Total Plasma Malondialdehyde Levels in 16 Taiwanese College Students Determined by Various Thiobarbituric Acid Tests and an Improved High-Performance Liquid Chromatography-based Method

Hong, Yu-Lun; Yeh, Shu-Lan; Chang, Chia Yu; Hu, Miao‐Lin

doi:10.1016/s0009-9120(00)00177-6

Cited by 138 publications

(112 citation statements)

References 27 publications

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“…Within physiological systems, MDA may exist in a free state or in a protein-bound form; various studies have looked at the relative proportions of MDA in human plasma and have reported that between 83% and 92% of the MDA is protein bound ( 17,18 ). Hydrolysis of the protein-bound MDA fraction may be achieved by either an alkaline treatment ( 18,19 ) or an acid treatment ( 20 ) of the sample prior to the acid-catalyzed TBA reaction.…”

Section: Validation Of the Tbars And Mda Assaysmentioning

confidence: 99%

A specific, accurate, and sensitive measure of total plasma malondialdehyde by HPLC

Moselhy

Reid

Yousef

et al. 2013

Journal of Lipid Research

140

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The process of lipid peroxidation results in a range of intermediates and end products including lipid hydroperoxides, aldyehydes, and malondialdehyde (MDA). These aldehydes and lipid hydroperoxides form DNA adducts and may result in extensive single-strand and doublestrand breaks ( 4 ). Various intermediates and end products generated during the lipid peroxidation cascade have been assayed, but the most-commonly employed approach continues to be the thiobarbituric acid (TBA) test ( 5 ).Although it is widely acknowledged that TBA reacts with a range of oxidized lipids, both saturated and unsaturated aldehydes, ( 6, 7 ) sucrose, and urea ( 8 ), to form various chromogens, referred to as TBA-reactive substances (TBARS), it is the reaction of TBA with MDA to produce a pink pigment ( 9 ) with an absorption maximum at 532 nm ( 10 ) and mass ion at 323 amu ( 11 ) that is a true indicator of lipid peroxidation. Historically, the TBARS test has been assayed by ultraviolet (UV) spectrophotometry or fl uorescence assays, but the specifi city of the TBA test, enabling the selective determination of MDA, may be achieved via chromatographic separation of the pink TBA 2 -MDA adduct ( 12 ). The adoption of HPLC techniques improves assay specifi city and the sensitivity of MDA determination, and we have previously confi rmed the validity of quantifying the TBA 2 -MDA adduct as a specifi c measure of lipid peroxidation in human biological fl uids ( 13 ).Despite the widely acknowledged limitations of the TBARS test ( 14 ), and in particular its lack of specifi city, it continues to be reported as a true measure of MDA in clinical disease ( 1,15,16 ). It is proposed that the poor assay specifi city associated with TBARS assays may lead to an overestimation of the levels of MDA in human plasma and other biological tissues and fl uids, and this, in turn, may Abstract Malondialdehyde (MDA) is one of the most commonly reported biomarkers of lipid peroxidation in clinical studies. The reaction of thiobarbituric acid (TBA) with MDA to yield a pink chromogen attributable to an MDA-TBA 2 adduct is a common assay approach with products being quantified by ultraviolet-Vis assay as nonspecific TBAreactive substances (TBARS) or chromatographically as MDA. The specifi city of the TBARS assay was compared with both chromatographic assays for total plasma MDA. The levels of total plasma MDA were signifi cantly lower than the plasma TBARS in each of the samples examined, and interestingly, the interindividual variation apparent in the level of plasma MDA was not evident in the plasma TBARS assay. Each of the four online chromatographic detectors yielded a precise, sensitive, and accurate determination of total plasma MDA, and selected-ion monitoring was the most-accurate assay (101.3%, n = 4). The online diode array detectors provided good assay specifi city (peak purity index of 999), sensitivity, precision, and accuracy. This research demonstrates the inaccuracy that is inherent in plasma TBARS assays, which claim to quantify MDA, and it is pro...

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Section: Validation Of the Tbars And Mda Assaysmentioning

confidence: 99%

A specific, accurate, and sensitive measure of total plasma malondialdehyde by HPLC

Moselhy

Reid

Yousef

et al. 2013

Journal of Lipid Research

140

View full text Add to dashboard Cite

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“…The HPLC-based method with fluoresence detection (515 nm excitation, 555 nm emission) of plasma TBA-MDA was reported in an earlier report 19) . First, 0.25 ml of plasma was added to 1.5 ml of methanol/acetic acid (3:2 v/v) that contained 1% KI and 0.025 ml of 0.2% BHT (in 95% ethanol).…”

Section: Plasma Mda Assaymentioning

confidence: 99%

Oxidative Damage of Workers in Secondary Metal Recovery Plants Affected by Smoking Status and Joining the Smelting Work

2008

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“…Bound malondialdehyde (MDA) in plasma was released from protein with 10 N NaOH, reacted with thiobarbituric acid (TBA) at 951C for 30 min, and the MDA-TBA complex analyzed by reverse-phase HPLC (Hewlett Packard 1100, Palo Alto, CA) with fluorescence detection as described. [15][16][17] Plasma total homocysteine, cysteine and cysteinylglycine Plasma concentrations of total homocysteine, cysteine and cysteinylglycine were determined with reverse-phase HPLC using a C18 column on a Waters HPLC instrument equipped with a WISP automatic injector and attached to a fluorimeter (Applied Sciences Inc., Dayton, OH). as described by Araki and Sako.…”

Section: Gpx 3 and Nitrotyrosinementioning

confidence: 99%

Plasma antioxidants in subjects before hematopoietic stem cell transplantation

White

Sousa

Blumberg

et al. 2006

Bone Marrow Transplant

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Chemo-irradiation induced oxidative damage to vascular endothelium may contribute to pulmonary complications of hematopoietic stem cell transplantation (HSCT). We measured antioxidants, markers of oxidative stress and plasma antioxidant capacity in plasma or serum from 24 subjects at day 7 before HSCT and 20 control subjects. The plasma concentration of extracellular glutathione peroxidase (GPX-3) was significantly reduced in the HSCT subjects compared with controls (HSCT: 98742 lg/ml, control: 169756 lg/ml, Po0.0001). The concentration of c-tocopherol was significantly higher in the HSCT subjects compared with controls (HSCT: 2077103 lg/dl; Control: 98752 lg/dl; P ¼ 0.0002). The plasma concentrations of protein carbonyl, nitrotyrosine, malondialdehyde, a-tocopherol, vitamin A, homocysteine, cysteine and cysteinylglycine did not differ between HSCT and control subjects. Plasma from HSCT subjects was as effective as control plasma in quenching menadione-induced intracellular reactive oxygen species production in human microvascular endothelial cells. In summary, subjects before HSCT have significantly reduced plasma concentrations of GPX-3, elevated plasma c-tocopherol yet retains the ability to quench an acute oxidative stress. These changes may play a role in chronic oxidative stress in the HSCT population.

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Total Plasma Malondialdehyde Levels in 16 Taiwanese College Students Determined by Various Thiobarbituric Acid Tests and an Improved High-Performance Liquid Chromatography-based Method

Cited by 138 publications

References 27 publications

A specific, accurate, and sensitive measure of total plasma malondialdehyde by HPLC

A specific, accurate, and sensitive measure of total plasma malondialdehyde by HPLC

Oxidative Damage of Workers in Secondary Metal Recovery Plants Affected by Smoking Status and Joining the Smelting Work

Plasma antioxidants in subjects before hematopoietic stem cell transplantation

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