The Construction and Performance of a Low Cost Automated HPLC System for Polyamine Assay

Shaw, Graham G.; Al-Deen, Ihsan H.S.; Elworthy, Paul

doi:10.1093/chromsci/18.4.166

Cited by 19 publications

(4 citation statements)

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“…Detection of products containing primary amino groups was achieved spectrophotometrically (at 214 nm) and following post-column derivatization by 5 mM o-phthalaldehyde (OPA)\12 mM 2-mercaptoethanol\0.4 mM potassium borate at pH 10. The combined flows were passed through a 1 ml coil at 40 mC, and OPA-derivatized products were detected by fluorescence (λ ex at 345 nm, λ em at 550 nm) [28]. Figure 1 shows a typical product profile obtained after radiolysis (approx.…”

Section: Identification Of Products Derived From Nhamentioning

confidence: 99%

Oxidative denitrification of Nω-hydroxy-l-arginine by the superoxide radical anion

Everett¹,

Dennis

Patel

et al. 1996

Biochemical Journal

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The superoxide radical anion (O2-.) produced during the catalytic activity of nitric oxide synthase (NOS) and cytochrome P-450 has been implicated in the oxidative denitrification of hydroxyguanidines ( > C = NOH). The reactivity of the radiolytically generated O2-. radical with N omega-hydroxy-L-arginine (NHA) is pH dependent and appears to parallel the prototropic equilibrium of the hydroxyguanidino group ( > C = NOH reversible > C = NO(-)+H+; pK = 8). The N omega-hydroxyguanidino group is more reactive towards O2-. when deprotonated but exhibits negligible reactivity when protonated. Based on a model, the rate constant for the reaction of the O2-. with NHA was estimated as kappa (O2-.+ > C = NO-) approximately 200-500 M-1.s-1, which is probably too low to compete with O2-. reactions with NO- or superoxide dismutase, which occur many orders of magnitude faster. The oxidative elimination of NO from NHA by O2-. was not accompanied by the formation of L-citrulline. Since only 21% of NHA will exist in the deprotonated > C = NO- form at physiological pH, it is unlikely that oxidative denitrification of NHA by cytochrome P-450 or NOS-derived O2-. radicals will prove a major free-radical pathway to NO. and L-citrulline.

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Section: Identification Of Products Derived From Nhamentioning

confidence: 99%

Oxidative denitrification of Nω-hydroxy-l-arginine by the superoxide radical anion

Everett¹,

Dennis

Patel

et al. 1996

Biochemical Journal

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“…The RBC count was determined and the samples stored at -20 ° C. Polyamines were extracted from the resuspended RBC by adding 1 ml of the suspension to 1 ml 0.8 M perchloric acid, shaking for 10 minutes and centrifuging for 5 minutes at 2000 G to remove the precipitated material [10]. High performance liquid chromatography was used to separate polyamines from 100 #1 aliquots of [16]. Polyamines were detected fluorimetrically after post-column derivatisation by orthophthalaldehyde with excitation and emission wave lengths of 345 nm and 455 nm [16][17][18].…”

Section: Polyamine Assaymentioning

confidence: 99%

“…High performance liquid chromatography was used to separate polyamines from 100 #1 aliquots of [16]. Polyamines were detected fluorimetrically after post-column derivatisation by orthophthalaldehyde with excitation and emission wave lengths of 345 nm and 455 nm [16][17][18].…”

Section: Polyamine Assaymentioning

confidence: 99%

Red blood cell polyamines as a diagnostic indicator of glioma presence and recurrence

Elworthy¹,

Hitchcock

1989

J Neuro-Oncol

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Red blood cell (RBC) polyamines, spermine and spermidine have been assayed from 103 patients with confirmed intracranial tumours and 87 controls. For spermine the test had a specificity of 97% and a sensitivity of 30% in preoperative patients, increasing to 40% in glioma patients. Tumour recurrence in postoperative patients gave a sensitivity of 48% increasing to 60% in glioma groups. For initial tumour presentation, therefore, this test is suggestive, though not unequivocal, of tumour presence. Its major role probably lies in monitoring the progress of the patient and its major importance in the detection of tumour recurrence.

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“…1 Many methods have been proposed for the determination of polyamines, such as thin layer chromatography, gas chromatography, ion-exchange chromatography and high performance liquid chromatography. 2,3 Several detection techniques, such as spectrophotometric, 4 electrochemical, 5 fluorescence, [6][7][8] chemiluminescence, 9 and mass spectrometric (MS) 10 detection have been applied. Among them, high performance liquid chromatography methods employing fluorescence, chemiluminescence or mass spectrometric detection are relatively sensitive and selective.…”

Section: Introductionmentioning

confidence: 99%

Determination of Polyamines by Flow Injection Analysis with a Chemiluminescence Detector Based on Their Complexation with Copper(II)

Chun

et al. 2006

ANAL. SCI.

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Through the flow injection analysis experiments, we discovered that an unsaturated complex of Cu(II) and polyamines (spermine, spermidine, putrescine) had a strongly catalytic effect on luminol-H2O2 chemiluminescence (CL) reaction, and that the CL intensity is proportional to the concentrations of polyamines. Based on the automatic formation of an unsaturated complex of polyamines and Cu(II) when the solution containing polyamines passed through a column packed with solid Cu(OH)2, a new flow injection chemiluminescence analysis method was proposed for the determination of polyamines. The effects of pH, buffer concentration, the concentration of chemiluminescence reagent, and the influence of mixing coil length were examined. Under optimal conditions, the linear range was from 1.0 × 10 -7 mol L -1 to 1.0 × 10 -5 mol L -1 , and the detection limits were 0.17, 0.38, 0.44 pmol for spermine, spermidine, and putrescine, respectively. Compared with other methods, the advantages of this method include convenience, time-saving and low cost.

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The Construction and Performance of a Low Cost Automated HPLC System for Polyamine Assay

Cited by 19 publications

References 0 publications

Oxidative denitrification of Nω-hydroxy-l-arginine by the superoxide radical anion

Oxidative denitrification of Nω-hydroxy-l-arginine by the superoxide radical anion

Red blood cell polyamines as a diagnostic indicator of glioma presence and recurrence

Determination of Polyamines by Flow Injection Analysis with a Chemiluminescence Detector Based on Their Complexation with Copper(II)

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