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Quantification of Purple Corn (Zea mays L.) Anthocyanins Using Spectrophotometric and HPLC Approaches: Method Comparison and Correlation

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Abstract

Purple corncob is a rich source of anthocyanins with great potential as food colorant. Accurate quantification of anthocyanin content in cereals such as purple corn is critical to evaluate their nutritional value and facilitate their application in foods. Our objective was to determine the advantages and disadvantages of four common spectrophotometric and chromatographic methods to quantify cereal anthocyanins. Anthocyanins from 14 purple corncob samples were extracted, identified by high-performance liquid chromatography (HPLC)-PDA-MS, and quantified different methods commonly cited in the literature or used by analytical laboratories: total anthocyanins, pH differential, and HPLC methods with intact or acid hydrolyzed pigments. Polymeric color was also determined as it affects color quality. Among the four methods, the total anthocyanin method produced the highest value, followed by the pH differential method and HPLC with intact pigments, which produced very similar results, while the HPLC with hydrolyzed pigments produced the lowest values. The quantitative differences among the four methods are likely due to the differences in their specificity. Despite the differences in quantitative results, three of the methods, pH differential and both HPLC methods, showed good linear correlation (R 2 ≥ 0.98). In addition, the effect of wavelength selection and the criteria for HPLC integration were also evaluated. In summary, anthocyanin quantification results were dependent on the method chosen to quantify the pigments in the material. So, reporting methodology is critical and recommended when interpreting the anthocyanin quantification results for better inter-literature comparison.

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References

  • Abdel-Aal ES, Hucl P (1999) A rapid method for quantifying total anthocyanins in blue aleurone and purple pericarp wheats. Cereal Chem 76(3):350–354

    Article  CAS  Google Scholar 

  • Abdel-Aal ESM, Young JC, Rabalski I (2006) Anthocyanin composition in black, blue, pink, purple, and red cereal grains. J Agric Food Chem 54(13):4696–4704

    Article  CAS  Google Scholar 

  • Ahmadiani N, Robbins RJ, Collins TM, Giusti MM (2014) Anthocyanins contents, profiles, and color characteristics of red cabbage extracts from different cultivars and maturity stages. J Agric Food Chem 62(30):7524–7531

    Article  CAS  Google Scholar 

  • Aoki H, Kuze N, Kato Y (2002) Anthocyanins isolated from purple corn (Zea mays L.). Foods & Food Ingredients J Japan,199:41–45

  • Awika JM, Rooney LW, Waniska RD (2004) Properties of 3-deoxyanthocyanins from sorghum. J Agric Food Chem 52(14):4388–4394

    Article  CAS  Google Scholar 

  • Cevallos-Casals BA, Cisneros-Zevallos L (2003) Stoichiometric and kinetic studies of phenolic antioxidants from Andean purple corn and red-fleshed sweet potato. J Agric Food Chem 51:3313–3319

    Article  CAS  Google Scholar 

  • Dandena A, Leimane I (2011) Validation of monomeric anthocianin determination method for bilberry juice and marc extracts. Foodbolt, 93-97

  • de Pascual-Teresa S, Santos-Buelga C, Rivas-Gonzalo JC (2002) LC–MS analysis of anthocyanins from purple corn cob. J Sci Food Agric 82:1003–1006

    Article  Google Scholar 

  • Escribano-Bailón MT, Santos-Buelga C, Rivas-Gonzalo JC (2004) Anthocyanins in cereals. J Chromatogr A 1054(1):129–141

    Article  Google Scholar 

  • FAO. 2013. Traditional High Andean Cuisine. http://www.fao.org/docrep/018/i1466e/i1466e.pdf. Accessed 26 May 2014

  • Finkel ML, Sanchez S, Mak T, Granstein J, Lefkowitz A (2013) Anthocyanin-rich purple corn extract and its effects on the blood pressure of adults. J Evid-Based Complement Altern Med 18:237–242

    Article  CAS  Google Scholar 

  • Fukamachi K, ImadaT OY, Xu J, Tsuda H (2008) Purple corn color suppresses Ras protein level and inhibits 7,12-dimethylbenz[α]anthracene-induced mammary carcinogenesis in the rat. Cancer Sci 99:1841–1846

    Article  CAS  Google Scholar 

  • Fuleki T, Francis FJ (1968a) Quantitative methods for anthocyanins: 1. Extraction and determination of total anthocyanin in cranberries. J Food Sci 33:72–77

    Article  CAS  Google Scholar 

  • Fuleki T, Francis FJ (1968b) Quantitative methods for anthocyanins: 2. Determination of total anthocyanin and degradation index for cranberry juice. J Food Sci 33:78–83

    Article  CAS  Google Scholar 

  • Giusti MM, Wrolstad RE (2001) Characterization and measurement of anthocyanins by UV-visible spectroscopy. In: Wrolstad RE, Acree TE, An H, Decker EA, Penner MH, Reid DS, Schwartz SJ, Shoemaker CF, Sporns P (eds) Current protocols in food analytical chemistry, 1st edn. Wiley, New York, pp F1.2.1–1.2.13

    Google Scholar 

  • González-Manzano S, Pérez-Alonso JJ, Salinas-Moreno Y, Mateus N, Silva AMS, de Freitas V, Santos-Buelga C (2008) Flavanol–anthocyanin pigments in corn: NMR characterisation and presence in different purple corn varieties. J Food Compos Anal 21:521–526

    Article  Google Scholar 

  • González-Paramás AM, Lopes da Silva F, Martín-López P, Macz-Pop G, González-Manzano S, Alcalde-Eon C, Pérez-Alonso JJ, Escribano-Bailón MT, Rivas-Gonzalo JC, Santos-Buelga C (2006) Flavanol–anthocyanin condensed pigments in plant extracts. Food Chem 94:428–436

    Article  Google Scholar 

  • Grotewold E, Drummond BJ, Bowen B, Peterson T (1994) The myb-homologous P gene controls phlobaphene pigmentation in maize floral organs by directly activating a flavonoid biosynthetic gene subset. Cell 76:543–553

    Article  CAS  Google Scholar 

  • Harborne JB (1967) The anthocyanin pigments. In: Comparative biochemistry of the flavonoids. Academic Press: New York. pp 1-30

  • Jing P, Giusti MM (2005) Characterization of anthocyanin-rich waste from purple corncobs (Zea mays L.) and its application to color milk. J Agric Food Chem 53:8775–8781

    Article  CAS  Google Scholar 

  • Jing P, Giusti MM (2007) Effects of extraction conditions on improving the yield and quality of an anthocyanin-rich purple corn (Zea mays L.) color extract. J Food Sci 72(7):C363–C368

    Article  CAS  Google Scholar 

  • Jing P, Noriega V, Schwartz SJ, Giusti MM (2007) Effects of growing conditions on purple corncob (Zea mays L.) anthocyanins. J Agric Food Chem 55:8625–8629

    Article  CAS  Google Scholar 

  • Jing P, Bomser JA, Schwartz SJ, He J, Magnuson BA, Giusti MM (2008) Structure-function relationships of anthocyanins from various anthocyanin-rich extracts on the inhibition of colon cancer cell growth. J Agric Food Chem 56:9391–9398

    Article  CAS  Google Scholar 

  • Kulling SE, Rawel HM (2008) Chokeberry (Aronia melanocarpa)—a review on the characteristic components and potential health effects. Planta Med 74(13):1625–1634

    Article  CAS  Google Scholar 

  • Kupic T (2004) Quality-control analytical methods: high-performance liquid chromatography. Int J Pharm Compd 8:223–227

    Google Scholar 

  • Lee J, Durst RW, Wrolstad RE (2005) Determination of total monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants, and wines by the pH differential method: collaborative study. J AOAC Int 88:1269–1278

    CAS  Google Scholar 

  • Lee J, Rennaker C, Wrolstad RE (2008) Correlation of two anthocyanin quantification methods: HPLC and spectrophotometric methods. Food Chem 110:782–786

    Article  CAS  Google Scholar 

  • Lee SG, Nam TG, Kim DO, Koo SI, Chun OK (2012) Comparison of analytical methods for anthocyanin quantification in berries: HPLC and pH differential methods. FASEB J. http://www.fasebj.org/cgi/content/meeting_abstract/26/1_MeetingAbstracts/lb316. Accessed 7 July 2014

  • Leja M, Mareczek A, Ben J (2003) Antioxidant properties of two apple cultivars during long-term storage. Food Chem 80:303–307

    Article  CAS  Google Scholar 

  • Li CY, Kim HW, Won SR, Min KJ, Park JY, Ahn MS, Rhee HI (2008) Corn husk as a potential source of anthocyanins. J Agric Food Chem 56:11413–11416

    Article  CAS  Google Scholar 

  • Long N, Suzuki S, Sato S, Naiki-Ito A, Sakatani K, Shirai T, Takahashi S (2012) Purple corn color inhibition of prostate carcinogenesis by targeting cell growth pathways. Cancer Sci 104:298–303

    Article  Google Scholar 

  • Lopez-Martinez LX, Oliart-Ros RM, Valerio-Alfaro G, Lee CH, Parkin KL, Garcia HS (2009) Antioxidant activity, phenolic compounds and anthocyanins content of eighteen strains of Mexican maize. LWT-Food Sci Technol 42(6):1187–1192

    Article  CAS  Google Scholar 

  • Matus-Cádiz MA, Daskalchuk TE, Verma B, Puttick D, Chibbar RN, Gray GR, Perron CE, Tyler RT, Hucl P (2008) Phenolic compounds contribute to dark bran pigmentation in hard white wheat. J Agric Food Chem 56(5):1644–1653

    Article  Google Scholar 

  • Mintel and Leatherhead Food Research (2013) Food Colours: Market, Technical and Regulatory Insights. Leatherhead Food Research, Leatherhead

    Google Scholar 

  • Montilla EC, Hillebrand S, Antezana A, Winterhalter P (2011) Soluble and bound phenolic compounds in different Bolivian purple corn (Zea mays L.) cultivars. J Agric Food Chem 59:7068–7074

    Article  CAS  Google Scholar 

  • Nyman NA, Kumpulainen JT (2001) Determination of anthocyanidins in berries and red wine by high-performance liquid chromatography. J Agric Food Chem 49(9):4183–4187

    Article  CAS  Google Scholar 

  • Pedreschi R, Cisneros-Zevallos L (2007) Phenolic profiles of Andean purple corn (Zea mays L.). Food Chem 100(3):956–963

    Article  CAS  Google Scholar 

  • Pinho C, Melo A, Mansilha C, Ferreira IM (2011) Optimization of conditions for anthocyanin hydrolysis from red wine using response surface methodology (RSM). J Agric Food Chem 59(1):50–55

    Article  CAS  Google Scholar 

  • Ramos-Escudero F, Muñoz AM, Alvarado-Ortíz C, Alvarado Á, Yáñez JA (2012) Purple corn (Zea mays L.) phenolic compounds profile and its assessment as an agent against oxidative stress in isolated mouse organs. J Med Food 15:206–215

    Article  CAS  Google Scholar 

  • Rodriguez-Saona LE, Wrolstad RE (2001) Extraction, isolation, and purification of anthocyanins. In: Wrolstad RE, Acree TE, An H, Decker EA, Penner MH, Reid DS, Schwartz SJ, Shoemaker CF, Sporns P (eds) Current protocols in food analytical chemistry, 1st edn. Wiley, New York, pp F1.1.1–1.1.11

    Google Scholar 

  • Selinger DA, Chandler VL (1999) A mutation in the pale aleurone color1 gene identifies a novel regulator of the maize anthocyanin pathway. Plant Cell 11:5–14

    Article  CAS  Google Scholar 

  • Thiraphatthanavong P, Wattanathorn J, Muchimapura S, Thukham-mee W, Wannanon P, Tong-un T, Suriharn B, Lertrat K (2014) Preventive effect of Zea mays L. (purple waxy corn) on experimental diabetic cataract. Biomed Res Int 3:1–8

    Article  Google Scholar 

  • Truong VD, Deighton N, Thompson RT, McFeeters RF, Dean LO, Pecota KV, Yencho GC (2010) Characterization of anthocyanins and anthocyanidins in purple-fleshed sweet potatoes by HPLC-DAD/ESI-MS/MS. J Agric Food Chem 58(1):404–410

    Article  CAS  Google Scholar 

  • Tsuda T, Horio F, Uchida K, Aoki H, Osawa T (2003) Dietary cyanidin 3-O-beta-D-glucoside-rich purple corn color prevents obesity and ameliorates hyperglycemia in mice. J Nutr 133:2125–2130

    CAS  Google Scholar 

  • Wallace TC (2011) Anthocyanins in cardiovascular disease. Adv Nutr 2(1):1–7

    Article  CAS  Google Scholar 

  • Wang SY, Lin HS (2000) Antioxidant activity in fruits and leaves of blackberry, raspberry, and strawberry varies with cultivar and developmental stage. J Agric Food Chem 48(2):140–146

    Article  CAS  Google Scholar 

  • Winkel-Shirley B (2001) Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol 126(2):485–493

    Article  CAS  Google Scholar 

  • Wu X, Beecher GR, Holden JM, Haytowitz DB, Gebhardt SE, Prior RL (2006) Concentrations of anthocyanins in common foods in the United States and estimation of normal consumption. J Agric Food Chem 54(11):4069–4075

    Article  CAS  Google Scholar 

  • Yang Z, Zhai W (2010) Optimization of microwave-assisted extraction of anthocyanins from purple corn (Zea mays L.) cob and identification with HPLC–MS. Innovative Food Sci Emerg Technol 11(3):470–476

    Article  CAS  Google Scholar 

  • Zhang Z, Kou X, Fugal K, McLaughlin J (2004) Comparison of HPLC methods for determination of anthocyanins and anthocyanidins in bilberry extracts. J Agric Food Chem 52(4):688–691

    Article  CAS  Google Scholar 

  • Zhao X, Corrales M, Zhang C, Hu X, Ma Y, Tauscher B (2008) Composition and thermal stability of anthocyanins from Chinese purple corn (Zea mays L.). J Agric Food Chem 56:10761–10766

    Article  CAS  Google Scholar 

  • Žilić S, Serpen A, Akıllıoğlu G, Gökmen V, Vančetović J (2012) Phenolic compounds, carotenoids, anthocyanins, and antioxidant capacity of colored maize (Zea mays L.) kernels. J Agric Food Chem 60:1224–1231

    Article  Google Scholar 

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Acknowledgments

We appreciate Zanaceutica E.I.R.L. (Lima, Peru), Alicorp S.A.A. (Lima, Peru), Agroindustrial S.A.C. (Lima, Peru), and Globenatural International S.A. (Chorrillos-Lima, Peru) for providing the purple corn samples.

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Fei Lao declares that she has no conflict of interest. M. Monica Giusti declares that she has no conflict of interest.

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  1. Department of Food Science and Technology, The Ohio State University, 110 Parker Food Science and Technology Building, 2015 Fyffe Road, Columbus, OH, 43210, USA

    Fei Lao & M. Monica Giusti

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  1. Fei Lao
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  2. M. Monica Giusti
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Correspondence to M. Monica Giusti.

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Lao, F., Giusti, M.M. Quantification of Purple Corn (Zea mays L.) Anthocyanins Using Spectrophotometric and HPLC Approaches: Method Comparison and Correlation. Food Anal. Methods 9, 1367–1380 (2016). https://doi.org/10.1007/s12161-015-0318-0

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