Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (05): 706-713.doi: 10.3724/SP.J.1006.2016.00706

• TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY • Previous Articles     Next Articles

Establishment of Ultra Performance Liquid Chromatography (UPLC) Protocol for Analyzing Carotenoids in Common Wheat

LI Wen-Shuang1,XIA Xian-Chun1,HE Zhong-Hu1,2,*   

  1. 1Institute of Crop Science / National Wheat Improvement Center, Chinese Academy of Agricultural Sciences (CAAS), Beijing 100081, China; 2CIMMYT-China Office, c/o CAAS, Beijing 100081, China
  • Received:2015-09-07 Revised:2016-03-02 Online:2016-05-12 Published:2016-03-11
  • Contact: He Zhonghu, E-mail: zhhecaas@163.com E-mail:lwshuang2012@163.com
  • Supported by:

    This study was supported by the Program of Introducing International Super Agricultural Science and Technology (2011-G(3)4) and Ministry of Science and Technology (2013DFG30530).

Abstract:

Carotenoids is an important criterion in the assessment of color and nutritional qualities of end-use products in common wheat. In this study, grain powders of Zhongmai 175 (soft wheat) and Zhongyou 206 (hard wheat) were used as samples to develop an optimal procedure for extraction and separation of carotenoids compositions extracted from flours using ultra performance liquid chromatography (UPLC). The most effective extraction of carotenoids extraction was obtained using the solvent system of N-hexane; acetone of 80:20 (v/v, 0.1% BHT w/v) under the oscillation condition of 300 rpm, 35ºC, and 1 h. The separation was conducted using YMC C30 Carotenoid column (100.0 mm × 4.6 mm, 3 µm) with photodiode array (PDA) detector, and the column was thermostated at 35°C. Under a gradient system consisting of acetonitrile: methanol: water: triethylamine (81:14:5:0.05, v/v/v/v) (A) and methanol: ethylacetate:triethylamine (68:32:0.05, v/v/v) (B) at a constant flow rate of 0.4 mL min–1, the carotenoids of common wheat flour samples can be well separated in less than 25 min. Carotenoids were detected at 450 nm. All the present results provided useful information for carotenoids compositions and quality improvement.

Key words: Triticum aestivum L., Carotenodis compositions, UPLC

[1]Fawzi W W, Hunter D J. Vitamins in HIV disease progression and vertical transmission. Epidemiology, 1998, 9: 457–466
[2]Ribaya-Mercado J D, Blumberg J B. Lutein and zeaxanthin and their potential roles in disease prevention. J Am Coll Nut, 2004, 23: 567S–587S
[3]Nishino H, Murakoshi M, Tokuda H, Yoshiko S. Cancer prevention by carotenoids. Arch Biochem Biophys, 2009, 483: 165–168
[4]Harjes C E, Rocheford T R, Bai L, Brutnell T P, Kandianis C B, Sowinski S G, Stapleton A E, Vallabhaneni R, Williams M, Wurtzel E T, Yan J B, Buckler E S. Natural genetic variation in lycopene epsilon cyclase tapped for maize biofortification. Science, 2008, 319: 330–333
[5]Cazzonelli C I, Pogson B J. Source to sink: regulation of carotenoid biosynthesis in plants. Trends Plant Sci, 2010, 15: 266–274
[6]Howitt C A, Pogson B J. Carotenoid accumulation and function in seeds and non-green tissues. Plant Cell Environ, 2006, 29: 435–445
[7]Matthews P D, Luo R B, Wurtzel E T. Maize phytoene desaturase and zeta-carotene desaturase catalyse a poly-Z desaturation pathway: implications for genetic engineering of carotenoid content among cereal crops. J Exp Bot, 2003, 54: 2215–2230
[8]Singh M, Lewis P E, Hardeman K, Bai L, Rose J K C, Mazourek M, Chomet P, Brutnell T P. Activator mutagenesis of the pink scutellum1/viviparous7 locus of maize. Plant Cell, 2003, 15: 874–884
[9]Gallagher C E, Matthews P D, Li F Q, Wurtzel E T. Gene duplication in the carotenoid biosynthetic pathway preceded evolution of the grasses. Plant Physiol, 2004, 135:1776–1783
[10]Li F, Murillo C, Wurtzel E T. Maize Y9 encodes a product essential for 15-cis-ζ-carotene isomerization. Plant Physiol, 2007, 144: 1181–1189
[11]Wong J C, Lambert R J, Wurtzel E T, Rocheford T R. QTL and candidate genes phytoene synthase and zeta-carotene desaturase associated with the accumulation of carotenoids in maize. Theor Appl Genet, 2004, 108: 349–359
[12]Chander S, Guo Y, Yang X, Zhang J, Lu X, Yan J, Song T, Rocheford T, Li J. Using molecular markers to identify two major loci controlling carotenoid contents in maize grain. Theor Appl Genet, 2008, 116: 223–233
[13]Ye X D, Al-Babili S, Kloti A, Zhang J, Lucca P, Beyer P, Potrykus I. Engineering the provitamin A (beta-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. Science, 2000, 287: 303–305
[14]Beyer P, Al-Babili S, Ye X D, Lucca P, Schaub P, Welsch R, Potrykus I. Golden rice: Introducing the beta-carotene biosynthesis pathway into rice endosperm by genetic engineering to defeat vitamin A deficiency. J Nutr, 2002, 132: 506S–510S
[15]Paine J A, Shipton C A, Chaggar S, Howells R M, Kennedy M J, Vernon G, Wright S Y, Hinchliffe E, Adams J L, Silverstone A L, Drake R. Improving the nutritional value of Golden Rice through increased pro-vitamin A content. Nat Biotechnol, 2005, 23: 482–487
[16]Gupta P K, Mir R R, Mohan A, Kumar J. Wheat genomics: present status and future prospects. Int J Plant Genomics, 2008, doi: 10.1155/2008/896451
[17]Jin H, Zhang Y, Li G Y, Mu P Y, Fan Z R, Xia X C, He Z H. Effects of allelic variation of HMW-GS and LMW-GS on mixograph properties and Chinese noodle and steamed bread qualities in a set of Aroona near-isogenic wheat lines. J Cereal Sci, 2013, 57: 146–152
[18]Mares D J, Campbell A. Mapping components of flour and noodle colour in Australian wheat. Aust J Agr Res, 2001, 52: 1297–1310
[19]何中虎, 晏月明, 庄巧生, 张艳, 夏先春, 张勇, 王德森, 夏兰芹, 胡英考, 蔡民华, 陈新民, 阎俊, 周阳. 中国小麦品种品质评价体系建立与分子改良技术研究. 中国农业科学 2006, 39: 1091–1101
He Z H, Yan Y M, Zhuang Q S, Zhang Y, Xia X C, Zhang Y, Wang D S, Xia L Q, Hu Y K, Cai M H, Chen X M, Yan J, Zhou Y. Establishment of quality evaluation system and utilization of molecular methods for the improvement of Chinese wheat quality. Sci Agric Sin, 2006, 39: 1091–1101 (in Chinese with English abstract)
[20]Symons S J, Dexter J E. Computer analysis of fluorescence for the measurement of flour refinement as determined by flour ash content, flour grade colour, and tristimulus colour measurements. Cereal Chem, 1991, 68: 454–460
[21]He X Y, Zhang Y L, He Z H, Wu Y P, Xiao Y G, Ma C X, Xia X C. Characterization of phytoene synthase 1 gene (Psy1) located on common wheat chromosome 7A and development of a functional marker. Theor Appl Genet, 2008, 116: 213–221
[22]Dong C H, Ma Z Y, Xia X C, Zhang L P, He Z H. Allelic variation at the TaZds-A1 locus on wheat chromosome 2A and development of a functional marker in common wheat. J Integr Agric 2012, 11: 1067–1074
[23]Abdel-Aal E S M, Young J C, Wood P J, Rabalski I, Hucl P, Fregeau-Reid J. Einkorn: A potential candidate for developing high lutein wheat. Cereal Chem, 2002, 79: 455–457
[24]Adom K K, Sorrells M E, Liu R H. Phytochemical profiles and antioxidant activity of wheat varieties. J Agric Food Chem, 2003, 51: 7825–7834
[25]Hung P V, Hatcher D W. Ultra-performance liquid chromatography (UPLC) quantification of carotenoids in durum wheat: Influence of genotype and environment in relation to the colour of yellow alkaline noodles (YAN). Food Chem, 2011, 125: 1510–1516
[26]周毅, 付志远, 李青, 徐淑兔, Chander Subhash, 杨小红, 李建生, 严建兵. 高油和普通玉米自交系类胡萝卜素和生育酚含量的比较. 作物学报, 2009, 35: 2073–2084
Zhou Y, Fu Z Y, Li Q, Xu S T, Chander S, Yang X H, Li J S, Yan J B. Comparative analysis of carotenoid and tocopherol compositions in high-oil and normal Maize (Zea mays L.) inbreds. Acta Agron Sin, 2009, 35: 2073–2084 (in Chinese with English abstract)
[27]Digesù A, Platani C, Cattivelli L, Mangini G, Blanco A. Genetic variability in yellow pigment components in cultivated and wild tetraploid wheats. J Cereal Sci, 2009, 50: 210–218
[28]Hidalgo A, Brandolini A, Pompei C, Piscozzi R. Carotenoids and tocols of einkorn wheat (Triticum monococcum ssp. monococcum L.). J Cereal Sci, 2006, 44: 182–193
[29]Ndolo V U, Beta T. Distribution of carotenoids in endosperm, germ, and aleurone fractions of cereal grain kernels. Food Chem, 2013, 139: 663–671
[30]Burkhardt S, Böhm V. Development of a new method for the complete extraction of carotenoids from cereals with special reference to durum wheat (Triticum durum Desf.). J Agric Food Chem, 2007, 55: 8295–8301
[31]Oliver J, Palou A, Pons A. Semi-quantification of carotenoids by high-performance liquid chromatography: saponification-induced losses in fatty foods. J Chromatogr A, 1998, 829: 393–399
[32]Konopka I, Czaplicki S, Rotkiewicz D. Differences in content and composition of free lipids and carotenoids in flour of spring and winter wheat cultivated in Poland. Food Chem, 2006, 95: 290–300
[33]Abdel-Aal El-SM, Young J C, Rabalski I, Hucl P, Fregeau-Reid J. Identification and quantification of seed carotenoids in selected wheat species. J Agric Food Chem, 2007, 55: 787–794
[34]Mashaba C S, Barros E. Screening South African potato, tomato, and wheat cultivars for five carotenoids. SAf J Sci, 2011, 107: 34–39
[35]Lookhart G L, Bean S R, Bietz J A. Reversed-phase high performance liquid chromatography in grain applications. Cereal Food World, 2002, 48: 9–16
[36]Kean E G, Hamaker B R, Ferruzzi M G. Carotenoid bioaccessibility from whole grain and degermed maize meal products. J Agric Food Chem, 2008, 56: 9918–9926

[1] NIU Xin-Ning,WANG Bu-Jun. Evaluation of matrix reference material of Fumonisins FB1 in corn flour [J]. Acta Agronomica Sinica, 2020, 46(7): 1128-1133.
[2] Li-Na WANG,Bu-Jun WANG. Analysis method of wheat germination metabolomics based on UPLC-QTOF/ MS [J]. Acta Agronomica Sinica, 2019, 45(12): 1899-1904.
[3] XU Wen,SHEN Hao,GUO Jun,YU Xiao-Cong,LI Xiang,YANG Yan-Hui,MA Xiao,ZHAO Shi-Jie,SONG Jian-Min. Drought Resistance of Wheat NILs with Different Cuticular Wax Contents in Flag Leaf [J]. Acta Agron Sin, 2016, 42(11): 1700-1707.
[4] ZHAO De-Hui,YAN Jun,HUANG Yu-Lian,XIA Xian-Chun,ZHANG Yan,TIAN Yu-Bing,HE Zhong-Hu,ZHANG Yong. Effect of 1BL/1RS Translocation on Gluten Protein Fraction Quantities and Dough Rheological Properties [J]. Acta Agron Sin, 2015, 41(11): 1648-1656.
[5] LIU Jin-Dong,YANG En-Nian,XIAO Yong-Gui,CHEN Xin-Min,WU Ling,BAI Bin,LI Zai-Feng,Garry M. ROSEWARNE,XIA Xian-Chun,HE Zhong-Hu. Development, Field and Molecular Characterization of Advanced Lines with Pleiotropic Adult-Plant Resistance in Common Wheat [J]. Acta Agron Sin, 2015, 41(10): 1472-1480.
[6] LIU Zheng-Shuai,LIU Gui-Fen,YANG Ming-Yu,JIA Xiao,LI Yun-Xiang,ZHAO Fa-Mao. Constitution and Spatiotemporal Expression of Starch Branching Enzyme in Developing Wheat Grain [J]. Acta Agron Sin, 2014, 40(12): 2176-2182.
[7] ZHU Yu-Lei,WANG Sheng-Xing,ZHAO Liang-Xia,ZHANG De-Xin,HU Jian-Bang,CAO Xue-Lian,YANG Ya-Jie,CHANG Cheng,MA Chuan-Xi,ZHANG Hai-Ping. Exploring Molecular Markers of Preharvest Sprouting Resistance Gene Using Wheat Intact Spikes by Association Analysis [J]. Acta Agron Sin, 2014, 40(10): 1725-1732.
[8] LIU Jin-Dong,CHEN Xin-Min,HE Zhong-Hu,WU Ling,BAI Bin,LI Zai-Feng,XIA Xian-Chun. Resistance of Slow Mildewing Genes to Stripe Rust and Leaf Rust in Common Wheat [J]. Acta Agron Sin, 2014, 40(09): 1557-1564.
[9] ZHANG Wei,SUN Hong,WEI Xiao-Jing,XING Li-Ping,WANG Hua-Zhong. Cloning of Autophagy-Related Genes, ATG10s, in Wheat and Their Expression Characteristics Induced by Blumeria graminis f. sp. tritici [J]. Acta Agron Sin, 2014, 40(08): 1392-1402.
[10] UN Juan,LI Wei-Xi,ZHANG Yan,SUN Li-Juan,DONG Xiao-Li,HU Xue-Xu,WANG Bu-Jun. Simultaneous Determination of Twelve Mycotoxins in Cereals by Ultra-high Performance Liquid Chromatography-Tandem Mass Spectrometry [J]. Acta Agron Sin, 2014, 40(04): 691-701.
[11] WANG Xuan,JU Li-Ping,LIU Fang-Jun,ZHANG Yu-Yu,ZHANG Fan,FU Xiao-Jie,FENG Yi,ZHANG Xiao-Ke. Vernalization Effects of Dominant Alleles Vrn-B1a and Vrn-B1b and Their Distributions in Cultivars from Yellow and Huai River Valleys Facultative Winter Wheat Zone [J]. Acta Agron Sin, 2014, 40(03): 439-446.
[12] LIU Ya-Nan,XIA Xian-Chun,HE Zhong-Hu. Characterization of Dense and Erect Panicle 1 Gene (TaDep1) Located on Common Wheat Group 5 Chromosomes and Development of Allele-Specific Markers [J]. Acta Agron Sin, 2013, 39(04): 589-598.
[13] XIA Jia-Ping,GUO Hui-Jun,XIE Yong-Dun,ZHAO Lin-Shu,GU Jia-Yu,ZHAO Shi-Rong,LI Jun-Hui,LIU Lu-Xiang. Differential Expression of Chloroplast Genes in Chlorophyll-Deficient Wheat Mutant Mt135 Derived from Space Mutagenesis [J]. Acta Agron Sin, 2012, 38(11): 2122-2130.
[14] SONG Su-Jie,GU Jia-Yu,GUO Hui-Jun,ZHAO Lin-Shu,ZHAO Shi-Rong,LI Jun-Hui, ZHAO Bao-Cun,LIU Lu-Xiang. Proteomic Analysis of Leaves of the Chlorophyll-Deficient Wheat Mutant Mt6172 and Its Wild-Type through 2D-Difference Gel Electrophoresis [J]. Acta Agron Sin, 2012, 38(09): 1592-1606.
[15] LI Wei-Yu,ZHANG Bin,ZHANG Jia-Nan,CHANG Xiao-Ping,LI Run-Zhi,JING Rui-Lian. Exploring Elite Alleles for Chlorophyll Content of Flag Leaf in Natural Population of Wheat by Association Analysis [J]. Acta Agron Sin, 2012, 38(06): 962-970.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!