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纸质出版日期:2018
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陈懿瑶, 罗阳, 闫燊, 等. HPAEC-PAD法测定石斛属植物单糖组分与主成分聚类分析[J]. 中国实验方剂学杂志, 2018,24(15):106-111.
CHEN Yi-yao, LUO Yang, YAN Shen, et al. Monosaccharide Composition and Principal Component Clustering Analysis of by HPACE-PAD[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(15): 106-111.
陈懿瑶, 罗阳, 闫燊, 等. HPAEC-PAD法测定石斛属植物单糖组分与主成分聚类分析[J]. 中国实验方剂学杂志, 2018,24(15):106-111. DOI: 10.13422/j.cnki.syfjx.20181046.
CHEN Yi-yao, LUO Yang, YAN Shen, et al. Monosaccharide Composition and Principal Component Clustering Analysis of by HPACE-PAD[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(15): 106-111. DOI: 10.13422/j.cnki.syfjx.20181046.
目的:建立一种高效阴离子交换色谱-脉冲安培(HPAEC-PAD)检测方法,直接用于检测石斛属植物单糖组分,并对其进行主成分聚类分析。方法:通过单因素试验筛选石斛多糖水解条件,使用DIONEX-ICS-5000+型离子色谱进行检测,梯度洗脱;并通过主成分聚类分析20种石斛属植物。结果:建立的HPACE-PAD法能在15 min之内实现7种单糖(甘露糖、鼠李糖、阿拉伯糖、葡萄糖、木糖、果糖、乳糖)的分离,精密度、稳定性、重复性RSD<5.0%,各成分平均加样回收率为141.95%,平均RSD 3.4%;对单糖组分分析结果表明,单糖组分差异显著,最高相差961倍,最低相差38.5倍;20种石斛均检测出甘露糖、鼠李糖、阿拉伯糖、葡萄糖和木糖,球花石斛、滇金石斛、尖刀唇石斛3种未检测出果糖和乳糖;长距石斛、长苏石斛未检测出果糖;景洪石斛、小黄花石斛、玫瑰石斛未检测出乳糖;通过主成分聚类分析,能将铁皮石斛、鼓槌石斛、金钗石斛、线叶石斛、姬竹叶石斛、小黄花石斛、玫瑰石斛、长苏石斛、长距石斛、尖刀唇石斛、蜻蜓石斛、滇金石斛、景洪石斛聚为一类,此类单糖组分含量较低,其他石斛种含量相对较高。结论:建立的HPAEC-PAD法灵敏度高、重复性好,样品处理简单,分析快速准确,仪器使用便利,可用于石斛单糖或其他植物、微生物单糖组分测定,为石斛多糖成分质控提供参考,通过组分分析也为高多糖种质选择、选育提供重点关注材料。
Objective: To develop a method for determination of monosaccharide compositions of Dendrobium by high performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD)
and conduct principal component clustering analysis. Method: The hydrolysis conditions of polysaccharide were screened by single factor experiment
and then DIONEX-ICS-5000+ ion chromatography was used for detection and gradient elution. Finally
20 species of dendrobium were analyzed by principal component clustering analysis. Result: The 7 kinds of monosaccharide (mannose
rhamnose
arabinose
glucose
xylose
fructose
and lactose) could be separated within 15 min with HPACE-PAD method
and the precision
stability
repeatability and accuracy of the established method were well-validated
RSD<5.0%. The recovery rate of each component was 141.95% with RSD of 3.4%. The results of monosaccharide components analysis showed that the difference of monosaccharide components was significant; the maximum difference was 961 times
and the minimum difference was 38.5 times. All of 20 species of dendrobium contained mannose
rhamnose
arabinose
glucose and xylose
but fructose and lactose were not detected in D. thyrsiflorum
Flickingeria albopurpurea and D. heterocarpum; fructose was not detected in D. longicornu and D. brymerianum; and lactose was not detected in D. exile
D. jenkinsii and D. crepidatum. By the method of principal component clustering analysis
D. officinale
D. chrysotoxum
D. nobile
D. aurantiacum
D. jenkinsii
D. crepidatum
D. brymerianum
D. longicornu
D. heterocarpum
D. pulchellum
F. albopurpurea
and D. exile were clustered into one class
as the content of the monosaccharide components was low in these species
and relatively high in the other dendrobium species. Conclusion: The established HPAEC-PAD method has high sensitivity
good reproducibility
easy processing of samples
accurate analysis and convenient use of instrument
so it can be used for component determination in Dendrobium monosaccharide or other plant monosaccharide
providing reference for the quality control of polysaccharides in dendrobiums. Through the component analysis
it also provides key points in the selection and breeding of high-polysaccharide germplasm.
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