Determination of astaxanthin in astaxanthin oil by high performance liquid chromatography
KONG Fanhua2, XU Jiajia2, GUO Qian2, BAI Shasha2, LI Dong2, FANG Congrong1, QIU Nannan1*, CUI Yajuan2*
1(NHC Key Laboratory of Food Safety Risk Assessment, Food Safety Research Unit of Chinese Academy of Medical Science, China National Center for Food Safety Risk Assessment, Beijing 100021, China) 2(Beijing Institute of Nutrition Resources, Beijing 100069, China)
Abstract: An analytical method was developed for the detection of astaxanthins in astaxanthin oil. The extraction reagent and enzymolysis time of astaxanthins in astaxanthin oil were selected by single factor experiment. The optimized pretreatment condition was bath shaking zymolytic for 60 min at 37 ℃ and petroleum ether extraction were selected for the extraction. The separation of astaxanthins were achieved on YMCTM C30 column(250 mm×4.6 mm, 5μm)using methanol-methyl tert-butyl ether as the mobile phase at a flow rate of 1.0 mL/min. The detection wavelength and column temperature were set at 472 nm and 30 ℃, respectively. The results showed that the linear range of all trans astaxanthin was 0.10-4.00 μg/mL. The relative standard deviation (RSD) of astaxanthin in astaxanthin oil was 1.36%. Moreover, the average recovery rate of astaxanthin was 93.58% and RSD was 2.84%, indicating high recovery rate and small relative standard deviation. The astaxanthin content in astaxanthin oil by this method is 92.9% which was 2.11 times of alkaline saponification content. Therefore, the method established can accurately quantify astaxanthin content in astaxanthin oil.
孔凡华,徐佳佳,郭倩,等. 高效液相色谱法测定虾青素油中虾青素的含量[J]. 食品与发酵工业, 2021, 47(6): 214-220.
KONG Fanhua,XU Jiajia,GUO Qian,et al. Determination of astaxanthin in astaxanthin oil by high performance liquid chromatography[J]. Food and Fermentation Industries, 2021, 47(6): 214-220.
AL-AMIN M M, AKHTER S, HASAN A T, et al.The antioxidant effect of astaxanthin is higher in young mice than aged:A region specific study on brain[J].Metabolic Brain Disease,2015, 30(5):1 237-1 246.
[2]
BOROWITZKA M A.High-value products from microalgae-their development and commercialisation[J].Journal of Applied Phycology,2013, 25(3):743-756.
[3]
KOLLER M, MUHR A, BRAUNEGG G.Microalgae as versatile cellular factories for valued products[J].Algal Research,2014, 6:52-63.
[4]
PÉREZ-LÓPEZ P, GONZÁLEZ-GARCÍA S, JEFFRYES C, et al.Life cycle assessment of the production of the red antioxidant carotenoid astaxanthin by microalgae:From lab to pilot scale[J].Journal of Cleaner Production,2014, 64(64):332-344.
[5]
ANDERSEN LP, HOLCK S, KUPCINSKAS L, et al.Gastric inflammatory markers and interleukins in patients with functional dyspepsia treated with astaxanthin[J].Fems Immunology & Medical Microbiology,2007, 50(2):244-248.
[6]
CHEW B P, PARK J S.Carotenoid action on the immune response[J].Journal of Nutrition,2004, 134(1):257-261.
[7]
MIYAWAKI H, TAKAHASHI J, TSUKAHARA H, et al.Effects of astaxanthin on human blood rheology[J].Journal of Clinical Biochemistry and Nutrition,2008, 43(2):69-74.
[8]
GRIMMIG B, DALY L, HUDSON C, et al.Astaxanthin attenuates neurotoxicity in a mouse model of Parkinson's disease[J].Cell Transplantation,2016, 25(4):759-759.
[9]
NAKAO R, NELSON OLPARK J S, MATHISON B D, et al.Effect of astaxanthin supplementation on inflammation and cardiac function in BALB/c mice[J].Anticancer Research,2010, 30(7):2 721-2 725.
[10]
AMBATI R R, MOI P S, RAVI S,et al.Astaxanthin:Sources, extraction, stability, biological activities and its commercial applications- a review[J].Marine Drugs,2014, 12(1):128-152.
[11]
YANG C, ZHANG H, LIU R H, et al.Bioaccessibility, cellular uptake, and transport of astaxanthin isomers and their antioxidative effects in human intestinal epithelial Caco-2 cells[J].Journal of Agricultural and Food Chemistry,2017, 65(47):10 223-10 232.
[12]
ZHAO Y Y, GUAN F F, WANG G L, et al.Astaxanthin preparation by Lipase-Catalyzed hydrolysis of its esters from Haematococcus pluvialis algal extracts[J].Journal of Food Science,2011, 76(4):643-650.
[13]
ANTONOIO M,JURI R,PATRIZIA C,et al.Extraction of astaxanthin from microalga Haematococcus pluvialis in red phase by using genrally recognized as safe solvents and accelerated extraction[J].Journal of Biotechnology,2018,283:51-61.
[14]
国家质量监督检验检疫总局,国家标准化管理委员会.GB/T 31520—2015 红球藻中虾青素的测定液相色谱法[S].北京:中国标准出版社, 2015.General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China,Standardization Administration.GB/T 31520—2015 Determination of astaxanthin in Haematococcus-High performance liquid chromatography method[S].Beijing:Standards Press of China,2015.
[15]
农业农村部.SC/T 3053—2019 水产品及其制品中虾青素含量的测定高效液相色谱法[S].北京:中国标准出版社, 2019.Ministry of Agriculture and Rural Affairs of the People's Republic of China.SC/T 3053—2019 Determination of astaxanthin in fish and fishery products by high performance liquid chromatography method[S].Beijing:Standards Press of China,2019.
[16]
国家质量监督检验检疫总局.SN/T 2327—2009 进出口动物源性食品中角黄素、虾青素的检测方法[S].北京:中国标准出版社, 2009.General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China.SN/T 2327—2009 Determination of canthaxanthin and astaxanthin in foods of animal origin food for import and export[S].Beijing:Standards Press of China,2019.
[17]
中国医药保健品进出口商会.T/CCCMHPIE 1.21—2016 植物提取物 虾青素油[S].北京:中国标准出版社, 2016.China Chamber of Commerce of Medicines & Health Products Importers & Exporters.T/CCCMHPIE 1.21—2016 Plant extract-Astaxanthin Oil[S].Beijing:Standards Press of China,2016.
[18]
YUAN J P, CHEN F.Purification of trans-astaxanthin from a high-yielding astaxanthin ester-producing strain of the microalga Haematococcus pluvialis[J].Food Chem,2000, 68(4):443-448.
[19]
孙伟红, 冷凯良, 邢丽红, 等.GPC-HPLC 法测定南极磷虾油中虾青素及校正因子计算[J].分析试验室, 2013, 32(2):26-30.SUN W H,LENG K L,XING L H, et al.Determination of astaxanthin in Antarctic krill oil by GPC-HPLC and calculation of correction factor[J].Chinese Journal of Analysis Laboratory, 2013, 32(2):26-30.
[20]
杨澍,张婷,徐杰,等.高效液相色谱手性拆分法分析生物体内虾青素光学异构体[J].食品科学,2015,36(8):139-144.YANG S, ZHANG T, XU J, et al.Chiral separation and analysis of astaxanthin stereoisomers in biological organisms by high-performance liquid chromatography[J].Food Science,2015,36(8):139-144.
[21]
HOLTIN K, KUEHNLE M, REHBEIN J, et al.Determination of astaxanthin and astaxanthin esters in the microalgae Haematococcus pluvialis by LC-(APCI)MS and characterization of predominant carotenoid isomers by NMR spectroscopy[J].Analytical and Bioanalytical Chemistry, 2009, 395(6):1 613-1 622.
[22]
LÓPEZ-CERVANTES J, SÁNCHEZ-MACHADO D I, GUTIÉRREZ-CORONADO M A, et al.Quantification of astaxanthin in shrimp waste hydrolysate by HPLC[J].Biomedical Chromatography, 2006, 20(10):981-984.
[23]
MACHMUDAH S, SHOTIPRUK A, GOTO M, et al.Extraction of Astaxanthin from Haematococcus pluvialis Using Supercritical CO2 and Ethanol as Entrainer[J].Industrial & Engineering Chemistry Research, 2006, 45(10):3 652-3 657.
[24]
PENG J, WIANG W Z, TANG Q M, et al.Comparative analysis of astaxanthin and its esters in the mutant E1 of Haematococcus pluvialis and other green algae by HPLC with a C30 column[J].Science in China Series C, 2008, 51(12):1 108-1 115.
[25]
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.GB/T 27404—2008 食品安全国家标准 实验室质量控制规范 食品理化检测[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.GB/T 27404—2008 Criterion on quality control of laboratories—Chemical testing of food[S].Beijing:Standards Press of China,2008.
[26]
苏芳. 类胡萝卜素在藻虾蟹鱼中的结构分布特征及虾青素异构化研究[D].青岛:中国科学院大学,2018.SU F.A study on the structure distribution of carotenoids in algae, shrimps, crabs and fishes and the isomerization of astaxanthin[D].Qingdao:University of Chinese Academy of Sciences,2018.
[27]
邢俊波,姜春来,靳守东,等.HPLC内标手性拆分法分析保健食品雨生红球藻片中虾青素光学异构体[J].解放军药学学报,2016,32(1):10-12.XING J B,JIANG C L,JIN S D,et al.Chiral separation and analysis of astaxanthin stereoisomers in health food yushenghongqiuzao tablets by HPLC internal standard method[J].Pharm J Chin PLA,2016,32(1):10-12.
[28]
吴伟. 红法夫酵母生产虾青素过程关键技术的研究[D].武汉:华中科技大学, 2011.WU W.The study on the key techniques for the production of astaxanthin by fermentation in xanthophyllomyces dendrorhous[D].Wuhan:Huazhong University of Science and Technology, 2011.
[29]
苗凤萍. 雨生红球藻(Haematococcus pluvialis)虾青素酯和脂肪酸的鉴定及差异表达基因的分析[D].武汉:中国科学院研究生院(武汉植物园),2007.MIAO F P.Characterization on astaxanthin esters and fatty acids of haematococcus pluvialis and analysis of differentially expressed genes[D].Wuhan:Wuhan Botanical Garden, Chinese Academy of Sciences,2007.
[30]
张丽瑶,张华敏,王志祥,等.超声处理对虾青素稳定性和抗氧化性的影响[J].食品与药品,2018,20(4):264-267.ZHANG L Y, ZHANG H M, WANG Z X, et al.Effect of ultrasonic treatment on stability and antioxidant activity of astaxanthin[J].Food and Drug,2018,20(4):264-267.
2021, Vol. 47 
