Development and Validation of a Liquid Chromatography Mass Spectrometry Method for Simultaneous Measurement of 25(OH)D3, epi-25(OH)D3, 25(OH)D2, Vitamin A, α-Tocopherol, and γ-Tocopherol

Yi Xiao, Edward Ki Yun Leung

Am J Clin Pathol . 2021 Aug 31;aqab088. doi: 10.1093/ajcp/aqab088. Online ahead of print.


Objectives: Fat-soluble vitamins are measured to identify deficiencies that may lead to rickets, osteomalacia, night blindness, and reversible motor and sensory neuropathies. We present a rapid liquid chromatography-mass spectrometry (LC-MS/MS) method that simultaneously measures 25-hydroxyvitamin D3 (25[OH]D3), epi-25(OH)D3, 25(OH)D2, vitamin A, α-tocopherol, and γ-tocopherol.

Methods: We mixed 100 µL serum with internal standard and extracted it by using supported liquid extraction plates. Reconstituted specimens were analyzed by LC-MS/MS with a 10-minute gradient.

Results: The method was linear, covering physiological levels with r2 > 0.99, and the total precision was less than 15% at all quality control levels. The lower limit of the measuring intervals for 25(OH)D3, epi-25(OH)D3, 25(OH)D2, vitamin A, α-tocopherol, and γ-tocopherol were 4 ng/mL, 4 ng/mL, 4 ng/mL, 1 µg/dL, 0.2 µg/mL, and 0.2 µg/mL, respectively, with coefficient of variation of 20% or less. Recoveries were between 92% and 111% for National Institute of Standards and Technology reference materials and 81% and 122% for spike-recovery studies. Comparison studies for vitamin D total, vitamin A, and α-tocopherol demonstrated slopes between 1.04 and 1.11 and r2 between 0.94 and 0.96. Minimal matrix effect was observed for all analytes.

Conclusions: We developed and validated a rapid LC-MS/MS method for the simultaneous measurement of 25(OH)D3, epi-25(OH)D3, 25(OH)D2, vitamin A, α-tocopherol, and γ-tocopherol.

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Vitamin E May Save Dental Enamel from Erosion, Study

SAO PAULO, Brazil: Researchers at the University of São Paulo have found that Vitamin E could protect dental enamel against erosion.

Daniela Rios and colleagues found this association by assessing the in-vitro effect of different components of palm oil on enamel in a short-term erosive challenge. To conduct a study, they use bovine enamel blocks treated with the solutions described in the results below separately and then immersed in citric acid. The following results have been analysed, which lead to the finding.

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Vitamin E may help protect dental enamel against erosion, study finds

The acquired enamel pellicle (AEP) was previously formed in situ for 2 h. Subsequently, the bovine enamel blocks were treated i vitro according to following solutions: G1- palm oil; G2- 85% tocotrienol solution; G3- oily vitamn E; G4- oily vitamin A; G5- deionized water (negative control); G6- stannous-containing solution (positive control).

The author concluded that ”Vitamn E presented similar preventive effect to a commercial mouthwash stannous-containing solution against initial erosion, and it can be considered as a promising alternative for the formulations of solutions aiming to prevent erosive tooth wear”.

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Tocotrienols as an Anti-Breast Cancer Agent

Madison Trujillo, Anupreet Kharbanda, Christa Corley, Pilar Simmons, Antiño R Allen

Antioxidants (Basel) . 2021 Aug 29;10(9):1383. doi: 10.3390/antiox10091383.


In the past few years, breast cancer has become the most prevalent type of cancer. The majority of patients receive combinatorial chemotherapy treatments, which may result in increased risk of developing drug resistance, a reduced quality of life, and substantial side effects. Treatment modalities that could lessen the physical toll of standard treatments or act in synergy with chemotherapeutic treatments would benefit women worldwide. Research into tocotrienols has thus far demonstrated their potential to be such an agent, with tocotrienols surpassing the pharmacological potential of tocopherols. Further research using in vitro and preclinical breast cancer models to support clinical trials is needed. This review uses bibliometric analysis to highlight this gap in research and summarizes the current and future landscape of tocotrienols as an anti-breast cancer agent.

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The effects of vitamin E on colistin-induced nephrotoxicity in treatment of drug-resistant gram-negative bacterial infections: A randomized clinical trial

Maryam Samsami, Minoosh Shabani, Mohammadreza Hajiesmaeili, Maria Tavakoli-Ardakani, Seyed Hossein Ardehali, Alireza Fatemi, Saghar Barati, Omid Moradi, Zahra Sahraei

J Infect Chemother . 2021 Aug;27(8):1181-1185. doi: 10.1016/j.jiac.2021.03.013. Epub 2021 Apr 15.


Introduction: Nephrotoxicity remains a major long-standing concern for colistin, and it is critical to find agents that can prevent it. The present study aims to investigate the effect of vitamin E on the prevention of colistin-induced nephrotoxicity based on its antioxidant and free radical scavenging properties.

Methods: A randomized clinical trial was designed for 52 patients taking colistin. These patients were categorized into two groups of equal size, receiving colistin or colistin plus vitamin E (α-Tocopherol). Vitamin E with doses of 400 units was administrated daily either orally or by a nasogastric tube if needed. The incidence of Acute Kidney Injury (AKI) and its duration was recorded based on RIFLE criteria.

Results: The Incidence of AKI based on RIFLE criteria was 42.3% and 46.2% in intervention and control groups, respectively. The analysis showed no significant difference in the prevalence of AKI for the two groups (P = 0.78). There was no significant difference in the duration of AKI neither (P = 0.83).

Conclusion: Although vitamin E is a powerful biological antioxidant, the effects of Vitamin E prophylaxis on colistin-induced nephrotoxicity was not taken into consideration in this study.

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The plasma antioxidant vitamin status of the INTAPP cohort examined: The unsuspected importance of β-carotene and γ-tocopherol in preeclampsia

Jean-François Bilodeau, Amélie Gagné, Karine Greffard, François Audibert, William D Fraser, Pierre Julien

Pregnancy Hypertens . 2021 Aug;25:213-218. doi: 10.1016/j.preghy.2021.06.009. Epub 2021 Jun 12.


Objective: Examine the levels of plasma antioxidant vitamins before and during a treatment with placebo or vitamin E + C supplement to prevent preeclampsia (PE).

Study design: Per-protocol analysis of a subset group of pregnant women (n = 295) from the International Trial of Antioxidants for the Prevention of PE (INTAPP) randomized case-control study. Normotensive receiving placebo or vitamins (n = 115 and 87 respectively) were compared to gestational hypertension (GH) without proteinuria (n = 30 and 27) and PE (n = 21 and 15). Vitamin quantification was performed at 12-18, 24-26 and 32-34 weeks of gestation.

Main outcome measures: Coenzyme (Co) Q10, β-carotene and vitamins E (α and γ forms) plasma levels.

Results: Vitamin E + C supplementation was found to increase the α-tocopherol levels by 40% but was associated with a 57% decrease in the γ-tocopherol isoform for all study groups (p < 0.001). The β -carotene was lower in the PE than in the normotensive and GH groups (p < 0.001) while the level of CoQ10 remained unaffected.

Conclusions: A more personalized approach that target the suboptimal levels of specific antioxidants without disturbing the α/γ-tocopherol ratio could be a more successful approach to counteract oxidative stress in PE.

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Vitamin E and selenium supplementation synergistically alleviate the injury induced by hydrogen peroxide in bovine granulosa cells

Meimei Wang, Yan Li, Adrian Molenaar, Qiufeng Li, Yufeng Cao, Yizhao Shen, Panliang Chen, Jinling Yan, Yanxia Gao, Jianguo Li

Theriogenology . 2021 Aug;170:91-106. doi: 10.1016/j.theriogenology.2021.04.015. Epub 2021 May 5.


Dairy cows are susceptible to reproductive disorders, which are thought to be associated with oxidative stress. In the study, we investigated the effects of vitamin E (VE) and selenium (Se) on the proliferation, apoptosis, and steroidogenesis in bovine ovarian granulosa cells under hydrogen peroxide (H2O2) – induced oxidative stress and elaborated the underlying mechanisms. Our results showed that VE or Se could stimulate the granulosa cell proliferation, possibly due to up-regulating the expression of CCND1 and decreasing the P21 levels under oxidative stress. VE or Se treatment also increased the secretion of estradiol (E2) and progesterone (P4), which could be owing to improving the expression of genes associated with steroidogenesis (StAR, HSD3β1, and CYP19A1) expression. VE or Se treatment down-regulated the apoptosis-related genes (BAX, CASP3) expression and decreased cell apoptosis. Furthermore, VE or Se treatment inhibited reactive oxidative species (ROS) and malondialdehyde (MDA) generation, increased total antioxidant capacity (T-AOC), and the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px). Additionally, VE or Se treatment also alleviated the endoplasmic reticulum stress, activated the nuclear factor erythroid 2-related factor 2 (NRF2), and up-regulated the expression of its downstream genes, including NQO1, HO-1, GCLM, GCLC. More importantly, compared with either VE or Se treatment alone, their combined treatment showed a better protective effect against oxidative damage. Overall, our results indicated that VE and Se synergistically stimulated the granulosa cell proliferation and steroidogenesis, decreased cell apoptosis, mitigated the endoplasmic reticulum stress by activating the NRF2 signal pathway.

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In vitro antiaging evaluation of sunscreen formulated from nanostructured lipid carrier and tocotrienol-rich fraction

Chee Chin Chu, Zafarizal Aldrin Bin Azizul Hasan, Chin Ping Tan, Kar Lin Nyam

J Pharm Sci . 2021 Aug 20;S0022-3549(21)00423-8. doi: 10.1016/j.xphs.2021.08.020.


Chronic exposure to ultraviolet (UV) radiation leads to photoaging. There is a tremendous rise in products having a dual activity of photoprotection and antiaging. In vitro analysis in dermal fibroblasts and their biological mechanisms involved are critical to determine antiaging potential. The study aimed to investigate the antiaging potential of sunscreen formulated from nanostructured lipid carrier and tocotrienol-rich fraction (NLC-TRF sunscreen). The antioxidant activity of the NLC-TRF sunscreen was evaluated by radical scavenging and hydrogen peroxide inhibition properties. Also, collagenase, elastase and matrix metalloproteinase-1 (MMP-1) inhibition activities, and type I collagen and elastin protein expression were studied. Quantitative real-time polymerase chain reaction (qPCR) was used to evaluate the mRNA expression of fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), transforming growth factor-β1 (TGF-β1), type I collagen (COL1A1), elastin (ELN), MMP-1, MMP-2, and tissue inhibitor matrix metalloproteinase-1 (TIMP-1). The results suggested that NLC-TRF sunscreen is effective in radical, anti-hydrogen peroxide, and collagenase, elastase and MMP-1 inhibition activities. Besides, a significant increase for type I collagen (3.47-fold) and elastin (2.16-fold) protein and fibroblast regeneration genes (FGF (2.12-fold), VEGF (1.91-fold), TGF-β1 (2.84-fold), TIMP-1 (1.42-fold), ELN (2.13-fold)) were observed after sample treatment. These findings support the therapeutic potential of NLC-TRF sunscreen in antiaging.

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Role of Vitamin E and the Orexin System in Neuroprotection

Maria Ester La Torre, Ines Villano, Marcellino Monda, Antonietta Messina, Giuseppe Cibelli, Anna Valenzano, Daniela Pisanelli, Maria Antonietta Panaro, Nicola Tartaglia, Antonio Ambrosi, Marco Carotenuto, Vincenzo Monda, Giovanni Messina, Chiara Porro

Brain Sci . 2021 Aug 20;11(8):1098. doi: 10.3390/brainsci11081098.


Microglia are the first line of defense at the level of the central nervous system (CNS). Phenotypic change in microglia can be regulated by various factors, including the orexin system. Neuroinflammation is an inflammatory process mediated by cytokines, by the lack of interaction of specific receptors such as the OX2-OX2R complex, caused by systemic tissue damage or, more often, associated with direct damage to the CNS. Chronic activation of microglia could lead to long-term neurodegenerative diseases. This review aims to explore how tocopherol (vitamin E) and the orexin system may play a role in the prevention and treatment of microglia inflammation and, consequently, in neurodegenerative diseases thanks to its antioxidant properties. The results of animal and in vitro studies provide evidence to support the use of tocopherol for a reduction in microglia inflammation as well as a greater activation of the orexinergic system. Although there is much in vivo and in vitro evidence of vitamin E antioxidant and protective abilities, there are still conflicting results for its use as a treatment for neurodegenerative diseases that speculate that vitamin E, under certain conditions or genetic predispositions, can be pro-oxidant and harmful.

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The effect of Vitamin E supplementation on treatment of chronic periodontitis

Parichehr Behfarnia, Mina Dadmehr, Seyedeh Negin Hosseini, Seyed Amir Mirghaderi

Dent Res J (Isfahan) . 2021 Aug 18;18:62. eCollection 2021.


Background: The purpose of this study was to evaluate the effect of Vitamin E supplements on chronic periodontitis based on the clinical parameters of pocket depth and clinical attachment level and total antioxidant capacity (TAC) of saliva.

Materials and methods: In this clinical trial, 16 patients with chronic periodontitis were selected and divided into two groups. The indices of pocket depth and attachment loss for 6 teeth per person were measured with a periodontal probe. A total of 41 teeth in the control group and 42 teeth in the case group were examined. Then, 2 ml nonstimulated saliva was collected from each patient. All patients were treated with scaling and root planing (SRP). The case group consumed 200 IU supplementary Vitamin E daily for up to 2 months. After 2 months, clinical indices were re-measured and 2 ml nonstimulated saliva was collected. The TAC of saliva samples was measured by using Zellbio’s TAC Kit. Data were analyzed by the SPSS software and were evaluated in each group between the first session and 2 months later with paired t-test. The differences between the two groups were evaluated through the independent t-test (α ≤ 0.05).

Results: Independent t-test showed that mean change in TAC (P = 0.14) and pocket depth changes (P = 0.33) was not significant between two groups 2 months after SRP, but mean attachment loss changes in the case group was significantly less than the control group (P = 0.03).

Conclusion: The results of this study indicate that Vitamin E supplementation with SRP can reduce the inflammatory process of periodontitis and improve periodontal clinical indices and decrease the amount of attachment loss.

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