ANTHOCYANINS FROM ROSE MAIZE (Zea mays L.) GRAINS Lucía Barrientos Ramírez, Hilda E. Ramírez Salcedo, María Fernanda Fernández Aulis, Mario Alberto Ruíz López, Arturo Navarro Ocaña and J. Jesús Vargas Radillo SUMMARY Here we present the anthocyanin composition of grains of Peonidin, and Pelargonidin, while HPLC profile revealed rose corn (Zea mays L.). Extraction was performed using that the most abundant anthocyanin was Cyanidin-3-(6-mal- methanol:water:HCl in an ultrasound bath for 20min. Antho- onyl-glucoside), a malonic derivative of Cyanidin. The re- cyanins were isolated from maize using the combination of sults increase available data regarding the presence of these column chromatography and HPTLC plates, whereas HPLC- compounds in varieties of pigmented corn grains, and thus UV/Vis and HPLC-MS were used for analysis and identifi- help to promote the consumption of these varieties for their cation. Compounds found include the glycosides Cyanidin, beneficial health effects. Introduction or more sugar molecules (gly- for cosmetic or pharmaceutical Traditionally, it is used to cosylated type), and may also applications (Ignat et al., make tortillas, pozole, gordi- The immediate predecessor contain acyl radicals in the 2011). They also have func- tas, and sweet atole (Ron of domesticated maize, the form of aliphatic organic acid tional properties, since they Parra et al., 2006). The pres- Teocintle, is considered native derivatives linked to sugar cause a slow digestion of ent study aims to assess the of Mesoamerica and, thus, (acylated type) which show starch that has been associat- types and characteristics of Mexico is considered to be the greater stability at different ed with beneficial health ef- anthocyanin metabolites in this origin of maize (Zea mays L., pH levels (Eng et al., 2017). fects like the control of hun- corn variety, which is abun- Poaceae family). Fifty nine of Moreover, they show substan- ger and an more evenly dant in many regions, with the 220-300 races of maize tial bioactivity, mainly as an- spaced food consumption large availability and interest- found in the American conti- tioxidants (Yang and Shai, (Bello-Pérez et al., 2016). ing biological properties due to nent grow in Mexico (Sánchez 2011). These metabolites ex- Bibliographic reports men- the presence of anthocyanin et al., 2000). In addition to hibit health benefits, promot- tion anthocyanin contents of and other polyphenols. white corn, there are over 45 ing reduction of inflammation 9-15.9mg·kg-1 dry matter (DM) races of pigmented maize (He and Giusti, 2010); with in white corn, 163.9mg·kg-1 Materials and Methods (Salinas-Moreno et al., 2005). properties such as prevention DM pink corn, 342.2 in Annual maize yield in 2014 of the chronic diseases diabe- blue corn, 1270 in red corn, Sample was ~23.27×106t of grain, pro- tes and obesity (Li et al., 1277 in purple corn, and duced on ~7.4×106ha, represent- 2015) and cancer (Chen et al., 5290mg·kg-1 DM in black corn Rose maize (Zea mays L.; ing an average yield of 3.14t·ha- 2015), among other. (Salinas-Moreno et al., 2013; National Registry Nº 05046) 1 (SIAP, 2014). The Mexican The Mexican races of maize Escalante-Aburto et al., 2016). was harvested in the summer state of Jalisco is the second present kernels with white, The rose western corn is a (rainy season) in the period of largest national producer of yellow, pink, red, blue, purple very productive variety, with June-August of 2015 in a corn, and harvested 3.47×106t/ and black coloration (Bello- cobs of 15 to 20cm in length, property located in the town year for a yield of 6.42ton·ha-1, Perez et al., 2016). The color cylindrical shape, and 12 to 14 of Nextipac, Zapopan, Jalisco, constituting 15% of national is related to the amounts and lines. The grain is jagged, Mexico (20º45’51”N; 103º31’ production (SIAP, 2014). types of anthocyanins. The pink or pink-red. This maize 44”W; 1634masl). Maize is Anthocyanins, a water-solu- pigmented maize varieties are is included in the variety of cultivated annually under the ble polyphenol, comprise one desirable as food, as well as cone shaped corn races. environmental conditions that KEYWORDS / Anthocyanin / Antioxidant Bioactivity / HPTLC / Maize / Natural Pigments / Received: 06/06/2017. Modified: 02/14/2018. Accepted: 02/16/2018. Lucia Barrientos Ramírez. Doc- María Fernanda Fernández Au- Arturo Navarro Ocaña. Doctor fessor, UdG, Mexico. Address: tor of Science in Biology, lis. Doctoral student in Chemis- in Biotechnology, CCH-UNAM, Departamento de Madera, Universidad de Guadalajara try, Universidad Nacional Autó- Mexico. Professor, UNAM, Celulosa y Papel (DMCyP), (UdeG), Mexico. Professor, noma de México (UNAM). e- Mexico e-mail: arturono@ UdeG. Zapopan, Jalisco, Mé- UdeG, Mexico. e-mail: lba- mail: [email protected] unam.mx xico, CP 45100, tel. (33)- [email protected] Mario Alberto Ruíz López. J. Jesús Vargas Radillo. Doctor 36820110. e-mail: jvargasr@ Hilda E. Ramírez Salcedo. Doc- Doctor of Science in Biolog), in Biosystemtics, Ecology and dmcyp.cucei.udg.mx. toral student, BEMARENA, UNAM, Mexico. Professor, Natural and Agricultueral UdeG, Mexico. e-mail: hildae- UdeG, Mexico. e-mail: mruiz Resources Management (BE- [email protected]. @cucba.udg.mx MARENA), UdeG, Mexico. Pro- 188 0378-1844/14/07/468-08 $ 3.00/0 MARCH 2018 • VOL. 43 Nº 3 ANTOCIANINAS DE GRANOS DE MAIZ (Zea mays L.) ROSA Lucía Barrientos Ramírez, Hilda E. Ramírez Salcedo, María Fernanda Fernández Aulis, Mario Alberto Ruíz López, Arturo Navarro Ocaña y J. Jesús Vargas Radillo RESUMEN Se presenta la composición de antocianinas en granos de ina-3-(6-malonilglucosido), un derivado malónico de la Cianidi- maíz Zea mays L., de la raza elotes occidentales tipo maíz rosa. na, el compuesto más abundante de acuerdo con su perfil cro- La extracción de estas fue con un sistema metanol:agua:HCl, us- matográfico. Estos resultados aumentan el acervo de datos so- ando un baño ultrasonido por 20min, para posteriormente ser bre estos compuestos presentes en las variedades de los granos analizado por HPLC-DAD y HPLC-MS. Se identificaron glucósi- pigmentados, y buscan fomentar el consumo de estas variedades dos de Cianidina, Pelargonidina y Peonidina, siendo la Cianid- por sus efectos benéficos en la salud. ANTOCIANINAS DE GRÃOS DE MILHO (Zea mays L.) ROSA Lucía Barrientos Ramírez, Hilda E. Ramírez Salcedo, María Fernanda Fernández Aulis, Mario Alberto Ruíz López, Arturo Navarro Ocaña e J. Jesús Vargas Radillo RESUMO Apresenta-se a composição de antocianinas em grãos de mi- na-3-(6-malonilglucosídeo), um derivado malônico da Cianidina, lho Zea mays L., da raça elote ocidental, tipo milho rosa. A ex- o composto mais abundante de acordo com seu perfil cromato- tração destas foi realizada com um sistema metanol:água:HCl, gráfico. Estes resultados aumentam o acervo de dados sobre es- usando um banho ultrassom por 20min, para posteriormente ser tes compostos presentes nas variedades dos grãos pigmentados, analisado por HPLC-DAD e HPLC-MS. Identificaram-se glicosí- e buscam fomentar o consumo destas variedades por seus efei- deos de Cianidina, Pelargonidina e Peonidina, sendo a Cianidi- tos benéficos na saúde. prevail in the area: average thanol acidified with 0.1% HCl, tially until there was no more UV/Vis detector (520nm) and temperature 20-22ºC; average in an ultrasonic bath (Branson separation of spots on the plate. a Hypersil Gold ODS column rainfall 980mm (Climate-data, model 8892; 42kHz), for (250×4.4mm, 5µm). We used a 2018; Sagarpa, 2015), applying 20min. The extract was filtered HPLC analysis gradient of solvent A (H2O: traditional fertilization consist- and concentrated by rotoevapo- ACN: HCOOH, 87:3:10) and ing of the incorporation of ni- ration at <40ºC and purified Crude extract was filtered solvent B (ACN), with the trogen and phosphorus, on a making use of a column through a 0.45µm membrane following elution program: Regosol type soil (sandy-loam (2.0×60cm) of amberlite (XAD- and analyzed using a Waters 97% A and 3% B from texture). About 30 pieces were 7HP, 20-60 mesh; Sigma- liquid chromatograph equipped 0-45min; 75% A and 25% B sampled, obtaining the grains Aldrich). Amberlite is an acryl- with a 2707 autosampler, a from 45-46min; 70% A and manually. A representative ic resin, nonionic, moderately 1525 binary pump, a 2478 30% B from 46-47min; and sample was evaluated for an- polar material that partially thocyanin content as follows: purified the anthocyanins the corn kernels were manually based on polar/non-polar inter- reeled off and washed, they actions (Wang et al., 2014). were then pulverized in a ham- The column was activated with mer mill (Retsch GmbH) using 250ml of methanol at a flow a screen with 0.5mm perfora- rate of 1ml·min-1 and so tions, and the obtained flour washed with 500ml of distilled was dehydrated to constant water in order to achieve a weight in an oven at 40ºC. greater adsorption of the or- ganic compounds and, then the Extraction and purification by extracts were loaded onto a combined column column and washed with abun- chromatography dant distilled water to remove most of sugars, organic acids, A schematic of the proposed proteins and ions (Wang et al., method is illustrated in Figu- 2014). The anthocyanin pig- re 1. Anthocyanins were ob- ment fraction was recovered tained with the ultrasound as- from amberlite with 300ml of sisted extraction (UAE) proce- methanol:acetic acid (19:1 v/v), dure (Vinatoru, 2001). The concentrated by rotary evapo- flour ground from whole corn rator at <40ºC, dryed (Labkept (10g) was extracted with 100ml in amber flasks at 4ºC. This Figure 1. Isolation and fragmentation scheme of anthocyanins of corn (10:1 w/v) of 80% (v/v) me- procedure was repeated sequen- grains.