HPLC-ELSD法同时测定党参休眠过程中5种可溶性糖含量

doi:10.16035/j.issn.1001-7283.2025.01.025

摘要/Abstract

摘要：

建立高效液相色谱―蒸发光散射（HPLC-ELSD）法同时测定党参5种可溶性糖含量的技术方法，探明可溶性糖在党参抗寒及休眠过程中发挥的作用。5种可溶性糖适宜的检测条件为Analysis Column（4.6 mm× 250 mm 5-Micron）色谱柱，流动相为乙腈:水（70:30）溶液，柱温35 ℃，流速1.0 mL/min；ELSD参数：雾化温度50 ℃，漂移管温度110 ℃，氮气流速2.2 mL/min，增益值1.0。在党参自然休眠过程中海藻糖只在休眠前期的根和休眠期的芽中检测到；棉子糖只在休眠前期的根中检测到；果糖和葡萄糖在休眠前期的叶中含量较高，蔗糖在休眠前期的根和休眠期的芽中含量较高，三者均在解除休眠后的芽中含量最高。本研究建立了高效的HPLC-ELSD法同时检测5种可溶性糖的技术体系，明确了海藻糖和棉子糖是党参抗寒的主要可溶性糖组分，其中海藻糖在根中合成并进一步转运至芽中。果糖、葡萄糖和蔗糖是党参根中储存的可溶性糖组分，也是萌发的物质和能量来源。

关键词: 党参, 可溶性糖组分, 高效液相色谱―蒸发光散射法, 休眠, 逆境生理

Abstract:

A method was developed for the simultaneous determination of contents of five soluble sugars in different organs of Codonopsis pilosula (Franch.) Nannf. during natural dormancy by HPLC-ELSD for investigating the role of soluble sugars in the cold resistance. The suitable detection conditions for the five soluble sugars were: Analysis Column (4.6 mm×250 mm 5-Micron), acetonitrile:water (70:30) solution as the mobile phase, column temperature of 35 °C, flow rate of 1.0 mL/min; ELSD parameters: nebulization temperature of 50 °C, drift tube temperature of 110 °C, nitrogen flow rate of 2.2 mL/min, gain value of 1.0. During the natural dormancy of C.pilosula, trehalose was only detected in roots (pre-dormant) and buds (dormant), raffinose was only detected in pre-dormant roots; fructose and glucose were found in higher levels in the leaves in the early stage of dormancy, and sucrose was found in higher levels in the roots in the early stage of dormancy and the buds in the dormant period. The highest levels of all three types of sugars were found in the buds after dormancy was released. This study established an efficient HPLC-ELSD technical system for the simultaneous detection of five soluble sugars, and confirmed that trehalose and raffinose are the main soluble sugar components of C.pilosula for cold resistance. Trehalose is synthesized in the roots and further transported to the buds. Fructose, glucose and sucrose are soluble sugar components stored in the roots of C.pilosula, and are also the source of substances and energy for germination.

Key words: Codonopsis pilosula (Franch.) Nannf., Soluble sugar component, HPLC-ELSD, Dormancy, Stress physiology

唐顺莉, 张延红, 杜弢, 陈晖, 何春雨, 董婉琦. HPLC-ELSD法同时测定党参休眠过程中5种可溶性糖含量[J]. 作物杂志, 2025 (1): 202–207

Tang Shunli, Zhang Yanhong, Du Tao, Chen Hui, He Chunyu, Dong Wanqi. Simultaneous Determination of Contents of Five Kinds of Soluble Sugars in Codonopsis pilosula during Dormancy by HPLC-ELSD Method[J]. Crops, 2025(1): 202–207

图/表 6

图1

图2

表1

图3

图4

图5

参考文献 34

[1]	Reyes-Diaz M, Alberdi M, Piper F, et al. Low temperature responses of Nothofagus dombeyi and Nothofagus nitida, two evergreen species from south central Chile. Tree Physiology, 2005, 25(11):1389-1398. pmid: 16105806
[2]	Benina M, Obata T, Mehterov N, et al. Comparative metabolic profiling of Haberlea rhodopensis, Thellungiella halophyla, and Arabidopsis thaliana exposed to low temperature. Frontiers in Plant Science, 2013, 4:499.
[3]	Ruan Y L. Sucrose metabolism: gateway to diverse carbon use and sugar signaling. Annual Review of Plant Biology, 2014, 65:33-67.
[4]	陈宏艳, 李小二, 李忠光. 糖信号及其在植物响应逆境胁迫中的作用. 生物技术通报, 2022, 38(7):80-89. doi: 10.13560/j.cnki.biotech.bull.1985.2021-1289
[5]	岳川. 茶树糖类相关基因的挖掘及其在茶树冷驯化中的表达研究. 北京: 中国农业科学院, 2015.
[6]	杨平, 李敏惠, 潘克俭, 等. 海藻糖的生物合成与分解途经及其生物学功能. 生命的化学, 2006, 26(3):233-236.
[7]	Sharma M, Banday Z Z, Shukla B N, et al. Glucose-regulated HLP1 acts as a key molecule in governing thermomemory. Plant Physiology, 2019, 180(2):1081-1100. doi: 10.1104/pp.18.01371 pmid: 30890662
[8]	Saksena H B, Sharma M, Singh D, et al. The versatile role of glucose signalling in regulating growth, development and stress responses in plants. Journal of Plant Biochemistry and Biotechnology, 2020, 29(4):687-699.
[9]	Cisse A. 蔗糖影响水稻耐热性的作用机理研究. 北京: 中国农业科学院, 2020.
[10]	黄紫嫣, 王婧, 丁华, 等. 全自动还原糖测定仪法与斐林试剂滴定法对比研究. 湖北农业科学, 2019, 58(增2):398-402.
[11]	刘胜辉, 魏长宾, 孙光明, 等. 高效液相色谱法测定台农6号菠萝果实中的糖分. 食品科学, 2009, 30(12):162-164. doi: 10.7506/spkx1002-6630-200912033
[12]	张淋洁, 王如伟, 何厚洪, 等. 高效液相色谱―蒸发光散射法测定铁皮石斛中单糖和双糖. 医药导报, 2013, 32(4):517-520.
[13]	Gibson S I. Plant sugar-response pathways. Part of a complex regulatory web. Plant Physiology, 2000, 124(4):1532-1539.
[14]	王静, 王睛, 向文胜. 色谱法在糖类化合物分析中的应用. 分析化学, 2001, 29(2):222-227.
[15]	施红林, 王保兴, 杨光宇, 等. 高效液相色谱法测定烟草中的糖. 分析化学, 2002, 30(3):384.
[16]	李忠, 杨光宇, 黄海涛, 等. 高效液相色谱法测定烟草料液中的糖、甘油和丙二醇. 分析化学, 2002, 30(6):687-689.
[17]	郑春英, 李庆勇, 祖元刚. HPLC-ELSD法测定甘草中单糖的含量. 东北林业大学学报, 2006, 34(2):109-110.
[18]	徐瑾, 张庆合, 张维冰, 等. 液相色谱荧光衍生法在糖类物质分析中的应用. 色谱, 2003, 21(2):115-120.
[19]	刘云惠. 高效液相色谱内标法分离和测定植物中的单糖. 色谱, 2000, 18(6):556-558.
[20]	魏泱, 丁明玉. 液相色谱/蒸发光散射测定转基因烟草提取液中的海藻糖. 色谱, 2001, 19(3):226-229.
[21]	罗进, 夏敏, 叶能胜, 等. HPLC-ELSD同时测定食品中5种糖含量. 食品科学, 2010, 31(8):226-229. doi: 10.7506/spkx1002-6300-201008051
[22]	辛秀兰, 李小萍, 马越, 等. HPLC-ELSD法测定红树莓果实中水溶性糖含量. 吉林农业大学学报, 2009, 31(5):624-627.
[23]	程勇, 李庆廷, 李剑政, 等. Prevail糖柱-HPLC-ELSD法测定烟草中水溶性糖. 烟草科技, 2010(3):32-37.
[24]	宋晓晖, 谢凯, 李艳丽, 等. HPLC-ELSD法测定梨果实中不同种类可溶性糖含量. 南京农业大学学报, 2012, 35(2):87-91.
[25]	李威涛, 郭建斌, 喻博伦, 等. 基于HPLC-RID的花生籽仁可溶性糖含量检测方法的建立. 作物学报, 2021, 47(2):368-375. doi: 10.3724/SP.J.1006.2021.04110
[26]	曹进, 徐燕, 张永知, 等. 清开灵注射液HPLC/ELSD指纹图谱建立及质量相关性研究. 分析化学, 2004, 32(4):469-473.
[27]	詹园凤, 贺滉, 党选民. HPLC-ELSD法测定礼品西瓜果实可溶性糖的种类和含量. 南方农业学报, 2014, 45(11):2005- 2008.
[28]	黄熊娟. 钾氮肥施用量影响微胚乳超高油玉米产量及含油率机理的研究. 南宁:广西大学, 2006.
[29]	Liu H L, Dai X Y, Xu Y Y, et al. Over-expression of OsUGE-1 altered raffinose level and tolerance to abiotic stress but not morphology in Arabidopsis. Journal of Plant Physiology, 2007, 164(10):1384-1390.
[30]	Li Y, Lee K K, Walsh S, et al. Establishing glucose- and ABA- regulated transcription networks in Arabidopsis by microarray analysis and promoter classification using a Relevance Vector Machine. Genome Research, 2006, 16(3):414-427.
[31]	王成君, 林建平, 岑沛霖. 高效液相色谱法快速检测海藻糖. 江南大学学报(自然科学版), 2005, 4(5):86-89.
[32]	Dos Santos T B, Budzinski I G F, Marur C J, et al. Expression of three galactinol synthase isoforms in Coffea arabica L. and accumulation of raffinose and stachyose in response to abiotic stresses. Plant Physiology and Biochemistry, 2011, 49(4):441- 448. doi: 10.1016/j.plaphy.2011.01.023 pmid: 21330144
[33]	Yue C, Cao H L, Wang L, et al. Effects of cold acclimation on sugar metabolism and sugar-related gene expression in tea plant during the winter season. Plant Molecular Biology, 2015, 88(6):591-608. doi: 10.1007/s11103-015-0345-7 pmid: 26216393
[34]	张丹. 木薯海藻糖合成酶基因MeTPS1-3的克隆与功能分析. 海口:海南大学, 2013.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

组分Composition	回归方程Regression equation	相关系数Correlation coefficient	线性范围Linearity range (mg/mL)
果糖Fructose	y=2055.4x-31.834	0.9998	0.016~10
葡萄糖Glucose	y=2326.7x-5.2971	0.9998	0.016~10
蔗糖Sucrose	y=2530.5x+120.21	0.9991	0.016~10
海藻糖Trehalose	y=2623.6x-57.838	1.0000	0.016~10
棉子糖Raffinose	y=2250.1x-256.03	0.9990	0.016~10