利用大孔吸附树脂与高速逆流色谱纯化脱氧雪腐镰刀菌烯醇-3-葡萄糖苷

doi:10.3864/j.issn.0578-1752.2025.08.013

中国农业科学 ›› 2025, Vol. 58 ›› Issue (8): 1627-1637.doi: 10.3864/j.issn.0578-1752.2025.08.013

利用大孔吸附树脂与高速逆流色谱纯化脱氧雪腐镰刀菌烯醇-3-葡萄糖苷

陈龙云¹^,²(), 胡俊强², 何灿², 史建荣², 徐剑宏¹^,², 王刚¹^,²()

¹ 江苏大学食品与生物工程学院，江苏镇江 212013
² 江苏省农业科学院农产品质量安全与营养研究所/江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地/农业农村部农产品质量安全控制技术与标准重点实验室/农业农村部农产品质量安全风险评估实验室（南京）/农业农村部农产品质量安全控制技术与标准重点实验室/江苏省现代粮食流通与安全协同创新中心，南京 210014

收稿日期:2024-08-24 接受日期:2025-01-21 出版日期:2025-04-16 发布日期:2025-04-21
通信作者:
王刚，E-mail：wanggang2015@jaas.ac.cn
联系方式: 陈龙云，E-mail：clyun99@163.com。
基金资助:
国家重点研发计划(2023YFD1301003); 国家自然科学基金(32161143034); 江苏省科技计划专项资金(BE2022786); 江苏省科技计划专项资金(3003)

Purification of Deoxynivalenol-3-Glucoside by Using Macroporous Adsorption Resin Combined with High-Speed Counter-Current Chromatography

CHEN LongYun¹^,²(), HU JunQiang², HE Can², SHI JianRong², XU JianHong¹^,², WANG Gang¹^,²()

¹ School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, Jiangsu
² Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology/Key Laboratory for Agro-Product Safety Risk Evaluation (Nanjing), Ministry of Agriculture and Rural Affairs/Key Laboratory for Control Technology and Standard for Agro-Product Safety and Quality, Ministry of Agriculture and Rural Affairs/Collaborative Innovation Center for Modern Grain Circulation and Safety/Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014

Received:2024-08-24 Accepted:2025-01-21 Published:2025-04-16 Online:2025-04-21

摘要/Abstract

摘要：

【目的】脱氧雪腐镰刀菌烯醇-3-葡萄糖苷（DON-3G）是粮食产品中最常见的隐蔽型真菌毒素，但高昂的底物价格限制了对其深入研究。开发联用大孔吸附树脂与高速逆流色谱（HSCCC）方法，以实现酶促转化产物中DON-3G的大量纯化。【方法】采用静态吸附试验筛选不同类型吸附树脂或离子交换树脂；利用单因素水平试验优化最佳吸附条件与洗脱条件并拟合吸附热力学模型；筛选合适的双相体系并采用HSCCC分离DON-3G纯物质；最终通过紫外吸收光谱与核磁共振波谱确证产物结构，通过高效液相色谱分析产物纯度。【结果】采用XAD-4大孔吸附树脂吸附反应体系中DON-3G，吸附容量达269.23 μg·g^-1；洗脱溶剂采用40%甲醇水溶液，可在5倍柱体积内洗脱DON-3G。采用正丁醇﹕三氟乙酸﹕水为1﹕0.01﹕1作为HSCCC双相体系，在流速1 mL·min^-1、转速1 000 r/min、柱温35 ℃、双向洗脱模式下可以实现底物DON与产物DON-3G的分离，DON-3G回收率79.7%，液相色谱分析表明产物纯度为97.46%。未反应DON可以通过反向洗脱回收。【结论】本方法可实现单批次100 mg以上的DON-3G分离纯化，为针对DON-3G的后续研究奠定了基础。

关键词: 脱氧雪腐镰刀菌烯醇-3-葡萄糖苷, 隐蔽型毒素, 分离纯化, 大孔吸附树脂, 高速逆流色谱

Abstract:

【Objective】 Deoxynivalenol-3-glucoside (DON-3G) is the most commonly found masked mycotoxin in cereals, however, the high cost of substrates hinders further studies on it. This study aimed to develop an efficient and affordable method for the large-scale purification of DON-3G from the enzymatic transformation product. 【Method】 Static adsorption experiments were utilized to screen various types of adsorption and ion-exchange resins, and a single-factor experimental design was employed to determine the optimal adsorption and desorption conditions. The adsorption thermodynamic model was established to fit the adsorption behavior. The pure material of DON-3G was then prepared using high-speed counter-current chromatography (HSCCC), and the biphasic systems were screened. The UV and NMR spectroscopy were used to confirm the product's structure, and high-performance liquid chromatography (HPLC) was used to assess its purity.【Result】With a maximal adsorption capacity of 269.23 μg·g^-1 resin, the XAD-4 resin was determined to be the most effective macroporous resin for adsorbing DON-3G from the reaction system. The 40% methanol solution was used as the desorption solvent, because it could elute DON-3G within 5 bed volume. DON-3G and unreacted DON were separated using a biphasic system with n-butanol/trifluoroacetic acid/water (1:0.01:1) at a flow rate of 1 mL·min^-1, a rotation speed of 1 000 r/min, and a column temperature of 35 ℃ under the bidirectional elution mode. With optimal conditions, the HSCCC process produced DON-3G with a purity of 97.46% and the recovery rate was 79.7%. Unreacted DON could be recovered by reverse elution.【Conclusion】Over 100 mg of DON-3G might be separated and purified using this approach in a single run, paving the way for the subsequent research targeting DON-3G.

Key words: deoxynivalenol-3-glucoside, masked mycotoxin, isolation and purification, macroporous adsorption resin, high-speed counter-current chromatography

陈龙云, 胡俊强, 何灿, 史建荣, 徐剑宏, 王刚. 利用大孔吸附树脂与高速逆流色谱纯化脱氧雪腐镰刀菌烯醇-3-葡萄糖苷[J]. 中国农业科学, 2025, 58(8): 1627-1637.

CHEN LongYun, HU JunQiang, HE Can, SHI JianRong, XU JianHong, WANG Gang. Purification of Deoxynivalenol-3-Glucoside by Using Macroporous Adsorption Resin Combined with High-Speed Counter-Current Chromatography[J]. Scientia Agricultura Sinica, 2025, 58(8): 1627-1637.

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参考文献 37

[1]	ABUDABOS A M, AL-ATIYAT R M, KHAN R U. A survey of mycotoxin contamination and chemical composition of distiller’s dried grains with solubles (DDGS) imported from the USA into Saudi Arabia. Environmental Science and Pollution Research, 2017, 24(18): 15401-15405.
[2]	ESKOLA M, KOS G, ELLIOTT C T, HAJŠLOVÁ J, MAYAR S, KRSKA R. Worldwide contamination of food-crops with mycotoxins:validity of the widely cited ‘FAO estimate’ of 25%. Critical Reviews in Food Science and Nutrition, 2020, 60(16): 2773-2789.
[3]	BERTHILLER F, CREWS C, DALL’ASTA C, DE SAEGER S, HAESAERT G, KARLOVSKY P, OSWALD I P, SEEFELDER W, SPEIJERS G, STROKA J. Masked mycotoxins: A review. Molecular Nutrition & Food Research, 2013, 57(1): 165-186.
[4]	DALL’ASTA C, BERTHILLER F. Masked mycotoxins in food: Formation, occurrence and toxicological relevance. Cambridge: Royal Society of Chemistry, 2015.
[5]	DONG F, WANG S, YU M, SUN Y, XU J, SHI J. Natural occurrence of deoxynivalenol and deoxynivalenol-3-glucoside in various wheat cultivars grown in Jiangsu province, China. World Mycotoxin Journal, 10(3): 285-294.
[6]	SCHWARTZ-ZIMMERMANN H E, HAMETNER C, NAGL V, SLAVIK V, MOLL W D, BERTHILLER F. Deoxynivalenol (DON) sulfonates as major DON metabolites in rats: From identification to biomarker method development, validation and application. Analytical and Bioanalytical Chemistry, 2014, 406(30): 7911-7924.
[7]	BRYŁA M, STĘPNIEWSKA S, MODRZEWSKA M, WAŚKIEWICZ A, PODOLSKA G, KSIENIEWICZ-WOŹNIAK E, YOSHINARI T, STĘPIEŃ Ł, URBANIAK M, ROSZKO M, GWIAZDOWSKI R, KANABUS J, PIERZGALSKI A. Dynamics of deoxynivalenol and nivalenol glucosylation in wheat cultivars infected with Fusarium culmorum in field Conditions-A 3 year study (2018-2020). Journal of Agricultural and Food Chemistry, 2022, 70(14): 4291-4302.
[8]	KOVALSKY PARIS M P, SCHWEIGER W, HAMETNER C, STÜCKLER R, MUEHLBAUER G J, VARGA E, KRSKA R, BERTHILLER F, ADAM G. Zearalenone-16-O-glucoside: A new masked mycotoxin. Journal of Agricultural and Food Chemistry, 2014, 62(5): 1181-1189. doi: 10.1021/jf405627d pmid: 24386883
[9]	BORZEKOWSKI A, DREWITZ T, KELLER J, PFEIFER D, KUNTE H J, KOCH M, ROHN S, MAUL R. Biosynthesis and characterization of Zearalenone-14-sulfate, Zearalenone-14-glucoside and Zearalenone- 16-glucoside using common fungal strains. Toxins, 2018, 10(3): 104.
[10]	PAN Y Y, LIU C D, YANG J G, TANG Y Q. Conversion of Zearalenone to β-Zearalenol and Zearalenone-14, 16-diglucoside by Candida parapsilosis ATCC 7330. Food Control, 2022, 131: 108429.
[11]	GAB-ALLAH M A, TAHOUN I F, YAMANI R N, REND E A, SHEHATA A B. Natural occurrence of deoxynivalenol, nivalenol and deoxynivalenol-3-glucoside in cereal-derived products from Egypt. Food Control, 2022, 137: 108974.
[12]	NIE D X, ZHU X T, LIU M H, CHENG M, FAN K, ZHAO Z H, HUANG Q W, ZHANG X L, HAN Z. Molecularly imprinted polymer-based electrochemical sensor for rapid detection of masked deoxynivalenol with Mn-doped CeO₂ nanozyme as signal amplifier. Journal of Hazardous Materials, 2024, 477: 135366.
[13]	BERTHILLER F, WERNER U, SULYOK M, KRSKA R, HAUSER M T, SCHUHMACHER R. Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) determination of phase II metabolites of the mycotoxin Zearalenone in the model plant Arabidopsis thaliana. Food Additives and Contaminants, 2006, 23(11): 1194-1200.
[14]	TROMBETE F, BARROS A, VIEIRA M, SALDANHA T, VENÂNCIO A, FRAGA M. Simultaneous determination of deoxynivalenol, deoxynivalenol-3-glucoside and nivalenol in wheat grains by HPLC-PDA with immunoaffinity column cleanup. Food Analytical Methods, 2016, 9(9): 2579-2586.
[15]	ZHANG Z Q, CAI Y L, FAN K, HUANG Q W, ZHAO X Y, CAO H J, ZHAO Z H, TANGNI E K, HAN Z. Development of a reliable UHPLC-MS/MS method for simultaneous determination of Zearalenone and Zearalenone-14-glucoside in various feed products. Frontiers in Chemistry, 2022, 10: 955266.
[16]	DE COLLI L, ELLIOTT C, FINNAN J, GRANT J, ARENDT E K, MCCORMICK S P, DANAHER M. Determination of 42 mycotoxins in oats using a mechanically assisted QuEChERS sample preparation and UHPLC-MS/MS detection. Journal of Chromatography B, 2020, 1150: 122187.
[17]	CASU A, CAMARDO LEGGIERI M, TOSCANO P, BATTILANI P. Changing climate, shifting mycotoxins: A comprehensive review of climate change impact on mycotoxin contamination. Comprehensive Reviews in Food Science and Food Safety, 2024, 23(2): e13323.
[18]	DONG F, QIU J B, XU J H, YU M Z, WANG S F, SUN Y, ZHANG G F, SHI J R. Effect of environmental factors on Fusarium population and associated trichothecenes in wheat grain grown in Jiangsu province, China. International Journal of Food Microbiology, 2016, 230: 58-63.
[19]	NAKAGAWA H, OHMICHI K, SAKAMOTO S, SAGO Y, KUSHIRO M, NAGASHIMA H, YOSHIDA M, NAKAJIMA T. Detection of a new Fusarium masked mycotoxin in wheat grain by high-resolution LC-Orbitrap™ MS. Food Additives & Contaminants: Part A, 2011, 28(10): 1447-1456.
[20]	KOVALSKY P, KOS G, NÄHRER K, SCHWAB C, JENKINS T, SCHATZMAYR G, SULYOK M, KRSKA R. Co-occurrence of regulated, masked and emerging mycotoxins and secondary metabolites in finished feed and maize: An extensive survey. Toxins, 2016, 8(12): 363.
[21]	SUZUKI T, IWAHASHI Y. Low toxicity of deoxynivalenol-3- glucoside in microbial cells. Toxins, 2015, 7(1): 187-200.
[22]	TITTLEMIER S A, BRUNKHORST J, CRAMER B, DEROSA M C, LATTANZIO V M T, MALONE R, MARAGOS C, STRANSKA M, SUMARAH M W. Developments in mycotoxin analysis: An update for 2019-2020. World Mycotoxin Journal, 2021, 14(1): 3-26.
[23]	MICHLMAYR H, MALACHOVÁ A, VARGA E, KLEINOVÁ J, LEMMENS M, NEWMISTER S, RAYMENT I, BERTHILLER F, ADAM G. Biochemical characterization of a recombinant UDP- glucosyltransferase from rice and enzymatic production of deoxynivalenol-3-O-β-D-glucoside. Toxins, 2015, 7(7): 2685-2700.
[24]	王刚, 胡俊强, 史建荣, 徐剑宏. 利用大孔吸附树脂与高速逆流色谱联用技术大量制备脱氧雪腐镰刀菌烯醇的方法. 江苏农业科学, 2018, 46(23): 207-211.
	WANG G, HU J Q, SHI J R, XU J H. Preparative purification of deoxynivalenol by combined use of macroporous resin and high-speed countercurrent chromatography. Jiangsu Agricultural Sciences, 2018, 46(23): 207-211. (in Chinese)
[25]	GUO X, WANG J L. Comparison of linearization methods for modeling the Langmuir adsorption isotherm. Journal of Molecular Liquids, 2019, 296: 111850.
[26]	ITO Y. Golden rules and pitfalls in selecting optimum conditions for high-speed counter-current chromatography. Journal of Chromatography A, 2005, 1065(2): 145-168. pmid: 15782961
[27]	HABLER K, FRANK O, RYCHLIK M. Chemical synthesis of deoxynivalenol-3-β-d-[¹³C6]-glucoside and application in stable isotope dilution assays. Molecules, 2016, 21(7): 838.
[28]	王丹丹, 刘芫汐, 左甜甜, 昝珂, 金红宇. 大孔吸附树脂及其在中药领域应用研究进展. 中国药事, 2022, 36(7): 826-835. doi: 10.16153/j.1002-7777.2022.06.012
	WANG D D, LIU Y X, ZUO T T, ZAN K, JIN H Y. Research progress of macroporous adsorption resin and its application in traditional Chinese medicines. Chinese Pharmaceutical Affairs, 2022, 36(7): 826-835. (in Chinese) doi: 10.16153/j.1002-7777.2022.06.012
[29]	杨楠, 阮国永, 王朝玉, 杨建波, 刘云, 谢飞燕, 李长军, 潘峰. 利用大孔吸附树脂同时清除云实根粗多糖色素和蛋白成分的研究. 贵州师范大学学报(自然科学版), 2023, 41(5): 107-113.
	YANG N, RUAN G Y, WANG C Y, YANG J B, LIU Y, XIE F Y, LI C J, PAN F. Study on simultaneous decoloration and deproteinization of crude polysaccharide from the root of Caesalpinia decapetala by using macroporous adsorption resin. Journal of Guizhou Normal University (Natural Sciences), 2023, 41(5): 107-113. (in Chinese)
[30]	赵沙沙, 何海, 张小荣, 郭玫, 崔治家, 邵晶. 红芪多糖 “脱蛋白-大孔吸附树脂” 的纯化富集工艺考察. 药物流行病学杂志, 2023, 32(6): 679-688.
	ZHAO S S, HE H, ZHANG X R, GUO M, CUI Z J, SHAO J. Study on purification and enrichment technology of Hedysarum polysaccharide “deproteinization-macroporous adsorption resin”. Chinese Journal of Pharmacoepidemiology, 2023, 32(6): 679-688. (in Chinese)
[31]	SPÓRNA-KUCAB A, JERZ G, KUMORKIEWICZ-JAMRO A, TEKIELI A, WYBRANIEC S. High-speed countercurrent chromatography for isolation and enrichment of betacyanins from fresh and dried leaves of Atriplex hortensis L. var. “Rubra”. Journal of Separation Science, 2021, 44(23): 4222-4236.
[32]	YUAN J J, TUO S, SHI X H, TU J L. A new combined technology: Macroporous adsorption resin and high speed counter current chromatography for hydroxytyrosol separation from olive leaf enzymatic hydrolysate. Journal of Chromatography B, 2024, 1235: 124058.
[33]	宋道光, 陈志. 高速逆流色谱法从小叶金钱草中分离四种黄酮苷类化合物. 食品工业科技, 2022, 43(24): 93-101.
	SONG D G, CHEN Z. Separation of four flavonoid glycosides from Hydrocotyle sibthorpioides Lam. by high-speed counter-current chromatography. Science and Technology of Food Industry, 2022, 43(24): 93-101. (in Chinese)
[34]	YANG X, SHEN C, LI H M, WANG N N, MA J L, WANG S, ZHAO J Y, CHEN J L, YANG L, CHEN T, LI Y L. Combined chromatographic strategy based on macroporous resin, high-speed counter-current chromatography and preparative HPLC for systematic separation of seven antioxidants from the fruit of Terminalia billerica. Journal of Separation Science, 2019, 42(20): 3191-3199.
[35]	CHEN T, LIU Y L, CHEN C, ZOU D L, YOU J M, SUN J, LI Y L. Application of high-speed counter-current chromatography combined with macroporous resin for rapid enrichment and separation of three anthraquinone glycosides and one stilbene glycoside from Rheum tanguticum. Journal of Chromatography B, 2014, 957: 90-95.
[36]	WANG G, CHEN W H, HU J Q, FAN B, SHI J R, XU J H. Preparative isolation and purification of Zearalenone from rice culture by combined use of macroporous resin column and high-speed counter-current chromatography. Journal of Chromatography B, 2019, 1110: 43-50.
[37]	WANG G, HE D, ZHAO F C, HU J Q, LEE Y W, SHI J R, XU J H. Extraction and purification of ustiloxin A from rice false smut balls by a combination of macroporous resin and high-speed countercurrent chromatography. Food Production, Processing and Nutrition, 2020, 2(1): 29.

树脂名称 Resin name	树脂类型 Resin type	平均孔径 Mean pore size (nm)	比表面积 Surface area (m²·g^-1)	孔隙容量 Pore volume (mL·g^-1)	基质 Matrix
XAD-2	非极性大孔吸附树脂 Nonpolar	9	300	0.65	苯乙烯－二乙烯基苯高聚物 Styrene-divinylbenzene
XAD-4	非极性大孔吸附树脂 Nonpolar	10	750	0.98	苯乙烯－二乙烯基苯高聚物 Styrene-divinylbenzene
XAD-7HP	弱极性大孔吸附树脂 Weakly polar	30-40	380	0.50	聚丙烯酸酯 Acrylic ester
XAD-1180	非极性大孔吸附树脂 Nonpolar	20	800	1.82	苯乙烯－二乙烯基苯高聚物 Styrene-divinylbenzene
XAD-1180N	非极性大孔吸附树脂 Nonpolar	30-40	450	1.40	苯乙烯－二乙烯基苯高聚物 Styrene-divinylbenzene
FPA91-OH	强碱性阴离子交换树脂 Strong basic anion exchange	30	NR	0.80	聚丙烯酸酯 Acrylic ester
IRA67	弱碱性阴离子交换树脂 Weakly basic anion exchange	NR	NR	1.60	聚丙烯酸酯 Acrylic ester

Langmuir吸附模型 Langmuir adsorption isotherm		Freundlich吸附模型 Freundlich adsorption isotherm
线性方程Linear equation	C_e/Q_e=0.0036Ce+0.081	线性方程Linear equation	ln(Q_e)=0.321ln(C_e)+3.821
Q_m	277.78	K_F	45.65
K_L	0.044	1/n	0.321
R²	0.9941	R²	0.9754

	溶剂组成 Solvent system	比例 Ratio（v/v）	DON K_d值	DON-3G K_d值
1	乙酸乙酯:水:正己烷:甲醇 Ethyl acetate:H₂O:n-hexane:methanol	8:8:2:2	0.62	0.004
2	乙酸乙酯:水 Ethyl acetate:H₂O	1:1	0.882	0.009
3	正己烷:正丁醇:甲醇:水 n-Hexane:n-butanol:methanol:H₂O	2:4:1:4	0.86	0.16
4	甲基叔丁基醚:乙腈:三氟乙酸:水 MTBE:acetonitrile:TFA:H₂O	2:2:0.003:3	0.71	0.06
5	正丁醇:TFA:水 n-Butanol:TFA:H₂O	1:0.005:1	1.13	0.28
6	正丁醇:TFA:水 n-Butanol:TFA:H₂O	1:0.01:1	1.27	0.47
7	正丁醇:TFA:水 n-Butanol:TFA:H₂O	1:0.015:1	1.23	0.48
8	正丁醇:TFA:水 n-Butanol:TFA:H₂O	1:0.02:1	1.20	0.44

利用大孔吸附树脂与高速逆流色谱纯化脱氧雪腐镰刀菌烯醇-3-葡萄糖苷

Purification of Deoxynivalenol-3-Glucoside by Using Macroporous Adsorption Resin Combined with High-Speed Counter-Current Chromatography

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[2]	李杰,贾栩超,张瑞芬,刘磊,池建伟,黄菲,董丽红,张名位. 黑芝麻黑色素的分离纯化、结构表征及体外抗氧化活性[J]. 中国农业科学, 2020, 53(12): 2477-2492.
[3]	贾丰，郭玉蓉，杨曦，刘冬，李洁. 发酵苹果渣多糖分离纯化、结构及其与加工特性的关系[J]. 中国农业科学, 2017, 50(10): 1873-1884.
[4]	陈思远，刘永祥，曹小舟，张逸婧，陈海娟，沈新春. 从麦胚清蛋白分离制备高活性抗氧化肽[J]. 中国农业科学, 2016, 49(12): 2379-2388.
[5]	苏东晓1, 2, 张瑞芬1, 张名位1, 黄菲1, 2, 魏振承1, 张雁1, 遆慧慧1, 邓媛元1, 唐小俊1. 荔枝果肉酚类物质大孔树脂分离纯化工艺优化[J]. 中国农业科学, 2014, 47(14): 2897-2906.
[6]	刘程程, 石波, 姚喜梅, 梁平, 李静梅. 制备型硅胶柱层析分离单一聚合度纤维寡糖及其测定[J]. 中国农业科学, 2011, 44(14): 2999-3006 .
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[9]	. 猪精液碱性N-乙酰-β-D-氨基葡萄糖苷酶的纯化与酶学性质研究[J]. 中国农业科学, 2008, (9): 2843-2849 .
[10]	张金林,徐扩,李川,马娟,董金皋. 灰葡萄孢诱变菌株毒素的除草活性研究[J]. 中国农业科学, 2005, 38(06): 1174-1181 .