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Carotenoid Contents in Different Millets Cultivars Collected from China and Korea

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Carotenoid Contents in Different Millets Cultivars Collected from China and Korea Asian Journal of Chemistry; Vol. 26, No. 2 (2014), 464-466 http://dx.doi.org/10.14233/ajchem.2014.15446A Carotenoid Contents in Different Millets Cultivars Collected from China and Korea 1 2 1 3 YEON BOK KIM , JAE KWANG KIM , MD ROMIJ UDDIN , CHEOL HO PARK , 4 5 5,* 1,* HAENG HOON KIM , EUNSOOK CHUNG , JAI-HEON LEE and SANG UN PARK 1Department of Crop Science, Chungnam National University, 99 Daehak-Ro, Yuseong-Gu, Daejeon, 305-764, Republic of Korea 2Division of Life Sciences, College of Life Sciences and Bioengineering, Incheon National University, Incheon, 406-772, Republic of Korea 3Department of Bio-Health Technology, College of Biomedical Science, Kangwon National Univesity, 192-1 Hyoja 2-Dong, Chuncheon 200- 701, Republic of Korea 4Department of Well-being Resources, Sunchon National University, 413 Jungangno, Suncheon, Jeollanam-do, 540-742, Republic of Korea 5Department of Genetic Engineering, Dong-A University, Busan 604-714, Republic of Korea *Corresponding authors: Fax: +82 51 2007505; Tel: +82 51 2007592; E-mail: [email protected]; [email protected] Received: 7 March 2013; Accepted: 25 July 2013; Published online: 15 January 2014; AJC-14567 Millet (Panicum miliaceum L.) cultivars from China and Korea were analyzed using HPLC to determine the levels of 5 different carotenoids: lutein, zeaxanthin, β-carotene, 9-cis-β-carotene and 13-cis-β-carotene. There was a significant difference in carotenoid content among cultivars from the 2 locations, with the Chinese cultivars generally having a higher content than the Korean cultivar. Lutein was present at the highest concentration in all of the cultivars, but was 34, 33 and 16 % higher in Joongback (China) than in Joongjukyumi (China), Hwangguem (Korea) and Joongjuk (China), respectively. Joongback also contained 58, 55 and 37 % higher concentrations of zeaxanthin than Joongjukyumi, Hwangguem and Joongjuk, respectively and a 33 % higher concentration of β-carotene than Hwangguem (Korea). The other two carotenoids (9-cis-β-carotene and 13-cis-β-carotene) were present at similar levels in all 4 cultivars. The results suggest that the millet cultivar Joongback from China could potentially be used as a source of carotenoids in the human diet. Keywords: Carotenoid, Cultivars, Millet, HPLC. INTRODUCTION black through to brown in pearl millet. Millet has a higher protein value than corn, as well as high fiber levels due to the Panicum is one of the largest genera of grasses, comprising attached hulls.1 Millet can be successfully grown in a wide more than 400 species. Within this genus, Panicum miliaceum range of environmental conditions, being better adapted than (proso millet) and Panicum sumatrense (little millet) are of most crops to hot, dry regions and is particularly valuable in economic importance. Proso millet, which is also known as semiarid regions because of its short growing season. It can common millet, hog millet or white millet is one of the world’s tolerate drought and intense heat, or avoids these conditions oldest cultivated crops. This cereal plant is cultivated for its by growing to maturity very quickly. Millet plays an impor- grain, mostly in Asia and North America, but may also be tant role in the economies of many developing countries in used for forage. Proso millet differs from other millets, such the world, where it is consumed directly as human food. In as pearl, foxtail, finger or barnyard millets. This annual grass contrast, in the Western world, millet is grown primarily for is found in central and eastern Asia, covering a wide area from birdseed, livestock feed, hay, or as an emergency catch crop2. the Caspian Sea, east to Xinjiang and Mongolia and is also Traditionally, Korea’s major crops were rice, barley, found in the mountains of the former U.S.S.R. and India at sorghum and millet, but cultivating area of these crops gradu- altitudes of up to 1,200 and 3,500 m, respectively1. The plant ally decreased due to grow other economic crops. However, can grow to heights of 50-100 cm and its leaves grow up to 30 recently minor crops such as barley, sorghum and millet have cm long. The stems are stout and erect and both the stems and attracted attention due to their high energy, protein, vitamin leaves are covered with small hairs. The seed heads grow in and mineral contents, which are believed to help prevent and bunches and can be divided into 3 types, based on the shape treat diseases associated with aging3-6. of the panicle: spreading, loose and one-sided and erect. The Carotenoids are isoprenoid pigments that are widely seeds are very small and oval in shape. Seed colour depends distributed in nature7. In plants, carotenoids play an important on species and can be white, cream, yellow, orange, red, or role in photosynthesis (where they contribute to the light- Vol. 26, No. 2 (2014) Carotenoid Contents in Different Millets Cultivars Collected from China and Korea 465 harvesting process), photomorphogenesis and photoprotection. Analysis System (SAS version 9.2). All data are given as They are also the precursors of an important plant hormone, the mean and standard deviation of triplicate experiments. abscisic acid, which is involved in regulating the plant stress Treatment mean comparisons were performed with the least response8,9. Carotenoids contain abundant pigments and are significant difference (LSD). accumulated in the plastids of leaves, flowers and fruits10. Carotenoids also have anticarcinogenic and antioxidant RESULTS AND DISCUSSION activities. For example, the provitamin A carotenoid β-carotene is a nutritional factor that is important for chronic disease Carotenoid contents of different cultivars of millet prevention and it has been shown that carotenoids can lower collected from China and Korea: Five different carotenoids the levels of bad cholesterol and triglycerides in the blood- were detected in the 4 millet cultivars collected from China stream11. Consequently, in recent years, several carotenoids and Korea: lutein, zeaxanthin, β-carotene, 9-cis-β-carotene have been used as colourants, nutritional supplements and and 13-cis-β-carotene. There was significant variation in nutraceuticals for food, cosmetic and pharmaceutical pur- carotenoid contents among the millet cultivars (Fig. 1), with poses12. In the present study, we analyzed the carotenoid the Chinese cultivars generally having a higher content than content of 4 different millet cultivars collected from Korea the Korean cultivar. Among the different cultivars, Joongback and China. from China was found to contain the highest concentration of carotenoids. Lutein was found to occur at the highest concen- EXPERIMENTAL tration in all 4 cultivars, but was 34, 33 and 16 % higher in Four millet cultivars were used in this study: 3 from China Joongback than in Joongjukyumi (China), Hwangguem and 1 from Korea. Millet seeds were germinated in a green- (Korea) and Joongjuk (China), respectively. There was much house and the seedlings were then transferred to the experi- greater variation in the content of zeaxanthin among cultivars mental farm at Chungnam National University (Daejeon, than in lutein, but once again, Joongback contained the highest Korea). All of the cultivars were harvested after 4 months, at amount, with concentrations that were 58, 55 and 37 % higher their ripening stage. They were then manually hulled and than those found in Joongjukyumi, Hwangguem and Joongjuk, ground to obtain a fine powder using a cyclone mixer mill respectively. The cultivar Joongback also contained 33 % more (HMF-590; Hanil, Seoul, Korea) and a mortar and pestle. The β-carotene than Hwangguem. The other two carotenoids (9- milled powders were immediately frozen in liquid nitrogen cis-β-carotene and 13-cis-β-carotene) were present at similar and stored at -80 ºC until required for use in the carotenoid levels in all 4 cultivars. analysis. This study showed that the levels of carotenoids varied Extraction and HPLC analysis of carotenoids: Caro- widely among 4 different cultivars of millet from 2 locations. tenoids were extracted from millet samples (0.1 g) with 3 mL In particular, the Joongback cultivar from China contained of ethanol containing 0.1 % ascorbic acid (w/v). This mixture the highest concentration of all 5 types of carotenoid sampled. was vortexed for 20 s and incubated in a water bath at 85 ºC These findings are consistent with previous findings, which for 5 min. Subsequently, 120 µL of potassium hydroxide (80 % have shown that the carotenoid contents of some vegetables w/v) was added to saponify any potentially interfering oils varies between cultivars and geographical locations. For example, Park et al.13 showed that carotenoid contents varied and the mixture was again vortexed and incubated at 85 ºC for 14 10 min. The samples were then placed on ice and 1.5 mL of in the skins and flesh of 2 types of kohlrabi. Habicht et al. cold deionized water and 0.05 mL of β-apo-8-carotenal (12.5 reported that the levels of saponin, linoleic acid and linolenic µg mL-1; an internal standard) were added. The carotenoids acid varied between cultivars of bitter melon, with white bitter were subsequently extracted twice with 1.5 mL of hexane and gourd varieties containing significantly lower concentrations of saponin (0.25 %) than green varieties (0.67 %). centrifuged at 1200 × g following each extraction in order to 15 separate the layers. Finally, the extracts were freeze dried Kim et al. reported that different coloured rice grains under a stream of nitrogen gas and resuspended in 50:50 (v/v) contained different levels of carotenoids, with black cultivars dichloromethane/methanol. containing mean levels of 0.482 mg/100 g, red cultivars For HPLC analysis, the carotenoids were separated using containing 0.052 mg/100 g and white cultivars containing an Agilent 1100 HPLC system using a C YMC column (250 0.021 mg/100 g.
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