Carotenoids in paprika

APCl in positive mode ionization and triple quadrupole detection was used for determination of free and bound carotenoids in paprika, obtaining the [M + H]+ and losses of fatty acids as neutral molecules from the [M + H]+ with MeOH, MTBE, and H2O as eluent from the C30 column. The positions of the fatty acids on unsymmetrical xanthophylls could not be established by the MS data. 

Breithaupt, D.E. and Schwack, W., Determination of free and bound carotenoids in paprika Capsicum annuum L.) by LC/MS, Eur. Food Res. TechnoL, 211, 52, 2000. Epler, K.S., Ziegler, R.G., and Craft, N.E., Liquid chromatographic method for the determination of carotenoids, retinoids and tocopherols in human serum and in food, J. Chrvmatogr, 619, 37, 1993. 

Rios, J. J. et al. (2008). Description of volatile compounds generated by the degradation of carotenoids in paprika, tomato and marigold oleoresins. Food Chem. 106(3) 1145-1153. 

Breithaupt, D.E. and Schwack, W. (2000) Determination of free and bound carotenoids in paprika (Capsicum annuum F.) by LC/MS. European Food Research and Technology 21 1(1), 52-55. 

Capsanthin (Figure 1) is the most abundant carotenoid in the paprika spice. The concentration of capsanthin is about IS90 mg per kg of dry matter and 41 54% of total carotenoids in paprika 18,19). The antiphotooxidative effect and kintics study of capsanthin on soybean oil has not been well studied. In addition, the study of the antiphotooxidative effect of carotenoids mostly focused on lipid or fatty acids. The antioxidative effect of carotenoids on the photooxidation of flavor compounds has not been well studied, either. 

FIGURE 4.2.2 Structures of carotenoids found in paprika (capsanthin and capsombin), saffron and gardenia (crocetin derivatives), and annatto (bixin and norbixin). 

Minguez-Mosquera MI and Homero-Mendez D. 1994. Comparative study of the effect of paprika processing on the carotenoids in peppers (Capsicum annuum) of the Bola and Agridulce varieties. J Agric Food Chem 42 1555-1560. 

Paprika contains capsombin and capsanthin (Fig. 8.3) which occur mainly as the lauric acid esters, and about 20 other carotenoid pigments. Paprika is produced in many countries which have developed their own specialties. Cayenne or cayenne pepper, produced from a different cultivar of C. annum, is usually more pungent. C. frutescens is the source of the very pungent Tabasco sauce. Paprika oleoresin is produced by solvent extraction of the ground powder. Obviously paprika supplies both flavor and color and its use is limited to those products compatible with the flavor. The recent rise in demand for tomato products in the form of pizza, salsa, etc., has increased the demand for paprika. Paprika is used in meat products, soups, sauces, salad dressings, processed cheese, snacks, confectionery and baked goods.1018... 

It was established that neither GLT nor JIP-/KT) are suitable for the enhancement of the stability of carotenoid pigments in paprika powders [32],... 

The effect of saponification on the concentration of carotenoids in fatty foods has also been investigated by RP-HPLC. Sausages containing 5.6 per cent powdered paprika were extracted exhaustively with chloroform-methanol (2 1, v/v). The extracting solvent contained 0.01 per cent butylated hydroxyanisole (BHA). An aliquot of the combined extracts was evaporated to dryness and saponified at 50°C for 5min with 10 per cent KOH in methanol in the presence of 0.01 per cent BHA. Free carotenoid pigments were extracted with diethyl ether, washed with water, dried over anhydrous NajSC and evaporated under... 

A. Levy, S. Harel, D. Palevitch, B. Akiri, E. Menagem and J. Kanner, Carotenoid pigments and / -carotene in paprika fruits (Capsicum spp.) with different genotypes. J. Agric. Food Chem. 43 (1995) 362-366. 

Deli J, Molnar P, Toth G (2001) Carotenoid composition in the fruits of red paprika (Capsicum annuum var. lycopersiciforme rubrum) during ripening biosynthesis of carotenoids in red paprika. J Agric Food Chem 49 1517-1523... 

Matsufuji H, Nakamura H, Chino M, Takeda M (1998) Antioxidant activity of capsanthin and the fatty acid esters in paprika (Capsicum annuum). J Agric Food Chem 46 3468-3472 WaU MM, WaddeU CA, Bosland PW (2001) Variation in P-carotene and total carotenoid content in fruits of Capsicum. HortScience 36 746-749... 

Coenen et al. [60,61] proposed a two-step extraction for the separation of pungent compounds and carotenoid fractions. Aroma- and pungent components were recovered at 120 bar and 40°C, and the paprika residue was re-extracted at 320 bar and 40°C to recover carotenoids. The solubility of capsaicin in carbon dioxide was relatively low at a pressure of 120 bar, so a great amount of solvent (for example 130 kg of CO2 per kg of paprika) was needed to recover the aroma components totally. The extraction time was 6.5 hour. In the separator the pressure was 56 bar and the temperature was 45°C. The orange, paste-like extract recovered in the first step was extremely pungent in taste. It contained water, and the yield was about 15%. In the second step, a relatively great amount of CO2 (approximately 50 kg/kg) was needed to recover the carotenoids in quantitative yield. The extraction time was 4 hours. The dark red, liquid colour-concentrate is without capsaicinoids. The yield was 2.5%. 

Ethoxyquin, a synthetic antioxidant, is not generally allowed for human consumption in foods, but it is being added to animal feed and to fruits as an antiscald agent (94,143). Ethoxyquin is also used in the spice industry to prevent carotenoid loss during postharvest handling. However, ethoxyquin-treated paprika is unacceptable for some markets and some consumers (129). Perfetti et al. (130) described a method for determination of ethoxyquin in paprika and chili powder. Ethoxyquin was extracted from the spice with hexane and partitioned into 0.3 N HC1. After adjusting the solution to pH 13-14, ethoxyquin was extracted into hexane, and the hexane layer was evaporated to dryness. An acetonitrile solution of the residue was then analyzed by reversed-phase HPLC, with detection at 254 nm. The mobile phase was water/acetonitrile with ammonium acetate buffer. Recoveries from samples fortified at 50, 100, and 200 ppm averaged 92%, with a coefficient of variation of 2.3%. The method was applied to a number of commercial samples of paprika and chili powder. Ethoxyquin was found in paprika samples at levels up to 63 ppm and in chili powder samples at levels up to 20 ppm. 

A method for the determination of ethoxyquin in paprika that avoided the previous separation steps from other colored substances was proposed by Vinas (133). Analysis is carried out by reverse-phase HPLC using the gradient elution technique and UV detection at 270 nm. Using fluorimetric detection with excitation at 311 nm and emission at 444 nm, a detection limit of 0.2 jig /ml was reached. The method can be applied to the determination of ethoxyquin in commercial samples in the presence of paprika (Capsicum annuum) carotenoids. 

The structure of a new carotenoid, isolated from fruits of the red tomato-shaped paprika, was elucidated to be (3S,5f ,6S,59P)-3,6-epoxy-5,6-dihydro-5-hydroxy-P, carotenes, 6 -dione by spectroscopic analyses and mass spectrometry and was designated as capsanthone 3,6-epoxide. Capsanthone 3,6-epoxide is assumed to be an oxidative metabolite of capsanthin 3,6-epoxide in paprika (Maoka et al., 2001a). 

Eleven apo-carotenoids (1-11), including five new compounds, 4, 6, 9,10 and 11, were isolated from the fruits of the red paprika collected from Japan by Maoka et al. (2001b). The structures of new apocarotenoids were determined to be apo-14 -zeaxanthinal (4), apo-13-zeaxanthinone (6), apo-12 -capsorubinal (9), apo-8 -capsorubinal (10) and 9,9 -diapo-10,9 -retro-carotene-9,9 -dione (11) by spectroscopic analysis. The other six known apocarotenoids were identified to be apo-8 -zeaxanthinal (1), apo-lO -zeaxanthinal (2), apo-12 -zeaxan-thinal (3), apo-15-zeaxanthinal (5), apo-11-zeaxanthinal (7) and apo-9-zeaxanthinone (8), which had not been found previously in paprika. These apocarotenoids were assumed to be oxidative cleavage products of C40 carotenoid, such as capsanthin in paprika. 

Molnar, P., Kawase, M., Satoh, K., Sohara, Y., Tanaka, T., Tani, S., Sakagami, H., Nakashima, H., Motohashi, N., Cyemant, N. and Molnar, J. (2005) Biological activity of carotenoids in red paprika, Valencia orange and golden delicious apple. Phytotherapy Research 19(8), 700-707. 

HORNERO-MENDEZ D and MINGUEZ-MOSQUERA MI. 2001. Rapid spectro-photometric determination of red and yellow isochromic carotenoid fractions in paprika and red pepper oleoresins. J. Agric. Food Chem. 49 3584-3588. 

Capsanthin (16) and capsorubin (16a), the colourants in paprika oleoresin, although not produced by commercial synthesis have been prepared in the course of carotenoid studies (ref. 58). Capsanthin has been synthesised from p-citraurin ( 3-hydroxy-p-apo-8 -carotenal ) which is available from zeaxanthin (3R, 3R )-p-carotene-3,3 -diol), by oxidation with potassium permanganate (ref. 59). 

The carotenoid activity during oxidation is strongly influenced by the oxygen pressure (PO2) of the experimental conditions. Kiokias and Oreopoulou have shown that certain natural carotenoid mixtures (paprika, bixin and tomato, and palm-oil preparations) inhibited the azo-initiated oxidation of sunflower oil-in-water emulsions (operated rapidly under low pOj) in terms of both primary and secondary oxidation products. However, other studies " concluded that carotenoids not only did not inhibit aerial lipid autoxidation (high PO2) but even exerted a prooxidant character, a phenomenon also observed at high carotenoid concentrations that could be due mainly to a more increased formation of carotene-peroxyl radicals, promoting the propagation of autoxidation. 

Capsorubin a carotenoid, M, 600.85, m.p. 218°C. It contains 2 identical cydopentanol rings. The secondary OH-groups have R configuration on C3, and S configuration on C3. C. is present in paprika fhiits (Capsicum annum), usually as esters, Ripe fruits contain about 1.5 mg per 100 g fresh weight. 

Zamilpa A, Tortoriello J, Navarro V, Delgado C, Alvarez L (2002) Five new steroidal saponins from Solanum chrysotrichum leaves and their antimycotic activity. J Nat Prod 65 1815-1819 Zechmeister L, Cholnoky LV (1927) tiber Paprika-Farbstoffe. Liebigs Ann 454 54-71 Zechmeister L, Cholnoky LV (1930) Zum Stand sauerstoffhaltiger Carotenoide in Pflanzen. 

Many spices also contain carotenoids. The composition and changes in concentration of different carotenoids during the ripening process in different species of pepper were studied by use of a binary gradient of water/acetone and a C18 RP column (245-247). An isocratic RP HPLC separation of capsanthin and capsorubin in Capsicum annum paprika and oleoresin on a Merck LiChros-pher 100 C18 5-p.m (0.4 X 25 cm) column with CH3CN/2-propanol/ethyl acetate (80 10 10) has been described (248). The carotenoids in saponified extracts of irradiated and ethylene oxide-treated red pepper were determined by HPLC on... 

Carotenoid composition of paprika fruit is shown in fig.2. The fruit distributed mono- and di-fatty acid esters of capsanthin and capsorubin in addition to free pigments, GLC analysis of fatty acids esterified with the carotenoid pigment indicated that capsanthin and capsorubin esterified mainly with saturated fatty acid to form diesters. Although unsaturated fatty acids are predominant in paprika, they were detected only with the fraction of capsanthin monoesters. 

Some of the methodologies used are outlined. For the separation of carotenes, interesting methods have been described. O Neil et al. evaluate the resolving capacity and quantihcation of Z/S-P-carotene isomers in four chromatographic columns and with various solvent systems, finding the Ca(OH)2 column the best. Biacs et al. employed the isocratic mixture acetonitrile/2-propanol/water (39 57 4) to separate esterified carotenoids of paprika. Khachik et al. quantify the major carotenoids and their esters in apricot, peach, melon, and grape. Heinonen et al. use this last method to analyze the carotenoid content of 69 types of vegetables and fruits, both fresh and processed. ... 

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