Measurement of free and bound malondialdehyde in plasma by high-performance liquid chromatography as the 2,4-dinitrophenylhydrazine derivative

Pilz, Jürgen; Meineke, Ingolf; Gleiter, Christoph H.

doi:10.1016/s0378-4347(00)00174-2

Cited by 219 publications

(170 citation statements)

References 34 publications

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“…We found mean 1.87±0.06 μmol/L of total MDA in human serum by using HPLC method as MDA-DNPH, This result is closed to the previous results reported by different authors with slight differences, these differences may be related to the factors such as age and nutrition of healthy volunteers [11].…”

Section: Resultscontrasting

confidence: 60%

“…The based determination of the MDA hydrazone, has proved to be precise and reproducible, as well as sensitive enough to reflect differences in the oxidative status in vivo [11]. In this method centrifugation of sample, followed by release of protein-bound MDA by alkaline hydrolysis of supernatants, and derivatization of MDA is required without any further extraction steps before direct injecting into the chromatographic system.…”

Section: Resultsmentioning

confidence: 99%

“…During the formation of this complex unsaturated fatty acids in biological sample also react with TBA to form colored complexes, which absorb light at or near 535 nm [5][6][7][8][9][10][11][12] Hence, spectrophotometric detection includes both MDA-TBA complex and other complexes which absorb at the same wavelength a higher amount of MDA is obtained.…”

Section: Resultsmentioning

confidence: 99%

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Comparison of two Methods for Malondialdehyde Measurement

2011

Ann Clin Anal Med

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Özet AmaçBu çalışmada; malondialdehid düzeylerinin iki ayrı metod ile ölçülmesi ve bu metodların karşılaştırılması amaçlanmıştır. Gereç ve Yöntemler 60 gönüllünün serum malondialdehid düzeyleri spektrofotometrik tiyobarbitürik asit testi ve 2,4-dinitrofenilhidrazin ile türevlendirilmesi sonrası yüksek basınçlı sıvı kromatografisi ile ölçülmüş ve bu iki metot karşılaştırılmıştır. Bulgular Yüksek basınçlı sıvı kromatografisi (1.85±0.09 µmol/L) ve spektrofotometre (2.47±0.18 µmol/L) ile ölçülen serum MDA seviyelerinde istatistiksel olarak anlamlı fark bulundu (p<0.001). İki ayrı yöntemle yapılan ölçümler arası tutarlılık değerlendirildi ve sınıflar arası korelasyon katsayısı 0.365 olarak bulundu (p=0.042). Ölçümler arasında istatistiksel olarak önemli, zayıf-orta derecede korelasyon bulunmuştur (r=0.284, P=0.028). Sonuç 2,4-dinitrofenilhidrazin ile türevlendirme sonrası yüksek basınçlı sıvı kromatografisi ile malondialdehid ölçüm metodu, konvansiyonel spektrofotometrik tiyobarbitürik asit metoduna göre lipit peroksidasyonunun daha doğru ve duyarlı değerlendirilmesini sağlamıştır. Anahtar KelimelerMalondialdehid, Lipid Peroksidasyonu, Oksidatif Stres, Yüksek Basınçlı Sıvı Kromatografisi, Spektrofotometrik Yöntem. Abstract AimIn this study, we aimed to measure malondialdehyde levels by two different methods and compare these methods. Material and MethodsSerum malondialdehyde levels of 60 volunteers were measured with thiobarbituric acid test and high pressure liquid chromatography after derivatization of malondialdehyde with 2,4 dinitrophenylhydrazine and these two methods were compared. Results A statistically significant difference (p<0.001) has been found in serum malondialdehyde levels measured by high pressure liquid chromatography (1.85±0.09 µmol/L) and spectrophotometry (2.47±0.18 µmol/L). Cohesion between measurements made by two different methods have been evaluated and the interclass correlation coefficient has been found as 0.365 (p=0.042). Statistically significant, weak-mild degree correlation has been found between measurements (r=0.284, p=0.028). Conclusions High pressure liquid chromatography method for malondialdehyde measurement after derivatization with 2,4 dinitrophenylhydrazine provided a more accurate and sensitive assessment of lipid peroxidation than the conventional spectrophotometric thiobarbituric acid method.

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Section: Resultscontrasting

confidence: 60%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Comparison of two Methods for Malondialdehyde Measurement

2011

Ann Clin Anal Med

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“…16 MDA was measured by RP-HPLC and UV detection after derivatisation with 2,4-dinitrophenylhydrazine. 17 Total antioxidant capacity was measured by chemiluminescence detection in a BMG Fluostar Optima plate reader, using ABEL wide-range antioxidant kit from Knight Scientific Limited, UK. 18 Protein carbonyls were measured by an enzyme-linked immunosorbent assay (ELISA) method, with absorbance detection, using Zentech PC Kit from Zenith Technologies, New Zealand.…”

Section: Blood Analysesmentioning

confidence: 99%

Associations between plasma antioxidants and hypertension in a community-based sample of 415 Australians aged 60–64

et al. 2004

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It has been suggested that increased oxidative stress may be both a cause as well as a consequence of hypertension. In vivo oxidation of low-density lipoproteins by oxygen-free radicals may increase hypertension-related atherogenesis, and antioxidants may be beneficial in this regard. Previous findings concerning associations between serum measures of antioxidants and hypertension have however been inconsistent. Plasma levels of b-carotene, Vitamin A, E, uric acid, homocysteine and total antioxidant capacity, as well as two markers of oxidative stress, malondialdehyde (MDA) and protein carbonyls, were measured in morning fasting blood samples provided by 415 Australians aged 60-64 years, selected randomly from the community. Participants also provided information on sociodemographic attributes, mental and physical health, and provided two measures of resting blood pressure, allowing a diagnosis of definite or borderline hypertension. Those with hypertension had lower levels of b-carotene and higher levels of uric acid and MDA compared to normotensive participants. The last two of these associations persisted when the analyses controlled for lifestyle and health factors. The findings from this study offer limited support for the proposition that lower antioxidant status and higher oxidative stress are associated with hypertension, and suggest the need for longitudinal studies to examine causality and intervention studies to determine the benefit of antioxidants in this group.

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“…A quantificação de MDA nos sistemas biológicos é um parâmetro importante para avaliação do estresse oxidativo celular (Pilz, Meineke, Gleiter, 2000;Sim et al, 2003). O método introduzido por Yagi (1976) tem sido amplamente utilizado.…”

Section: Introductionunclassified