Characteristic odor components of Citrus sphaerocarpa Tanaka (Kabosu) cold-pressed peel oil

The volatile components of Citrus sphaerocarpa Tanaka (Kabosu) cold-pressed peel oil were investigated by chemical and sensory analyses. Monoterpene hydrocarbons (more than 94.6%) were predominant in Kabosu peel oil, with limonene and myrcene accounting for the major proportions (70.5% and 20.2%, respectively). The Kabosu oxygenated fraction was characterized by quantitative abundance in aldehydes and a relatively wide variety of alcohols. The weight percentages of aldehydes, alcohols, and esters in Kabosu cold-pressed oil were 1.3%, 0.1%, and 0.1%, respectively. Aroma extract dilution analysis was employed for determination of the odors of Kabosu volatile components, flavor dilution factors, and relative flavor activities. Gas chromatography/olfactometry using Kabosu cold-pressed oil and its oxygenated fraction completed by a chiral analysis revealed that (R)-(+)-citronellal is a characteristic element of Kabosu peel oil odor. Careful sniff testing demonstrated that aqueous solutions of both 0.25% and 0.016% (R)-(+)-citronellal gave an odor similar to that of Kabosu.     

Characterization of Citrus unshiu (C. unshiu Marcov. forma Miyagawa-wase) blossom aroma by solid-phase microextraction in conjunction with an electronic nose

The volatile composition of the headspace from Citrus unshiu Marcov. forma Miyagawa-wase blossom was investigated. The volatile constituents were absorbed by a solid-phase microextraction (SPME) fiber and directly transferred to a GC-MS. Volatile compositional changes of C. unshiu blossom prepared via different drying methods (shade, microwave, and freeze-drying methods) were also determined. A total of 96 volatile constituents were confirmed in the headspace from these samples. Monoterpene hydrocarbons were prominent in the headspace volatiles of C. unshiu blossom: fresh, 84.1%; shade-dried, 60.0%; microwave-dried, 88.4%; and freeze-dried, 29.9%. p-Cymene (23.3%) was the most abundant component in the headspace of fresh C. unshiu blossom; gamma-terpinene was the most abundant in shade- and microwave-dried samples (26.8 and 31.2%, respectively) and beta-caryophyllene (10.5%) in freeze-dried sample. By using an electronic nose consisting of six metal oxide sensors, principal component analysis of the volatile compounds showed a clear aroma discrimination of the fresh and all dried blossom samples.     

Juice quality of two new mandarin-like hybrids (Citrus clementina Hort. ex Tan x Citrus sinensis L. Osbeck) containing anthocyanins

Tacle and Clara [Monreal clementine (Citrus clementina Hort. ex Tan) x Tarocco orange (Citrus sinensis L. Osbeck)] are two new triploid citrus hybrids developed by the CRA-Istituto Sperimentale per l'Agrumicoltura (Acireale, Italy). The fruits are easy-peeling and juicy, have a pleasant taste like the Tarocco orange, and are sweet like the Monreal clementine. In addition, a distinctively attractive characteristic of these mandarin-like fruits is the red-pigmented flesh caused by the presence of anthocyanins. This study reports the juice quality attributes of fresh fruits harvested at different ripening stages and of cold-stored fruits kept for 104 days at 6+/-1 degrees C and 90-95% relative humidity. Physico-chemical analyses showed that the fresh-fruit juice yield ranged between 39 (Tacle) and 41% (Clara); these values were 11-14% lower after 104 days of storage. Vitamin C content in the Clara juice was decisively higher than that in the Tacle juice. Juice anthocyanins and other polyphenols increased during cold storage. These results show that low-temperature storage enhances the functional attributes of Tacle and Clara fruit juices.     

Juice components of a new pigmented citrus hybrid Citrus sinensis (L.) Osbeck x Citrus clementina Hort. ex Tan

Fruit juice of a new pigmented citrus hybrid named Omo-31 and those of its parents clementine cv. Oroval (Citrus clementina Hort. ex Tan.) and Moro orange [Citrus sinensis (L.) Osbeck] were analyzed during fruit maturation to determine juice yield, total soluble solids (TSS), total acidity (TA), TSS/TA ratio (classical parameters of quality), and potential health beneficial components, such as vitamin C, flavanones, anthocyanins, and phenolic acids. Results showed that juice yield, TA, TSS, and TSS/TA ratio values of Omo-31 were similar to those of the Moro orange. Vitamin C content of the new hybrid was slightly higher than that of clementine and lower than that of the Moro orange, but at maturity stage no differences were observed among the three genotypes. The phenolic compounds content of the new hybrid and those of the parents and their evolution during maturation were studied. At maturity stage the amount of anthocyanins, flavanones, and hydroxycinnamic acids in Omo-31 was found to be notably higher than those of the parents. The high level of antioxidant substances makes this new fruit important for its nutritional benefits.     

Changes of the volatile profile and artifact formation in Daidai (Citrus aurantium) cold-pressed peel oil on storage

The chemical changes and artifact formation in daidai (Citrus aurantium L. var. Cyathifera Y. Tanaka) cold-pressed peel oil upon storage at 20, 5, and -21 degrees C for 3, 6, and 12 months were investigated using capillary gas chromatography (GC) and GC-MS. Major changes occurred in the oil stored at 20 and 5 degrees C. No changes were found at -21 degrees C. Monoterpene hydrocarbons decreased from 98.0 to 66.4% upon 12 months at 20 degrees C, while sesquiterpene hydrocarbons and alcohols increased from 0.1 to 2.4% and from 0.3 to 7.9%, respectively. Notable decreases of germacrene D, myrcene, linalyl acetate, and limonene occurred. Prominent increases of cis-carveol, trans-beta-farnesene, trans-p-2,8-menthadien-1-ol, linalool, and beta-caryophyllene were found. Thirty-four artifact compounds constituting 17.0% of the total volatile compounds were formed upon 12 months at 20 degrees C. The artifacts consisted of 13 alcohols (6.0%), five carbonyl compounds (5.3%), seven esters (4.9%), three epoxides (0.4%), four hydrocarbons (0.3%), and two unidentified. The prominent artifact compounds were (+)-carvone, trans,trans-farnesyl acetate, sabinene hydrate, 1-octen-3-ol, cis,cis-farnesyl acetate, and dihydrocarveol acetate. The results could be applied for monitoring and control of the flavor quality of daidai essential oil and related products.     

Ultrafiltration performance of heat-treated shamouti orange [Citrus sinensis (L.) Osbeck] juice.

During ultrafiltration of orange juice with inorganic membranes, heating of the juice prior to the filtration experiment resulted in a significant increase of the fouling indices. The effect of the irreversible fouling (Rif) was always high, whereas the reversible fouling (Rrf) depended on the treatment. It was clearly seen that fouling was reduced after pectin degradation, but the heat treatment applied to the juice before filtration still resulted in reduced fluxes. It is suggested that pectins and proteins that undergo flocculation/coagulation during the heat treatment tend to interact with the membrane-filtering layer and to cause reduction of permeation flux. To clean the membrane to restore its pure water flux, close to the initial one, a proteolitic enzyme detergent wash was needed.     

Involvement of proteins in cloud instability of valencia orange [Citrus sinensis (L.) Osbeck] juice.

The present study examined the involvement of proteins in cloud flocculation of Valencia orange juice. Marked differences in cloud instability were found between juices of different harvest dates. Heating of enzymatic pectin degraded juice from April and June harvests resulted in development of clumps and their precipitation. Although the juice from both harvesting dates remained hazy, the juice of April harvest was more turbid than that from June. Usually clarification increases as the temperature increases from ambient to 125 degrees C. Clarification occurred at pH 2.5-4.5 and was maximal at pH 3.5. The clarification of the April harvest juice was markedly lower than that of the June harvest. The fresh juice contained about 5.2 and 1.7 mg mL(-1) insoluble cloud matter (ICM) and alcohol-insoluble serum solids (AISS), respectively. The ICM and the AISS, respectively, contained: proteins (244.5+/-8.7 and 132+/-1.8 microg mg(-1)), galacturonic acid (40+/-0 and 120+/-0 microg mg(-1)) and neutral sugars (270+/-39 and 329+/-23 microg mg(-1)). Enzymatic pectin degradation resulted in removal of a marked portion of the pectin, and was accompanied by partial removal of neutral sugars (mainly glucose and galactose) and some proteins from the pectic polymer in both AISS and ICM. Proteins of the AISS included major bands at 10-14, 20, and 28 kDa and those of the ICM bands at 22, 24, 26, and 45 kDa.     

Involvement of proteins in cloud instability of shamouti orange [Citrus sinensis (L.) Osbeck] juice.

The present study provides evidence for the involvement of protein in cloud instability of natural orange juice. No heat-coagulable proteins were found in the serum. Insoluble cloud matter (ICM) was heat-flocculated following enzymatic pectin degradation (EPD). The degree of flocculation depended on temperature (from approximately 50 to 75 degrees C) and was highest at pH 3.5. The fresh juice contained about 6.5 and 1.8 mg mL(-1) of ICM and alcohol-insoluble solids of the serum (AISS), respectively. The ICM and the AISS contained, respectively, proteins (182+/-14 and 119+/-3 microg mg(-1)), galacturonic acid (37+/-6.6 and 175+/-1 microg mg(-1)), and neutral sugars (350+/-44 and 338+/-22 microg mg(-1)). EPD resulted in removal of a marked portion of the pectin and was accompanied by partial removal of neutral sugars (mainly glucose and galactose) and some proteins from the pectic polymer in both AISS and ICM. Under electrophoresis, proteins of the AISS included bands in the range of 20-52 kDa and 10-14 kDa and those of the ICM at 22 and 50 kDa.     

Characterization of the most odor-active volatiles of orange wine made from a Turkish cv. Kozan (Citrus sinensis L. Osbeck)

The aroma-active compounds of cv. Turkish Kozan orange wine were analyzed by sensory and instrumental analyses. Liquid-liquid extraction with dichloromethane was used for extraction of volatile components. According to sensory analysis, the aromatic extract obtained by liquid-liquid extraction was representative of orange wine odor. A total of 63 compounds were identified and quantified in orange wine. The results of the gas chromatography-olfactometry analysis showed that 35 odorous compounds were detected by the panelists. Of these, 28 aroma-active compounds were identified. Alcohols followed by terpenes and esters were the most abundant aroma-active compounds of the orange wine. Among these compounds, ethyl butanoate (fruity sweet), 3-methyl-1-pentanol (roasty), linalool (floral citrusy), gamma-butyrolactone (cheesy burnt sugar), 3-(methylthio)-propanol (boiled potato, rubber), geraniol (floral citrusy), and 2-phenylethanol (floral rose) were the most important contributors to the aroma of the orange wine because they were perceived by all eight panelists.     

Gene isolation, analysis of expression, and in vitro synthesis of glutathione S-transferase from orange fruit [Citrus sinensis L. (Osbeck)

Glutathione S-transferases (GSTs) (EC 2.5.1.18) are ubiquitous enzymes that have a defined role in xenobiotic detoxification, but a deeper knowledge of their function in endogenous metabolism is still lacking. In this work, we isolated the cDNAs as well as the genomic clones of orange GSTs. Having considered gene organization and homology data, we suggest that the isolated GST gene is probably involved in the vacuolar import of anthocyanins. We also found that the blood and blond orange GSTs shared the same nucleotide sequences, but as expected, the GST expression in the nonpigmented orange cultivar [Citrus sinensis L. (Osbeck)] (Navel and Ovale) was strongly reduced as compared to that of the pigmented orange (Tarocco). Interestingly, in the crude extracts of pigmented orange fruits, the GST activity was reproducibly detected by providing either 1-chloro-2,4 dinitrobenzene (CDNB) or cyanidin-3-O-glucoside (C-3-G) as substrates; moreover, we have shown that cyanidin-3-O-glucoside acted as a powerful competitive inhibitor of 1-chloro-2,4 dinitrobenzene conjugation to reduced glutathione (GSH) in the pigmented orange, confirming that this molecule might easily bind to the active site of the enzyme and functions as a putative substrate. In addition, we have reported here the successful in vitro expression of orange GST cDNAs leading to a GST enzyme that is active against cyanidin-3-O-glucoside, thus suggesting the probable involvement of the isolated gene in the tagging of anthocyanins for vacuolar import. This last result will help to study the kinetic and structural properties of orange fruit GST avoiding time-consuming protein purification procedures.     

Metabolism of [(1)(4)C]prometryn in rats.

[(1)(4)C]Prometryn, 2, 4-bis(isopropylamino)-6-(methylthio)-s-triazine, was orally administered to male and female rats at approximately 0.5 and 500 mg/kg; daily urine and feces were collected. After 3 or 7 days rats were sacrificed, and blood and selected tissues were isolated. The urine and feces extracts were characterized for metabolite similarity as well as for metabolite identification. Over 30 metabolites were observed, and of these, 28 were identified mostly by mass spectrometry and/or cochromatography with available reference standards. The metabolism of prometryn was shown to occur by N-demethylation, S-oxidation, S-S dimerization, OH substitution for NH(2) and SCH(3), and conjugation with glutathione or glucuronic acid. Rat liver microsomal incubations of prometryn were conducted and compared to the in vivo metabolism. Both in vivo and in vitro phase I metabolisms of prometryn were similar, with S-oxidation and N-dealkylation predominating. The involvement of cytochrome P-450 and flavin-containing monooxidase in the in vitro metabolism of prometryn was investigated.     

Degradation of [(14)C]carfentrazone-ethyl under aerobic aquatic conditions.

Carfentrazone-ethyl (CF-E) is an aryl triazolinone reduced-risk herbicide for use on corn, wheat, and soybean. As part of the assessment of its metabolic fate, the aerobic aquatic metabolism of [(14)C]CF-E at a concentration of 0.22 microg/g was investigated. Two separate aquatic sediments (silty clay loam and clay loam soils, flooded with water) were used in the study. At each of eight samplings throughout the 30-day study, the distribution of radioactivity between surface water, sediment, and volatile fractions was assessed. At zero time, the majority of the applied radioactivity was contained in the water layer (83-90%), declining to 70-80% after 30 days. This was coupled with an increase in the percent radioactivity in the soil layer from 4-6% at day 0 to 13-19% after 30 days. Nonextractable soil residues and volatile degradation products were formed in negligible amounts. Analysis of the incubation extracts from either aquatic sediment indicated a rapid conversion (<2 days) of the parent CF-E ester to carfentrazone-chloropropionic acid. Over time, increasing amounts of a cascade of acidic degradation products comprising >90% of the applied radioactivity were formed. Identification of these degradation products was initially achieved through chromatographic comparison with reference synthetic standards and subsequently confirmed using LC-MS analysis. A degradation pathway for CF-E under aerobic aquatic conditions is proposed.     

Evaluation of Sweet Orange (Citrus sinensis L. Osbeck) cv. Sathgudi Budded on Five Rootstocks for Differential Behavior in Relation to Nutrient Utilization in Alfisol

The experiment was conducted to evaluate the nutrient utilization ability of sweet orange (Citrus sinensis L. Osbeck) budded on five rootstocks (viz., Sathgudi, Rangpur lime, Cleopatra mandarin, Troyer citrange, and Trifoliate orange) in Alfisols at the experimental farm of the Citrus Improvement Project, S. V. Agricultural College Farm, Tirupati, Andhra Pradesh, India. Results of the study revealed that all the five rootstocks showed differential behaviors in terms of nutrient absorption from the soil. Rootstocks exhibited significant variation in the leaf content of potassium (K), copper (Cu), manganese (Mn), and boron (B) at all the three stages of sampling. Concentrations of the following key nutrient elements significantly varied: phosphorus (P), calcium (Ca), magnesium (Mg), zinc (Zn), and Cu at stage 1; K, Ca, Mg, Zn, iron (Fe), and Mn at stage 2; and nitrogen (N), P, Zn, Fe, and B at stage 3. The performances of rootstocks in terms of relative nutrient accumulation indices (RNAIs) were in the order of Sathgudi (1.00) > Rangpur lime (0.98) > Cleopatra mandarin (0.96) > Trifoliate orange (0.76) > Troyer citrange (0.69). The present study clearly demonstrated that citrus rootstocks employed had differential nutritional behavior and different abilities to utilize plant nutrient elements. Thus, the findings of the present study and the methodology adopted can help the horticultural breeders and nutritionists choose the best rootstock/scion combination having the desirable traits of nutrient utilization ability and also to plan effective fertilizer schedule programs for achieving greater yields.