LC-MS/MS assay for the simultaneous determination of tocopherols, polyunsaturated fatty acids and their metabolites in human plasma and serum

Giusepponi, Danilo; Galarini, Roberta; Barola, Carolina; Torquato, Pierangelo; Bartolini, Desirée; Moretti, Simone; Saluti, Giorgio; Gioiello, Antimo; Libetta, Carmelo; Galli, Francesco

doi:10.1016/j.freeradbiomed.2019.04.017

Cited by 23 publications

(21 citation statements)

References 34 publications

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“…Besides, these results may be secondary to LCMS, a more sensitive and precise separating method than traditional detection of high-performance liquid chromatography (HPLC) [24].…”

Section: Discussionmentioning

confidence: 99%

Plasma omega-3 polyunsaturated fatty acid and tocopherol statuses and their relationships with aging-related diseases

Liu¹,

Liu²,

Chen³

et al. 2020

Preprint

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Background: The beneficial effects of omega-3 polyunsaturated fatty acids (PUFAs) and tocopherols remain controversial. This study was conducted to examine the plasma levels of different analogues of omega-3 PUFAs and tocopherols, and to evaluate their relationships with aging-related diseases. Methods: 136 consecutive men with the median age of 70 (ranging from 50 to 97) years old were recruited. Plasma eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), α-, γ-, and δ-tocopherol were measured by liquid chromatography mass spectrometry. Results: Plasma omega-3 PUFAs were positively correlated with α- and γ-tocopherol (p<0.01), while the relationships between omega-3 PUFAs and δ-tocopherol were not significant (p>0.05). Increasing age was the most profound risk factor for coronary artery disease (CAD), carotid atherosclerosis (CAS), hypertension and these comorbidities (p<0.01). Age had positive associations with certain atherosclerotic parameters, including carotid intima-media thickness (p=0.046), carotid artery plague area (p<0.001) and brachial-ankle pulse wave velocity (p<0.001). In these observed isoforms, only γ-tocopherol had a mild inverse correlation with age (p=0.002). High plasma α-tocopherol concentration served as a potential protective factor for CAD (odds ratio [OR] 0.65; 95% CI 0.48-0.89; p=0.006), and was inversely associated with maximum systolic internal carotid artery (ICA) velocity (p=0.011). Plasma DHA concentration was negatively associated with carotid artery plague area (p=0.028) and ICA velocity (p=0.006), while its correlations with atherosclerosis diseases, including CAD and CAS, were not significant (p>0.05). Conclusions: Age is positively correlated with atherosclerosis, hypertension and comorbidities. Alpha-tocopherol may be a protective factor for atherosclerosis, while the correlations between omega-3 PUFAs and atherosclerosis were not significant.

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“…Besides, these results may be secondary to LCMS, a more sensitive and precise separating method than traditional detection of high-performance liquid chromatography (HPLC) [24].…”

Section: Discussionmentioning

confidence: 99%

Plasma omega-3 polyunsaturated fatty acid and tocopherol statuses and their relationships with aging-related diseases

Liu¹,

Liu²,

Chen³

et al. 2020

Preprint

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“…To avoid the auto-oxidation of α-TOH and α-TQ extracted from leaves, we performed the liquid extraction of α-TOH and α-TQ in chilled methanol. We considered the preanalytical methods mentioned in Giusepponi et al 28 . α-TOH (30 µM) in the presence of rose bengal (5 µM ) and from thylakoid membranes (750 µg Chl ml −1 ) was extracted in methanol by vortexing (5 min) and centrifuged at 2000 g for 60 s at 4 °C, the supernatant was transferred to HPLC vial using a syringe with needle.…”

Section: Determination Of α-Tocopherol and α-Tocopherol Quinone By Hplcmentioning

confidence: 99%

Formation of α-tocopherol hydroperoxide and α-tocopheroxyl radical: relevance for photooxidative stress in Arabidopsis

Kumar

Prasad

Pospı́šil

2020

Sci Rep

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Tocopherols, lipid-soluble antioxidants play a crucial role in the antioxidant defense system in higher plants. The antioxidant function of α-tocopherol has been widely studied; however, experimental data on the formation of its oxidation products is missing. In this study, we attempt to provide spectroscopic evidence on the detection of oxidation products of α-tocopherol formed by its interaction with singlet oxygen and lipid peroxyl radical. Singlet oxygen was formed using photosensitizer rose bengal and thylakoid membranes isolated from Arabidopsis thaliana. Singlet oxygen reacts with polyunsaturated fatty acid forming lipid hydroperoxide which is oxidized by ferric iron to lipid peroxyl radical. The addition of singlet oxygen to double bond carbon on the chromanol head of α-tocopherol forms α-tocopherol hydroperoxide detected using fluorescent probe swallow-tailed perylene derivative. The decomposition of α-tocopherol hydroperoxide forms α-tocopherol quinone. The hydrogen abstraction from α-tocopherol by lipid peroxyl radical forms α-tocopheroxyl radical detected by electron paramagnetic resonance. Quantification of lipid and protein hydroperoxide from the wild type and tocopherol deficient (vte1) mutant Arabidopsis leaves using a colorimetric ferrous oxidation-xylenol orange assay reveals that α-tocopherol prevents formation of both lipid and protein hydroperoxides at high light. Identification of oxidation products of α-tocopherol might contribute to a better understanding of the protective role of α-tocopherol in the prevention of oxidative damage in higher plants at high light.

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“…There are many methods for detecting and quantifying tocochromanols in the literature already; each has its advantages and disadvantages [20,21]. GCMS (Gas chromatography-mass spectrometry) requires derivatization of samples and ideally inclusion of heavy internal standards added to each sample.…”

Section: Conclusion and Remarksmentioning

confidence: 99%

“…We selected HPLC fluorescent detection for its combination of robustness and relatively moderate cost of implementation (compared to costs associated with LCMS systems). GCMS and LCMS are certainly superior for more broadly assessing the relative levels of a wide range of compounds, some of which may be tocochromanols or their metabolites [20,21]. Indeed, and our lab has used GCMS in the past to identify and assess tocopherol recycling metabolites in plants but have found the method to not be amenable or robust for high throughput extraction, derivatization and analysis of crude extracts [9].…”

Section: Conclusion and Remarksmentioning

confidence: 99%

High throughput profiling of tocochromanols in leaves and seeds of Arabidopsis and Maize

et al. 2020

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Background Tocochromanols are a group of lipid-soluble antioxidants produced by all plants and include the tocopherols, which are ubiquitous in the plant kingdom, and the biosynthetically-related compounds tocotrienols, which along with tocopherols commonly occur in seed of monocots. Most tocochromanols have some level of vitamin E activity, with α-tocopherol being the highest, and as such are essential nutrients in the human diet. Tocochromanols are particularly abundant in seeds and are critical for maintaining seed longevity and proper germination and as seed oils are a major component of the human diet, they are an important source of dietary vitamin E. In vegetative tissues, tocochromanols are important components in plant responses to stressful environments and can accumulate to high levels in response to various stresses including high light, heat, and dark. Results We report a robust, high throughput extraction and HPLC analysis method to quantify the levels of tocopherols and tocotrienols in leaves and seeds of plants, using Arabidopsis and maize tissues as examples. Conclusion The described method provides a rapid, high-throughput, cost-effective approach to quantifying the composition and content of tocopherols, and if needed simultaneously tocotrienols, in vegetative tissues and seeds. Optimized extraction methods are described for the two tissue types and have been used to study tocochromanol (vitamin E) natural variation in seed of large Arabidopsis and maize diversity panels, to assess gene function in T-DNA and Mu-tagged populations of Arabidopsis and maize, respectfully, and study the impact of environmental stresses, including high light stress, heat stress, and dark on tocopherols content and composition of vegetative tissue.

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LC-MS/MS assay for the simultaneous determination of tocopherols, polyunsaturated fatty acids and their metabolites in human plasma and serum

Cited by 23 publications

References 34 publications

Plasma omega-3 polyunsaturated fatty acid and tocopherol statuses and their relationships with aging-related diseases

Plasma omega-3 polyunsaturated fatty acid and tocopherol statuses and their relationships with aging-related diseases

Formation of α-tocopherol hydroperoxide and α-tocopheroxyl radical: relevance for photooxidative stress in Arabidopsis

High throughput profiling of tocochromanols in leaves and seeds of Arabidopsis and Maize

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