Storage stability assessment of freeze-dried royal jelly by furosine determination

The effect of freeze-drying and the assessment of the storage stability of freeze-dried royal jelly (RJ) were investigated by the determination of furosine and blocked lysine. The level of furosine in the RJ samples collected from cells at different times (1, 2, and 3 days after grafting) showed that the Maillard reaction had already occurred in the hive as indicated by the increase in furosine: from 9.6 to 20.8 mg/100 g of protein. Freeze-dried RJ was more prone to the early stage of the Maillard reaction than fresh RJ, as confirmed by the significantly higher furosine values found after 12 months, both at 4 degrees C (253.4 versus 54.9 mg/100 g of protein) and at room temperature (884.3 versus 332.5 mg/100 g of protein). After 18 months at room temperature, the lyophilized samples reached a furosine level of 1440.4 mg/100 g of protein, which corresponded to the blocked lysine levels, amounting to 24% of total lysine.     

Hydroxymethylfurfural and furosine reaction kinetics in tomato products

The reaction kinetics of two heat damage indices, HMF and furosine, were examined in four tomato products with different dry matter contents (10.2, 25.5, 28.6, and 34.5%) over a temperature-time range of 80-120 degrees C and 0-255 min. The reactions followed pseudo-zero order kinetics. E(a) and z-value were, respectively, 139. 9 kJ/mol and 19.2 degrees C for HMF, and 93.9 kJ/mol and 28.4 degrees C for furosine. The analyses of both indices in several samples of commercial and industrial tomato products showed very low levels of HMF (from 1 to 42 ppm) and a lack of correlation between HMF and furosine mainly because of the different evolution of the two indices during storage. The HMF level of a tomato paste sample stored at 25 degrees C decreased from 609 to 17 ppm after 98 days, while furosine increased from 458 to 550 mg/100 g of protein.     

Presence of 2-furoylmethyl derivatives in hydrolysates of processed tomato products

Acid hydrolysis of Amadori compounds yields the corresponding 2-furoylmethylamino acids (2-FM-AA) that can be analyzed by ion-pair HPLC. The relative proportions of the different 2-FM-AA present in the hydrolysates of tomato products were determined to assess their usefulness as indicators of quality. In the lyophilized tomato samples stored at 50 degrees C and a(w) = 0.44 the formation of 2-FM derivatives of alanine, gamma-aminobutyric acid (GABA), asparagine, aspartic acid, glutamic acid, lysine, serine, and threonine was detected. In commercial tomato products the most abundant 2-FM-AA was 2-FM-GABA (from traces to 26.4 mg/100 g of dry matter) followed by 2-FM-lysine (furosine). Differences in 2-FM-AA contents among samples may be related to processing and storage conditions.     

Chemical indicators of heat treatment in fortified and special milks

Carbohydrate and furosine contents in 12 commercial fortified and special milk samples (pasteurized goat's and ewe's milks; ultrahigh-temperature (UHT) goat's milk, UHT milks fortified with calcium, magnesium, fiber, or royal jelly and honey; and lactose-hydrolyzed milks) were analyzed. Except for lactose-hydrolyzed milks, furosine, lactose, lactulose, galactose, glucose, N-acetylgalactosamine, N-acetylglucosamine, and myo-inositol contents were similar to the previously reported values for UHT or pasteurized milk samples. In lactose-hydrolyzed milks, lactulose was not detectable and lactose was present in low amount; high levels of glucose, galactose, fructose, tagatose, and furosine were also detected in this type of milk. Results found in commercial milks were compared to those obtained in laboratory-prepared UHT milks with lactose hydrolyzed prior to heating. Hydrolysis of lactose before thermal treatments promoted elevated accumulation of reducing sugars (galactose and glucose) that could be partially converted to the corresponding isomers (tagatose and fructose) during heating. In addition, the reducing sugars could also react with the amino groups of proteins, giving rise to the corresponding Amadori compound. According to the obtained results, heating prior to hydrolysis of lactose is suggested to avoid a considerable loss of available lysine.     

Generation of furosine and color in infant/enteral formula-resembling systems

The extent of the Maillard reaction was studied by measuring furosine and color formation in infant and enteral formula-resembling model systems prepared by mixing calcium caseinate, laboratory-obtained or commercial whey protein with lactose or dextrinomaltose (ingredients similar to those used in infant and enteral formula manufacture) and heating the mixture at 100, 120, or 140 degrees C for 0-30 min. The furosine determination was performed by HPLC and the color determination by measuring colorimetric parameters L, a, and b in a reflection photometer. The first steps of the Maillard reaction could be followed by furosine determination when initial ingredients had low thermal damage. Hence, furosine may be an indicator of low thermal damage in ingredients with <100 mg/100 g of protein. At the concentrations used in these model systems, similar to those in infant and enteral formulas, furosine values (indirect measure of lysine losses) were higher in lactose than in dextrinomaltose systems, in which only glucose, maltose, maltotriose, and maltotetraose among all of the sugars present showed reactivity with casein. Finally, the advanced steps could be followed by color determination when the initial ingredients had high thermal damage or the model systems were heated at high temperature or for a long time. Among the parameters assayed, b was the most sensitive.     

Studies on N-terminal glycation of peptides in hypoallergenic infant formulas: quantification of alpha-N-(2-furoylmethyl) amino acids

To obtain information about the extent of the early Maillard reaction between the N-termini of peptides and lactose, alpha-N-(2-furoylmethyl) amino acids (FMAAs) were quantified together with epsilon-N-(2-furoylmethyl)lysine (furosine) in acid hydrolyzates of hypoallergenic infant formulas, conventional infant formulas, and human milk samples using RP-HPLC with UV-detection. FMAAs are formed during acid hydrolysis of peptide-bound N-terminal Amadori products (APs), and furosine is formed from the Amadori products of peptide-bound lysine. Unambiguous identification was achieved by means of LC/MS and UV-spectroscopy using independently prepared reference material. The extent of acid-induced conversion of APs to FMAAs was studied by RP-HPLC with chemiluminescent nitrogen detection (CLND). Depending on the corresponding alpha-N-lactulosyl amino acid, between 6.0% and 18.1% of FMAAs were formed during hydrolysis for 23 h at 110 degrees C in 8 N HCl. From epsilon-N-lactulosyllysine, 50% furosine is formed under these conditions. Whereas furosine was detectable in all assayed samples, five different FMAAs, alpha-FM-Lys, alpha-FM-Ala, alpha-FM-Val, alpha-FM-Ile, and alpha-FM-Leu, were exclusively detected in acid hydrolyzates of hypoallergenic infant formulas in amounts ranging from 35 to 396 mumol/100 g protein. Taking the conversion factors into account, modification of N-terminal amino acids in peptides by reducing carbohydrates was between 0.3% and 8.4%. This has to be considered within the discussion concerning the nutritional quality of peptide-containing foods.     

Browning indicators in bread

Bread is the most important food in the Spanish household and represents the largest proportion of products produced by commercial bakeries. The browning indicators furosine, hydroxymethylfurfural (HMF), and color were determined to evaluate heat effects induced during manufacture of these foods. The breads analyzed were common, special, sliced toasted, and snack breads. Identical sample preparation and HPLC conditions were used to determine HMF in all breads. The precision tested at high and low HMF concentration in breads was 2.60% and 1.57%, respectively. Recovery of HMF was 96.2%. The HMF values ranged from 2.2 to 68.8 mg/kg. Color index (100 - L) ranged from 17.0 to 38.2. The linear correlations (r(2)) between 100 - L/HMF were above 0.70 for common, special, and snack breads. Similar correlation was obtained between 100 - L/HMF in a dough baking at different times. The furosine content in common bread ranged between 125 and 208 mg/100 g of protein. No linear correlation was found between furosine and HMF. Moreover, HMF and furosine were also determined in crumb and crust. Levels of HMF had a wide range (0.9-1.76 mg/kg) and furosine was between 43 and 221 mg/100 g of protein.     

Formation of Amadori compounds in dehydrated fruits.

The presence of Amadori compounds in commercial dehydrated fruits has been shown through HPLC analysis of the corresponding 2-furoylmethyl-amino acids obtained by acid hydrolysis. Furosine (2-furoylmethyl-lysine) was the main 2-furoylmethyl derivative observed in dried figs and apricot samples, whereas in prunes and dates similar amounts of furosine and 2-furoylmethyl-gamma-aminobutyric acid were detected. A considerable variation of 2-furoylmethyl-amino acid contents among commercial raisin samples was observed. 2-Furoylmethyl-gamma-aminobutyric acid and 2-furoylmethyl-arginine, the most abundant 2-furoylmethyl-amino acids, ranged between 9.9 and 75.8 mg/100 g sample and 10.0 and 62.5 mg/100 g sample, respectively. Most of the Amadori compounds present in raisins seem to have originated during the commercial shelf life period rather than during processing. Determination of 2-furoylmethyl-amino acids could be used as a method of controlling commercial dehydrated fruit and selecting storage conditions.     

New capillary electrophoresis method for the determination of furosine in dairy products

A new capillary electrophoresis (CE) method was established for the quantitative determination of furosine in dairy products. Sample preparation and suitable electrophoretic conditions allowed accurate and reproducible quantitation of furosine in dairy products. Sample preparation consisted of drying hydrolyzed samples, redissolving them in 0.2 M NaOH, and purifying them by solid-phase extraction. The electrophoretic separation was carried out in an uncoated capillary maintained at 30 degrees C using 0.1 M phosphate buffer containing the additive hexadecyl trimethylammonium bromide (HDTAB, 1.2 mM) (pH 7.0) under 10 kV voltage and reverse polarity. Coefficients of variation of less than 2.25% for migration time and 5.80% for peak areas indicated that the technique was reproducible. The calibration curve followed a linear relationship with a highly significant (p < 0.01) coefficient of multiple determination (R (2) = 0.997). The limit of quantitation was 0.5 ppm, a concentration that corresponds to 4.5 mg/100 g of protein in milk samples. Furosine concentration (mg/100 g of protein) ranges of different dairy products (raw, pasteurized, UHT, and evaporated milks and yogurt) agreed with ranges previously reported. Therefore, the CE method presented is a suitable technique for the routine assessment of furosine in dairy products.     

不同采收时期蜂王浆抗氧化活性研究

为研究不同采收时间对蜂王浆品质的影响,以4-9月份采收的蜂王浆为研究对象,通过比较不同采收时间蜂王浆中蜂王浆主蛋白(MRJPs)、总水溶性蛋白和多酚的含量,分析了MRJPs与自由基清除能力和总抗氧化能力的相关性。结果表明,不同采收期蜂王浆中MRJPs含量存在显著差异(P<0.05),且6月份采收的MRJPs含量最低。水溶性蛋白及总酚含量差异不大(P>0.05)。蜂王浆自由基清除能力与MRJP1和 MRJP3含量的相关系数达0.828和0.847;总抗氧化能力与MRJP1和 MRJP3含量的相关系数分别为0.680和0.743。蜂王浆的抗氧化活性与其MRJPs存在一定程度的正相关,但与多酚含量相关性不明显,说明其自由基清除能力与总抗氧化活性可能是多种抗氧化性活性物质共同作用的结果。本研究为蜂王浆的抗氧化活性研究提供了参考。     

Effects of thermal processing and storage on available lysine and furfural compounds contents of infant formulas

The Maillard reaction-related effects that thermal treatments during the manufacturing process and storage (at 20 and 37 degrees C) have on powdered adapted and follow-up milk-based infant formulas were estimated by measuring the available lysine and furfural compounds contents of raw cow milk used in manufacturing, intermediate products and formulas. A fluorimetric method was used to measure the available lysine contents, and free and total furfural compounds were determined by HPLC. Statistically significant losses in available lysine (about 20%) in the infant formulas with respect to raw milk were found. The storage period did not affect the available lysine contents of adapted formulas but reduced (16%) the contents of the follow-up ones (from 6.61 to 5.33 g/100 g of protein). No furfural compounds were detected in raw milk, and free and total furyl methyl ketone (FMC) and methylfurfural (MF) were not observed in the analyzed samples. After 6 months of storage, an increase in free hydroxymethylfurfural (HMF) (from 0.34 to 0.77 mg/100 g of protein) and furfural (F) (from nondetectable to 0.1 mg/100 g of protein) in adapted formulas and free HMF (from 1.84 to 2.62 mg/100 g of protein) in follow-up formulas was observed.     

两蜂种蜂王浆蛋白质组分析

采用蛋白质组双向电泳技术和已鉴定蜂王浆蛋白质组的比对,对王浆高产蜜蜂（浆蜂）和中华蜜蜂（中蜂）的蜂王浆蛋白质组进行了差异比较。结果表明,浆蜂蜂王浆的总蛋白质点数（93个）显著高于中蜂蜂王浆总蛋白质点数（70个）,它们的等电点主要集中在5-8之间,分子量在50-80kDa之间,其中共有蛋白质点为30个;通过与已鉴定的浆蜂蜂王浆蛋白质组比较,这些共有蛋白质点推断为蜂王浆主蛋白1、2、3和葡萄糖氧化酶。同时两蜂种蜂王浆的蛋白质丰度也存在较大差异,共有点中有4个蛋白质的丰度中蜂显著高于浆蜂,7个浆蜂显著高于中蜂中。结果表明浆蜂和中蜂蜂王浆蛋白质种类及丰度均有较大差异。     

Effects of yam starch films on storability and quality of fresh strawberries (Fragaria ananassa)

Yam starch films, formulated with yam starch (4.00 g/100 g of solution) and glycerol (1.30 and 2.00 g/100 g of solution) in filmogenic solution, were employed as packaging to extend storage life of strawberries stored at 4 degrees C and 85% RH. The effects of yam starch films on fruits were compared to the effect of PVC (poly(vinyl chloride)) packaging. Starch and PVC films significantly reduced decay of the fruits compared to control. Compared to starch films, PVC presented the better behavior on weight and firmness retention of fruits, especially in the last 7 days of storage. Considering microbiological counts, the shelf life of control fruits was 14 days, and of all packaged samples, stored at same conditions, was 21 days. Two different formulations of yam starch film were tested and had different mechanical properties as a function of glycerol content (1.30 and 2.00 g/100 g of solution) but showed no difference when employed as strawberries packaging.     

2-Furoylmethyl amino acids and hydroxymethylfurfural as indicators of honey quality

Determination of changes in 2-furoylmethyl amino acids and hydroxymethylfurfural during the storage of four honey samples at 25 and 35 degrees C during 12 months was achieved to assess the potential use of both parameters, singly or in combination, as quality indicators. 2-Furoylmethyl amino acids increased during storage at both temperatures, whereas hydroxymethylfurfural only presented slight variations during storage at 25 degrees C but increased noticeably at 35 degrees C. The study of 2-furoylmethyl amino acids in 49 commercial honeys revealed that 2-furoylmethyl lysine (furosine) was present in all samples, whereas 2-furoylmethyl derivatives of arginine, GABA, and proline were only present in seven samples. Hydroxymethylfurfural can be considered as a good indicator of heat treatments applied to honey samples, whereas 2-furoylmethyl amino acids can be used as suitable markers of the storage period. The use of both parameters can be useful to detect adulteration with invert syrups, excessive heat treatments, or prolonged storage of honey samples.     

Comparison of Nutritional Compounds in Premature Green and Mature Yellow Whole Wheat in Korea

Until now few comparisons of nutritional compounds in premature green and mature yellow wheat have been reported. In this study, the contents of amino acids, vitamins, mineral compounds, phytosterols, and fatty acids as well as the proximate composition of premature green and mature yellow wheat were investigated. Premature green wheat had lower protein content (12.0 g/100 g db) and higher dietary fiber content (19.3 g/100 g db) than mature yellow wheat (13.6 and 14.3 g/100 g db for protein and dietary fiber, respectively). Despite a small difference in total amino acids, protein in premature wheat had a significantly greater proportion of essential amino acids: 16.1, 39.9, and 32.7 mg/g of protein for methionine, lysine, and threonine, respectively. Furthermore, the protein digestibility‐corrected amino acid scores of whole grain premature green and mature yellow wheat were 62.8 and 46.4, respectively, showing significant difference (P < 0.05). Total fatty acids content was 2.66 g/100 g db for premature green wheat and 2.21 g/100 g db for mature yellow wheat. Vitamin C, β‐carotene (provitamin A), α‐ and γ‐tocopherol, and niacin were the major vitamins in premature green wheat, whereas vitamin C and β‐carotene were not detected in mature yellow wheat. The results obtained indicated that premature green wheat has potential for the human diet because of its desirable nutritional value.     

Total chemically available (free and intrachain) lysine and furosine in pea, bean, and lentil sprouts

The effect of the germination of peas, beans, and lentils under differing conditions of illumination for different times on parameters linked to the Maillard reaction (chemically available free and intrachain lysine, lysine availability, and furosine) was evaluated. The chemically available free lysine content in the raw seeds of the three legumes was quite small compared to the chemically available intrachain lysine content, and furosine was detectable only in the beans and the lentils. The effect of germination was to increase lysine availability compared with levels in the raw seeds in all of the germinated samples, the smallest increase taking place in the lentils. In addition, furosine became detectable in all of the germinated samples. Quantities varied depending on the germination conditions but in all cases were higher than the quantities observed in the raw seeds. Linear correlations were observed to exist between some of the parameters considered in the three legumes tested.     

Kinetics of formation of three indicators of the maillard reaction in model cookies: influence of baking temperature and type of sugar

The Maillard reaction (MR), despite its impact on flavor, color, and texture of cereal products, must be controlled for possible deleterious effects on protein nutritional quality. The present study aims to simultaneously monitor three indicators of the MR reaction (acid-released lysine, furosine, and carboxymethyllysine (CML)) by GC/MS in model cookies and evaluate the effect of formulation and baking temperature. Whereas furosine followed a bell-shape kinetic, indicative of an intermediary compound, CML linearly accumulated, proving to be a good indicator of the advanced MR. Acid-released lysine continuously decreased during baking. A reference baking level was defined to compare differently processed cookies using fluorescence synchronous spectra, highly sensitive to the dough physicochemical properties. Furosine was maximal in glucose-containing cookies, but only accounted for 5-50% lysine blockage, depending on the sugar and baking temperature. High oven temperatures and the use of fructose as the sugar source were associated with lowest the lysine damage and CML formation.     

Effect of antioxidants, citrate, and cryoprotectants on protein denaturation and texture of frozen cod (Gadus morhua)

To investigate the role of antioxidants and cryoprotectants in minimizing protein denaturation in frozen lean fish, cod fillets were treated with either antioxidants (vitamin C (500 mg kg(-1)) or vitamin C (250 mg kg(-1)) + vitamin E (250 mg kg(-1))), antioxidants (vitamins C + E 250 mg kg(-1)each) with citrate (100 mg kg(-1)), cryoprotectants (4% (w/w) sucrose + 4% (w/w) sorbitol), or a mixture of antioxidants (vitamins C + E 250 mg kg(1)), citrate (100 mg kg(-1)), and cryoprotectants (sucrose 40 g kg(-1) + sorbitol 40 g kg(-1)). Untreated and treated fish samples were stored at -10 degrees C; cod fillets stored at -30 degrees C were used as a control. Stored frozen samples were analyzed at intervals for up to 210 days for changes in protein extractability, thermodynamic parameters (transition temperature T(m) and enthalpy DeltaH), structure by FT-Raman spectroscopy, and rheological properties by large and small deformation tests. Results indicated that protein denaturation and texture changes were minimized in the presence of cryoprotectants, as well as in the presence of antioxidants with citrate, antioxidants alone, or the mixture of antioxidants, citrate, and cryoprotectants. In the presence of increased formaldehyde levels in fish treated with vitamin C, toughening was still lower compared to that of the -10 degrees C control due to the antioxidant property of vitamin C. Thus, ice crystal formation and lipid oxidation products are the major factors that cause protein denaturation in lean frozen fish, and antioxidants in addition to cryoprotectants can be used to minimize toughness.     

Stability of sterols in phytosterol-enriched milk under different heating conditions

Commercially available phytosterol-enriched milk was subjected to usual and drastic heating conditions to evaluate the stability of the sterols at different treatments. Products showed 422.2 mg of phytosterols/100 g of milk and 132 microg of sterol oxidation products (SOPs)/g of fat (277 microg of SOPs/100 g of milk). Schaal oven conditions (24 h/65 degrees C, equivalent to 1 month of storage at room temperature) reduced the phytosterol content by only 4%. Drastic heating treatments (2 min of microwave heating at 900 W or 15 min of electrical heating at 90 degrees C) led to a 60% decrease of total phytosterol content, with a significant increase of TBARs. The oxysterol amount under those conditions (which was higher in microwave-treated samples) was lower than expected, probably because of the degradation of the oxidation products. Usual heating conditions (1.5 min of microwaves) maintained phytosterol content on physiologically active values (301 mg/100 g of milk) with oxidation percentages around 0.12-0.40% for phytosterols and 1.13% for cholesterol.     

Identification and quantification of epsilon-(gamma-glutamyl)lysine in digests of enzymatically cross-linked leguminous proteins by high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS)

A rapid and convenient method for the precise quantification of epsilon-(gamma-glutamyl)lysine isopeptide in lyophilized proteolytic digests of cross-linked plant protein samples was developed. The isopeptide was baseline-separated from three other isomers containing lysyl and glutamyl residues by reverse-phase high-performance liquid chromatography after exhaustive proteolytic digestion of the samples cross-linked by a microbial transglutaminase (MTG). Highly selective detection was performed by electrospray mass spectrometry in MS/MS mode. Demonstrating the applicability of the suggested analytical procedure, enzymatic cross-linking of protein isolates from soy [Glycine max (L.) Merr.], pea [Pisum sativum L.], and the sweet lupin species Lupinus albus L. and Lupinus angustifolius L. was investigated after incubation with 0.01 g of MTG/100 g of protein for 0-240 min at 40 degrees C. The liquid chromatography-mass spectrometry (LC-MS) method was successfully applied to monitor the kinetics of epsilon-(gamma-glutamyl)lysine isopeptide formation. Since the calculated initial levels of epsilon-(gamma-glutamyl)lysine in the genuine leguminous protein isolates were between 40 and 77 micromol/100 g, an isopeptide detection limit of 0.5 microg/mL, corresponding to approximately 50 micromol/100 g of protein, was shown to suffice for quantifying the cross-linking rate enzymatically induced by MTG. Concentrations of epsilon-(gamma-glutamyl)lysine in the texturized proteins ranged from 100 to 500 micromol/100 g of protein.