摘要
目的:揭示亳芍多糖的动态积累规律,为亳芍的科学采收、质量评价及临床用药提供依据。方法:采用蒽酮比色法,以葡萄糖为标准对照品,检测波长628 nm,测定亳芍多糖的含量;亳芍多糖溶于0.5 mol·L^(-1)盐酸甲醇溶液,80℃水浴酸解16 h,氮气吹干后加入吡啶-六甲基二硅胺烷-三甲基氯硅烷(10∶2∶1),80℃水浴反应30 min,氮气吹干,残渣溶于正己烷,采用TG-5熔融石英毛细管柱、1.0 m L·min^(-1)氦气为载气、电子电离离子源(EI)、质量扫描范围为50~350 amu、三甲基硅烷(TMS)柱前衍生化-GC-MS分析法测定单糖组成及其含量。结果:栽培年限影响亳芍多糖积累,1~7年亳芍中多糖的含量分别为(2.7±0.03)%、(3.1±0.23)%、(4.1±0.10)%、(2.2±0.33)%、(1.5±0.23)%、(1.6±0.10)%和(1.6±0.07)%;栽培1~7年亳芍多糖均为葡萄糖、果糖、甘露糖、半乳糖、半乳糖醛酸、木糖、阿拉伯糖、鼠李糖组成的杂多糖,但比例随栽培年限不同而变化。结论:从多糖含量角度,栽培3年的亳芍多糖含量最高,品质最佳;栽培年限对亳芍多糖的单糖种类没有影响,但对单糖比例及含量产生明显影响,提示不同株龄的亳芍多糖的结构可能不同。
Objective： To reveal the accumulation of polysaccharide in Bozhou Peony P. lactiflora and provide references for its scientific harvesting, quality evaluating and clinical medication. Methods： The contents of the total polysaccharide in different ages of Bozhou Peony P. lactiflora were determined by an anthrone-sulfuric acid colorimetric method at 628 nm using glucose as standard. The polysaccharide of each Dendrobium sample was treated with methanolysis reagent （ MeOH containing HC1 0.5 mol. L^-1 ） in nitrogen atmosphere at 80 ℃ for 16 h to hydrolyze them to their corresponding methyl monosaccharides, and thereafter the excess of reagent was removed using a stream of nitrogen gas. The conversion of the methyl glycosides to their trimethylsilyl （ TMS ） derivatives was performed by adding a mixed pyridine-hexamethyldisilazane-trimethylclorosilane （ 10 ： 2 ： 1, v/v ） to the dried material. The reaction was carried out at 80 ~C for 30 min and the reagent removed using a stream of nitrogen gas and the derived residues were extracted with hexane. The trimethylsilyl-ester O-methyl monosaccharides were separated with Trace 1300 Gas chromatograph coupled to ISQ mass spectrometerseries equipments including a TriPLUS RSH autosampler on a TG-5 fused-silica capillary column. The carrier gas was Helium, 1.0 mL. min^-1. The MS transfer line and ion source were at 250 ℃ and 280 ℃, respectively. The MS mode was electron ionization （ El ）. The mass range scanned was 50-350 amu. Different standard carbohydrates were also converted to their corresponding TMS derivatives and analyzed by GC-MS in order to obtain patterns for identification and the standard calibration curves. Results： The accumulation of polysaccharide varied remarkably as their ages increased. The polysaccharide contents in 1-7 cultivated-years of Bozhou Peony P. lactiflora were （ 2.7 ± 0.03 ） %, （3.1±0.23）%, （4.1±0.10）%, （2.2±0.33）%, （1.5±0.23）%, （1.6±0.10）% and（1.6＋0.07）%, respectively. The polysaccharides in different ages of Bozhou Peony P. lactiflora were all heteropolysaccharides composed of glucose, fructose, mannose, galactose, galacturonic acid, xylose, arabinose and rhamnose while the ratios of the mono-saccharides changed remarkable as the age varied. Conclusion： The kinds of the monosaccharides were the same while the ratios of them were quite different in the plants at 1-7 ages. This suggested that the structure of polysaccharide in Bozhou Peony P. lactiflora might be varied as the plant ages changes.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2016年第8期1349-1356,共8页
Chinese Journal of Pharmaceutical Analysis
基金
国家自然科学基金(NSFC81274021
NSFC81573536)
中国博士后基金(第9批)特别资助(2016T90559)
中国博士后基金面上项目(2014M551791)
安徽省自然科学基金(1608085MH221)
安徽省人事厅博士后研究项目
关键词
亳芍
栽培年限
多糖
气相色谱-质谱联用
三甲基硅烷衍生化
Bozhou Peony Paeonia lactiflora Pall.
cultivation age
polysaccharides
GC-MS
TMS hanges derivatized
