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.     

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.     

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%.     

Capillary electrophoresis analysis of orange juice pectinesterases

Pectinesterase (PE) was extracted from orange juice and pulp with 1 M NaCl, desalted, and separated using capillary electrophoresis (CE) gel procedures (CE-SDS-CGE) and isoelectric focusing (CE-IEF). PE resolved as a single peak using noncoated fused silica columns with CE-SDS-CGE. CE-IEF separation of PE required acryloylaminoethoxyethanol-coated columns, which had limited stability. Thermal stability of PE extracts before and after heating at 75 degrees C for 30 min and at 95 degrees C for 5 min established heat labile and heat stabile fractions with identical PE migration times by CE-SDS-CGE or CE-IEF. Peak magnitude decreased to a constant value as heating time increased at 75 degrees C. Regression analysis of CE-SDS-CGE peak migration times of molecular weight (MW) standards estimated both heat labile and heat stable PE at MW approximately 36 900. Traditional SDS-PAGE gel separation of MW standards and active PE isolated by IEF allowed estimation of MW approximately 36 000. CE-SDS-CGE allowed presumptive, but not quantitative, detection of active PE in fresh juice.     

Flavonoids in pigmented orange juice and second-pressure extracts

Flavanone glycosides (FGs) and polymethoxylated flavones (PMFs) have been studied in pigmented orange (Citrus sinensis) juices and second-pressure extracts (SPEs) by high-performance liquid chromatography and diode array detector. Detection was performed simultaneously at two different wavelengths: 278 nm (for determination of FGs) and 325 nm (for determination of PMFs). Qualitative distribution patterns of FGs and PMFs in juices and SPEs were similar, although the quantitative results are quite different. An increased narirutin/hesperidin ratio after centrifugation and the presence of high amounts of PMFs in SPEs, which remain unchanged after centrifugation, were observed. Therefore, a simple and affordable procedure to distinguish an orange juice from SPEs was proposed.     

Degradation of monoterpenes in orange juice by gamma radiation

Single-strength orange juice was irradiated with 0, 0.89, 2.24, 4.23, and 8.71 gGy of gamma radiation at 5 degrees C and then stored at 7 degrees C for 21 days. Volatile compounds, isolated by solid-phase microextraction, were separated and identified using a gas chromatograph equipped with a mass selective detector. The majority of the volatile compounds were terpenes, and the most abundant volatile compounds were ethanol and limonene. Most volatile compounds were stable during the 21-day storage period except geranial and neral which decreased over time. Irradiation reduced the concentration of acyclic monoterpenes, such as geranial, neral, myrcene, and linalool 1 and 7 days after irradiation, but did not affect other monoterpenes, sesquiterpenes, or other volatile compounds. The reduction of acyclic monterpenes increased linearly with radiation dose, and correlated with an increase in thiobarbituric acid reactive substrates (TBARS) content. Reduction in the concentration of monoterpenes induced by irradiation was not significant 21 days after irradiation. Our results indicate that acyclic monoterpenes are sensitive to irradiation whereas most other volatile compounds are resistant.     

Identification of synthetic regioisomeric lutein esters and their quantification in a commercial lutein supplement

Synthetic mixtures of 24 mono- and diesters of the asymmetric hydroxylated carotenoid lutein with lauric, myristic, palmitic, and stearic acids were analyzed by liquid chromatography-ultraviolet/visible spectroscopy (LC-UV-vis) and characterized by LC-mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR). These compounds were then used for identifying the composition of a commercial lutein supplement. This is the first report of chromatographic separation of mixed fatty acid lutein diesters. Preferential MS loss of fatty acids or water occurred initially at the 3'-hydroxy position in the epsilon-ionone ring and subsequently at the 3-hydroxy position in the beta-ionone ring. This selective fragmentation leads to facile assignment of the specific fatty acids to the appropriate regioisomeric ionone ring. A commercial lutein supplement contained low levels of two pairs of regioisomeric monoesters and nearly equal levels of three homogeneous diesters and five pairs of mixed diesters. Palmitic acid was the predominant fatty acid, with lower amounts of myristic, stearic, and lauric acids.     

Solid phase microextraction (SPME) of orange juice flavor: odor representativeness by direct gas chromatography olfactometry (D-GC-O)

The sensorial quality of solid phase microextraction (SPME) flavor extracts from orange juice was measured by direct gas chromatogrphy-olfactometry (D-GC-O), a novel instrumental tool for evaluating odors from headspace extracts. In general, odor impressions emerging from SPME extracts poorly resembled that of the original orange juice. In an attempt to improve the sensorial quality of extracts, sample equilibration and exposure times were varied on Carboxen/polydimethylsiloxane (CAR/PDMS) and divinylbenzene/Carboxen/polydimethylsiloxane (DVB/CAR/PDMS) SPME fibers. Best sensorial results were obtained with the DVB/CAR/PDMS fiber exposed for the shortest time; a trained panel of eight assessors judged its odor as the most representative of the reference orange juice. The analysis of odor active compounds by classical GC-O accounted for odor characteristics revealed by D-GC-O. A principal component analysis (PCA) was applied on SPME and headspace extracts using flavor recoveries as variables. Interestingly, PCA discriminated samples according to their odor representations described by D-GC-O analysis. This paper provides the first comprehensive methodology to "smell" SPME extracts and "evaluate" their sensorial quality. This method will enable future investigations to further improve SPME performance.     

橙汁冷冻浓缩动力学模型的研究

为了降低果汁在冷冻浓缩过程中由于冰晶夹带造成的的损失并提高浓缩速度,需要获得冷冻浓缩过程中冰晶的增长规律。以橙汁为原料,采用二级冷冻浓缩方法,利用冰与水溶液之间的固液相平衡的原理,将水分从液态转变为固态的冰,使橙汁的浓度从13°Bx浓缩到34°Bx。该试验中,测定了橙汁的浓度及其对应的冻结点温度的关系,结果表明:在试验范围内,冻结点与浓度呈线性关系并随浓度的升高而降低。该文还研究了在冰晶中所夹带的可溶性固形物与橙汁浓度的关系,所获得分配系数K表明,冰晶中所夹带的可溶性固形物随浓度的提高而增加。根据Fick扩散方程式建立了冰晶增长动力学模型,模型显示冰晶增长速率与冰晶质量成正比,同时也受到最大冰晶量的抑制,采用Powell法,对试验数据回归模型参数、采用龙格—库塔法进行数值积分求得模型解,计算结果与试验结果吻合,该模型有助于为建立冰晶增长的质量、热量传递之间的平衡提供一定的理论依据和试验参考,加深对橙汁冷冻浓缩过程的认识。     

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.     

Linalool in orange juice: origin and thermal stability

Linalool concentrations were determined in juice from three groups of 60 Valencia oranges using pentane:ether extraction and high-resolution capillary GC. The outer peel (flavedo) was removed from one group. The other two groups retained their peel intact. Juice was extricated from the halved fruits of the flavedo-less group and from one of the peel-intact groups using a hand reamer. A peel-cutting/macerating juice extractor was used for the other peel-intact group. Linalool concentrations were 0.004 mg/L in peeled fruit juice and 0.020 and 0.106 mg/L for hand-reamed and mechanically extracted peel-intact juice, respectively. Juice from peeled fruit contained significantly (P < 0.05) less linalool than peel-intact juice. Approximately 80% of the total juice linalool content was associated with peel using reamer design, and 96% was associated with peel-cutting/macerating design. Linalool increased with increasing peel oil levels; however, the increases were not proportionate. Since all commercial juices are mechanically extracted, the vast majority of linalool in commercial orange juice originates from the peel and not from the juice vesicle cytoplasm. Juice from peel-macerated, mechanically extracted fruit increased from 0.106 to 0.134 mg/kg after thermal processing, whereas juice from reamer extraction was essentially unchanged.     

Bioavailability and antioxidant effects of orange juice components in humans

Seven healthy females and six males consumed daily 256 mg of vitamin C, 271 mg of flavanones (mainly as glycosides), 6 mg of carotenoids (mainly xanthophylls and cryptoxanthins), and 0.16 mg of folate by incorporation of daily three times 236 mL of not from concentrate orange juice (OJ) into their habitual diet. At the end of 3 weeks, mean vitamin C, folate, carotenoid, and flavanone plasma concentrations increased significantly relative to baseline by 59% (p < 0.001), 46% (p = 0.018), and 22% (p < 0.001), and 8-fold (p = 0.045), respectively. Flavanones were excreted in urine 9-fold more at the end of the intervention (p = 0.01) but returned to baseline 2 days after study completion. After the 3 week intervention, plasma concentrations of vitamins A and E did not change. 8-Hydroxydeoxyguanosine in white blood cells declined by 16% (p = 0.38; n = 11), and in individuals with high baseline concentrations by 29% (p = 0.36; n = 7), respectively. Low-density lipoprotein (LDL)-/high-density lipoprotein (HDL)-cholesterol ratios decreased but cholesterol (HDL, LDL, total) and thiobarbituric acid reactive substance plasma concentrations did not change significantly. We conclude from this pilot study that OJ is an excellent food source to enhance circulating concentrations of valuable hydrophilic as well as lipophilic phytochemicals.     

Early stages of maillard reaction in dehydrated orange juice.

The formation of furoylmethyl derivatives of amino acids as indicators of the early stages of Maillard reaction in dehydrated orange juices and model systems was studied. In stored dehydrated orange juices, the presence of furoylmethyl derivatives of arginine, asparagine, proline, alanine, glutamic acid, and GABA was detected. Their formation increased with temperature of storage. After 2 weeks at 30 degrees C and a(w) = 0.44, the reconstituted orange juice contained 94 mg/L furoylmethyl derivatives, whereas up to 1215 mg/L was detected in samples stored at 50 degrees C.     

Anaerobic digestion of wastewater derived from the pressing of orange peel generated in orange juice production

A study of the anaerobic digestion of wastewater from the pressing of orange peel generated in orange juice production was carried out in a laboratory-scale completely stirred tank reactor at mesophilic temperature (37 degrees C). Prior to anaerobic treatment the raw wastewater was subjected to physicochemical treatment using aluminum sulfate as a flocculant and to pH reduction using a solution of sulfuric acid. The reactor was batch fed at COD loads of 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, and 5.0 g of COD. The process was very stable for all of the loads studied, with mean pH and alkalinity values of 7.5 and 3220 mg of CaCO3/L, respectively. The anaerobic digestion of this substrate was found to follow a first-order kinetic model, from which the specific rate constants for methane production, K(G), were determined. The K(G) values decreased considerably from 0.0672 to 0.0078 L/(g h) when the COD load increased from 1.5 to 5.0 g of COD, indicating an inhibition phenomenon in the system studied. The proposed model predicted the behavior of the reactor very accurately, showing deviations of <5% between the experimental and theoretical values of methane production. The methane yield coefficient was found to be 295 mL of CH4 STP/g of COD removed, whereas the mean biodegradability of the substrate (TOC) was 88.2%. A first-order kinetic model for substrate (TOC) consumption allowed determination of the specific rate constants for substrate uptake, K(C), which also decreased with increasing loading, confirming the above-mentioned inhibition process. Finally, the evolution of the individual volatile fatty acid concentrations (acetic, C2; propionic, C3; butyric, C4; isobutyric, iC4; valeric, C5; isovaleric, iC5; and caproic, C6) with digestion time for all loads used was also studied. The main acids generated were acetic and propionic for all loads studied, facilitating the conversion into methane.     

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.     

Identification of sulfur volatiles in canned orange juices lacking orange flavor

The purpose of this study was to understand why some canned orange juices are not perceived as orange juice. Sensory flavor profile data indicated that the primary odor (orthonasal) attributes were tropical fruit/grapefruit, cooked/caramel, musty, and medicine. By comparison fresh-squeezed juice lacked these odor attributes. GC-O analysis found 43 odor-active components in canned juices. Eight of these aroma volatiles were sulfur based. Four of the 12 most intense aroma peaks were sulfur compounds that included methanethiol, 1-p-menth-1-ene-8-thiol, 2-methyl-3-furanthiol, and dimethyl trisulfide. The other most intense odorants included 7-methyl-3-methylene-1,6-octadiene (myrcene), octanal, 2-methoxyphenol (guaiacol), 2-ethyl-4-hydroxy-5-methyl-3(2H)-furanone (homofuraneol), (E)-non-2-enal, (E,E)-deca-2,4-dienal, 4-hydroxy-3-methoxybenzaldehyde (vanillin), and alpha-sinensal. Odorants probably responsible for the undesirable sensory attributes included grapefruit (1-p-menth-1-ene-8-thiol), cooked [2-ethyl-4-hydroxy-5-methyl-3(2H)-furanone, 4-hydroxy-2,5-dimethyl-3(2H)-furanone (Furaneol), and 3-(methylthio)propanal (methional)], musty [7-methyl-3-methylene-1,6-octadiene and (E)-non-2-enal], and medicine (2-methoxyphenol). The canned juices also lacked several aldehydes and esters normally found in fresh orange juice.     

Liquid chromatographic methodology for the characterization of orange juice

Liquid chromatographic (LC) methodology potentially useful for the characterization of orange juice, with particular regard to detecting adulteration of orange juice by computer pattern recognition analysis, has been developed. After dilution with methanol the juice is extracted with hexane to remove the carotenoids, which are chromatographed on a C18 column with an acetonitrile-methanol-methylene chloride mobile phase and detection at 450 nm. Further extraction of the juice with methylene chloride isolates the methoxylated flavones, which are chromatographed by reverse phase LC with an acetonitrile-methanol-water mobile phase and detection at 280 nm. The flavanone glycosides remaining in solution are chromatographed on a C18 column with an acetonitrile-water mobile phase and detection at 280 nm. The precisions of the heights of the 32 LC peaks selected for pattern recognition analysis were determined from 5 replicate analyses of a single juice. Coefficients of variation of the replicates ranged from 0.3 to 4.5%, with an average of 2.1%. Adulteration of products with sodium benzoate-fortified pulpwash or grapefruit juice can be detected by this method. Pattern recognition analysis of the data obtained for 80 authentic and 19 adulterated orange juices showed that the method is potentially useful for distinguishing between authentic and adulterated products.     

Immunoanalytical method for quality control of orange juice products

The main goal of the present study is to develop an immunoanalytical method for the quality control of orange juice products. Peptides from various parts (juice, albedo, and flavedo) of citrus fruits (orange, mandarin, grapefruit, and lemon) were analyzed and isolated by SDS-PAGE. Antisera were developed in mice against the protein pool of orange juice and peel and tested by Western blot analysis. Using these antisera, some juice- and peel-specific peptides were detected. One of the antibodies in the antiserum developed against peel proteins recognized a single peel-specific peptide with a molecular mass of 28 kDa in 10000-fold dilution. It did not give any positive reactions against the sample prepared from the juice. The 24 and 27 kDa juice-specific peptides were isolated in electrophoretically pure form, and polyclonal antibodies were developed against them in mice. The anti-27 kDa antibody reacted with a 29 kDa protein in the peel sample, and it gave a positive reaction against the 27 kDa peptide of the juice. The antibodies developed in the course of the present work seem to be useful for determining the juice content in commercial citrus beverages and for evaluating the peel contamination in them.     

Identification of (poly)phenolic compounds in concord grape juice and their metabolites in human plasma and urine after juice consumption

Analysis of Concord grape juice by HPLC with ESI-MS(n), PDA, and fluorescence detection resulted in the identification and quantification of 60 flavonoids and related phenolic compounds, which were present at an overall concentration of 1508 ± 31 μmol/L. A total of 25 anthocyanins were detected, which were mono- and di-O-glucosides, O-acetylglucosides, O-p-coumaroyl-O-diglucosides, and O-p-coumaroylglucosides of delphinidin, cyanidin, petunidin, peonidin, and malvidin. The anthocyanins represented 46% of the total phenolic content of the juice (680 μmol/L). Tartaric esters of hydroxycinnamic acids, namely, trans-caftaric and trans-coutaric acids, and to a lesser extent trans-fertaric acid accounted for 29% of the phenolic content, with a total concentration of 444 μmol/L, of which 85% comprised trans-caftaric acid. Free hydroxycinnamic acids were also quantified but contributed to <1% of the total phenolic content (8.4 μmol/L). The other groups of polyphenolic compounds present in the juice, accounting for 24% of the total, comprised monomeric and oligomeric units of (epi)catechin and (epi)gallocatechin (248 μmol/L), flavonols (76 μmol/L), gallic acid (51 μmol/L), and trans-resveratrol (1.5 μmol/L). The bioavailability of the (poly)phenolic compounds in 350 mL of juice was investigated following acute intake by healthy volunteers. Plasma and urine were collected over 0-24 h and analyzed for parent compounds and metabolites. In total, 41 compounds, principally metabolites, were identified.