Free and Total Malondialdehyde Assessment in Biological Matrices by Gas Chromatography–Mass Spectrometry: What Is Needed for an Accurate Detection

Cighetti, Giuliana; Debiasi, Sandra; Paroni, Rita; Allevi, Pietro

doi:10.1006/abio.1998.2952

Cited by 107 publications

(79 citation statements)

References 41 publications

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“…Similar results were reported in rats administered with Phenylhydrazine to induce anemia [20]. Administration of PHZ to rats has been reported to result in the production of both aryl and hydroxyl radicals which produce oxidative stress on the red cell membrane resulting in haemolysis by lipid peroxidation [21,22]. Results are expressed as Mean ± standard deviation (SD) of five animals in each group.…”

Section: Discussionsupporting

confidence: 73%

In Vivo Antianaemic Effect and Safety of Aqueous Extracts of Erythrina abyssinica and Zanthoxylum usambarensis in Mice Models

TM¹,

NW²

2015

J Hematol Thrombo Dis

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This study was carried out to determine the hematinic effects and long term safety of Zanthoxylum usambarensis and Erythrina abyssinica in mice. Aqueous stem extracts of Z. usambarensis and E. abyssinica were screened for their haematinic activity in Phenylhydrazine induced anemic mice using the oral route. Hematological parameters were analysed as indices of anemia. The safety of these plant extracts was studied by orally administering 1 g/kg body weight of aqueous extracts daily in mice for thirty days and determining changes in body and organ weight, hematological and biochemical parameters. The mineral content of the extracts was estimated using Total Reflection X-Ray Fluorescence system (TRXF) while phytochemicals were assessed using standard procedures. Phenylhydrazine (PHZ) treatment induced macrocytic anemia. Administration of Z. usambarensis and E. abyssinica extracts at 100 mg/kg body weight daily for three weeks increased the red blood cell count, hemoglobin, and packed cell volume and decreased the mean cell volume and mean cell hemoglobin though E. abyssinica returned these parameters to normal. Administration of Z. usambarensis and E. abyssinica extracts at 1 g/kg body weight daily for four weeks significantly increased the activities of alkaline phosphatase, gamma glutamyltransferase, lactate dehydrogenase and amylase and the levels of urea and creatine in the treated mice. Extracts from both plants had alkaloids and flavonoids and minerals potassium, calcium, chromium, iron, copper, zinc, manganese, nickel, arsenic, and lead. This study has confirmed in vivo haematinic activity and safety of aqueous stem bark extracts of Z. usambarensis and E. abyssinica. The observed haematinic activity could be attributed to the phytonutrients present in these plants. The study recommends continued use of Z. usambarensis and E. abyssinica in the management of anaemia at the right doses as high doses are toxic.

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Section: Discussionsupporting

confidence: 73%

In Vivo Antianaemic Effect and Safety of Aqueous Extracts of Erythrina abyssinica and Zanthoxylum usambarensis in Mice Models

TM¹,

NW²

2015

J Hematol Thrombo Dis

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“…Plasma MDA was determined using an adaptation of the selected ion-monitoring gas chromatography-mass spectrometry (GC-MS) analysis of phenylhydrazine-derivatized plasma, as described by Cighetti et al 11,12 Specifically, the sample was prepared by the mixture of 0.2 ml plasma, 0.26 nmol methyl malondialdehyde (MMDA), 5 nmol butylated hydroxytoluene (10 ml of 0.5 mM), 0.2 ml citrate buffer (0.4 M, pH 4.0) and deionized water to a final volume of 480 ml. Then 20 ml 50 mM phenylhydrazine (1 mmol) was added as the derivatizing agent and incubated for 30 min at 25 1C.…”

Section: Mda Assaymentioning

confidence: 99%

Effect of mode of birth on purine and malondialdehyde in umbilical arterial plasma in normal term newborns

Calderon

Rawson

et al. 2008

J Perinatol

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Objective: To examine the effect of mode of birth on plasma purine and malondialdehyde levels in normal term infants.Study Design: Umbilical arterial cord blood was obtained immediately after birth from a convenience sample of 119 normal term newborns born by vaginal delivery, with or without oxytocin augmentation or by elective cesarean delivery. Plasma was analyzed for purine and/or malondialdehyde levels. Numeric data were analyzed utilizing independent samples t-test and ordinal data were analyzed using MannWhitney test. Correlation coefficients were obtained using Spearman's r.Result: Uric acid levels were significantly elevated (P<0.001) in neonates undergoing vaginal birth, compared to neonates born by elective cesarean delivery. When the effect of oxytocin and length of labor was analyzed, neonates born to mothers on oxytocin had lower hypoxanthine, significantly lower xanthine (P ¼ 0.05) and higher uric acid levels. In addition, malondialdehyde levels were significantly higher (P<0.006) in neonates born to mothers who received oxytocin compared to neonates born to mothers without oxytocin augmentation. We also found significant correlations between malondialdehyde (MDA) and hypoxanthine (r ¼ À0.465, P<0.039) and between MDA and xanthine (r ¼ À0.753, P ¼ 0.003) in neonates born via oxytocin-augmented birth. Mode of birth had no statistically significant effect on clinical outcomes, although infants born by elective cesarean had higher incidence of acute respiratory distress and transient tachypnea of the newborn compared to those born vaginally. Conclusion:Neonates born by elective cesarean had the lowest purine levels in cord blood compared to neonates born vaginally. Oxytocin augmentation is associated with some degree of uterine hyperstimulation which may enhance the ATP degradation pathway resulting in the rapid conversion of hypoxanthine to xanthine and xanthine to uric acid. Significantly higher MDA levels in neonates whose mothers received oxytocin as well as significant correlation between MDA and the purines hypoxanthine and xanthine, suggest free-radical production, most likely due to xanthine oxidase activation. However, despite differences in plasma purine and malondialdehyde levels, no significant differences were seen in neonatal outcome. Further studies are required to fully characterize the effect of mode of birth on purine metabolism and free-radical production.

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“…GC/MS based quantification of MDA was adapted from Paroni et al and Cighetti et al. 16,17 . Xanthine oxidase activity was measured by HPLC by quantifying the conversion of pterin to isoxanthopterin 18 .…”

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confidence: 99%

Biochemical Measurement of Neonatal Hypoxia

Plank

Calderon

Asmerom

et al. 2011

JoVE

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Neonatal hypoxia ischemia is characterized by inadequate blood perfusion of a tissue or a systemic lack of oxygen. This condition is thought to cause/exacerbate well documented neonatal disorders including neurological impairment 1-3 . Decreased adenosine triphosphate production occurs due to a lack of oxidative phosphorylation. To compensate for this energy deprived state molecules containing high energy phosphate bonds are degraded 2 . This leads to increased levels of adenosine which is subsequently degraded to inosine, hypoxanthine, xanthine, and finally to uric acid. The final two steps in this degradation process are performed by xanthine oxidoreductase. This enzyme exists in the form of xanthine dehydrogenase under normoxic conditions but is converted to xanthine oxidase (XO) under hypoxia-reperfusion circumstances 4, 5 . Unlike xanthine dehydrogenase, XO generates hydrogen peroxide as a byproduct of purine degradation 4, 6 . This hydrogen peroxide in combination with other reactive oxygen species (ROS) produced during hypoxia, oxidizes uric acid to form allantoin and reacts with lipid membranes to generate malondialdehyde (MDA) [7][8][9] . Most mammals, humans exempted, possess the enzyme uricase, which converts uric acid to allantoin. In humans, however, allantoin can only be formed by ROS-mediated oxidation of uric acid. Because of this, allantoin is considered to be a marker of oxidative stress in humans, but not in the mammals that have uricase.We describe methods employing high pressure liquid chromatography (HPLC) and gas chromatography mass spectrometry (GCMS) to measure biochemical markers of neonatal hypoxia ischemia. Human blood is used for most tests. Animal blood may also be used while recognizing the potential for uricase-generated allantoin. Purine metabolites were linked to hypoxia as early as 1963 and the reliability of hypoxanthine, xanthine, and uric acid as biochemical indicators of neonatal hypoxia was validated by several investigators [10][11][12][13] . The HPLC method used for the quantification of purine compounds is fast, reliable, and reproducible. The GC/MS method used for the quantification of allantoin, a relatively new marker of oxidative stress, was adapted from Gruber et al 7 . This method avoids certain artifacts and requires low volumes of sample. Methods used for synthesis of MMDA were described elsewhere 14,15 . GC/MS based quantification of MDA was adapted from Paroni et al. and Cighetti et al. 16,17 . Xanthine oxidase activity was measured by HPLC by quantifying the conversion of pterin to isoxanthopterin 18 . This approach proved to be sufficiently sensitive and reproducible.

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Free and Total Malondialdehyde Assessment in Biological Matrices by Gas Chromatography–Mass Spectrometry: What Is Needed for an Accurate Detection

Cited by 107 publications

References 41 publications

In Vivo Antianaemic Effect and Safety of Aqueous Extracts of Erythrina abyssinica and Zanthoxylum usambarensis in Mice Models

In Vivo Antianaemic Effect and Safety of Aqueous Extracts of Erythrina abyssinica and Zanthoxylum usambarensis in Mice Models

Effect of mode of birth on purine and malondialdehyde in umbilical arterial plasma in normal term newborns

Biochemical Measurement of Neonatal Hypoxia

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