Comparative determination of purine compounds in carotid plaque by capillary zone electrophoresis and high-performance liquid chromatography

Terzuoli, Lucia; Porcelli, Brunetta; Setacci, Carlo; Giubbolini, M.; Cinci, Giuliano; Carlucci, Filippo; Pagani, Roberto; Marinello, Enrico

doi:10.1016/s0378-4347(99)00119-x

Cited by 28 publications

(18 citation statements)

References 23 publications

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“…As can be seen, the integration of the chromatographic signals registered for the four monitored compounds indicates that allantoin and hypoxanthine are always present in higher amount whereas uric acid and xanthine are generally less abundant. This behavior is in agreement with the data obtained by Terzuoli et al: 8 the detected higher levels of allantoin and the lower levels of uric acid have already been explained with the occurrence of the rapid oxidation of uric acid to allantoin. 8,11,12 Plaques 1 and 9 were also pretreated according to the well established sample preparation method 8 and, as a comparison test, analyzed under the same experimental conditions.…”

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“…This behavior is in agreement with the data obtained by Terzuoli et al: 8 the detected higher levels of allantoin and the lower levels of uric acid have already been explained with the occurrence of the rapid oxidation of uric acid to allantoin. 8,11,12 Plaques 1 and 9 were also pretreated according to the well established sample preparation method 8 and, as a comparison test, analyzed under the same experimental conditions. From the results in Table 1, the more complex sample preparation procedure seems to cause a general decrease in the recovery of the four bases, being more pronounced for uric acid and xanthine.…”

supporting

confidence: 93%

“…The supernatant, filtered through 0.45 µm PTFE, was injected directly into the HPLC/MS system. Samples 1 and 9 were treated in parallel following the 'classical' sample preparation method, 8 i.e. adding to the supernatant obtained after a first centrifugation 0.1 ml of 4 N perchloric acid (for protein precipitation) and centrifuging the resulting suspension at 12 000 g for 10 min; the supernatant was collected and, to re-suspend the sample and neutralize the pH, 40 µl of 5 N KOH were added.…”

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Determination of purine compounds in carotid artery plaque by liquid chromatography electrospray ionization tandem mass spectrometry

et al. 2001

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Determination of purine compounds in carotid artery plaque by liquid chromatography electrospray ionization tandem mass spectrometry † In recent years, the oxidation theory of atherogenesis has been attracted considerable attention. 1 -4 This theory envisages oxidation of low-density lipoprotein (LDL) as a key and early event responsible for the lipid loading into atherosclerotic foam cells and for progression of atherosclerotic disease: however, in carotid artery plaque, oxidative stress has never been directly correlated with the presence of other compounds except ox-LDL.It is well known that some purine compounds, such as hypoxanthine, xanthine, uric acid and allantoin, in the presence of situations such as experimental tumors, oxidative stress and hypoxia, change their plasmatic or urinary levels owing to the occurrence of nucleic acid turnover alterations. 5 -7 In a recent publication, 8 using high-performance liquid chromatography (HPLC) and capillary zone electrophoresis (CZE) techniques, the presence of the four reported compounds in carotid artery plaques was already described to demonstrate, through the presence of hypoxanthine, xanthine, uric acid and allantoin, the occurrence of oxidative stress in the arterial wall. Owing to the complexity of the substrate under investigation, in order to achieve good specificity a careful sample preparation procedure proved to be essential.Liquid chromatography/mass spectrometry (LC/MS) with application of multiple reaction monitoring (MRM) experiments has been widely demonstrated to be a good choice in qualitative and quantitative complex mixture analysis: 9 the higher specificity and sensitivity achievable by monitoring the occurrence of a selected molecule fragmentation often allow the application of simpler and less time-consuming sample preparation procedures. For this reason, we decided to apply this kind of approach to the qualitative determination of the four already reported purine bases in carotid plaque with the aim of developing a more rapid and selective analytical assay.We have previously used off-line liquid chromatography/electron ionization tandem mass spectrometry to examine purine bases in urine extracts from both healthy and colo-rectal cancer patients, but some difficulties were encountered in the structural assignment of a number of observed ions and in some isomers differentiation for the absence of an on-line chromatographic separation. 10 Ten carotid artery plaque samples (¾50 mg each) obtained from patients (eight males and two females, mean age 74 years) who had undergone carotid endoarterectomy were frozen with liquid nitrogen, pulverized, suspended in 500 µl of water and centrifuged at 12 000 g for 10 min. The supernatant, filtered through 0.45 µm PTFE, was injected directly into the HPLC/MS system. Samples 1 and 9 were treated in parallel following the 'classical' sample preparation method, 8 i.e. adding to the supernatant obtained after a first centrifugation 0.1 ml of 4 N perchloric acid (for protein precipitation) and centrifuging...

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Determination of purine compounds in carotid artery plaque by liquid chromatography electrospray ionization tandem mass spectrometry

et al. 2001

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“…The only separation of purine compounds using CE was proposed by Pizzichini et al [5], but the separation of these compounds in the serum was not clearly performed. Terzuoli et al [12] investigated these purine compounds in carotid plaque.…”

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confidence: 99%

Simultaneous determination of allantoin, hypoxanthine, xanthine, and uric acid in serum/plasma by CE

et al. 2007

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Allantoin (All) is an oxidative end product of purines in mammals. The small amount of All present in human plasma or serum results from free radical action on urate and may provide a stable marker of in vivo free radical activity. Because free radicals have been implicated in the development and progression of atherosclerosis, this study focused on the metabolic compounds of the All pathway. We propose a new fast CE (CE/UV) method for the simultaneous determination of All, uric acid (UA), hypoxanthine (HX), and xanthine (X) in human plasma. These products were quantified in the plasma of patients with chronic renal failure before hemodialysis (n = 6), patients with chronic heart failure (n = 6) and controls (n = 6). The filtered plasma were diluted ten-fold before the direct injection in CE/UV (195 nm), which allows separating the four compounds in less than 13 min. The metabolites were detectable at concentrations of 0.3-0.6 micromol/L. The method was linear over the range 0.5-150 micromol/L for All, HX, and X and 10-1500 micromol/L for UA (r > 0.99). The analytical performance of this method is satisfactory with intra-assay CV < 3.4%, inter-assay CV < 5% (HX and X < 7%), and recovery (93-101%). The proposed CE-UV method appears to be a useful tool for studying physiological and pathological changes of HX, UA, and All levels in plasma samples, the latter being a possible indicator of free radical damage in vivo.

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“…Methods for quantitative analysis of allantoin were reported using HPLC (Terzuoli et al 1999) and capillary zone electrophoresis Terzuoli et al 1999). On the other hand, there have been few reports on methods using LC-MS coupled with an APCI mode for allantoin and allantoic acid.…”

Section: Resultsmentioning

confidence: 99%

Quantitative and isotopic analysis of amino acids, allantoin, and allantoic acid in soybeans by LC-MS using the atmospheric pressure chemical ionization method

Ohtake

Takano

Ito

et al. 2004

Soil Science and Plant Nutrition

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Comparative determination of purine compounds in carotid plaque by capillary zone electrophoresis and high-performance liquid chromatography

Cited by 28 publications

References 23 publications

Determination of purine compounds in carotid artery plaque by liquid chromatography electrospray ionization tandem mass spectrometry

Determination of purine compounds in carotid artery plaque by liquid chromatography electrospray ionization tandem mass spectrometry

Simultaneous determination of allantoin, hypoxanthine, xanthine, and uric acid in serum/plasma by CE

Quantitative and isotopic analysis of amino acids, allantoin, and allantoic acid in soybeans by LC-MS using the atmospheric pressure chemical ionization method

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