Simultaneous HPLC analysis of biogenic amines, amino acids, and ammonium ion as aminoenone derivatives in wine and beer samples

A method has been developed for the simultaneous analysis of biogenic amines, amino acids, and the ammonium ion in wine and beer. Aminoenones formed by the reaction of amino acids, biogenic amines, and the ammonium ion with the derivatization reagent diethyl ethoxymethylenemalonate are separated by HPLC. Reaction takes place in methanolic alkaline medium for 30 min in an ultrasonic bath. Further heating at 70 degrees C for 2 h produces complete degradation of excess derivatization reagent and byproducts. Comparison of the results of ammonium analysis and enzymatic analysis showed a good correlation (r = 0.953). The proposed analytical method has the following advantages: easy derivatization of wines and beers; quantification of 24 amino acids, nine biogenic amines, and the ammonium ion in a single injection; use of the photodiode array detector; complete degradation of excess derivatization reagent during sample preparation; and detection limits below 0.40 mg/L for amino acids and below 0.06 mg/L for biogenic amines.     

Precolumn phenylisothiocyanate derivatization and liquid chromatography of amino acids in food

A precolumn phenylisothiocyanate derivatization method is described for the determination of amino acids in protein hydrolysates from a wide variety of complex food matrixes, with and without performic acid oxidation pretreatment. Analysis of samples that were not pretreated with performic acid was necessary since this pretreatment destroyed an average of 25% of the histidine and 87% of the tyrosine present in the food samples. This method is rapid and reproducible; coefficients of variation between duplicate analyses of the same food item were less than 5% for a majority of the amino acids. Occasionally, variation between duplicate analyses for histidine and tyrosine was greater than 10%. Recoveries of amino acids added to samples were in the 100% range.     

Detection and quantification of protein residues in food grade amino acids and nucleic acids using a dot-blot fluorescent staining method

Food allergies represent an important health problem in industrialized countries, such that detection and quantitative analysis of the protein considered to be the main allergen is crucial. A dot-blot fluorescent staining method for the detection and quantitative analysis of protein residues in food grade amino acids and nucleic acids is presented. This method combines fluorescence staining with dot-blotting onto PVDF membrane. Several standard proteins, such as bovine serum albumin (66 kDa), lysozyme (14 kDa), ubiquitin (8.6 kDa), bovine insulin (5.7 kDa), and oxidized insulin B chain (3.5 kDa), were detectable at 0.1 ppm using SYPRO Ruby blot stain. Twenty-five different amino acids and two different nucleic acids of food grade were analyzed using this method combined with an internal standard addition method using BSA as an internal standard. All amino acids and nucleic acids were dissolved in 3.6% aqueous HCl and dot-blotted onto PVDF membrane before a large amount of amino acids and nucleic acid were removed. Protein residues and the internal standard protein immobilized on the membrane were stained using SYPRO ruby blot stain. The internal standard in all samples was detectable at 0.1 ppm. Samples were dissolved at 120 or 70 mg/mL, according to their solubility under acidic conditions. The detection limit of protein residues per weight was 0.8-1.4 ppm in amino acids and nucleic acids; residual protein was not detected in any sample.     

A system for the rapid analysis of organic phosphorus in water samples or fractions from chromotographic columns

Summary

An automatic method for the colorimetric determination of Al in soil extracts using catechol violet was developed. The method is very reproducible and precise. The manifold arrangement covers a range of 0–1 μg ml^?1. By using a dilution loop, solutions containing up to 10 μg ml^?1 can be analysed. The method compares favourable with a manual catechol violet method and the alumin on method.     

Capillary gas chromatographic analysis of amino acids by enantiomer labeling

The optical isomers of amino acids can be easily separated by gas chromatography using capillary columns coated with the chiral polysiloxane peptide, Chirasil-Val. Quantitative trace amino acid analysis in complex mixtures such as biological fluids, sea water, or protein hydrolysates can be achieved by enantiomer labeling: The D-amino acid enantiomers, which do not occur naturally, are added to the sample prior to analysis as internal standards. Because the D-enantiomers show the same physical and chemical properties as the natural L-enantiomers, they are ideal standard references. In routine analysis, the derivatization is achieved with a new automated derivatization robot. The D-standard serves as overall internal standard for the whole analytical procedure from sample enrichment to derivatization, chromatography, and response of the detector.     

Oxidation and hydrolysis determination of sulfur amino acids in food and feed ingredients: collaborative study

Samples of 6 food and feed ingredients and a purified protein, beta-lactoglobulin, were analyzed by 7 laboratories to determine the concentrations of cysteine as cysteic acid and methionine as methionine sulfone. Samples were oxidized by reaction with performic acid before hydrolysis with 6N HCl. The free amino acids were then separated and measured by ion-exchange chromatography on dedicated amino acid analyzers. Each laboratory was provided with a detailed method as well as sealed vials containing solutions of standards. For the determination of cysteine as cysteic acid, the coefficients of variation between laboratories for duplicate samples ranged from 7.13 to 10.8% for the 6 ingredients. For the determination of methionine as methionine sulfone, the coefficients of variation between laboratories for duplicate samples ranged from 1.18 to 12.8% for the 6 ingredients. Cysteine and methionine recoveries were determined by analysis of beta-lactoglobulin and were based on expected levels of each amino acid from amino acid sequence data. The mean recovery of cysteine was 95% with a range of 91-101%. The mean recovery of methionine was 101% with a range of 98-106%. This method has been adopted official first action.     

Profile of trans fatty acids (FAs) including trans polyunsaturated FAs in representative fast food samples

The content of trans fat in foods is most commonly determined by summing the levels of individual trans fatty acids (FAs), analyzed as FA methyl esters (FAME) by gas chromatography. Current Official Methods of the American Oil Chemists' Society (AOCS) enable quantitation of total trans fat in foods but were not designed for the determination of transFA isomeric compositions. In the present study, the content of trans fat in 32 representative fast food samples ranged from 0.1 to 3.1 g per serving, as determined according to AOCS Official Method Ce 1j-07. Further analysis of FAME using the 200 m SLB-IL111 ionic liquid column yielded quantitative results of total, trans, saturated, and cis unsaturated fat that were comparable to those of Method Ce 1j-07 and also allowed for the complementary determination of individual trans 18:1, trans 18:2, and trans 18:3 FA isomeric compositions under conditions suitable for routine sample analysis.     

Determination of sulfur amino acids and tryptophan in foods and food and feed ingredients: collaborative study

Samples of 4 foods, 1 animal feed, isolated soy protein, and beta-lactoglobulin were analyzed by 9 laboratories to determine concentrations of cysteine as cysteic acid, methionine as methionine sulfone, and tryptophan. Sulfur amino acids were determined by AOAC method 43.A08-43.A13 for food and feed ingredients, in which samples are oxidized with performic acid before protein hydrolysis with 6N HCl. Tryptophan was determined after protein hydrolysis with 4.2N NaOH. In both methods, free amino acids were separated by ion-exchange or reverse-phase chromatography. Each laboratory was provided with detailed methods and with sealed vials containing solutions of standards. Samples were analyzed in duplicate, and variation between laboratories was determined. Coefficients of variation between laboratories for the 6 samples ranged from 5.50 to 11.8% for methionine as methionine sulfoxide, 8.59 to 17.3% for cysteine as cysteic acid, and 3.87 to 16.1% for tryptophan. Amino acid recoveries were determined by analysis of beta-lactoglobulin and were based on expected levels of each amino acid obtained from amino acid sequence data. The mean recovery of cysteine was 97% with a range of 88-119%. For methionine, mean recovery was 98% (range 89-115%) and for tryptophan, 85% (range 59-102%). Method 43.A08-43.A13 for food and feed ingredients has been adopted official first action for determination of cysteine and methionine in processed foods. The alkaline hydrolysis method has been adopted official first action for determination of tryptophan in foods and food and feed ingredients.     

Gas chromatographic analysis of amino acids as the N-heptafluorobutyryl isobutyl esters

The N-heptafluorobutyryl isobutyl derivatives of proteic amino acids are well resolved by gas chromatography and form the basis of a convenient, rapid assay. The derivatives are prepared by acid-catalyzed esterification at 120 degrees C for 20 min in 3N HCl-isobutanol followed by acylation with heptafluorobutyric anhydride at 150 degrees C for 10 min. The reaction sequence is performed without any transfers or extractions and thus is compatible with microscale analysis. A complete proteic amino acid profile can be completed in less than 20 min by using a packed column or less than 10 min by using a capillary column in combination with an elevated oven temperature program rate. Physiological sample matrixes, which frequently contain a complex mixture of components, and thus require maximum resolution, can be assayed in less than 1 h using a program rate of 4 degrees C/min. A capillary column is recommended for this application. Capillary column chromatography, in combination with a nitrogen-specific detector, is useful for identifying and assaying nonproteic amino acids in physiological sample matrixes. Frequently, a prior cleanup of the sample can be avoided.     

Comparative analysis of meat samples prepared with food chopper and bowl cutter

Analyses of meat samples after preparation with either a bowl cutter or by the official procedure with a food chopper were compared for homogeneity of comminution and for differences in fat, moisture, and protein content. Cutting time in the bowl cutter was limited to minimize temperature rise in samples. Beef chuck, pork shoulder, and beef shank, cheek, and tongue were used in the study. Variances of replicate analysis data for the 5 meat types were pooled for either cutter or chopper treatment and for each analyzed component. Sample portions cut and mixed by using the bowl cutter were more homogeneous than those ground with a food chopper. Comparative accuracy was indicated by fat and moisture means: 5 were in good agreement and 5 differed significantly; 3 of 5 paired protein means differed significantly but were within 0.3% protein. Results on precision and accuracy as well as the simplicity and convenience of the bowl cutter procedure favor its use as an alternative to a food chopper for preparing meat samples for analysis.     

Gas chromatographic analysis of amino acids from the action of proteolytic enzymes on soil humic acids

Six proteases released more amino compounds from albumin than from a humic acid. Pronase and thermolysin released about one-sixth of the acid-hydrolysable amino acids from a humic acid, papain had slight activity while chymotrypsin, trypsin and subtilopeptidase did not attack humic acid. Thermolysin hydrolysates of four humic acids contained peptides which on acid hydrolysis broke down to increase three fold the yield of α-amino N. Net amounts of amino compounds released on incubation of thermolysin and humic acids were directly related to incubation time, substrate concentration and enzyme concentration.     

Composition of amino acids,organic acids,and sugars in the peribacteroid space of soybean root nodules

In leguminous root nodules, bacteroids are differentiated from rhizobia and are surrounded by a peribacteroid membrane (PBM) forming an intracellular structure designated as symbiosome. Through the peribacteroid space (PBS) between the PBM and bacteroids, metabolic substances and signal compounds are exchanged between two symbionts. In this study, organic compounds with low molecular weight in the PBS were collected from isolated symbiosomes of soybean (Glycine max L.) root nodules, and their composition was analyzed and compared with that of the organic compounds in whole root nodules and bacteroids. Major differences were detected in the molar percentages of amino and organic acids, and sugars, to the total low molecular weight organic compounds among whole root nodules, PBS, and bacteroids. The PBS composition was characterized by abundant sugars and poor amino acids. Also the composition of the amino acids, organic acids, and sugars in the PBS was clearly different from that in whole root nodules and bacteroids. The PBS sugar composition was characterized by the predominance of inositols, especially myo-inositol at the 5th and 7th weeks of the host plant growth stages. Changes in the myo- and D-chiro- inositol balance at the host plant growth stages occurred and a syntony was observed between the PBS and bacteroids. The localization of myo-inositol in the PBS accounted for almost 70% of the total myo-inositol in root nodules. A small difference in the PBS composition between two soybean cultivars was recorded but it varied with the growth stages. It was tentatively concluded that the PBS sugar composition affected the bacteroidal sugar composition in soybean plants, and that inositol utilization in the bacteroids could be a factor controlling the bacteroidal function level which varied with the host plant growth stages.     

Identification of amino acids in humic acid

Davidson, Sowden and Atkinson^l) in 1951 reported on the amino acids in the three soil organic matter fractions, and Stevenson, Marks, Varner and Martin~) in 1952 reported on the identification of eight amino acids from clay-adsorbed organic colloids in Brookstom slity clay loam. Also, some investigations on these amino acids have been carried out in our laboratory.     

A field method to store samples from temperate mountain grassland soils for analysis of phospholipid fatty acids

The storage of soil samples for PLFA analysis can lead to shifts in the microbial community composition. We show here that conserving samples in RNAlater, which is already widely used to store samples for DNA and RNA analysis, proved to be as sufficient as freezing at?-20?°C and preferable over storage at 4?°C for temperate mountain grassland soil. The total amount of extracted PLFAs was not changed by any storage treatment. Storage at 4?°C led to an alteration of seven out of thirty individual biomarkers, while freezing and storage in RNAlater caused changes in the amount of fungal biomarkers but had no effect on any other microbial group. We therefore suggest that RNAlater could be used to preserve soil samples for PLFA analysis when immediate extraction or freezing of samples is not possible, for example during sampling campaigns in remote areas or during transport and shipping.     

Bound amino acids in humic acids from arable cropping systems

We investigated the varying concentrations of bound amino acids in humic acids (HA) extracted from soils under both crop rotation and continuous cropping of rye. The experiment was created in 1957. Since then, winter rye had been grown continuously and also the sequence of the 7 yr rotation had been started: potato, spring barley, alfalfa, alfalfa, oil seed rape, winter rye, and winter rye. Soils were fertilized with NPK and manure. Continuous cropping of rye increased total acidity of soils and the contents of carboxylic and phenolic groups in HA. The total amounts of the bound amino acids in HA from soils under crop rotation were higher than from continuous cropping of rye. Fertilization with NPK increased the contents of bound amino acids more than manure. Neutral amino acids dominated in all samples of HA, and basic amino acids had the lowest concentrations. In both types of cultivation, glutamic acids, glycine, alanine, valine, and lysine dominated. The proline contents in HA from continuous rye cropping were higher than in HA from soils under crop rotation. The concentrations of β‐alanine and lysine were higher in HA from crop rotation indicating a higher microbial biomass since these compounds are typical constituents of bacteria cell walls.     

植物对氨基酸的吸收利用及氨基酸在农业中的应用

植物对氨基酸的吸收、转运、代谢以及氨基酸在肥料和农药上的应用国内外已有报道。已有研究证明,植物可直接吸收土壤中的氨基酸分子,其吸收后的转运、分配、代谢因氨基酸种类而异,产生的生理效应也不相同;氨基酸农药易被日光分解或被自然界微生物降解,在土壤中、植物体内不留残毒,其降解产物还可作为农作物的营养物质,提高农作物的品质和产量,施用这类农药,人畜安全,没有公害;氨基酸肥具有促进植株生长发育、增强抗逆性、改善土壤状况和提高作物产量的作用。     

HPLC method for analysis of free amino acids in fish using o-phthaldialdehyde precolumn derivatization

Precolumn derivatization applying o-phthaldialdehyde (OPA) was used to analyze free lysine, histidine, and ornithine, precursors of the respective biogenic amines cadaverine, histamine, and putrescine, which are considered indicators of fish quality and safety. This method uses 75% methanol to eliminate the use of strong acids as the extraction solution. Each analysis took 35 min, was reproducible, and allowed separation of primary amino acids in fish samples. A binary solvent delivery system coupled with a fluorescence detector and an Ultrasphere ODS column were utilized for HPLC separation. Linearity of the calibration curves was very good (r(2) = 0.99) for the amino acids of interest. Minimum concentrations of detection were 40 pmol/mL for histidine and lysine and 70 pmol/mL for ornithine. Average recoveries were 72% for lysine, 93% for histidine, and 98% for ornithine. This method used solvent gradient elution to study the levels of these analytes in mahi-mahi, bigeye tuna, and flounder.     

基于核主成分分析(KPCA)的一种食品评价模型

该文提出了一种新的食品评价模型核主成分分析(KPCA).它通过一个非线性变换,首先将原变量空间映射到高维特征空间,然后在这个高维特征空间中进行线性主成分分析.通过核技巧,不必知道非线性变换的具体形式,只要选取适当的核及其参数,就可使第一主成分的贡献率达到85%以上,从而能有效地避免主成分分析(PCA)只能处理线性问题和降维效果不明显的弊端.在对豆腐乳的品质进行评价时,得到了较理想的评价效果.