酚酸类及二氢黄酮类成分[J].中国药学杂志,2010,45(13):975-979.
[6]魏刚,顺庆生,黄月纯,等. 3种铁皮石斛种源HPLC特征图谱比较研究[J].中药新药与临床药理,2014,25(4):467-471.
[7]叶子,卢叶,薛亚甫,等.铁皮石斛专属性成分的分离制备及质量标准研究[J].中国中药杂志,2016,41(13):2481-2486.
[8]黄月纯,谢镇山,任晋,等. 3种种源铁皮石斛叶黄酮类成分HPLC特征图谱比较[J].中国实验方剂学杂志,2015,21(24):37-40.
[9]黄月纯,杨丽娥,刘宏源,等.鲜铁皮石斛茎、叶高效液相色谱特征图谱比较[J].广州中医药大学学报,2012,29(5):561-565.
[10]韩宝来,李元元,张玲玲,等.芦丁的光谱分析[J].化学与黏合,2017,39(1):72-76.
[11] Velozo L S M,Ferreira M J P,Santos M I S,et al. Cglycosyl flavones from Peperomia blanda[J].Fitoterapia,2008,80(2):119-122.
[12]唐海明,陈建南,徐香琴,等.狭基线纹香茶菜水溶性成分及其抗肿瘤活性研究[J].中草药,2014,45(22):3219-3223.
[13] ZHANG Y,LUO J,HAN C et al. Bioassay-guided preparative separation of angiotensin-converting enzyme inhibitory C-flavone glycosides from Desmodium styracifolium by recycling complexation high-speed counter-current chromatography[J]. J Pharm Biomed Anal,2015,102:276-281.
[14]刘晖晖,于永明,张涛,等.半边铁角蕨化学成分研究[J].中国药学杂志,2017,52(2):100-104.
[15] Flamini G. Flavonoids and other compounds from the aerial parts of Viola etrusca[J]. Chem Biodivers,2007,4(2):139-144.
[16]周桂芬,吕圭源.铁皮石斛叶中黄酮碳苷类成分的高效液相指纹图谱及指标成分的含量测定[J].中国药学杂志,2012,47(11):889-893.