Gas chromatography-mass spectrometry method for the determination of free amino acids as their dimethyl-tert-butylsilyl (TBDMS) derivatives in animal source food

The suitability of a one-step derivatization procedure using N-methyl-N-(tert-butyldimethylsilyl)trifluoroacetamide for the simultaneous assay of 22 free amino acids and its application for their analysis in six animal source foods (pork, dry cured ham, chicken stock, fresh cheese, ripened cheese, and dry salted sardine) by GC-MS were studied. All 22 free amino acid derivatives were correctly detected and resolved. Reproducibility (%RSD) of the method was in the range of 1.9-12.2%. Detection and quantitation limits of the analytical procedure ranged from 0.01 to 0.46 mg/100 g dry weight and from 0.02 to 1.55 mg/100 g dry weight, respectively. The calibration curves were linear within the range 0.1-15.0 mg/100 g with correlation coefficient values (R(2)) from 0.9891 to 0.9983. All analyzed food products showed free amino acid contents similar to those found in the scientific literature. The proposed GC-MS method for the determination of free amino acids in animal source food can be used in routine for both analytical and research purposes.     

Free amino acid and cysteine sulfoxide composition of 11 garlic (Allium sativum L.) cultivars by gas chromatography with flame ionization and mass selective detection

Two garlic subspecies (n = 11), Allium sativum L. var. opioscorodon (hardneck) and Allium sativum L. var. sativum (softneck), were evaluated for their free amino acid composition. The free amino acid content of garlic samples analyzed ranged from 1121.7 to 3106.1 mg/100 g of fresh weight (mean = 2130.7 +/- 681.5 mg/100 g). Hardneck garlic had greater methiin, alliin, and total free amino acids contents compared to softneck garlic. The major free amino acid present in all but one subspecies was glutamine (cv. Mother of Pearl had aspartic acid as the major free amino acid). Cv. Music Pink garlic (a rocambole hardneck variety) contained the most methiin, alliin, and total free amino acids. The solid-phase extraction, alkylchloroformate derivatization, GC-FID, and GC-MS methods used in this study were simple and rapid, allowing 18 free amino acids in garlic to be separated within 10 min.     

Characteristics of salt-fermented sauces from shrimp processing byproducts

A salt-fermented sauce from shrimp processing byproducts (heads, shells, and tails) was prepared and characterized. Three types of sauces were prepared; sauce C, with 30 g of salt/100 g of byproduct (high salt); sauce E, with 30 g of salt and 0.2 g of sodium erythorbate (high salt); and sauce L, with 20 g of salt, 0.2 g of sodium erythorbate, 6 g of sorbitol, 0.5 mL of lactic acid, and 5 mL of ethanol (low salt). Sauces C and E showed higher exopeptidase activities than sauce L, whereas sauce L showed the highest endopeptidase activity. After 3 months of fermentation, the amino N content of sauce increased from 150-200 to 500-600 mg/100 g and the nonprotein nitrogen content increased from 300 to 950-1050 mg/100 g. Volatile basic nitrogen content increased significantly from 18 to 60 mg/100 g. The total carotenoids retained in sauces C, E, and L were 26.3, 76.2, and 73%, respectively, thus indicating that the addition of sodium erythorbate to sauces E and L retarded oxidation. Water activities of sauces C, E, and L were 0.753, 0.751, and 0.773, respectively. According to the omission test, the taste of sauces was influenced by the content of free amino acids, mainly glutamic acid and aspartic acid. All three sauces examined showed a 35% higher total amino acid content than commercial salt-fermented shrimp sauces. Therefore, shrimp processing byproducts may lend themselves to the preparation of high-quality salt-fermented sauces.     

D-amino acid formation in sterilized alkali-treated olives

The occurrence of d-amino acids in commercial ripe olives, a well-known sterilized alkali-treated product, was investigated by high-performance liquid chromatography (HPLC) with precolumn automatic derivatization. Absolute amounts of D-amino acids were in total 18.6-38.2 mg/100 g edible portion. The major D-amino acids were D-aspartic acid, D-glutamic acid, D-serine, and D-leucine. Furthermore, to evaluate the effects of sterilization time and olive pH on amino acid racemization, a simulated processing of green ripe olives was carried out. Serine (both free and bound form) was the most-racemized amino acid after heat treatment. Sterilization (15-35 min at 121 degrees C) increased the racemization values of both free and protein-bound amino acids, although in case of protein-bound phenylalanine the increase was not statistically significant. With an increase of pH from 8 to 10 units, the racemization values of all amino acids increased significantly, except for free forms of aspartic and glutamic acids. In general, the effects of the sterilization time and olive pH on total concentration (L + D enantiomers) of each amino acid were also significant.     

不同干燥方式对中国对虾风味组分的影响

为了研究不同干燥方式对中国对虾风味组分的影响,该文采用热风(温度:(50±2)℃,风速:1.5 m/s,时间:8 h)、冷风(温度:18~20℃,风速:1.5 m/s,时间:56 h)、微波真空(功率500 W,真空度70 kPa,时间:40 min)、微波真空-冷风联合(先温度为18~20℃,时间27h的冷风干燥,后功率500 W,真空度70 kPa,时间10 min的微波真空干燥)4种干燥方式对其干制品的游离氨基酸组成、呈味核苷酸、等鲜量以及挥发性成分进行研究。结果表明,热风干燥后的中国对虾总游离氨基酸质量分数为63.31 mg/g,相对于对照组鲜虾(72.04 mg/g)有明显损失(P0.05);呈味核苷酸质量分数为7.9 mg/g,较对照组(9.05 mg/g)损失严重(P0.05);其等鲜量(127 g/(100 g))较鲜虾(180 g/(100 g))显著降低(P0.05);对虾产生以烤肉香味和海鲜风味为主的挥发性成分。冷风干燥使中国对虾总游离氨基酸质量分数较对照组损失偏大,其值为63.70 mg/g(P0.05);等鲜量(155(g/100 g))损失较大(P0.05);挥发性成分以烃类化合物为主,风味较寡淡。微波真空干燥后的中国对虾呈味核苷酸和等鲜量分别为9.17 mg/g和176 g/(100 g),总游离氨基酸质量分数较对照组损失较严重,为55.81 mg/g(P0.05);挥发性成分以肉香味和烤香味为主。微波真空-冷风联合干燥后的中国对虾呈味核苷酸含量最高,其值是9.90 mg/g;等鲜量值为189 g/(100 g),相对于鲜虾有所提高(P0.05);总游离氨基酸质量分数为62.84 mg/g呈现降低(P0.05);产生以烤肉香味和海鲜风味为主的挥发性成分。因此,微波真空-冷风联合干燥方式对中国对虾风味变化影响最小,是一种具有发展前景的干燥方式。     

Determination of nonprotein amino acids and betaines in vegetable oils by flow injection triple-quadrupole tandem mass spectrometry: a screening method for the detection of adulterations of olive oils

A novel screening method using an automated flow injection electrospray ionization tandem mass spectrometry system is proposed for the simultaneous determination of five nonprotein amino acids (β-alanine, alloisoleucine, ornithine, citrulline, pyroglutamic acid) and three betaines (glycine betaine, trigonelline, proline betaine) after derivatization with butanolic HCl. MS/MS experiments were carried out in a triple-quadrupole instrument using multiple reaction monitoring mode in <2 min. The proposed method provided high fingerprinting power to identify the presence of five of the studied compounds in different types of vegetable oils (soybean, sunflower, corn, olive) with LODs at parts per billion levels. The method was validated, and different mixtures of extra virgin olive oil with seed oils were analyzed, achieving the typification for the detection of adulterations in extra virgin olive oils up to 2% w/w. The nonprotein amino acid ornithine was confirmed as a marker for adulteration in the olive oils analyzed.     

鲜牛奶与豆浆的氨基酸营养比较

用氨基酸自动分析仪分析了鲜牛奶和5%豆浆的蛋白质氨基酸组成及含量。必需氨基酸(含胱氨酸和酪氨酸)占氨基酸总量牛奶为45.47%,豆浆为37.93%;必需氨基酸指数(EAAI)牛奶为89.72,豆浆为74.92;赖氨酸含量牛奶为479mg/gN,豆浆为351mg/gN;第一限制氨基酸均为蛋氨酸+胱氨酸,其含量鲜牛奶为208mg/gN,豆浆为190mg/gN;氨基酸分(AAS)和化学分(CS),鲜牛奶为0.95和0.54,豆浆为0.86和0.49。     

Phosphorous and Mung Bean Residue Incorporation Improve Soil Fertility and Crop Productivity in Sorghum and Mungbean-Lentil Cropping System

ABSTRACT

In sorghum and mungbean – lentil cropping system, field experiments were conducted for three successive years to assess the effect of mung bean residue incorporation on sorghum and succeeding lentil productivity along with different doses of phosphorus (P; 0, 30, 60 kg ha^{? 1}) applied to these crops. The level of soil fertility was also tested with or without incorporation of mung bean residue. The interaction of phosphorus to mungbean residue incorporation was thus studied in relation to improve crop productivity with balancing fertilizer requirements through an eco-friendly approach. Sorghum grain yield increased significantly when 60 kg P₂O₅ ha^{? 1} was applied and mungbean residue incorporated. The response was reduced to 30 kg P₂O₅ ha^{? 1} when mungbean residue was not incorporated. The succeeding lentil crop responded up to 60 kg P₂O₅ ha^{? 1} only when preceding sorghum crop received 0 or 30 kg P₂O₅ ha^{? 1}. Response to applied P₂O₅ to lentil reduced to 30 kg ha^{? 1} when preceding sorghum crop received 60 kg P₂O₅ ha^{? 1} and mungbean residue incorporated. Available soil nitrogen, phosphorus, and organic carbon content increased when mungbean residue was incorporated; however, available potassium (K) of the soil decreased from its initial value.     

Topography of the Cotyledon Surfaces and Adjoining Seed Coat of Chickpea (Cicer arietinum L.) Genotypes Differing in Milling Performance

Milling of pulse seeds generally refers to dehulling (decortication or seed coat removal) and splitting (division of the two cotyledons) to produce split seeds, known as dhal. Reduction of whole seeds or dhal to flour, in comparison, is generally termed “grinding” for pulses. Many pulses are consumed as dhal, including desi chickpea (Cicer arietinum L.), faba bean (Vicia faba L.), lentil (Lens culinaris L.), field pea (Pisum sativum L.), pigeon pea (Cajanus cajan L.), mungbean (Vigna radiata L.), black gram (Vigna mungo L.), and hyacinth bean (Lablab purpureus L.). Hence, ease‐of‐milling is an important quality attribute of pulse species that are commonly dehulled or split prior to consumption. Seed structure and the surface topography at the junctions between seed coat and cotyledons have the potential to influence the varietal differences observed in the ease of milling. The epicuticular wax surfaces of six genotypes of chickpea differing in ease of milling were examined by scanning electron microscopy to evaluate whether the physical attributes of seed coat and cotyledon surfaces could be involved in adhesion. Differing epicuticular wax patterns were observed for each of the six genotypes. The possible roles these patterns play in genotypic differences in ease of milling are discussed.     

Seed storage protein deficiency improves sulfur amino acid content in common bean (Phaseolus vulgaris L.): redirection of sulfur from gamma-glutamyl-S-methyl-cysteine

The contents of sulfur amino acids in seeds of common bean ( Phaseolus vulgaris L.) are suboptimal for nutrition. They accumulate large amounts of a gamma-glutamyl dipeptide of S-methyl-cysteine, a nonprotein amino acid that cannot substitute for methionine or cysteine in the diet. Protein accumulation and amino acid composition were characterized in three genetically related lines integrating a progressive deficiency in major seed storage proteins, phaseolin, phytohemagglutinin, and arcelin. Nitrogen, carbon, and sulfur contents were comparable among the three lines. The contents of S-methyl-cysteine and gamma-glutamyl-S-methyl-cysteine were progressively reduced in the mutants. Sulfur was shifted predominantly to the protein cysteine pool, while total methionine was only slightly elevated. Methionine and cystine contents (mg per g protein) were increased by up to ca. 40%, to levels slightly above FAO guidelines on amino acid requirements for human nutrition. These findings may be useful to improve the nutritional quality of common bean.     

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.     

Comparison of antioxidant activity, anthocyanins, carotenoids, and phenolics of three native fresh and sun-dried date (Phoenix dactylifera L.) varieties grown in Oman

Fresh and sun-dried dates of three native varieties from Oman, namely, Fard, Khasab, and Khalas, were examined for their antioxidant activity and total contents of anthocyanins, carotenoids, and phenolics, as well as free and bound phenolic acids. All results are expressed as mean value +/- standard deviation (n = 3) on a fresh weight basis. Fresh date varieties were found to be a good source of antioxidants (11687-20604 micromol of Trolox equiv/g), total contents of anthocyanins (0.24-1.52 mg of cyanidin 3-glucoside equiv/100 g), carotenoids (1.31-3.03 mg/100 g), phenolics (134-280 mg of ferulic acid equiv/100 g), free phenolic acids (2.61-12.27 mg/100 g), and bound phenolic acids (6.84-30.25 mg/100 g). A significant (p < 0.05) amount of antioxidants and carotenoids was lost after sun-drying of dates, whereas the total content of phenolics and free and bound phenolic acids increased significantly (p < 0.05). Anthocyanins were detected only in fresh dates. Date varieties had different levels and patterns of phenolic acids. Four free phenolic acids (protocatechuic acid, vanillic acid, syringic acid, and ferulic acid) and nine bound phenolic acids (gallic acid, protocatechuic acid, p-hydroxybenzoic acid, vanillic acid, caffeic acid, syringic acid, p-coumaric acid, ferulic acid, and o-coumaric acid) were tentatively identified. Of the date varieties studied, Khalas, which is considered to be premium quality, had higher antioxidant activity, total carotenoids, and bound phenolic acids than other varieties. These results suggest that all date varieties serve as a good source of natural antioxidants and could potentially be considered as a functional food or functional food ingredient, although some of their antioxidant constituents are lost during sun-drying.     

发芽绿豆苗的生理指标和抗氧化特性对邻苯二甲酸酯的响应特征

Single phytotoxicity of two representative phthalate esters （PAEs）, di-n-butyl phthalate （DnBP） and bis（2-ethylhexyl） phthalate （DEHP）, was tested in mung bean （Vigna radiata） seedlings germinated for 72 h in soils spiked with varying concentrations （0-500 mg kg-1 soil） of DnBP or DEHP. PAEs added at up to 500 mg kg-1 soil exerted no significant effect on germination but both pollutants significantly inhibited root elongation （P 〈 0.01）; DEHP inhibited shoot elongation （P 〈 0.01） and DnBP depressed biomass on a fresh weight basis （P 〈 0.05）. Seedling shoot and root malondialdehyde （MDA） Contents tended to be stimulated by DnBP but inhibited by DEHP. However, increases in superoxide dismutase, peroxidase, ascorbate peroxidase and polyphenol oxidase activities, as well as glutathione （GSH） content, were induced at higher concentrations （e.g., 20 mg kg-1） of both compounds. Accumulation of proline in both roots and shoots and the storage compounds, such as free amino acids and total soluble sugars, in whole plant was induced under the stress exerted by both PAEs. The general responses of mung bean seedlings indicated higher toxicity of DnBP than DEHP on primary growth, during which root elongation was a more responsive index. MDA and GSH were more sensitive parameters in the roots than in the shoots and they might be recommended as physiologically sensitive parameters to assess the toxicity of PAE compounds in soils in future long-term studies.     

Effect of starter fertilizer in seed-row on emergence,biomass and nutrient uptake by six pulse crops grown under controlled environment conditions

Abstract

Legumes have a unique ability to obtain a significant portion of atmospheric nitrogen (N₂) through a symbiotic relationship with Rhizobia spp of bacteria but it takes time, thus, an early supply of N to the plant may positively influence growth and development. However, too much fertilizer in close proximity to the seed can damage the seedling. Therefore, this study was conducted to determine the maximum safe rates for starter seed-row fertilizer application under low seedbed utilization conditions (15%). Emergence, biomass yield and nitrogen (N), phosphorus (P) and sulfur (S) uptake responses to starter fertilizer products and blends applied at 0, 10, 20 and 30?kg?N?ha^?1 in the seed-row were investigated for six different pulse crops: soybean, pea, faba bean, black bean, lentil and chickpea. The general sensitivity (injury potential) for starter N, P, S fertilizer was lentil?≥?pea?≥?chickpea?>?soybean?≥?black bean?>?faba bean. Lentil, pea and chickpea could generally only tolerate the 10?kg?N?ha^?1 rates while soybean and black bean could tolerate 10–20?kg?N?ha^?1. Faba bean emergence appeared relatively unaffected by all three rates of N and showed least sensitivity to seed row placed fertilizer. In terms of 30-day biomass response, soybean and black bean were most responsive to fertilization, while pea, faba bean, lentil and chickpea were least responsive to the starter fertilizer applications, with no benefit increasing above the 10?kg?N?ha^?1 rate.     

外源聚γ-谷氨酸对水稻幼苗耐旱性和渗透调节的影响

在水培条件下添加聚乙二醇-6000 (PEG-6000) 模拟干旱胁迫,研究聚γ-谷氨酸 (γ-PGA）对杂交稻品种金优38幼苗耐旱性和渗透调节的影响,探讨γ-PGA在提高水稻耐干旱中的效果和生理作用机制。结果表明,在12.5% PEG-6000胁迫下,使用100、200mg/L外源γ-PGA后,水稻幼苗的叶枯率降低,叶鲜重、干重和叶片活力明显提高,水稻幼苗的耐旱性较未添加γ-PGA处理（CK）增强。经外源γ-PGA处理后,水稻幼苗叶片中活性氧的累积量减少,脯氨酸、游离氨基酸和可溶性总糖的含量较对照分别提高1.3%~8.6%、0.2%~10.7%、13%~28.6%,差异达显著水平 (P< 0.05);外源γ-PGA可明显减缓干旱胁迫下钾离子含量降低的速率。外源γ-PGA可能通过参与水稻幼苗的渗透调节而提高水稻的耐旱能力。     

Simple method of isolation and purification of alpha-galactosides from legumes

A simple method for the isolation and purification of alpha-galactosides, raffinose family oligosaccharides (RFOs), from legumes has been developed. The method includes (i) imbibition of seeds, (ii) extraction with 50% ethanol, (iii) precipitation of RFOs, (iv) purification of RFOs on diatomaceous earth and charcoal, and (v) cation-exchange chromatography. The described method allows one to obtain high purity RFO preparations (90% for lentil and 80% for pea seeds, determined by HPLC-RI analysis) in the form of white, fine powder. Yields of alpha-galactosides isolated from 100 g of seeds of lentil and pea were 5.6 and 4.3 g, respectively.     

多效唑和三碘苯甲酸对大叶黄杨根系和叶片氮积累的影响

目的多效唑 (PP333) 和三碘苯甲酸 (TIBA) 是植物生长延缓剂,因其对植株矮化的显著效果,广泛应用于绿篱植物的化学修剪中,本研究使用此两种激素对苗木进行叶面喷施,探讨其对大叶黄杨根系和叶片氮代谢的影响。方法以三年生扦插大叶黄杨 (Buxus megistophylla) 苗为试验材料,在北京林业大学实验林场苗圃地内进行了叶面喷施田间试验。采用双因素随机区组设计,设置PP333 0、20、50、80 mg/L 四个浓度水平,TIBA为0、50、100 mg/L 三个浓度水平,共12个处理组合。自2017年4月5日起,每隔25天在每个小区喷施1 L备好的溶液。第三次喷施25天后取扦插苗全株样,分别测定了根系、叶片全氮、硝态氮、游离氨基酸含量以及叶片可溶性蛋白和光合色素含量。结果1) 单施高浓度PP333对细、中根全氮、硝态氮和游离氨基酸含量有显著提升作用,而单施TIBA仅对细根全氮、硝态氮和游离氨基酸含量有显著影响。PP333和TIBA及二者的交互效应对大叶黄杨细根的全氮、硝态氮和游离氨基酸含量均有极显著提升作用。2) PP333、TIBA及其交互作用均能极显著促进叶片全氮、可溶性蛋白含量和光合色素含量提升,50或80 mg/L PP333能够促进游离氨基酸含量提升,且仅在80 mg/L时对硝态氮含量有显著影响。3) PP333对不同径级根系全氮、游离氨基酸、硝态氮以及叶片全氮、硝态氮、游离氨基酸、可溶性蛋白和光合色素含量的促进作用较TIBA更为显著。结论PP333和TIBA对大叶黄杨氮代谢有显著影响,且相对于TIBA, PP333更能影响大叶黄杨氮的生理代谢过程。在生产应用中,80 mg/L PP333与100 mg/L TIBA结合喷施会加速根系中细根的氮代谢过程,并对叶片中蛋白质和光合色素的提升有显著促进作用,单施80 mg/L PP333显著促进叶片硝态氮和游离氨基酸的含量。     

Phenolic compounds and antioxidant capacity of Georgia-grown blueberries and blackberries

Blueberries and blackberries grown at various locations in Georgia were collected and analyzed for flavonoids, total anthocyanins, total polyphenols, and Trolox-equivalent antioxidant capacity (TEAC). Each sample was analyzed for phenolic acids (gallic acid, p-hydroxybenzoic acid, caffeic acid, ferulic acid, and ellagic acid) and flavonoids (catechin, epicatechin, myricetin, quercetin, and kaempferol). A high-performance liquid chromatographic (HPLC) method with photodiode array detection was used for analysis. Compounds were analyzed as aglycons after acid hydrolysis with 1.2 M HCl. Identification of each compound was based on retention time and UV spectra by comparison with pure commercial standards. Phenolic acids ranged from 0.19 to 258.90 mg/100 g fresh weight (FW), and flavonoids ranged from 2.50 to 387.48 mg/100 g FW. Total polyphenols ranged from 261.95 to 929.62 mg/100 g FW, and total anthocyanins ranged from 12.70 to 197.34 mg/100 g FW. TEAC values varied from 8.11 to a maximum of 38.29 microM/g FW. A linear relationship was observed between TEAC values and total polyphenols or total anthocyanins. The data indicate that blueberries and blackberries are rich sources of antioxidants.     

酵母发酵法改善沙丁鱼蛋白酶解液风味与品质的研究

为研究长孢洛德酵母对鱼蛋白酶解液风味与品质的改善作用,本试验以远东拟沙丁鱼酶解液为原料进行发酵,采用电子鼻主成分分析(PCA)法判断不同发酵条件下产物风味物质的差异,以感官评价及氨基酸态氮含量为指标,通过正交试验优化其发酵条件,对发酵产物进行游离氨基酸组成分析和必需氨基酸营养评价,并利用气相色谱-离子迁移谱(GC-IM...     

Determination of epsilon-N-pyrrolylnorleucine in fresh food products.

epsilon-N-Pyrrolylnorleucine was determined in different fresh food products to study its presence as a normal component of food proteins. Twenty-two different products were screened: cod, cuttlefish, salmon, sardine, trout, beef, chicken, pork, broad bean, broccoli, chickpea, garlic, green pea, lentil, mushroom, soybean, spinach, sunflower, almond, hazelnut, peanut, and walnut. Foods were homogenized, their proteins were precipitated with trichloroacetic acid and hydrolyzed with 2 N NaOH for 20 h, and the epsilon-N-pyrrolylnorleucine content was determined by capillary electrophoresis. The epsilon-N-pyrrolylnorleucine, which was identified by HPLC/MS in sardine muscle hydrolysate, ranged in the 22 foods analyzed from 0.24 to 6.36 micromol/g. This concentration was correlated with the protein content of the food (r = 0.687, p = 0.00041). In addition, the epsilon-N-pyrrolylnorleucine/lysine ratio was found to be a function of the lipid, iron, and protein contents of the food (r = 0.881, p < 0.0001) and was directly correlated with lipid and iron contents and inversely correlated with the protein content. These results are in agreement with the oxidative stress origin proposed for epsilon-N-pyrrolylnorleucine and suggest that the epsilon-N-pyrrolylnorleucine/lysine ratio is a characteristic of each food. In addition, epsilon-N-pyrrolylnorleucine seemed to be a normal component of many fresh food products, in which it may be acting as a natural antioxidant.