Comparative study on pressure and temperature stability of 5-methyltetrahydrofolic acid in model systems and in food products

A comparative study on the pressure and temperature stability of 5-methyltetrahydrofolic acid (5-CH(3)-H(4)folate) was performed in model/buffer systems and food products (i.e., orange juice, kiwi puree, carrot juice, and asparagus). Effects of pH and ascorbic acid (0.5 mg/g) on 5-CH(3)-H(4)folate stability in buffer systems were studied on a kinetic basis at different temperatures (from 65 to 160 degrees C) and different pressure/temperature combinations (from 100 to 700 MPa/from 20 to 65 degrees C). These studies showed that (i) the degradation of 5-CH(3)-H(4)folate in all model systems could be described by first-order reaction kinetics, (ii) the thermostability of 5-CH(3)-H(4)folate was enhanced by increasing pH up to 7, (iii) 5-CH(3)-H(4)folate was relatively pressure stable at temperatures lower than 40 degrees C, and (iv) ascorbic acid enhanced both the thermo- and barostabilities of 5-CH(3)-H(4)folate. In food products, temperature and pressure stabilities of 5-CH(3)-H(4)folate were studied at different temperatures (70-120 degrees C) and different pressure/temperature combinations (from 50 to 200 MPa/25 degrees C and 500 MPa/60 degrees C). 5-CH(3)-H(4)folate in orange juice and kiwi puree was relatively temperature (up to 120 degrees C) and pressure (up to 500 MPa/60 degrees C) stable in contrast to carrot juice and asparagus. Addition of ascorbic acid (0.5 mg/g) in carrot juice resulted in a remarkable protective effect on pressure (500 MPa/60 degrees C/40 min) and temperature degradation (120 degrees C/40 min) of 5-CH(3)-H(4)folate.     

Total folate content and retention in rosehips (Rosa ssp.) after drying

Folate concentrations in rosehips and commercial rosehip products and factors affecting folate retention during drying were investigated. On the basis of the raw material studied during 3 years, rosehips were shown to be a rich folate source, 400-600 microg/100 g based on dry matter and 160-185 microg/100 g based on the fresh weight (edible part). Rosehips are not often consumed fresh; therefore, drying to produce stable semimanufactures is a crucial step. The degradation of folate was shown to be dependent on the drying time until the water activity was below 0.75. The required drying time was reduced by cutting the rosehips in slices and to some extent also by increasing the temperature. Retention of folate and ascorbic acid was affected by the same factors, and high content of ascorbic acid could provide a possible protection for folate degradation.     

3-Hydroxy-4,5-dimethyl-2(5H)-furanone (Sotolon) causing an off-flavor: elucidation of its formation pathways during storage of citrus soft drinks.

Gas chromatography/olfactometry (GCO) and gas chromatography-mass spectrometry (HRGC-MS) revealed 3-hydroxy-4, 5-dimethyl-2(5H)-furanone (sotolon) to be responsible for the "burnt" and "spicy" off-flavor observed in citrus soft drinks during storage. Among the ingredients of citrus soft drinks, ethanol and ascorbic acid were identified as the essential precursors of sotolon. Two formation pathways were postulated by studies using (2)H (D)- and (13)C-labeled ethanol and ascorbic acid; i.e., sotolon is formed from two molecules of ethanol and carbons 2 and 3 of ascorbic acid (pathway 1), or it is generated from one molecule of ethanol and carbons 3-6 of ascorbic acid (pathway 2).     

Folate content in strawberries (Fragaria x ananassa): effects of cultivar,ripeness, year of harvest,storage, and commercial processing

Folate concentrations in strawberries and folate retention during storage and commercial processing of strawberries were investigated. No previous study has focused on the effects of cultivar, ripeness, and year of harvest of strawberries with respect to the folate content. This study showed the folate concentration in strawberries to significantly depend on all of these different factors. Total folate was quantified using a modified and validated radioprotein-binding assay with external calibration (5-CH(3)-H(4)folate). Folate content in 13 different strawberry cultivars varied from 335 microg/100 g of dry matter (DM) for cv. Senga Sengana to 644 microg/100 g of DM for cv. Elsanta. Swedish harvests from 1999 and 2001 yielded higher folate concentrations than did the harvest from 2000, and the grade of ripeness affected the folate content in strawberries. This study indicated high folate retention in intact berries during storage until 3 or 9 days at 4 degrees C (71-99%) and also in most tested commercial products (79-103%). On the basis of these data fresh strawberries as well as processed strawberry products are recommended to be good folate sources. For instance, 250 g (fresh weight) of strawberries ( approximately 125 microg of folate) supplies approximately 50% of the recommended daily folate intake in various European countries (200-300 microg/day) or 30% of the U.S. recommendation (400 microg/day).     

Quantitation of total folate in whole blood using LC-MS/MS

An accurate method for measuring whole blood total folate using liquid chromatography with tandem mass spectrometry is described and compared to GC/MS and a chemiluminescence assay. Whole blood from normal adults (n = 15) was fortified with a [(13)C(6)]para-aminobenzoic acid (pABA) internal standard and treated with 12.1 N hydrochloric acid at 110 degrees C for 4 h to hydrolyze all folates to pABA. Contaminants in the hydrolysate were adsorbed onto a C18 SPE cartridge. The eluate containing the folate catabolite pABA was partitioned into ethyl acetate and methylesterified with trimethylsilyldiazomethane. The methyl-pABA derivatives were quantified by positive-ion atmospheric pressure chemical ionization (APCI)LC-MS/MS. An isocratic mobile phase of acetonitrile-water (70:30) (v/v) on a C18 analytical column was used with a postcolumn reagent of 0.025% formic acid. The limit of quantitation for folate was 56.6 nmol/L RBC, and the limit of detection was 22.6 nmol/L RBC. Folate levels as determined by LC-MS/MS correlated well with the chemiluminescence assay and a GC/MS method. This new LC-MS/MS method provides enhanced sample throughput (n = 36 per day) as compared to GC/MS methods. LC-MS/MS will enable accurate measurements of red blood cell (RBC) folate in nutrition surveys and clinical trials.     

Stability and sensory shelf life of orange juice pasteurized by continuous ohmic heating

Electrical heating of food products provides rapid and uniform heating, resulting in less thermal damage to the product. The objective of this research was to examine the effects of ohmic heating on the stability of orange juice with comparison to conventional pasteurization. During storage at 4 degrees C, degradation curves of ascorbic acid followed a linear decrease pattern in both ohmic-heated and conventionally pasteurized orange juices. For five representative flavor compounds (decanal, octana, limonene, pinene, and myrcene), higher concentrations were measured during storage in the ohmic-heated orange juice than in conventionally pasteurized juice. Although residual pectin esterase activity remained negligible in both types of juices, particle size was lower in the ohmic-heated orange juice. The sensory shelf life was determined by using the Weibull-Hazard method. Although both thermal treatments prevented the growth of microorganisms for 105 days, the sensory shelf life of ohmic-treated orange juice was >100 days and was almost 2 times longer than that of conventionally pasteurized juice.     

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.     

Effect of Baking Method and Fermentation on Folate Content of Rye and Wheat Breads

The effect of baking method on folates of rye and wheat breads, as well as the effect of sourdough fermentation of rye, were examined. Sourdough fermentations were performed both with and without added yeast, and samples were taken throughout the baking process. Samples were analyzed microbiologically for their total folate content after trienzyme extraction. Individual folate vitamers were determined by HPLC after affinity chromatographic purification. The lowest folate contents for both rye and wheat breads were found from breads baked without added yeast. Total folate content increased considerably during sourdough fermentation due to increased amounts of 10‐HCO‐H₂folate, 5‐CH₃‐H₄folate, and 5‐HCO‐H₄folate. Baker's yeast contributed markedly to the final folate content of bread by synthesizing folates during fermentation. Proofing did not influence total folate content but changes in vitamer distribution were observed. Folate losses in baking were ≈25%. The variety of sourdoughs and baking processes obviously lead to great variation in folate content of rye breads. The possibilities to enhance natural folate content of rye bread by improving folate retention in technological processes and by screening and combining suitable yeasts and lactic acid bacteria should be further investigated.     

Stabilization of ascorbic acid and its measurement by liquid chromatography in nonfat dry milk

The determination of ascorbic acid by liquid chromatography (LC) was improved by performing the analysis in the presence of solvents that had been purged with argon to reduce the concentration of oxygen. This methodological modification eliminated the oxidation of ascorbic acid during the chromatographic procedure and reduced the minimum detection level to 1 microgram. Solutions of ascorbic acid have been successfully stabilized for 67 days by addition of dithiothreitol to a deaerated solution of water-acetonitrile (25 + 75 v/v), sealed under argon in amber vials and stored at -20 degrees C. In a second independent study, a procedure for the extraction of ascorbic acid from nonfat dry milk in a single step was developed. The ascorbic acid content of Nonfat Dry Milk (SRM 1549) was determined by LC, using the method of standard additions. The mean ascorbic acid content was 54 +/- 5 micrograms/g of sample. Analysis of variance of the analytical results indicates that there is a significant continual increase in the content of the ascorbic acid in each bottle from first to last sample.     

Changes of folate and other potential health-promoting phytochemicals in legume seeds as affected by germination

Folate deficiency associated with low dietary intake is a well-documented public health problem, resulting in serious health and socioeconomic burdens. Therefore, optimization of the germination process of different cultivars of legume seeds in relation to the content and composition of folate, vitamin C, and total phenolics and total antioxidant capacity was carried out to maximize the health-promoting properties. The content and composition of folate, vitamin C, and total phenolic and total antioxidant capacities varied between species, among cultivars, and with germination time. During germination, total folate content was maximum at 815.2 μg/100 g fresh weight in soybean sprout and at 675.4 μg/100 g fresh weight in mungbean sprout on the fourth day, which were equivalent to, respectively, 3.5- and 3.9-fold increases in the seed's content, and total folate content strongly decreased thereafter. 5-CH(3)-H(4)folate was the most abundant folate species in legume sprouts and reached a maximum on the fourth day. Vitamin C was not detected in raw seeds, and its content increased sharply in soybean and mungbean sprouts and reached a maximum at the fourth day of germination (29 and 27.7 mg/100 g fresh weight, respectively). Germination of soybean and mungbean for 4 days provided the largest amount of total folate as well as the more stable species 5-CH(3)-H(4)folate and also brought about large amounts of vitamin C and total phenolics and substantial antioxidant capacities.     

Submerged citric acid fermentation on orange peel autohydrolysate

The citrus-processing industry generates in the Mediterranean area huge amounts of orange peel as a byproduct from the industrial extraction of citrus juices. To reduce its environmental impact as well as to provide an extra profit, this residue was investigated in this study as an alternative substrate for the fermentative production of citric acid. Orange peel contained 16.9% soluble sugars, 9.21% cellulose, 10.5% hemicellulose, and 42.5% pectin as the most important components. To get solutions rich in soluble and starchy sugars to be used as a carbon source for citric acid fermentation, this raw material was submitted to autohydrolysis, a process that does not make use of any acidic catalyst. Liquors obtained by this process under optimum conditions (temperature of 130 degrees C and a liquid/solid ratio of 8.0 g/g) contained 38.2 g/L free sugars (8.3 g/L sucrose, 13.7 g/L glucose, and 16.2 g/L fructose) and significant amounts of metals, particularly Mg, Ca, Zn, and K. Without additional nutrients, these liquors were employed for citric acid production by Aspergillus niger CECT 2090 (ATCC 9142, NRRL 599). Addition of calcium carbonate enhanced citric acid production because it prevented progressive acidification of the medium. Moreover, the influence of methanol addition on citric acid formation was investigated. Under the best conditions (40 mL of methanol/kg of medium), an effective conversion of sugars into citric acid was ensured (maximum citric acid concentration of 9.2 g/L, volumetric productivity of 0.128 g/(L.h), and yield of product on consumed sugars of 0.53 g/g), hence demonstrating the potential of orange peel wastes as an alternative raw material for citric acid fermentation.     

Quenching mechanisms and kinetics of Trolox and ascorbic acid on the riboflavin-photosensitized oxidation of tryptophan and tyrosine

The effects of 0, 0.3, 0.6, and 0.9 mM Trolox and ascorbic acid on the singlet oxygen oxidation of tryptophan and tyrosine containing 25 ppm of riboflavin were determined by measuring tryptophan and tyrosine concentration by high-performance liquid chromatography analysis. The samples were stored in the a 1000 lx light storage box for 4 h at 30 degrees C. As the concentration of Trolox and ascorbic acid increased, the degradation of tryptophan and tyrosine decreased significantly at p < 0.05. Trolox reduced tryptophan and tyrosine degradation by quenching both singlet oxygen and excited triplet riboflavin, whereas ascorbic acid quenched singlet oxygen only. The total singlet oxygen quenchings of Trolox in the presence of tryptophan and tyrosine were 1.55 x 10(7) and 1.32 x 10(7) M(-1) s(-1), respectively. The total singlet oxygen quenchings of ascorbic acid in the presence of tryptophan and tyrosine were 1.16 x 10(7) and 1.10 x 10(7) M(-1) s(-1), respectively. Trolox was more effective than ascorbic acid in preventing the degradation of tryptophan and tyrosine.     

Effects of pulsed electric fields on the quality of orange juice and comparison with heat pasteurization

Effects of pulsed electric fields (PEF) at 35 kV/cm for 59 micros on the quality of orange juice were investigated and compared with those of heat pasteurization at 94.6 degrees C for 30 s. The PEF treatment prevented the growth of microorganisms at 4, 22, and 37 degrees C for 112 days and inactivated 88% of pectin methyl esterase (PME) activity. The PEF-treated orange juice retained greater amounts of vitamin C and the five representative flavor compounds than the heat-pasteurized orange juice during storage at 4 degrees C (p < 0.05). The PEF-treated orange juice had lower browning index, higher whiteness (L), and higher hue angle (theta) values than the heat-pasteurized orange juice during storage at 4 degrees C (p < 0. 05). The PEF-treated orange juice had a smaller particle size than the heat-pasteurized orange juice (p < 0.05). degrees Brix and pH values were not significantly affected by processing methods (p > 0. 05).     

Pineapple juice and its fractions in enzymatic browning inhibition of banana [Musa (AAA group) Gros Michel

The effectiveness of pineapple juice in enzymatic browning inhibition was evaluated on the cut surface of banana slices. After storage of banana slices at 15 degrees C for 3 days, pineapple juice showed browning inhibition to a similar extent as 8 mM ascorbic acid but less than 4 mM sodium metabisulfite. Fractionation of pineapple juice by a solid-phase C18 cartridge revealed that the directly eluted fraction (DE fraction) inhibited banana polyphenol oxidase (PPO) about 100% when compared to the control. The DE fraction also showed more inhibitory effect than 8 mM ascorbic acid in enzymatic browning inhibition of banana puree during storage at 5 degrees C for 24 h. Further identification of the DE fraction by fractionation with ion exchange chromatography and confirmation using model systems indicated that malic acid and citric acid play an important role in the enzymatic browning inhibition of banana PPO.     

Inactivation of heat-resistant pectinmethylesterase from orange by manothermosonication

Pectinmethylesterase of navel oranges shows two fractions greatly differing in thermostability. The most thermostable fraction accounts for approximately 10% of total activity. The thermal inactivation of this fraction follows first-order kinetics both in 5 mM, pH 3.5, citrate buffer and in orange juice at the same pH, showing a z value of 5.1 degrees C and an activation energy (E(a)) of 435 kJ mol(-)(1) K(-)(1). The heat resistance of the enzyme is approximately 25-fold higher in the juice than in citrate buffer. When ascorbic acid, sucrose, glucose, and fructose are added to the citrate buffer at the concentrations found in orange juice, the heat resistance of the enzyme increases 3-fold. The addition of pectin at 0.01% concentration multiplies it by a factor of 50. Manothermosonication (MTS), the simultaneous application of heat and ultrasound under moderate pressure (200 kPa), at 72 degrees C, increases the inactivation rate 25 times in buffer and >400 times in orange juice. MTS inactivation shows a higher z value (35.7 degrees C) and lower E(a) (56.9 kJ mol(-)(1) K(-)(1)) than simple heating.     

Storage temperature effects on blood orange fruit quality.

Orange fruits of two blood varieties (Tarocco and Moro) were stored at 8 degrees C and 22 degrees C for 85 and 106 days, respectively, and analyzed periodically for standard quality parameters (total soluble solids, total acidity, ascorbic acid, juice yield, and rind color) and sensory influencing parameters (anthocyanins, and total and free hydroxycinnamic acids). A decrease in total acidity (TA) and juice yield during storage was observed for both cultivars; total soluble solids (TSS) increased only in the Tarocco oranges stored at 8 degrees C. The increase in TSS observed for Tarocco and the simultaneous decrease in TA in both varieties resulted in a higher maturity index (TSS/TA) for the two cultivars. No loss of vitamin C was noted in Tarocco orange at either temperature, whereas a sharp reduction in vitamin C occurred in the first 50 days of storage for Moro. A significant increase in anthocyanin content was observed in Tarocco and Moro stored at 8 degrees C. Overlong storage induces extensive hydrolysis of hydroxycinnamic derivatives to free acids in Moro orange and these, in turn, could develop the malodorous vinylphenols.     

Difference in folate content of green and red sweet peppers (Capsicum annuum) determined by liquid chromatography-mass spectrometry

Folic acid (pteroylmonoglutamic acid) is used in enriched foods; however, very little folic acid occurs naturally in fruits and vegetables. For the U.S. Department of Agriculture's National Food and Nutrient Analysis Program, a number of fruits and vegetables have been assayed for endogenous folates, by a liquid chromatography-mass spectrometry method, to evaluate the accuracy of existing data for total folate determined by standard microbiological analysis. Folate in red and green sweet peppers (Capsicum annuum) differed notably (70.2 and 20.7 microg/100 g, respectively) and exceeded existing values determined by microbiological assay (18 and 11 microg/100 g, respectively). 5-Methyltetrahydrofolate was the predominant vitamer, but a significant amount of 5-formyltetrahydrolfolate and some 10-formylfolate were present. These findings may assist in making dietary recommendations or developing research diets related to folate. The data from this study have been used to update the folate values in release 19 of the USDA Nutrient Database for Standard Reference.     

Model studies on the stability of folic acid and 5-methyltetrahydrofolic acid degradation during thermal treatment in combination with high hydrostatic pressure

Stability of folic acid and 5-methyltetrahydrofolic acid in phosphate buffer (0.2 M; pH 7) toward thermal (above 65 degrees C) and combined high pressure (up to 800 MPa)/thermal (20 up to 65 degrees C) treatments was studied on a kinetic basis. Residual folate concentration after thermal and high pressure/thermal treatments was measured using reverse phase liquid chromatography. The degradation of both folates followed first-order reaction kinetics. At ambient pressure, the estimated Arrhenius activation energy (E(a)) values of folic acid and 5-methyltetrahydrofolic acid thermal degradation were 51.66 and 79.98 kJ mol(-1), respectively. It was noticed that the stability of folic acid toward thermal and combined high pressure thermal treatments was much higher than 5-methyltetrahydrofolic acid. High-pressure treatments at room temperature or higher (up to 60 degrees C) had no or little effect on folic acid. In the whole P/T area studied, the rate constant of 5-methyltetrahydrofolic acid degradation was enhanced by increasing pressure, and a remarkable synergistic effect of pressure and temperature on 5-methyltetrahydrofolic acid degradation occurred at temperatures above 40 degrees C. A model to describe the combined pressure and temperature effect on the 5-methyltetrahydrofolic acid degradation rate constant is presented.     

Pressure and temperature stability of 5-methyltetrahydrofolic acid: a kinetic study

Detailed kinetic studies of [6S] and [6RS] 5-methyltetrahydrofolic acid (5-CH3-H4folate) degradation during thermal (from 60 to 90 degrees C) and high pressure/thermal (from 30 to 45 degrees C; from 200 to 700 MPa) treatments were carried out. The results confirmed that the temperature and pressure induced degradation kinetics of [6S] 5-CH3-H4folate were identical (within 95% confidence interval) with those of [6RS] 5-CH3-H4folate. Under equal processing conditions, the estimated degradation rate constants (k), activation energy (E(a)), and activation volume (V(a)) values of [6S] and [6RS] 5-CH3-H4folate were the same (95% confidence interval). The modified thermodynamic model proposed by Nguyen and co-workers (J. Agric. Food Chem. 2003, 51, 3352-3357) to describe the pressure and temperature dependence of the rate constant for folate degradation was reevaluated.     

Impact of high pressure and pulsed electric fields on bioactive compounds and antioxidant activity of orange juice in comparison with traditional thermal processing

Bioactive compounds (vitamin C, carotenoids, and flavanones) and DPPH* radical scavenging capacity (RSC) were measured in orange juice (OJ) subjected to different technologies. High pressure (HP) (400 MPa/40 degrees C/1 min), pulsed electric fields (PEF) (35 kVcm(-1)/750 micros), low pasteurization (LPT) (70 degrees C/30 s), high pasteurization (HPT) (90 degrees C/1 min), HPT plus freezing (HPT+F) (-38 degrees C/15 min), and freezing (F) were studied. Among the treatments assayed, even though the losses in total vitamin C were < 9%, treatments with the higher temperatures tended to show the higher decrease in the content of both forms of vitamin C. HP treatment led to an increased (P < 0.05) carotenoid release (53.88%) and vitamin A value (38.74%). PEF treatment did not modify individual or total carotenoids content. Traditional thermal treatments did not exert any effect on total carotenoid content or vitamin A value. With regard to individual carotenoid extraction, HPT and HPT+F led to different releases of carotenoids. With respect to flavanones, HP treatment led to increased (P < 0.05) naringenin (20.16%) and hesperetin (39.88%) contents, whereas PEF treatment did not modify flavanone content. In general, pasteurization and freezing process led to a diminished (P < 0.05) naringenin content (16.04%), with no modification in hesperetin. HP and PEF treatments did not modify DPPH* RSC. In the case of traditional thermal technologies, HPT treatment showed a decrease (P < 0.05) in RSC (6.56%), whereas LPT, HPT+F, and F treatments did not modify RSC. Vitamin C modulated RSC, in terms of antioxidant concentration (EC50) and kinetics (AE = 1/EC50TEC50), in the treated and untreated OJ. In summary, HP and PEF technologies were more effective than HPT treatment in preserving bioactive compounds and RSC of freshly squeezed orange juice.