Gas chromatographic determination of fatty acids and sterols in orange juice

A gas chromatographic (GC) method has been developed for the simultaneous quantitation of fatty acids and sterols in orange juice, using a bonded phase fused silica capillary column of intermediate polarity, splitless automatic injection, and flame ionization detection. Sample preparation has been simplified by using 1 g C-18 adsorbent in a disposable minicolumn to extract 2 mL orange juice. Methylation of fatty acids and silylation of the sterols were carried out in the eluted extract (low polarity lipid fraction). The method precision was 7%; recoveries ranged from 83 to 113%. The precision of the injection technique was 2%. Seven major fatty acids and 5 sterols in orange juice were quantitated by the GC method and identified by GC/mass spectrometry. Quantitative data for several orange juice samples indicated that the levels of the compounds of interest were in the 1.3-72.0 mg/L range. The results demonstrate that bonded phase fused silica capillary GC has great versatility and potential for the quantitative determination of fatty acids and sterols.     

Identification and aroma impact of norisoprenoids in orange juice

Four norisoprenoids, alpha-ionone, beta-ionone, beta-cyclocitral, and beta-damascenone, along with their putative carotenoid precursors, were identified in Valencia orange juice using time-intensity GC-O, GC-MS, and photodiode array HPLC. alpha-Ionone and beta-cyclocitral are reported in orange juice for the first time. GC-O aroma peaks were categorized into seven groups with similar sensory qualities: citrus/minty, metallic/mushroom/geranium, roasted/cooked/meaty/spice, fatty/soapy/green, sulfury/solventy/medicine, floral, and sweet fruity. The four norisoprenoids contributed approximately 8% of the total aroma intensity and 78% of the total floral aroma category. The putative carotenoid norisoprenoid precursors, alpha- and beta-carotene, alpha- and beta-cryptoxanthin, and neoxanthin, were identified in the same orange juice using photodiode array HPLC retention times and spectral characteristics.     

Identification of isolutein (lutein epoxide) as cis-antheraxanthin in orange juice

The carotenoid profile of orange juice is very complex, a common characteristic for citrus products in general. This fact, along with the inherent acidity of the product, which promotes the isomerization of some carotenoids, makes the correct identification of some of these pigments quite difficult. Thus, one of the carotenoids occurring in orange juice has been traditionally identified as isolutein, a term used to refer to lutein epoxide, although enough evidence to support that identification has not been given. In this study, the carotenoid previously identified as isolutein/lutein epoxide in orange juice has been isolated and identified as a 9 or 9'-cis isomer of antheraxanthin as a result of different tests. To support this identification, a mixture of geometrical isomers of lutein epoxide isolated from petals of dandelions was analyzed under the same conditions used for orange juice carotenoids to check that neither their retention times nor their spectroscopic features matched with those of the orange juice carotenoid now identified as a cis isomer of antheraxanthin.     

Characterization of carotenoids in juice of red navel orange (Cara Cara)

High-performance liquid chromatography with photodiode array detection was applied for the separation and characterization of carotenoids from a red-fleshed navel orange (Cara Cara). Carotenoid pigments were extracted using hexane/acetone/ethanol and saponified using 10% methanolic potassium hydroxide. More than 29 carotenoid pigments were separated within 60 min using a ternary gradient (75% acetonitrile/25% methanol, methyl tert-butyl ether, and water) elution on a C(30) reversed-phase column. The presence of lycopene (3.9 +/- 1.1 ppm) and a relatively large percentage of beta-carotene were distinct differences in the pigment profile of this red navel orange juice as compared to the profile of standard navel orange juice. The juice color in the red navel was much deeper orange than that from navel orange; hue angle ranged from 84.1 to 89.4 for red navel compared to 98.2 to 100.5 for standard navel.     

Identification of zeinoxanthin in orange juices

The monohydroxycarotenoid fraction of orange juice has been isolated by TLC and studied to determine whether the carotenoid accompanying beta-cryptoxanthin was alpha-cryptoxanthin or zeinoxanthin. The provitamin A carotenoid alpha-cryptoxanthin has been widely reported in orange juice, although its identification has been carried out mainly on the basis of its spectral features, which are virtually identical with those of its non-provitamin A isomer, zeinoxanthin. As a result of a study of the UV-vis and mass spectra of the monohydroxycarotenoid fraction and of the methylation test, it was concluded that the carotenoid accompanying beta-cryptoxanthin was the non-provitamin A carotenoid zeinoxanthin.     

Supercritical CO(2) and subcritical propane extraction of pungent paprika and quantification of carotenoids, tocopherols, and capsaicinoids.

Ground paprika (Capsicum annuum L.) was extracted with supercritical carbon dioxide (SC-CO(2)) and subcritical propane at different conditions of pressure and temperature to estimate the yield and variation in carotenoid, tocopherol, and capsaicinoid contents and composition. The yield of paprika extract was found to be affected by the extraction conditions with SC-CO(2) but fairly constant at different conditions with subcritical propane. The maximum yields of oleoresin were 7.9 and 8.1% of ground paprika by SC-CO(2) and subcritical propane, respectively. The quantitative distribution of carotenoids, tocopherols, and capsaicinoids between paprika extract and powder was influenced by extraction conditions. SC-CO(2) was inefficient in the extraction of diesters of xanthophylls even at 400 bar and 55 degrees C, whereas tocopherols and capsaicinoids were easy to extract at these conditions. Under mild conditions subcritical propane was superior to SC-CO(2) in the extraction of carotenoids and tocopherols but less efficient in the extraction of capsaicinoids.     

Carotenoid profile modification during refrigerated storage in untreated and pasteurized orange juice and orange juice treated with high-intensity pulsed electric fields

A comparative study was made of the evolution and modification of various carotenoids and vitamin A in untreated orange juice, pasteurized orange juice (90 degrees C, 20 s), and orange juice processed with high-intensity pulsed electric fields (HIPEF) (30 kV/cm, 100 micros), during 7 weeks of storage at 2 and 10 degrees C. The concentration of total carotenoids in the untreated juice decreased by 12.6% when the juice was pasteurized, whereas the decrease was only 6.7% when the juice was treated with HIPEF. Vitamin A was greatest in the untreated orange juice, followed by orange juice treated with HIPEF (decrease of 7.52%) and, last, pasteurized orange juice (decrease of 15.62%). The decrease in the concentrations of total carotenoids and vitamin A during storage in refrigeration was greater in the untreated orange juice and the pasteurized juice than in the juice treated with HIPEF. During storage at 10 degrees C, auroxanthin formed in the untreated juice and in the juice treated with HIPEF. This carotenoid is a degradation product of violaxanthin. The concentration of antheraxanthin decreased during storage, and it was converted into mutatoxanthin, except in the untreated and pasteurized orange juices stored at 2 degrees C.     

Orange, mandarin, and hybrid classification using multivariate statistics based on carotenoid profiles

A study was undertaken to differentiate citrus on the basis of a carotenoid profile obtained from the HPLC determination of 12 carotenoids found in saponified fresh juice. Mandarin, orange, and various hybrid varieties were analyzed to determine their carotenoid profiles. The resulting peak areas were analyzed using principal component analysis (PCA), canonical discriminate analysis (CDA), and Mahalanobis distances. These were used to develop models for classifying the juices into appropriate groups. Thirty-two samples were analyzed to determine classification techniques. Mandarin and orange juices were quite distinct, with the hybrids scattered throughout the mandarin and orange clusters using PCA. CDA resulted in three distinct groups with only a few of the hybrids in the orange grouping.     

Effects of blood orange juice intake on antioxidant bioavailability and on different markers related to oxidative stress

Orange juice is a source of antioxidants that might afford in vivo protection from oxidative stress. To test this hypothesis, we carried out a human intervention study with blood orange juice containing high amounts of vitamin C, anthocyanins, and carotenoids. Sixteen healthy female volunteers were enrolled in a crossover study and were given 600 mL/day of blood orange juice or a diet without juice for 21 days. Before and after each intervention period, plasma vitamin C, cyanidin-3-glucoside, and carotenoids were quantified. Furthermore, plasma antioxidant capacity, malondialdehyde concentration in plasma, 11-dehydrotromboxane B(2) urinary excretion, and lymphocyte DNA damage were evaluated as biomarkers of oxidative stress. Blood orange juice consumption determined a significant increase in plasma vitamin C, cyanidin-3-glucoside, beta-cryptoxanthin, and zeaxanthin. Also, lymphocyte DNA resistance to oxidative stress was improved whereas no effect was observed on the other markers that we analyzed. In turn, these results suggest that blood orange juice is a bioavailable source of antioxidants, which might moderately improve the antioxidant defense system; however, the long-term effects of its consumption are to be further investigated.     

Characterization of carotenoid high-producing Capsicum annuum cultivars selected for paprika production

Twelve selected pepper (Capsicum annuum L.) cultivars, bred for mechanical harvesting (grouped ripeness) and adaptation to different cultivation cycles (short to long), have been characterized by their carotenoid pigment content and composition with the aim of producing high-quality paprika. A detailed analysis of the carotenogenesis was performed throughout the ripening process, but with special emphasis on the ripe stage, with the aim of selecting the best cultivar for paprika production. The MA1 cultivar (with grouped ripeness and very short cultivation cycle) showed the highest carotenoid content (12697.58 mg/kg dwt), followed by DN5 and RN2 cultivars with 11086.88 and 10393.29 mg/kg dwt, respectively. Most of the cultivars (MA3, RN1, LR2, LR7, DN3, DR6, Datler, and Mulato) showed a total carotenoid content in the range of 7000-9700 mg/kg dwt. In general, chlorophyll-retaining character was related to high carotenoid content (cultivars DN3, DN5, MA3, Mulato, RN1, and RN2). The general trend of the cultivation cycle was that the shorter the cycle, the higher the total carotenoid content (as exemplified by the cultivar MA1). The lowest total carotenoid content was found for the RR1 cultivar (4856.77 mg/kg dwt), which showed the longest cultivation cycle. Carotenogenic capacity of the cultivars has been discussed relative to total carotenoid content and the R/Y and Caps/Zeax ratios, the main quality traits for breeding cultivars for production of high-quality paprika. The cultivar MA1, with the highest total carotenoid content, high R/Y (2.11) ratio, and highest Caps/Zeax (9.85) ratio, was found to be the most suitable cultivar for paprika production in terms of carotenoid pigment biosynthesis capacity. Moreover, this cultivar has a short cultivation cycle and grouped ripeness, which are both important characteristics for a proper application of mechanical harvesting. The potential improvement of other varieties is also discussed.     

Profiling of carotenoids in tomato juice by one- and two-dimensional NMR

Epidemiological data have shown a link between dietary intake of tomatoes and tomato products (rich in carotenoids) and a decreased risk of chronic diseases. The carotenoid profile in tomato products depends on tomato variety as well as the thermal conditions used in processing. The final carotenoid profile may affect the bioaccessibility and bioavailability of these biomolecules. Therefore, nondestructive, reliable methods are needed to characterize the structural and stereochemical variation of carotenoids. CDCl(3) rapid extraction was used to extract carotenoids from tomato juice as an alternative rapid procedure that minimizes solvents and time consumption prior to NMR analysis. The profile of these biomolecules was characterized by application of high-resolution multidimensional NMR techniques using a cryogenic probe. The combination of homonuclear and heteronuclear two-dimensional NMR techniques served to identify (all-E)-, (5Z)-, (9Z)-, and (13Z)-lycopene isomers and other carotenoids such as (all-E)-beta-carotene and (15Z)-phytoene dissolved in the extracted lipid mixture. The use of one-dimensional NMR enabled the rapid identification of lycopene isomers, thereby minimizing further isomerization of (all-E)-lycopene as compared to HPLC data. On the basis of the assignments accomplished, the carotenoid profile of typical tomato juice was successfully determined with minimal purification procedures.     

Carotenoid esters in vegetables and fruits: a screening with emphasis on beta-cryptoxanthin esters

Carotenoids are found in food plants in free form or as fatty acid esters. Most studies have been carried out after saponification procedures, so the resulting data do not represent the native carotenoid composition of plant tissues. Therefore, nonsaponified extracts of 64 fruits and vegetables have been screened to determine the amount of carotenoid esters in food plants. Because one of the major problems in the quantitation of carotenoids is the availability of pure standard material, the total carotenoid ester content was calculated as lutein dimyristate equivalents. Lutein dimyristate was independently synthesized from lutein and myristoyl chloride. The highest ester concentrations were found in red chili (17.1 mg/100 g) and orange pepper (9.2 mg/100 g); most of the investigated fruits and vegetables showed concentrations up to 1.5 mg/100 g. Special attention was dedicated to beta-cryptoxanthin esters. To enable an accurate detection of the beta-cryptoxanthin ester content, beta-cryptoxanthin was purified from papaya and used for synthesis of beta-cryptoxanthin laurate, myristate, and palmitate, representing the major beta-cryptoxanthin esters in food plants. The study proved tropical and subtropical fruits to be an additional source of beta-cryptoxanthin esters in the human diet. The contents ranged from 8 microg/100 g beta-cryptoxanthin laurate in Tunisian orange to 892 microg/100 g beta-cryptoxanthin laurate in papaya.     

Carotenoids, color, and ascorbic acid content of a novel frozen-marketed orange juice

A recently developed food, the so-called ultrafrozen orange juice (UFOJ), has been characterized in terms of carotenoid pigments, ascorbic acid, and color. The juice, obtained from Valencia late oranges, is frozen immediately after the squeezing of the oranges, which makes it a product showing good organoleptic and nutritional quality. In relation to the carotenoid profile, it was observed that the 5,6-epoxy carotenoids violaxanthin and antheraxanthin (specifically (9Z)-violaxanthin and (9Z)- or (9'Z)-antheraxanthin), were by far the major pigments and that dihydroxycarotenoids predominate over monohydroxycarotenoids. As far as color was concerned, it was seen that there were little differences among the juices analyzed. The hue of the samples, ranging from 77.19 degrees to 80.15 degrees and from 79.99 degrees to 83.04 degrees depending on the kind of instrumental measurement, and their chroma (ranging from 63.06 to 72.25 and from 44.40 to 58.38) revealed readily that the juice surveyed exhibited a deep orangeish coloration, the color coordinate best correlated with the total carotenoid content being b*. The levels of ascorbic acid ranged from 332.64 to 441.44 mg/L, with an average content of 391.06 +/- 28.86 mg/L.     

Differences in the carotenoid content of ordinary citrus and lycopene-accumulating mutants

High-performance liquid chromatography, coupled with photodiode array detection, was used to analyze the carotenoid composition of peel and juice vesicle tissues of ordinary and lycopene-accumulating mutants (referred to as red mutants in this article) of orange, pummelo, and grapefruit. Thirty-six major carotenoids, including some cis-trans isomers, were separated on a C30 reversed phase column, and 23 of them were identified on the basis of retention times and spectral characteristics with authentic standards. Carotenoid profiles varied with tissue types, citrus species, and mutations. beta-Citraurin occurred in the peel of oranges but not in juice vesicles, whereas the reverse was found for violaxanthin, 9-cis-violaxanthin, and luteoxanthin. The diversity of carotenoids in peel and juice vesicle tissues and the fact that there was over 250 times higher content of total carotenoids in peels of Yuhuan pummelo than juice vesicles suggested that the biosynthesis of carotenoids in these two tissues was independent and exchange of carotenoids between the tissues was not likely. Lutein was observed in peels of pummelos and grapefruits and juice vesicles of ordinary pummelo but not in orange tissues. Accumulation of lycopene and beta-carotene was observed in red mutant citrus, except for the peel of Cara Cara red orange. Additionally, phytoene accumulated in all tissues except for the peel of Chuzhou Early Red pummelo. No obvious change in the total content of xanthophylls was observed in the Cara Cara red orange. Ordinary grapefruit (Marsh) tissues and pummelo (Yuhuan) juice vesicles were almost devoid of carotenoids, and in red mutants, the content of total carotenoids increased dramatically up to 790-fold. The different changes in carotenoid content and profiles in mutant(s) of different citrus species suggest that the underlying mechanisms for the mutations might be different.     

Seasonal changes of carotenoid pigments and color in Hamlin, Earlygold, and budd blood orange juices

The developmental patterns of carotenoids in Hamlin, Earlygold (an early-maturing selection), and Budd Blood sweet orange juices were studied during the September to mid-January period of the 1996-97 and 1997-98 seasons. The carotenoid concentration of Earlygold increased by as much as 4.9 times during the color development compared to 3.9 times in Hamlin and 4.5 times in Budd Blood juice in the same period. For the profiles of carotenoid pigment, dramatic changes occurred among the pigments that were present in high concentrations at the beginning of the season, with lutein and violaxanthin noted as the predominant pigments in Hamlin fruit. A marked increase in the percentage of beta-cryptoxanthin allowed it to become a major pigment in the late stage of maturation. The color development in the new cultivar Earlygold was especially notable, reaching the 36 color number, which is grade A, by late October to mid-November whereas Hamlin juice did not reach this grade A color number until January. Budd Blood juice was similar in carotenoid pigment content and seasonal changes to Hamlin juice, but also, the development of red anthocyanin pigment in January significantly increased juice color.     

Capsanthone 3,6-epoxide, a new carotenoid from the fruits of the red paprika Capsicum annuum L

The structure of a new carotenoid, isolated from the fruits of the red tomato-shaped paprika Capsicum annuum L., was elucidated to be (3S,5R,6S,5'R)-3,6-epoxy-5,6-dihydro-5-hydroxy-beta,kappa-carotene-3',6'-dione by spectroscopic analyses, including fast atom bombardment collision-induced dissociation-mass spectrometry/mass spectrometry (FAB CID-MS/MS) and was designated capsanthone 3,6-epoxide. Capsanthone 3,6-epoxide is assumed to be an oxidative metabolite of capsanthin 3,6-epoxide in paprika.     

2-Methyl-3-furanthiol and methional are possible off-flavors in stored orange juice: aroma-similarity, NIF/SNIF GC-O, and GC analyses.

The occurrence of methional in fresh orange juice, and possible occurrence of beta-damascenone in heated orange juice, has been previously suggested. Here we report on the occurrence of 2-methyl-3-furanthiol in the headspace, collected by solid-phase micro-extraction, of fresh, pasteurized, and stored orange juice. The contents of 2-methyl-3-furanthiol and methional were quantified, and the relative level of beta-damascenone was estimated, in the headspace of fresh, pasteurized, and stored orange juices using the nasal impact frequency (NIF) and surface of NIF (SNIF) GC-Olfactometry procedure. 2-Methyl-3-furanthiol concentrations were 2 ng/L in fresh and pasteurized Shamuti orange juice, and 270 ng/L in stored juice of the same variety. Methional concentrations were 550, 830, and 11,550 ng/L in fresh, pasteurized, and stored pasteurized juices, respectively. beta-Damascenone content appeared to have increased during pasteurization and storage. Aroma-similarity experiments strongly suggest that 2-methyl-3-furanthiol and methional, at the levels found in stored orange juice (21 days at 35 degrees C), contribute to stored orange juice off-flavor.     

Flavor, color, and vitamin C retention of pulsed electric field processed orange juice in different packaging materials

Effects of packaging materials, storage temperature, and time on the stability of pulsed electric field (PEF) processed orange juice were investigated. Single-strength orange juice was treated with PEF at an electric field strength of 35 kV/cm for 59 micros using an integrated pilot plant scale PEF processing and glovebox packaging system. The retention of eight orange juice aroma compounds, color, and vitamin C in glass, polyethylene terephthalate (PET), high-density polyethylene, and low-density polyethylene were evaluated at 4 and 22 degrees C for 112 days. Packaging material had a significant effect (p < or = 0.05) on the retention of orange juice aroma compounds, color, and vitamin C. PEF-treated orange juice had a shelf life of >16 weeks in glass and PET at 4 degrees C.     

Liquid chromatographic determination of naringin and neohesperidin as a detector of grapefruit juice in orange juice

Naringin/neohesperidin ratios can be used to differentiate orange juice which may contain added grapefruit juice from orange juice which may include juices from other naringin-containing cultivars. The naringin/neohesperidin ratios in juice vary from 14 to 83 in grapefruit (C. grandis) and from 1.3 to 2.5 in sour orange (C. aurantium) cultivars; the ratio is always less than 1 for the K-Early tangelo. Concentrations of both naringin and neohesperidin can be determined in orange juice by using a single liquid chromatographic isocratic reverse-phase system with a C-18 column. The detection limit for both compounds is 1 ppm with a linear working range to 500 ppm. Concentration relative standard deviations range from 0.47 to 1.06% for naringin and from 0.4 to 1.27% for neohesperidin. Naringin and neohesperidin recoveries ranged from 93 to 102% at concentrations of 5 and 50 ppm. Naringin values from blind duplicate samples of orange/grapefruit juice blends could be duplicated to +/- 3%.