实验方法与实验动物 EXPERIMENTAL METHOD AND ANIMAL

固相萃取-反相高效液相色谱法测定蛋黄中的斑蝥黄

  • 王凤芹 ,
  • 刘鑫 ,
  • 程远之 ,
  • 郭冠群 ,
  • 路则庆 ,
  • 汪以真
展开
  • 浙江大学饲料科学研究所, 农业农村部(华东)动物营养与饲料重点实验室, 浙江省饲料与动物营养重点实验室, 杭州 310058
王凤芹(1976-),女,湖北黄梅人,高级实验师,博士,主要研究方向为饲料与动物产品质量与安全评价。E-mail:wangfq@zju.edu.cn

收稿日期: 2020-12-13

  网络出版日期: 2021-08-11

基金资助

现代农业生猪产业体系(CARS-36);浙江省高校实验室工作研究项目(ZD202002)

Determination of Canthaxanthin in Egg Yolk Based on Solid-Phase Extraction and Reversed-Phase High-Performance Liquid Chromatography Method

  • WANG Fengqin ,
  • LIU Xin ,
  • CHENG Yuanzhi ,
  • GUO Guanqun ,
  • LU Zeqing ,
  • WANG Yizhen
Expand
  • Key Laboratory of Animal Feed and Nutrition of Zhejiang Province, Key Laboratory of Animal Nutrition and Feed Science (Eastern of China), Ministry of Agriculture and Rural Affairs, Feed Science Institute of Zhejiang University, Hangzhou 310058, China

Received date: 2020-12-13

  Online published: 2021-08-11

摘要

本试验旨在建立一种定量测定蛋黄中斑蝥黄的固相萃取-反相高效液相色谱法。蛋黄用乙腈在超声波辅助下提取,提取液经过Oasis PRiME HLB固相萃取柱净化,再由C18色谱柱分离,流动相为乙腈-水(95 : 5,体积比),流速为1.2 mL/min,等度洗脱,在紫外-可见吸收波长474 nm处检测,外标法定量。结果显示:蛋黄中斑蝥黄的检出限为0.05 μg/g,定量限为0.1 μg/g。斑蝥黄在0.1~10.0 μg/mL的浓度范围内线性关系良好,相关系数(R2)为0.999 9。在3个加标浓度(0.1、1.0和4.0 μg/g)下,斑蝥黄的回收率为85.9%~98.6%,相对标准偏差(RSD)为1.7%~8.0%。应用上述方法测得市售新鲜鸡蛋蛋黄中斑蝥黄含量高达8.3 μg/g。本试验建立的固相萃取-反相高效液相色谱法能够满足蛋黄中斑蝥黄及其异构体快速定量的需要。

本文引用格式

王凤芹 , 刘鑫 , 程远之 , 郭冠群 , 路则庆 , 汪以真 . 固相萃取-反相高效液相色谱法测定蛋黄中的斑蝥黄[J]. 动物营养学报, 2021 , 33(8) : 4675 -4683 . DOI: 10.3969/j.issn.1006-267x.2021.08.046

Abstract

A simplified solid-phase extraction (SPE) and reversed-phase high-performance liquid chromatography (RP-HPLC) method for the quantitative determination of canthaxanthin in egg yolk was established in this experiment. The egg yolk sample was extracted using acetonitrile and cleaned up on Oasis PRiME HLB SPE cartridges, the separation was achieved on a C18 column using acetonitrile and water (95:5, v/v) at a flow rate of 1.2 mL/min of isocratic elution. Canthaxanthin was quantified by an external standard method coupled to a diode array detector at 474 nm. The results showed that the limit of detection was 0.05 μg/g and the limit of quantification was 0.1 μg/g. Within the range of canthaxanthin concentration at 0.1 to 10.0 μg/mL, the linear relationship was good, and the correlation coefficient (R2) was 0.999 9. Under the three spiked concentrations (0.1, 1.0 and 4.0 μg/g), the recovery of canthaxanthin was 85.9% to 98.6%, and the relative standard deviation (RSD) was 1.7% to 8.0%. Measured by the above method, the canthaxanthin content in egg yolk of commercial fresh eggs up to 8.3 μg/g. The SPE and RP-HPLC method developed in this experiment can be used for the fast analysis of canthaxanthin isomers in egg yolk.

参考文献

[1] BRULC L,SIMONOVSKA B,VOVK I,et al.Determination of egg yolk xanthophylls by isocratic high-performance liquid chromatography[J].Journal of Chromatography A,2013,1318:134-141.
[2] WENZEL M,SEUSS-BAUM I,SCHLICH E.Influence of pasteurization,spray- and freeze-drying,and storage on the carotenoid content in egg yolk[J].Journal of Agricultural and Food Chemistry,2010,58(3):1726-1731.  
[3] CHOI C,KOO S.Efficient syntheses of the keto-carotenoids canthaxanthin,astaxanthin,and astacene[J].Journal of Organic Chemistry,2005,70(8):3328-3331.  
[4] WEBER G M,MACHANDER V,SCHIERLE J,et al.Tolerance of poultry against an overdose of canthaxanthin as measured by performance,different blood variables and post-mortem evaluation[J].Animal Feed Science and Technology,2013,186(1/2):91-100.
[5] SURAI P F.The antioxidant properties of canthaxanthin and its potential effects in the poultry eggs and on embryonic development of the chick.Part 2[J].World's Poultry Science Journal,2012,68(4):717-726.  
[6] FURUSAWA N.A simple and small-scale sample preparation technique to determine canthaxanthin in hen egg yolk[J].Food Chemistry,2011,124(4):1643-1646.  
[7] FURUSAWA N.A simplified and small-scale sample preparation technique for determining astaxanthin,canthaxanthin,and β-apo-8'-carotenoic acid ethyl ester in hen's egg yolk[J].Acta Chromatographica,2014,26(4):587-597.  
[8] BONONI M,COMMISSATI I,LUBIAN E,et al.A simplified method for the HPLC resolution of alpha-carotene and beta-carotene (trans and cis) isomers[J].Analytical and Bioanalytical Chemistry,2002,372(2):401-403.  
[9] SCHLATTERER J,BREITHAUPT D E.Xanthophylls in commercial egg yolks:quantification and identification by HPLC and LC-(APCI)MS using a C30 phase[J].Journal of Agricultural and Food Chemistry,2006,54(6):2267-2273.  
[10] NIMALARATNE C,LOPES-LUTZ D,SCHIEBER A,et al.Effect of domestic cooking methods on egg yolk xanthophylls[J].Journal of Agricultural and Food Chemistry,2012,60(51):12547-12552.  
[11] TSIAKA T,LANTZOURAKI D Z,SIAPI E,et al.Macular carotenoids in lipid food matrices:DOE-based high energy extraction of egg yolk xanthophylls and quantification through a validated APCI(+) LC-MS/MS method[J].Journal of Chromatography B:Analytical Technologies in the Biomedical and Life Sciences,2018,1096:160-171.
[12] WENZEL M,SEUSS-BAUM I,SCHLICH E.Influences of storage time and temperature on the xanthophyll content of freeze-dried egg yolk[J].Food Chemistry,2011,124(4):1343-1348.  
[13] LI H X,TYNDALE S T,HEATH D D,et al.Determination of carotenoids and all-trans-retinol in fish eggs by liquid chromatography-electrospray ionization-tandem mass spectrometry[J].Journal of Chromatography B:Analytical Technologies in the Biomedical and Life Sciences,2005,816(1/2):49-56.
[14] VENUGOPALAN V,TRIPATHI S K,NAHAR P,et al.Characterization of canthaxanthin isomers isolated from a new soil Dietzia sp. and their antioxidant activities[J].Journal of Microbiology and Biotechnology,2013,23(2):237-245.  
[15] LARSEN E,CHRISTENSEN L P.Simple saponification method for the quantitative determination of carotenoids in green vegetables[J].Journal of Agricultural and Food Chemistry,2005,53(17):6598-6602.  
[16] HE K H,ZOU X L,LIU X,et al.Determination of canthaxanthin and astaxanthin in egg yolks by reversed phase high performance liquid chromatography with diode array detection[J].Journal of Sichuan University (Medical Science Edition),2012,43(1):113-117.
[17] 中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会.河豚鱼、鳗鱼和烤鳗中角黄素残留量的测定液相色谱-紫外检测法:GB/T 22958-2008[S].北京:中国标准出版社,2008. General Administration of Quality Supervision,Inspection and Quarantine of the People's Republic of China,Standardization Administration of the People's Republic of China.Determination of canthaxanthin residues in fugu,eel and baked eel-HPLC-UV method:GB/T 22958-2008[S].Beijing:Standards Press of China,2008.(in Chinese)
[18] 中华人民共和国国家质量监督检验检疫总局.饲料添加剂10%β,β-胡萝卜-4,4-二酮(10%斑蝥黄):GB/T 18970-2003[S].北京:中国标准出版社,2003. General Administration of Quality Supervision,Inspection and Quarantine of the People's Republic of China.Feed additive-10% 4,4'-diketo-β-carotene (10% canthaxanthin):GB/T 18970-2003[S].Beijing:Standards Press of China,2003.(in Chinese)
文章导航

/