Simultaneous determination of 16 purine derivatives in urinary calculi by gradient reversed-phase high-performance liquid chromatography with UV detection

Safranow, Krzysztof; Machoy, Z

doi:10.1016/j.jchromb.2004.11.013

Cited by 34 publications

(22 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Detection of their presence in stones by use of traditional analytical techniques (x-ray diffraction, infrared spectroscopy, and semiquantitative wet tests) has not been reported. We have recently developed a gradient HPLC method for separation of 16 purines, including 10 methyl derivatives of Xan and UA, in urinary calculi (5 ). We present here results of analysis of purine derivatives in urinary calculi by this method.…”

Section: 27) Deficiency (3 )mentioning

confidence: 99%

“…The extracts were diluted in 50 mmol/L KH 2 PO 4 and analyzed by HPLC (5 ). Using this method, we separated 16 compounds: UA; Xan; hypoxanthine (Hyp); 2,8-DHA; allopurinol; oxypurinol; 1-, 3-, 7-, and 9-methyluric acid (1-, 3-, 7-, and 9-MUA); 1,3-, 1,7-, and 3,7-dimethyluric acid (1,3-, 1,7-, and 3,7-DMU); and 1-, 3-, and 7-methylxanthine (1-, 3-, and 7-MX).…”

Section: Clinical Chemistry 51:8 1493-1498 (2005)mentioning

confidence: 99%

See 1 more Smart Citation

Methylated Purines in Urinary Stones

Safranow¹,

Machoy²

2005

Clinical Chemistry

Self Cite

View full text Add to dashboard Cite

Background: The aim of the study was to measure the content of methylated purines that appear as admixtures in uric acid stones. Methods: We analyzed urinary calculi from 48 residents of Western Pomerania who underwent surgery at the urology ward in Szczecin. Stone samples were dissolved in 0.1 mol/L NaOH. Extracts were diluted in 50 mmol/L KH 2 PO 4 and analyzed by reversed-phase HPLC with ultraviolet detection and use of a gradient of methanol concentration and pH. Results: Uric acid was the main component of 9 stones. All 9 showed admixtures of 9 other purine derivatives: endogenous purine breakdown products (xanthine, hypoxanthine, and 2,8-dihydroxyadenine) and exogenous methyl derivatives of uric acid and xanthine (1-, 3-, and 7-methyluric acid; 1,3-dimethyluric acid; and 3-and 7-methylxanthine). Amounts of these purine derivatives ranged from the limit of detection to 12 mg/g of stone weight and showed a strong positive correlation (Spearman rank correlation coefficients, 0.63-0.94) with the uric acid content of the samples. The main methylated purine in the stones was 1-methyluric acid. Conclusions: Urinary purines at concentrations below their saturation limits may coprecipitate in samples supersaturated with uric acid and appear as admixtures in urinary stones. The amount of each purine depends on its average urinary excretion, similarity to the chemical structure of uric acid, and concentration of the latter in the stone. These findings suggest that purines in stones represent a substitutional solid solution with uric acid as solvent. Methylxanthines, which are ubiquitous components of the diet, drugs, and uric acid calculi, may be involved in the pathogenesis of urolithiasis. © 2005 American Association for Clinical ChemistryMany purines are poorly soluble in aqueous solutions and may precipitate from urine, forming urinary calculi, if their concentrations exceed the limit of saturation. Uric acid (UA) 1 is the end product of human purine metabolism and is excreted mainly in the urine. Acidic urine is often a supersaturated solution of uric acid, even when the daily output remains in the normal range, and uric acid stones may form. Crystallization is usually prevented by inhibitors (citrates, glycosaminoglycans, and some proteins) (1 ), although not always effectively enough, particularly at low urine pH. Calculi composed of UA also form in males with hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8) deficiency (complete or partial). Xanthine (Xan) may appear in xanthinuria as well as during treatment with allopurinol (2 ). 2,8-Dihydroxyadenine (2,8-DHA) is excreted in urine and can precipitate in renal tissue and the urinary tract in patients with dihydroxyadeninuria attributable to adenine phosphoribosyltransferase (EC 2.4.2.7) deficiency (3 ).Limited attention has been paid to the possible role of metabolites of methylxanthines (caffeine, theophylline, and theobromine) in the pathogenesis of urolithiasis, although their average daily output in urine, depending on dietary intake, is ϳ500...

show abstract

Section: 27) Deficiency (3 )mentioning

confidence: 99%

Section: Clinical Chemistry 51:8 1493-1498 (2005)mentioning

confidence: 99%

Methylated Purines in Urinary Stones

Safranow¹,

Machoy²

2005

Clinical Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…1 Their determination is very important in many areas of research, like clinical analysis, [2][3][4][5][6][7] or food and beverage analysis. In food analysis some of these compounds have been related to off-flavors in food, being a marker for its freshness, on the other hand the end product of purine catabolism in the human body is uric acid, which is known to be a major trigger for gout, a common disease in developed countries.…”

Section: Introductionmentioning

confidence: 99%

“…The most widely used technique for biological samples is reversed phase (RP) or RP ion-pairing 15 high performance liquid chromatography (HPLC) because it gives reproducible results, and is sensitive, selective and easily automated. [2][3][5][6][7]10,[12][13][14] Capillary electrophoresis is also widely used for the separation of these analytes because of its minimal sample volume, short analysis time and high separation efficiency. [16][17][18] For both techniques UV/VIS, 5,18 electrochemical 19 or MS detection is used.…”

Section: Introductionmentioning

confidence: 99%

“…[2][3][5][6][7]10,[12][13][14] Capillary electrophoresis is also widely used for the separation of these analytes because of its minimal sample volume, short analysis time and high separation efficiency. [16][17][18] For both techniques UV/VIS, 5,18 electrochemical 19 or MS detection is used. [13][14]18,[20][21][22] Since nucleobases are electroactive, in some papers voltammetric determination with static mercury drop electrode, by modified graphite/carbon electrodes, [23][24] and on the boron-doped carbon nanotubes are also reported.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Optimization of High Performance Liquid Chromatography Method for Simultaneous Determination of Some Purine and Pyrimidine Bases

Markelj

Župančič

Pihlar

2016

ACSi

View full text Add to dashboard Cite

Nine purine and pyrimidine bases were separated and determined simultaneously using reversed phase (RP) high performance liquid chromatography (HPLC) in some food samples and biological fluids. Chromatographic behavior of these ionizable compounds highly depends on the interactions with the solvent as confirmed experimentally and by calculation of distribution of this species as a function of pH. Chromatograms show the optimal separation of five purine (uric acid, hypoxanthine, xanthine, adenine, and guanosine), and four pyrimidine (cytosine, uracil, cytidine and tymine) bases at pH around four. Accordingly, acetate buffer was selected due to high buffer capacity in this region. By variation of pH, concentration of buffer and volume ratio between buffer and methanol, we found that a mixture of 50 mM acetate buffer of pH 4.0 ± 0.1 with 3% of methanol ensures reproducibility, complete separation in less than 15 minutes and compatibility with UV and MS detection. Developed screening method was validated and applied for the analysis of complex clinical and beverage samples.

show abstract

Simultaneous Determination of Uric Acid and Ascorbic Acid Using Edge Plane Pyrolytic Graphite Electrodes

2006

View full text Add to dashboard Cite

Edge plane pyrolytic graphite electrodes have been applied for the determination of uric acid and ascorbic acid. The separate determination of uric acid was found to produce three linear ranges from 100 nM to 3400 mM with a detection limit of 30 nM found to be possible. Uric acid detection was also explored in the presence of 200 mM ascorbic acid where a detection limit of 52 nM was found to be possible. The detection of ascorbic acid in the presence of uric acid was also explored over three linear ranges of ascorbic acid with a limit of detection of 80 nM. Last the simultaneous determination of both uric acid and ascorbic acid is investigated over the range 100 nM to 1000 mM where detection limits of 50 nM and 120 nM were obtained respectively. Analysis of uric acid in a growth tissue medium was found to be successful, confirming the applicability of the methodology to real matrices. This protocol is shown to provide low detection limits, easy handling (no electrode modification), good voltammetric peak separation of uric acid and ascorbic acid and a wide linear dynamic range.

show abstract

Simultaneous determination of 16 purine derivatives in urinary calculi by gradient reversed-phase high-performance liquid chromatography with UV detection

Cited by 34 publications

References 27 publications

Methylated Purines in Urinary Stones

Methylated Purines in Urinary Stones

Optimization of High Performance Liquid Chromatography Method for Simultaneous Determination of Some Purine and Pyrimidine Bases

Simultaneous Determination of Uric Acid and Ascorbic Acid Using Edge Plane Pyrolytic Graphite Electrodes

Contact Info

Product

Resources

About