Evaluation of liquid chromatographic analysis of nutritionally important amino acids in food and physiological samples

Sarwar, Ghulam; Botting, H. G.

doi:10.1016/0378-4347(93)80286-d

Cited by 88 publications

(52 citation statements)

References 35 publications

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“…In fact, in the last several years some characteristics of OPA/ESH and OPA/E(OH)SH amino acid derivatives have been identified Kowalczyk, 1998, 1999;Molnar-Perl, 2001). Exhaustive investigations have demonstrated that OPA/E(OH)SH and OPA/ESH amino acid derivatives can be successfully resolved by gradient RP-HPLC with fluorescence detection (Sarwar and Botting, 1993;Kowalczyk, 1998, 1999;Kutlan and Molnar-Perl, 2001;Molnar-Perl, 2001). Therefore in the presented paper, on the basis of our earlier experience Kowalczyk, 1998, 1999), a new gradient HPLC system with UV detection for primary amino acids has been developed.…”

Section: Resultsmentioning

confidence: 99%

“…The separation and quantification of oxidized proline, hydroxyproline and cyst(e)ine, as their OPA/ESH derivatives, can be achieved applying a Symmetry C 18 column (5 |um, 250 x 4.6 mm, I.D., Waters) and UV detection at -337 nm (Czauderna and Kowalczyk, 1998). Obviously, secondary amino acids may be derivatized with other reagents (e.g., 4-chloro-7-nitrobenzofurazan or 9-fluorenylmethyl chloroformate), and then separated using the HPLC systems reported by Umagat (1982) or by Sarwar (1993). Moreover, methionine and cyst(e)ine estimation in biological materials hydrolysed with hydrochloric acid show that certain amount of cyst(e)ine and methionine can be oxidized.…”

Section: Discussionmentioning

confidence: 99%

“…Although there are many techniques for this analysis, the application of commercial amino acids analyzers using ion-exchange liquid chromatography is by far the most popular (Ng et al, 1991;Moller, 1993;Sarwar and Botting, 1993;Williams, 1994). However, many high performance liquid chromatography (HPLC) methods and gas chromatography techniques (GC) for quantification of amino acids have also been published in recent years (Sarwar and Botting, 1993;Cohen and Michaud, 1998;Czauderna and Kowalczyk, 1998;Peter et al, 1998;Albin et al, 2000;Polak and Golkiewicz, 2000;Kutlan and Molnar-Perl, 2001;Molnar-Perl, 2001). The major shortcomings of automatic amino acid analyzers for amino acids observed are long analysis time and inadequate detection limits, dedication of the analyzers to only amino acid analysis, and high cost of the instruments.…”

Section: Introductionmentioning

confidence: 99%

“…On the other hand, the use of reversed-phase HPLC with pre-column derivatization for the analysis of protein-and free amino acids is becoming established as a cheaper alternative to commercial amino acid analyzers. Five main pre-column derivatization (o-phthaldialdehyde (OPA), phenylthiohydantoin, phenylthiocarbamyl, dansyl chloride and dabsyl chloride) HPLC methods were compared in terms of detection limit, precision, resolution, stability and duration of analysis (Sarwar and Botting, 1993 ). The rapid OPA derivatization procedure has become popular because the reagent itself does not fluoresce, the instrumentation in pre-column derivatization is simpler and the cost of such systems is lower compared with post-column derivatization or amino acid analyzers (Rattenbury, 1981;Sarwar and Botting, 1993;Williams, 1994).…”

Section: Introductionmentioning

confidence: 99%

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Determination of free- and protein primary amino acids in biological materials by high-performance liquid chromatography and photodiode array detection

Czuderna¹,

Kowalczyk²,

Niedźwiedzka³

et al. 2002

J. Anim. Feed Sci.

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Work from this laboratory resulted in improved high-performance liquid chromatography methods for quantification of free-and protein primary amino acids in biological materials. Biological samples were hydrolyzed with 6 M HC1 for 20 h at 104±2°C. Primary amino acids, with the exception of tryptophan, were separated after pre-column derivatization with o-phthaldialdehyde (OPA) in the presence of ethanethiol (ESH). Derivatized amino acids were analyzed using a Nova Pak C 18 column (4 jum, 250 x 4.6 mm I.D., Waters) by quaternary gradient system I. Detection was carried out simultaneously using UV monitoring at 337 nm and fluorescence detection (\J^c m -336/425 nm). Derivatized amino acids were completely resolved from all interfering species in about 50 min. HPLC system I with two detection modes can also be used for quantification of free primary amino acids in ovine blood plasma. A trace amount of cysteine as its OPA/ESH derivative can be quickly quantified (~ 3.5 min) using an isocratic HPLC system with UV detection at 274 nm. Separation and quantification of tryptophan in alkaline hydrolysates was achieved without derivatization using the same HPLC column by ternate gradient elution system II with UV monitoring at 219 nm or fluorescence detection (^e x /^e m = 280/360 nm). The obtained average recoveries of assayed amino acids added to biological samples were near 100% when UV and fluorescence detection were applied. Generally, fluorescence detection in comparison with UV monitoring offers lower limits of detection (0.04-1.25 vs 0.05-12.7 jumoM" 1 ) and quantification (0.12-11.7 vs 0.18-42.0 jumol-l" 1 ), however, the sensitivity of the UV detection mode is satisfactory for accurate and precise quantification of free-and protein amino acids in biological materials. The HPLC systems with UV detection assured better resolution of amino acid peaks in comparison with fluorescence detection. Satisfactory purity of analytical amino acid peaks (near 100%) and precision of HPLC systems with both detection modes renders these procedures suitable for routine analysis of amino acid concentrations in large numbers of 144 METHODS OF AMINO ACIDS DETERMINATION biological samples. HPLC system I enabled quantification of 2,6-diaminopimelic acid, so the current chromatographic system can also be applied for the estimation of bacterial protein production in ruminants.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Determination of free- and protein primary amino acids in biological materials by high-performance liquid chromatography and photodiode array detection

Czuderna¹,

Kowalczyk²,

Niedźwiedzka³

et al. 2002

J. Anim. Feed Sci.

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“…The amino acids present in a crude plant extract were derivatized with mercaptoethanol and O-phthaldialdehyde yielding a fluorescent isoindole (Roth, 1971;Sarwar and Botting, 1993), a method inappropriate for the detection of Cys (weakly fluorescent derivative) and Pro (contains no primary amino group for derivatization reaction). Rosettes of individual 3-to 4-week-old individual plants were harvested, and individually frozen in liquid nitrogen, lyophilized, and pulverized.…”

Section: Amino Acid Analysesmentioning

confidence: 99%

Two Arabidopsis Genes (IPMS1 and IPMS2) Encode Isopropylmalate Synthase, the Branchpoint Step in the Biosynthesis of Leucine

et al. 2006

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Heterologous expression of the Arabidopsis (Arabidopsis thaliana) IPMS1 (At1g18500) and IPMS2 (At1g74040) cDNAs in Escherichia coli yields isopropylmalate synthases (IPMSs; EC 2.3.3.13). These enzymes catalyze the first dedicated step in leucine (Leu) biosynthesis, an aldol-type condensation of acetyl-coenzyme A (CoA) and 2-oxoisovalerate yielding isopropylmalate. Most biochemical properties of IPMS1 and IPMS2 are similar: broad pH optimum around pH 8.5, Mg 21 as cofactor, feedback inhibition by Leu, K m for 2-oxoisovalerate of approximately 300 mM, and a V max of approximately 2 3 10 3 mmol min 21 g 21 . However, IPMS1 and IPMS2 differ in their K m for acetyl-CoA (45 mM and 16 mM, respectively) and apparent quaternary structure (dimer and tetramer, respectively). A knockout insertion mutant for IPMS1 showed an increase in valine content but no changes in Leu content; two insertion mutants for IPMS2 did not show any changes in soluble amino acid content. Apparently, in planta each gene can adequately compensate for the absence of the other, consistent with available microarray and reverse transcription-polymerase chain reaction data that show that both genes are expressed in all organs at all developmental stages. Both encoded proteins accept 2-oxo acid substrates in vitro ranging in length from glyoxylate to 2-oxohexanoate, and catalyze at a low rate the condensation of acetyl-CoA and 4-methylthio-2-oxobutyrate, i.e. a reaction involved in glucosinolate chain elongation normally catalyzed by methylthioalkylmalate synthases. The evolutionary relationship between IPMS and methylthioalkylmalate synthase enzymes is discussed in view of their amino acid sequence identity (60%) and overlap in substrate specificity.

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Liquid Chromatography in Food Analysis

Nollet

2000

Encyclopedia of Analytical Chemistry

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Evaluation of liquid chromatographic analysis of nutritionally important amino acids in food and physiological samples

Cited by 88 publications

References 35 publications

Determination of free- and protein primary amino acids in biological materials by high-performance liquid chromatography and photodiode array detection

Determination of free- and protein primary amino acids in biological materials by high-performance liquid chromatography and photodiode array detection

Two Arabidopsis Genes (IPMS1 and IPMS2) Encode Isopropylmalate Synthase, the Branchpoint Step in the Biosynthesis of Leucine

Liquid Chromatography in Food Analysis

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