Variation in Â-Carotene and Total Carotenoid Content in Fruits Of

HORTSCIENCE 36(4):746–749. 2001. the food industry, but also provides nutritional data for the range of fruit types within the β Capsicum genus. Most vitamin A values re- Variation in -Carotene and Total ported in food composition tables are over- estimated, because they are based on method- Carotenoid Content in Fruits of ology in which β-carotene was not separated from other provitamin A pigments with lower Capsicum activity (Haytowitz and Matthews, 1984). β- Carotene concentrations now can be accu- Marisa M. Wall1, Cynthia A. Waddell2, and Paul W. Bosland3 rately determined with HPLC (Bushway and Wilson, 1982). Department of Agronomy and Horticulture, New Mexico State University, Las Cruces, NM 88003-8003 Materials and Methods Additional index words. peppers, pigments, human nutrition, vegetable breeding, chiles, β-Carotene and total carotenoid content Capsicum annuum, Capsicum chinense, Capsicum baccatum, Capsicum frutescens, were evaluated in different Capsicum fruit Capsicum chacoense, Capsicum eximium types and species using fresh fruit from 25 Abstract. The β-carotene and total carotenoid content of either fresh or dried tissue of fruits cultivars and dried fruit from 40 cultivars of a total of 57 cultivars of six Capsicum species were analyzed using high performance selected to represent a range of fruit types and liquid chromatography (HPLC). β-Carotene levels in ripe fruit varied from 0 to 166 uses (fresh or processed). Plots were estab- µg·g–1 fresh weight, and carotenoid levels were from 1 to 896 µg·g–1 in ripe fruit in 1996. The lished from transplants on a sandy loam soil at range of values for β-carotene was similar in 1997, but that for total carotenoids was wider the Fabian Garcia Agricultural Science Center (4 to 1173 µg·g–1 fresh weight). Fresh fruit of the cultivars Greenleaf Tabasco, Pulla, in Las Cruces, N.M., in 1996 and 1997. The beds were on 1-m centers and furrow-irrigated. Guajillo, NuMex Conquistador, Ring-O-Fire, and Thai Dragon contained greater amounts –1 –1 of β-carotene per 100 g fresh weight than the recommended dietary allowance (RDA) for Fertilizer (112 kg·ha P and 168 kg·ha N) vitamin A for the average adult. For dried Capsicum entries, New Mexican, aji, pasilla, and was applied preplant as diammonium phos- ancho types had the highest levels of β-carotene. In 1996, β-carotene levels among the dried phate and in the irrigation water as urea and Capsicum germplasm ranged from 2 to 739 µg·g–1 dry weight, and carotenoid levels from ammonium nitrate. Either fully ripened, suc- 111 to 6226 µg·g–1. Values were higher in 1997, ranging from 24 to 1198 µg·g–1 dry weight culent fruit or ripened, partially dry fruit were for β-carotene and from 187 to 10,121 µg·g–1 for total carotenoids. A pasilla type (C. harvested for analysis. The number of fruit annuum L.) had the highest total carotenoid content among the dried entries in both years. harvested per plant varied according to fruit type, and ranged from three for the larger bell, cubanelle, and pimiento types, to 150 for the Chiles (Capsicum sp.) are one of the oldest prika carotenoids are used by the food industry smaller piquin and tepin types. Fruit were cultivated plants of the Americas, have been as natural red colorants, but also have im- harvested separately from four plants and part of the human diet for thousands of years mense nutritional value as provitamin A and destemmed in the field. Fruit were deseeded (Bosland, 1994), and are rich in vitamins A antioxidants. Evidence for antitumor activity for analysis of fresh fruit, except for the small- and C (Bureau and Bushway, 1986; Howard et of carotenoids has renewed interest in foods fruited tabasco, guajillo, Asian hot, serrano, al., 1994; Mejia et al., 1988; Simonne et al., containing these compounds. The antioxidant and datil pod types, but not of dry fruit. Mois- 1997). Worldwide, vitamin A deficiency is the properties of carotenoids appear to protect ture content at harvest was determined from a most common form of malnutrition after pro- against heart disease (Kritchevsky, 1999) and subsample of each entry using an IR-100 mois- tein deficiency (World Bank, 1993). In parts cancer of the lungs, stomach, cervix, and throat ture balance (Denver Instruments Co., Arvada, of Asia, Africa, and Latin America, vitamin A (Byers and Perry, 1992; Machlin, 1995). Re- Colo.). Fruit harvested from individual plants deficiency occurs in millions of children ports on the carotenoid content of chiles are were kept separate in the laboratory, and one (Sommer, 1989), and chiles could be an im- primarily for bell, paprika, and jalapeño types sample from each accession was extracted and portant source of provitamin A for the people (Bureau and Bushway, 1986; Howard et al., analyzed at the same time. There were four of those regions. Chile production and con- 1994; Levy et al., 1995, Mejia et al., 1988; samples per accession. sumption is increasing globally. Worldwide Simonne et al., 1997). More than 30 caro- Carotenoid extraction from fresh fruit production is 11 million tons, with more than tenoids have been isolated from chiles (Gre- samples was according to the methodology of 48% produced in Asia, and per capita con- gory et al., 1987). The red capsanthin, Bureau and Bushway (1986). Chile samples sumption is estimated at 10 g per day (Food capsorubin, and cryptocapsin pigments are were chopped into small pieces, and a 10-g and Agriculture Organization, 1995). In north- valued mostly as natural colorants, whereas β- subsample was immediately combined with 1 ≈ α γ β ern Mexico, consumption is 40 g per day carotene, -carotene, -carotene and -cryp- g MgCO3, 5 g Na2SO4, and 125 mL tetrahydro- (Valencia et al., 1983). toxanthin have provitamin A activity, with β- furan (THF) stabilized with 0.01% butylated The yellow, orange, and red colors of Cap- carotene having the highest activity. hydroxytoluene (BHT). Each sample was ho- sicum fruit originate from carotenoid pigments Improvement of chiles (New Mexican type, mogenized with a Tekmar tissumizer (Cincin- produced during ripening. Red chile and pa- C. annuum L.) for carotenoid and nutrient nati) for 5 min on medium speed. Samples content is the goal of several breeding pro- were vacuum filtered (Whatman #42), and the Received for publication 31 Mar. 2000. Accepted grams. To assist in identifying sources of high residue was re-extracted with an additional for publication 2 Oct. 2000. This research was levels of carotenoids, 57 Capsicum cultivars 125 mL THF. A 20-mL portion of the sample supported by the Chile Improvement Project of the from six species and 25 different fruit types was concentrated under nitrogen gas and stored New Mexico State Univ. Agricultural Experiment were evaluated for variation in β-carotene and at –20 °C until HPLC analysis. Samples were Station. We thank Kathleen Dixon, Kelly Callahan, total carotenoid content. β-Carotene is the resuspended in 0.4 mL THF and vortexed. and Li Fang for excellent technical assistance. The precursor for the predominant orange and red Acetonitrile and methanol were added in equal cost of publishing this paper was defrayed in part by pigments in chiles, and genotypes with high amounts to a final volume of 4 mL. the payment of page charges. Under postal regula- concentrations of β-carotene proved to be rich- Prior to extraction of dry samples, whole tions, this paper therefore must be hereby marked β ° advertisement solely to indicate this fact. est in total carotenoid content. Therefore, - fruit were dried in a 55 C oven and ground in 1Associate Professor. E-mail address: mawall@ carotene may be a selection criterion for devel- a Wiley mill to pass through a 1-mm sieve. nmsu.edu oping highly colored chiles (Levy et al., 1995). Carotenoids were extracted from dried samples 2Research Assistant, Senior. This information is useful not only for by placing 0.2 g of powder in 50 mL stabilized 3Professor. breeding highly colored paprika and chiles for THF. After 16 h in the dark, a 10-mL aliquot was concentrated under nitrogen gas and stored Standard solutions of β-carotene were pre- (–1.7 °C), allowing less time for pigment ac- at –20 °C. Samples were resuspended in 0.2 pared in 20 mL THF, and capsanthin standards cumulation and crop dry-down before the mL THF, and acetonitrile and methanol were in methanol:acetonitrile (1:1). Aliquots were freeze. In 1997, a light frost occurred on 27 added in equal amounts for a final volume of diluted in methanol: acetonitrile (1:1) to pro- Oct. (–0.6 °C), but the first hard freeze did not 4 mL. vide standard concentrations ranging from 2 to occur until 30 Nov. (–2.2 °C). Fruit allowed to All samples were filtered through a nylon 10 µg·L–1. The detection limit was 0.1 µg·L–1. dry on the plant accumulate more pigments Magna 0.22-µm filter into HPLC vials for Data were analyzed using the generalized (Kanner et al., 1977; Palevitch et al., 1975), injection. Analysis followed a modified caro- linear models procedure of SAS (SAS Insti- and fruit harvested after a freeze generally tenoid separation method of Ittah et al. (1993) tute, Cary, N.C.) for a randomized complete- have less color in the dried product (Isidoro et and Levy et al. (1995) for nonsaponified block design, with samples blocked based on al., 1990). pigments. A Hewlett-Packard 1090 series time of laboratory extraction and analysis. The interaction between years and culti- liquid chromatograph (Hewlett-Packard, vars was significant for all analyses (P ≤ Wilmington, Del.) fitted with a Hewlett- Results and Discussion 0.0001).

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