Capillary Gas Chromatographic Analysis of Amino Acids by Enantiomer Labeling

1987 ◽  
Vol 70 (2) ◽  
pp. 234-240
Author(s):  
Ernst Bayer ◽  
Hartmut Frank ◽  
Jürgen Gerhardt ◽  
Graeme Nicholson
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Sea Water ◽  
Biological Fluids ◽  
Chemical Properties ◽  
Internal Standard ◽  
Optical Isomers ◽  
Internal Standards ◽  
Sample Enrichment ◽  
Physical And Chemical

Abstract 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.

scholarly journals Determination of amino acids in human biological fluids by high-performance liquid chromatography: critical review

Amino Acids ◽  
2021 ◽  
Author(s):  
Grażyna Gałęzowska ◽  
Joanna Ratajczyk ◽  
Lidia Wolska
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
High Performance ◽  
Hplc Analysis ◽  
Limit Of Detection ◽  
Biological Fluids ◽  
Analytical Techniques ◽  
Epidemiological Studies ◽  
Preparation Conditions

AbstractThe quantitation and qualification of amino acids are most commonly used in clinical and epidemiological studies, and provide an excellent way of monitoring compounds in human fluids which have not been monitored previously, to prevent some diseases. Because of this, it is not surprising that scientific interest in evaluating these compounds has resurfaced in recent years and has precipitated the development of a multitude of new analytical techniques. This review considers recent developments in HPLC analytics on the basis of publications from the last few years. It helps to update and systematize knowledge in this area. Particular attention is paid to the progress of analytical methods, pointing out the advantages and drawbacks of the various techniques used for the preparation, separation and determination of amino acids. Depending on the type of sample, the preparation conditions for HPLC analysis change. For this reason, the review has focused on three types of samples, namely urine, blood and cerebrospinal fluid. Despite time-consuming sample preparation before HPLC analysis, an additional derivatization technique should be used, depending on the detection technique used. There are proposals for columns that are specially modified for amino acid separation without derivatization, but the limit of detection of the substance is less beneficial. In view of the fact that amino acid analyses have been performed for years and new solutions may generate increased costs, it may turn out that older proposals are much more advantageous.

STUDY OF AMINO ACID MODIFIERS IN GUAYULE NATURAL RUBBER

2015 ◽  
Vol 88 (2) ◽  
pp. 310-323 ◽  
Author(s):  
Colleen McMahan ◽  
Dhondup Lhamo
Keyword(s):  
Amino Acids ◽  
Natural Rubber ◽  
Oxidative Stability ◽  
Chemical Properties ◽  
The United States ◽  
Thermal Oxidative Stability ◽  
Green Strength ◽  
Strain Induced Crystallization ◽  
New Perspective ◽  
Physical And Chemical

ABSTRACT Guayule, a desert shrub indigenous to the United States, is under development as a source of natural rubber that can be used in place of petroleum-based rubber or Hevea rubber. In natural rubbers, physical and chemical properties can be strongly affected by nonrubber constituents, typically proteins and lipids, present in the material, depending on the plant species and postharvest processing. Hevea natural rubber typically contains high levels of nonrubber constituents that contribute to thermal-oxidative stability, cure acceleration, and especially strain-induced crystallization. The latter has been attributed to compound properties that render Hevea natural rubber uniquely suited for the most demanding rubber applications (e.g., aircraft tires). Hevea proteins are susceptible to hydrolysis, releasing free amino acids into the latex, which can affect rubber and compound properties. Here, low-protein guayule latex was blended with a series of amino acids varying in chemical structure. Bulk viscosity was reduced, thermal-oxidative stability was improved, and cure rate was influenced by the addition of amino acids. Generally, gel formation, green strength, and tensile strength were not affected. The results introduce a new perspective for amino acids as biobased rubber compound additives and provide insights into naturally occurring nonrubber constituents' interaction with natural rubber polymers.

Rabbit Cardiac Myosin. II. Proteolytic Fragmentation with Insolubilized Papain

1975 ◽  
Vol 53 (2) ◽  
pp. 175-188 ◽  
Author(s):  
William T. Wolodko ◽  
Cyril M. Kay
Keyword(s):  
Amino Acid ◽  
Chemical Properties ◽  
Single Type ◽  
Myosin Molecule ◽  
Cardiac Myosin ◽  
Dialdehyde Starch ◽  
Parent Molecule ◽  
Documented Information ◽  
Physical And Chemical ◽  
Helical Molecules

The substructure of the cardiac myosin molecule was examined by the limited proteolytic digestion of the parent molecule with (dialdehyde starch)-methylenedianiline–mercuripapain, S-MDA–mercuripapain, at low temperatures and neutral pH, using moderate enzyme to myosin ratios. Pertinent properties of the insoluble enzyme complex were also examined. Kinetic, ultracentrifugal, and chromatographic observations of the fragmentation process revealed that a single type of lytic reaction occurs during the early stages, predominantly releasing heavy meromyosin subfragment 1 (HMM-S1) and myosin rods. With further time of digestion, the rods are additionally cleaved yielding light meromyosin and HMM-S2, and HMM-S1 is found to be partially degraded. The major proteolytic subfragments were isolated, purified, and characterized with respect to their enzymatic, optical, amino acid, and physicochemical properties. Only HMM-S1 exhibited Ca2+-activated ATPase activity, and at a level three- to fourfold higher than that of native myosin. Moreover, its hydrodynamic properties suggest that it is globular in structure. On the other hand, light meromyosin-A (LMM-A) (which consists mainly of rods), and HMM-S2 appear to be highly asymmetric, rigid, α-helical molecules devoid of the amino acid proline. Strong similarities were evident in all aspects upon comparison of these results with documented information concerning the skeletal system. On the basis of the physical and chemical properties of the proteolytic subfragments relative to that of native myosin, it was further concluded that the cardiac myosin molecule is a double-stranded, α-helical rod ending in two subfragment 1 globules, of which only one may be enzymatically active at a time.

The Composition of the Seminal Plasma of Balanus Balanus

1962 ◽  
Vol 39 (3) ◽  
pp. 345-352
Author(s):  
H. BARNES
Keyword(s):  
Amino Acids ◽  
Ascorbic Acid ◽  
Amino Acid ◽  
Seminal Plasma ◽  
Free Amino ◽  
Reducing Sugars ◽  
Sea Water ◽  
Protective Action ◽  
Inorganic Ions ◽  
Chemical Analyses

1. The results of some chemical analyses for inorganic and organic constituents of the seminal plasma of Balanus balanus are presented. 2. The inorganic ions show a cation deficit of 178 m-equiv./l., which is probably made up by free amino-acids. 3. Cystine is a prominent amino acid present. 4. Potassium and calcium are present in excess of their quantities in sea water. 5. Reducing sugars, compared with the amount found in sea-urchin spermatozoa, are found in moderate quantities, 1 mg./ml. 6. Phosphorus of all kinds is present in only small quantities (total of o.14 mg./ml.). 7. Some phosphatases are present. 8. There are 21 /µg./ml. of ascorbic acid; the function of this is discussed in relation to its possible contribution to the protective action against the poisoning of -SH groups by thiol-reactive agents.

Free Amino Acid and Haemolymph Concentration Changes in Sphaeroma Rugicauda (Isopoda) During Adaptation to a Dilute Salinity

1969 ◽  
Vol 50 (2) ◽  
pp. 319-326
Author(s):  
R. R. HARRIS
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Free Amino Acid ◽  
Free Amino Acids ◽  
Free Amino ◽  
Sea Water ◽  
Protein Nitrogen ◽  
Body Sodium ◽  
Concentration Changes ◽  
Total Body

1. Non-protein and protein nitrogen fractions of the isopod Sphaeroma rugicauda were measured in animals adapted to 100 and 2% sea water. 2. The non-protein nitrogen component was reduced in animals acclimatized to the lower salinity. 3. Free amino acids accounted for 88 and 74% respectively of the non-protein nitrogen in the two salinities. 4. In 2% sea water taurine, proline, glycine, alanine and glutamic acid showed the greatest decreases in concentration compared to the levels measured in animals adapted to 100% sea water. 5. The decrease in total free amino acids of animals acclimatized to 100% sea water and transferred to 2% sea water was measured. 6. The total free amino acid concentration is reduced to the 2% sea water level within 12 hr. after transfer. 7. Free amino acid, haemolymph sodium and total body sodium levels after transfer to 2% sea water were compared. 8. The asymmetry between the fall in haemolymph sodium concentration and the decrease in total body sodium under these conditions is thought to be due to a water shift from the haemolymph into the tissues. 9. It is suggested that the osmotic pressure of the cells falls at a slower rate than that of the haemolymph.

Gas Chromatography in Diagnostic Biochemistry of Abnormal Valine Metabolism

1973 ◽  
Vol 19 (9) ◽  
pp. 1040-1044 ◽  
Author(s):  
Hugh N Tucker ◽  
Samuel V Molinary
Keyword(s):  
Amino Acids ◽  
Gas Chromatography ◽  
Amino Acid ◽  
Biological Fluids ◽  
Sample Sizes ◽  
Diagnostic Aid ◽  
Chromatographic Techniques ◽  
Biochemical Evaluation ◽  
Amino Acid Concentrations ◽  
Chromatographic Evidence

Abstract Amino acids in biological fluids can be rapidly and accurately quantitated by gas chromatography. Modification of existing gas-chromatographic techniques allow as little as 25 µl of biological fluids to be analyzed, compatible with sample sizes easily obtained in pediatric studies. Simplified diagnostic biochemical evaluation of a suspected aminoaciduria can be begun at birth, or shortly after. Normal amino acid patterns are compared with those found in a patient with abnormal amino acid concentrations. Gas chromatographic evidence is presented as a diagnostic aid in the evaluation of a patient with abnormal valine metabolism.

Determination of total proteinogenic amino acids and taurine by pre-column derivatisation and UHPLC: Single Laboratory Validation, First Action 2019.09

2020 ◽  
Author(s):  
George Joseph ◽  
Asha Varughese ◽  
Ann Daniel
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Infant Formula ◽  
Retention Time ◽  
Sulfonic Acid ◽  
Internal Standard ◽  
The Other ◽  
Method Performance ◽  
Single Laboratory

Abstract Background A method has been developed and validated for selective, accurate and precise determination of total proteinogenic amino acids and taurine from Infant Formula and Adult/Pediatric Nutritional Formulas (powders, ready-to-feed liquids, and liquid concentrates). The method was reviewed by the AOAC INTERNATIONAL SPIFAN Expert Review Panel (ERP) during the 133rd AOAC Annual Meeting & Expo on September 7, 2019 in Denver, CO, USA and was recommended to First Action Official MethodsSM status. Objective The method involves protein hydrolysis to amino acids, a simple pre-column derivatisation of amino acids and separation of derivatised amino acids by UHPLC. The quantification of amino acids is performed by multi-point calibration using norvaline as the internal standard. The analytical method is capable of quantitative determination for 22 proteinogenic amino acids, but cannot be used to quantitate tryptophan, which is destroyed during the acid hydrolysis step. Asparagine is determined as aspartic acid and glutamine as glutamic acid. The cystine and cysteine are converted to S-2-carboxyethylthiocysteine (CYSx) and the derivative is separated from the other amino acids. Citrulline which is present in some matrices and it is separated from other amino acids is not included in the method performance evaluation in the single laboratory validation (SLV). Method The proposed method met all the performance requirement limits set in standard method performance requirements (SMPR) 2014.013 for total proteinogenic amino acids and taurine. The correlation coefficient of multi-point calibration was not less than 0.999 for any amino acids at any point in the SLV study confirming the validity of linear dynanic range (LDR) and linearity of the method. The individual amino acids in the chromatogram were identified by absolute retention time and relative retention time (RRT) with respect to the internal standard norvaline. There were no significant (S/N Ratio <10) interferences from the reagents or by-products of derivatisation and targeted matrices. The method demonstrated high selectivity. Result Accuracy of the method was validated using standard reference materials (NIST SRM 1869 and 1849a) and spike recovery studies. The amino acid results in the SRMs were within the ranges of Reference Mass Fraction Values. The accuracy of the method was corroboratively validated by spike recovery studies. The average spike recovery range between 93 to 107% ensure the accuracy of the method for amino acids and compliance to the AOAC SMPR 2014.013. Conclusions Precision data of the method demonstrate that it meets the stakeholder requirements as per the SMPR. The mean RSDr for all the amino acids for 17 matrices selected for the SLV were not more than 4%. The method is very sensitive and the LOQ can go down to approximately ten times lower than the SMPR requirements. The sensitivity of method is a direct reflection of its signal to noise ratio which ensures guaranteed method performance at the lower levels of amino acids in these matrices. Highlights Taurine (aminoethane sulfonic acid) unlike the other amino acids is a beta-sulfonic amino acid that is not used in protein synthesis but is found as a free amino acid in tissues. The acidic functional group (-COOH) in common amino acid is replaced with a sulfonic acid (-SO3H) group in Taurine. The method offers baseline separation of citrulline which is an alpha amino acid generally present in Infant Formula and Adult/Pediatric Nutritional products. The separation of citrulline eliminates the risk of interference of this compound with other amino acids. The method can also separate and quantitate hydroxyproline, an important component of collagen that is often used to quantitate collagen. The method is simple and does not include any proprietary chemicals or instruments and can be performed on any basic reverse phase UHPLC system with UV detection.

EFFECT OF PROTEIN ADDITION ON PROPERTIES OF GUAYULE NATURAL RUBBER

2017 ◽  
Vol 90 (2) ◽  
pp. 387-404 ◽  
Author(s):  
Dhondup Lhamo ◽  
Colleen McMahan
Keyword(s):  
Amino Acids ◽  
Natural Rubber ◽  
Oxidative Stability ◽  
Chemical Properties ◽  
Green Strength ◽  
Truck Tires ◽  
Oxidative Resistance ◽  
Strain Induced Crystallization ◽  
General Protein ◽  
Physical And Chemical

ABSTRACT Parthenium argentatum, commonly known as guayule, is a desert shrub cultivated as a domestic source of natural rubber in the semi-arid southwestern United States. Guayule natural rubber (GR) may be used to replace petroleum-based rubber or in place of Hevea natural rubber (NR), but substitution must take into consideration differences in physical and chemical properties. Currently, Hevea NR is required in tire applications, especially aircraft and truck tires, because of its high oxidative resistance, rapid cure rate, and exceptional stress–strain response. These outstanding features are attributed to the presence of nonrubber constituents, mainly proteins and lipids, which cause the rubber to gel, and they contribute to strain-induced crystallization. In contrast, GR is low in proteins and is thus deprived of some attributes of Hevea. Addition of amino acids and proteins to guayule could potentially improve performance and thereby widen the range of applications for use. In a previous study, amino acids blended with GR latex improved thermo-oxidative stability, served as plasticizers and cure accelerators, and enhanced green strength slightly, but tensile strength was not improved. Here, a series of bio-based commercial proteins (gelatin, soy, albumin, casein, zein, gliadin, and gluten) were added to GR as a latex blend. In general, protein addition reduced bulk viscosity and improved thermo-oxidative stability. The gel content and green strength of the polymer–protein blends were increased, with the exception of gliadin, but not to levels observed for Hevea. Effects on vulcanization and mechanical properties in compounds were surprisingly influenced by the choice of antioxidants used. Our results demonstrate the potential of proteins as bio-based rubber compounding additives.

scholarly journals Porin from Marine Bacterium Marinomonas primoryensis KMM 3633T: Isolation, Physico-Chemical Properties, and Functional Activity

Molecules ◽  
2020 ◽  
Vol 25 (14) ◽  
pp. 3131
Author(s):  
Olga D. Novikova ◽  
Valentina A. Khomenko ◽  
Natalia Yu. Kim ◽  
Galina N. Likhatskaya ◽  
Lyudmila A. Romanenko ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Lipid Membranes ◽  
Marine Bacterium ◽  
Chemical Properties ◽  
Cell Permeability ◽  
Oligomeric Structure ◽  
Black Lipid Membranes ◽  
Physico Chemical ◽  
Coastal Sea

Marinomonas primoryensis KMM 3633T, extreme living marine bacterium was isolated from a sample of coastal sea ice in the Amursky Bay near Vladivostok, Russia. The goal of our investigation is to study outer membrane channels determining cell permeability. Porin from M. primoryensis KMM 3633T (MpOmp) has been isolated and characterized. Amino acid analysis and whole genome sequencing were the sources of amino acid data of porin, identified as Porin_4 according to the conservative domain searching. The amino acid composition of MpOmp distinguished by high content of acidic amino acids and low content of sulfur-containing amino acids, but there are no tryptophan residues in its molecule. The native MpOmp existed as a trimer. The reconstitution of MpOmp into black lipid membranes demonstrated its ability to form ion channels whose conductivity depends on the electrolyte concentration. The spatial structure of MpOmp had features typical for the classical gram-negative porins. However, the oligomeric structure of isolated MpOmp was distinguished by very low stability: heat-modified monomer was already observed at 30 °C. The data obtained suggest the stabilizing role of lipids in the natural membrane of marine bacteria in the formation of the oligomeric structure of porin.

Effect of Deproteinating Agents (Picric Acid and Sulfosalicylic Acid) on Analysis for Free Amino Acids in Swine Blood and Tissue

1969 ◽  
Vol 52 (5) ◽  
pp. 981-984 ◽  
Author(s):  
J E Knipfel ◽  
D A Christensen ◽  
B D Owen
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Ion Exchange ◽  
Free Amino ◽  
Biological Fluids ◽  
Picric Acid ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Sulfosalicylic Acid ◽  
Coefficients Of Variation

Abstract Amino acid analyses were performed on samples of blood, liver tissue, loin muscle, and ham muscle by ion exchange chromatography after deproteination of the samples with picric acid or sulfosalicylic acid (SSA). Resolution of threonine and serine from the ion exchange column was poor when SSA was used as the deproteinating agent. Twelve of sixteen amino acids were higher (P < 0.05) in serum deproteinated with picric acid as compared to concentrations determined after SSA deproteination. Amino acid values for ham muscle tended to be higher after deproteination with picric acid; however, with liver and loin muscle samples, the values were somewhat higher after SSA deproteination. In both serum and tissue analyses, coefficients of variation were lower for niGSt amino acids when picric acid was utilized as the deproteinating agent. The latter observation, in particular, suggests that picric acid is preferable to SSA as a deproteinating agent before amino acid analyses of biological fluids. Standardization of methods of deproteination is needed to allow meaningful comparisons of data.

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