Rapid quantitative determination of fat-soluble vitamins and coenzyme Q-10 in human serum by reversed phase ultra-high pressure liquid chromatography with UV detection

Paliakov, Ekaterina; Crow, Brian S.; Bishop, Michael J.; Norton, Dean; George, Joe; Bralley, J. Alexander

doi:10.1016/j.jchromb.2008.11.012

Cited by 38 publications

(37 citation statements)

References 29 publications

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“…The assay of fat-soluble vitamins was based on a modified method from Paliakov et al [18]. Briefly, 200 mL of plasma was deproteinized with 0.5 mL methanol, then was vortex mixed and extracted twice with 0.75 mL hexane for 5 min, and centrifuged at 1500 g for 15 min.…”

Section: Analysis Of Fat-soluble Vitaminsmentioning

confidence: 99%

Effects of cold‐pressed and vitamin E‐enriched flaxseed oils on lipid profile and antioxidant status in high‐fat fed rats

Deng

et al. 2012

Euro J Lipid Sci & Tech

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In this study we evaluate whether cold-pressed flaxseed oil (CFO), vitamin E-enriched hot pressed and totally refined flaxseed oils (HRFOs) characterized by different qualities of micronutrients have beneficial effects on lipid profile and antioxidant status in high-fat fed rats. Male Wistar rats were fed synthetic diets containing 10% of HRFO, CFO, low-dose (100 mg/100 g oil) vitamin E-enriched HRFO (L-HRFO) or high-dose (200 mg/100 g oil) vitamin E-enriched HRFO (H-HRFO), respectively for 4 wk. The results showed that plasma triglycerides (TG), total cholesterol (TC), and low-density lipoprotein cholesterol levels in CFO group decreased by 12, 21, and 18% compared with HRFO group ( p < 0.05), the total hepatic lipids, TC and TG levels in CFO group were also lower significantly than those in HRFO group, but no significant differences were observed in plasma and hepatic lipid profiles among HRFO, L-HRFO, and H-HRFO groups. T-AOC levels significantly increased by 11, 12, and 22% in CFO, L-HRFO and H-HRFO groups compared with HRFO group, which was negatively correlated with the MDA levels and similar changes were observed for GSH and vitamin E levels in plasma. The CFO had a significantly higher cholesterol secretion than HRFO, L-HRFO, and H-HRFO. These results indicated that CFO had a more beneficial effect on improving lipid status and preventing oxidative stress than HRFO, and vitamin E-enriched HRFO decreased the lipid peroxidation concentration though there was no significant effect on lipid profiles.Practical applications: The cold-pressed and vitamin E-enriched HRFOs characterized by different qualities of micronutrients reduced the risk factors of cardiovascular disease. This could provide a theoretical basis for the development of high value-added functional food ingredients based on flaxseed oil or a-linolenic acid.

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Section: Analysis Of Fat-soluble Vitaminsmentioning

confidence: 99%

Effects of cold‐pressed and vitamin E‐enriched flaxseed oils on lipid profile and antioxidant status in high‐fat fed rats

Deng

et al. 2012

Euro J Lipid Sci & Tech

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“…The development of columns of very small particle size allows for the use of much lower flows of mobile phase at very high pressures. UHPLC has been used for the determination of fat-soluble vitamins but less so for hydrosoluble ones (apart from some mentions of manufacturers brochures) [18,19]. Xingnan and Adrian [20] developed and validated an UHPLC method for the quantification of serum L-AA and DHAA, by the subtraction method.…”

Section: Introductionmentioning

confidence: 99%

An improved and fast UHPLC-PDA methodology for determination of L-ascorbic and dehydroascorbic acids in fruits and vegetables. Evaluation of degradation rate during storage

et al. 2012

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This study provides a versatile validated method to determine the total vitamin C content, as the sum of the contents of L-ascorbic acid (L-AA) and dehydroascorbic acid (DHAA), in several fruits and vegetables and its degradability with storage time. Seven horticultural crops from two different origins were analyzed using an ultrahigh-performance liquid chromatographic-photodiode array (UHPLC-PDA) system, equipped with a new trifunctional high strength silica (100% silica particle) analytical column (100 mm×2.1 mm, 1.7 μm particle size) using 0.1% (v/v) formic acid as mobile phase, in isocratic mode. This new stationary phase, specially designed for polar compounds, overcomes the problems normally encountered in HPLC and is suitable for the analysis of large batches of samples without L-AA degradation. In addition, it proves to be an excellent alternative to conventional C18 columns for the determination of L-AA in fruits and vegetables. The method was fully validated in terms of linearity, detection (LOD) and quantification (LOQ) limits, accuracy, and inter/intraday precision. Validation experiments revealed very good recovery rate of 96.6±4.4% for L-AA and 103.1±4.8 % for total vitamin C, good linearity with r 2 -values >0.999 within the established concentration range, excellent repeatability (0.5%), and reproducibility (1.6%) values. The LOD of the method was 22 ng/mL whereas the LOQ was 67 ng/mL. It was possible to demonstrate that L-AA and DHAA concentrations in the different horticulture products varied oppositely with time of storage not always affecting the total amount of vitamin C during shelf-life. Locally produced fruits have higher concentrations of vitamin C, compared with imported ones, but vegetables showed the opposite trend. Moreover, this UHPLC-PDA methodology proves to be an improved, simple, and fast approach for determining the total content of vitamin C in various food commodities, with high sensitivity, selectivity, and resolving power within 3 min of run analysis.

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“…Other groups have reported LOQs of 2.9 nmol/L (2.5 ng/mL) to 58 nmol/L (20 ng/mL) (14 -17, 19 ) for LC-MS CoQ10 assays. Most previously reported assays for CoQ10 in biological samples are based on HPLC with ultraviolet (8,21,(25)(26)(27)(28) or electrochemical (29 -32 ) detection. The reported LOD of the HPLC assays is at best 1.6 nmol/L (30 ).…”

Section: Discussionmentioning

confidence: 99%

Mitochondrial Coenzyme Q10 Determination by Isotope-Dilution Liquid Chromatography–Tandem Mass Spectrometry

2013

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BACKGROUND Coenzyme Q10 (CoQ10) is an essential part of the mitochondrial respiratory chain. Unlike most other respiratory chain disorders, CoQ10 deficiency is potentially treatable. We aimed to develop and validate an accurate liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for the determination of mitochondrial CoQ10 in clinical samples. METHODS We used mitochondria isolated from muscle biopsies of patients (n = 166) suspected to have oxidative phosphorylation deficiency. We also used fibroblast mitochondria from 1 patient with CoQ10 deficiency and 3 healthy individuals. Samples were spiked with nonphysiologic CoQ10-[2H6] internal standard, extracted with 1-propanol and with ethanol and hexane (2 mL/5 mL), and CoQ10 quantified by LC-MS/MS. The method and sample stability were validated. A reference interval was established from the patient data. RESULTS The method had a limit of quantification of 0.5 nmol/L. The assay range was 0.5–1000 nmol/L and the CVs were 7.5%–8.2%. CoQ10 was stable in concentrated mitochondrial suspensions. In isolated mitochondria, the mean ratio of CoQ10 to citrate synthase (CS) activity (CoQ10/CS) was 1.7 nmol/U (95% CI, 1.6–1.7 nmol/U). We suggest a CoQ10/CS reference interval of 1.1–2.8 nmol/U for both sexes and all ages. The CoQ10/CS ratio was 5-fold decreased in fibroblast mitochondria from a patient with known CoQ10 deficiency due to recessive prenyl (decaprenyl) diphosphate synthase, subunit 2 (PDSS2) mutations. CONCLUSIONS Normalization of mitochondrial CoQ10 concentration against citrate synthase activity is likely to reflect most accurately the CoQ10 content available for the respiratory chain. Our assay and the established reference range should facilitate the diagnosis of respiratory chain disorders and treatment of patients with CoQ10 deficiency.

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Rapid quantitative determination of fat-soluble vitamins and coenzyme Q-10 in human serum by reversed phase ultra-high pressure liquid chromatography with UV detection

Cited by 38 publications

References 29 publications

Effects of cold‐pressed and vitamin E‐enriched flaxseed oils on lipid profile and antioxidant status in high‐fat fed rats

Effects of cold‐pressed and vitamin E‐enriched flaxseed oils on lipid profile and antioxidant status in high‐fat fed rats

An improved and fast UHPLC-PDA methodology for determination of L-ascorbic and dehydroascorbic acids in fruits and vegetables. Evaluation of degradation rate during storage

Mitochondrial Coenzyme Q10 Determination by Isotope-Dilution Liquid Chromatography–Tandem Mass Spectrometry

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