维生素K1生理功能及其在农产品中的检测方法

doi:10.3864/j.issn.0578-1752.2019.18.013

中国农业科学 ›› 2019, Vol. 52 ›› Issue (18): 3207-3217.doi: 10.3864/j.issn.0578-1752.2019.18.013

维生素K₁生理功能及其在农产品中的检测方法

徐月清,汪丹丹,赵新楠,喻理,王秀嫔,张良晓(),张奇,张文,李培武()

中国农业科学院油料作物研究所/农业农村部油料作物生物学与遗传改良重点实验室/农业农村部油料及制品质量监督检验测试中心/农业农村部油料产品质量安全风险评估实验室(武汉)/农业农村部生物毒素检测重点实验室,武汉 430062

收稿日期:2019-03-20 接受日期:2019-06-18 出版日期:2019-09-16 发布日期:2019-09-23
通讯作者: 张良晓,李培武
作者简介:徐月清,E-mail：280465976@qq.com。
基金资助:
国家重点研发计划(2017YFC1601700);国家自然科学基金面上项目(31871886);现代农业产业体系建设专项资金(CARS-12);国家农产品质量安全风险评估重大专项(GJFP2019003)

Advances in Research on Physiological Functions of Vitamin K₁and Its Detection Methods in Agricultural Products

XU YueQing,WANG DanDan,ZHAO XinNan,YU Li,WANG XiuPin,ZHANG LiangXiao(),ZHANG Qi,ZHANG Wen,LI PeiWu()

Oil Crops Research Institute, Chinese Academy of Agricultural Science/Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs/Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture and Rural Affairs/Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture and Rural Affairs/Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture and Rural Affairs, Wuhan 430062

Received:2019-03-20 Accepted:2019-06-18 Online:2019-09-16 Published:2019-09-23
Contact: LiangXiao ZHANG,PeiWu LI

摘要/Abstract

摘要：

农产品中含有丰富的营养物质,对维持人体物质代谢和生理功能具有重要作用。维生素K₁在自然界中广泛存在,在绿叶蔬菜和油料等农产品中含量丰富。作为人和动物必需的脂溶性维生素,维生素K₁在日常膳食中起着不可或缺的作用,越来越受到人们的关注。最新研究进展表明,农产品中维生素K₁不仅具有促进血液正常凝固、预防新生婴儿出血疾病的生理功能,还具有抑制癌症、预防血管钙化、参与骨骼代谢、抑制糖尿病性白内障、治疗急慢性肝炎等生理功能。维生素K₁对碱、强酸以及紫外线照射敏感,因此对检测前处理方法要求较高,目前主要的检测前处理方法有皂化法、酶解法、有机溶剂萃取法、超临界流体萃取法以及净化效果良好的固相萃取法,但往往由于检测前处理基质不同,需要在实际操作中选择合适的前处理方法;维生素K₁的检测方法主要采用荧光分光光度法、液相色谱法、色谱质谱联用法等方法,筛选准确、高效、快速的检测方法也是当今的研究热点。本文在大量文献基础上,综述了近年来国内外不同农产品基质中维生素K₁的前处理以及检测方法,旨在比较前处理和检测技术的优劣,为农产品中维生素K₁高灵敏检测技术开发、高维生素K₁农产品开发与评价提供参考。最后,提出今后维生素K₁检测技术发展的方向：(1)研制绿色溶剂提高提取效率;(2)研制萃取材料,提高提取选择性和回收率;(3)建立农产品中维生素K₁与其他营养功能成分同步检测技术。

关键词: 农产品, 维生素K₁, 生理功能, 前处理, 检测方法

Abstract:

Agricultural products are rich in nutrients, which play important roles in maintaining the normal metabolism and physiological functions of human beings. Vitamin K₁ is widely found in agricultural products such as green vegetables and oilseeds. As a fat-soluble vitamin, vitamin K₁ plays indispensable roles in daily diet and draws more and more attentions. The recent researches showed that vitamin K₁ in agricultural products had the physiological functions including not only promoting normal blood coagulation, preventing bleeding disorders in newborn babies, but also inhibiting cancer, preventing vascular calcification, participating in bone metabolism, inhibiting diabetic cataract, and treating acute and chronic hepatitis. Since vitamin K₁ is sensitive to oxidation, alkali, strong acid and ultraviolet radiation, it is necessary to conduct pre-treatment methods, including saponification, enzymatic hydrolysis, organic solvent extraction, supercritical fluid extraction and purification and solid-phase extraction method with good effect. With the advantages and disadvantages of different pre-treatment methods, it is necessary to select a suitable pre-treatment method according to real samples. Meanwhile, vitamin K₁ was mainly detected by fluorescence spectrophotometry, liquid chromatography, liquid chromatographic mass spectrometry and other methods. An accurate, efficient and rapid detection method is also a hot research topic. This review summarized the pretreatment and detection methods of vitamin K₁ in different agricultural products in recent years. Finally, the trends and prospect of vitamin K₁ detection methods were proposed as follows (1) developing green solvent to improve extraction efficiency; (2) developing extraction materials to improve extraction selectivity and recovery rate; (3) Establishing simultaneous detection methods of vitamin K₁ and other nutrient functional components in agricultural products. This review systematically summarized the research progress of the physiological functions and detection methods of vitamin K₁ in agricultural products, and aimed to promote the in-depth study of nutrients in agricultural products and comparisons of advantages and disadvantages of pretreatment and detection methods.

Key words: agricultural products, Vitamin K₁, physiological function, pretreatment, detection method

徐月清,汪丹丹,赵新楠,喻理,王秀嫔,张良晓,张奇,张文,李培武. 维生素K₁生理功能及其在农产品中的检测方法[J]. 中国农业科学, 2019, 52(18): 3207-3217.

XU YueQing,WANG DanDan,ZHAO XinNan,YU Li,WANG XiuPin,ZHANG LiangXiao,ZHANG Qi,ZHANG Wen,LI PeiWu. Advances in Research on Physiological Functions of Vitamin K₁and Its Detection Methods in Agricultural Products[J]. Scientia Agricultura Sinica, 2019, 52(18): 3207-3217.

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图/表 1

表1

农产品食品中维生素K1的前处理和检测技术"

前处理方法 Pretreatment method	检测技术 Detection Technology	样品 Sample	优点 Advantage	缺点 Disadvantage	参考文献 References
有机溶剂（丙酮、石油醚）提取,氧化铝柱净化 Extraction of organic solvents (acetone, petroleum ether), purification by alumina column	HPLC	小葱、小白菜等 Shallots, Chinese cabbage, etc.	特异性强,分离效果好,基线稳定。灵敏、精密度高,分析速度快 Strong specificity, good separation and stable baseline. High sensitivity and precision, fast analysis	前处理时间长且处理过程复杂 Long pre-processing time and complicated processing	[12]
固相萃取 Solid phase extraction	HPLC	豆油、黄豆等 Soybean oil, soybeans, etc.	SPE柱对样品的富集和净化效果好,操作简便 The SPE column has good enrichment and purification effect on the sample and is easy to operate.	回收率偏低,有机溶剂用量大 Low recovery rate and large amount of organic solvent	[72]
酶解法,皂化法 Enzymatic hydrolysis, saponification	HPLC	小米、大米、玉米、面粉、麦片 Millet, rice, corn, flour, cereal	分离效果好,基线稳定,精密度和准确度高 Good separation, stable baseline, high precision and accuracy	维生素K₁遇碱易分解,皂化处理容易破坏样品中的维生素K₁ Vitamin K₁ is easily decomposed by alkali, and saponification treatment easily destroys vitamin K₁ in the sample	[52]
超临界萃取精馏塔提取 Supercritical extraction distillation column extraction	HPLC	维生素K₁粗品 Vitamin K₁ crude	步骤简单,无溶剂残留 Simple steps, no solvent residue	仪器设备昂贵 Expensive equipment	[57]
三氯甲烷提取 Chloroform extraction	反相HPLC Reversed phase HPLC	刺儿菜 Stinger	操作简便,分析速度快 Easy to operate, fast analysis	使用大量溶剂,溶剂回收成本高,样品未经净化或净化不完全 The use of a large amount of solvent, solvent recovery cost is high, the sample is not purified or incomplete purification	[54]
丙醇﹕正己烷（3﹕2）提取,SPE柱净化 Propanol﹕n-hexane (3﹕2) extraction, SPE column purification	GC-MS	西兰花 Broccoli	提取效果好,检测速度快 Good extraction and fast detection	有机溶剂用量多,仪器昂贵 Large amount of organic solvent, expensive instrument	[73]
甲醇﹕水（2﹕8） Methanol﹕water (2﹕8)	紫外光诱导荧光检测 Ultraviolet light induced fluorescence detection	血清 Serum	前处理简单,有机试剂用量少 Simple pretreatment and low dosage of organic reagents	某些荧光持续时间短,因此实验速度要快 Some fluorescences have a short duration, so the experiment is faster	[13]
异丙醇和正己烷提取,中性氧化铝固相萃取柱净化 Isopropanol and n-hexane extraction, neutral alumina column solid phase extraction purification	HPLC-MS/MS	四季豆、茄子、草莓等31种水果蔬菜 31 kinds of fruits and vegetables such as green beans, eggplant and strawberries	HPLC-MS/MS具有更高的选择性和灵敏度,净化效果好,直接测定 HPLC-MS/MS has higher selectivity and sensitivity, and the purification effect is good	分析步骤繁琐,精密度不高,仪器设备昂贵 The analysis steps are cumbersome, the precision is not high, and the equipment is expensive	[50]
异丙醇和正己烷提取,中性氧化铝柱净化 Isopropanol and n-hexane extraction, neutral alumina column purification	HPLC-MS/MS	菠菜、莲藕、葡萄 Spinach, lotus root, grapes	基线稳定,灵敏、精密度高,分析速度快 Stable baseline, sensitive, high precision, fast analysis	前处理时间长且处理过程复杂 Long pre-processing time and complicated processing	[71]
正庚烷﹕乙酸乙酯（4﹕1）萃取,固相萃取柱净化 N-heptane﹕ethyl acetate (4﹕1) extraction, solid phase extraction column purification	LC-APCI-MS/MS	菠菜、豌豆、苹果、香蕉、甜菜根和蘑菇 Spinach, peas, apples, bananas, beetroots and mushrooms	提取和净化效果好,回收率高,重现性好 Good extraction and purification, high recovery rate and good reproducibility	固相萃取柱成本较高 Solid phase extraction cartridges cost more	[66]

表1

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维生素K₁生理功能及其在农产品中的检测方法

Advances in Research on Physiological Functions of Vitamin K₁and Its Detection Methods in Agricultural Products

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