Determination of coenzyme Q10, α-tocopherol and cholesterol in biological samples by coupled-column liquid chromatography with coulometric and ultraviolet detection

doi:10.1016/0378-4347(88)80009-4

Journal of Chromatography B: Biomedical Sciences and Applications

Volume 425, 1988, Pages 87-97

https://doi.org/10.1016/0378-4347(88)80009-4 Get rights and content

Abstract

Coenzyme (Co) Q₁₀, Co Q₁₀H₂, α-tocopherol and cholesterol were dissociated from lipoproteins in plasma by treatment with 1-propanol. The supernatant obtained was injected directly for determination of Co Q₁₀ and Co Q₁₀H₂. Precolumn reduction with borohydride was used for determination of total Co Q₁₀ simultaneously with α-tocopherol and cholesterol. Total Co Q₁₀ in freeze-dried myocardial biopsies was determined after extraction with 1-propanol and oxidation of Co Q₁₀H₂ with ferric chloride. The chromatographic system comprised two reversed-phase columns and a three-electrode coulometric detector and a UV detector coupled in series. A pre-fractionation on the first column protected the coulometric detector from contamination and reduced the time for analysis by eliminating strongly retained solutes. The coulometric electrodes were operated in the oxidation—reduction—oxidation mode, and the last electrode was used for detection of α-tocopherol, Co Q₁₀ and Co Q₁₀H_2, while cholesterol was detected by UV at 215 nm. The fast isolation procedure made it possible to determine the reduced and oxidized forms of Co Q₁₀ in plasma. Quantitative recoveries were obtained for all the analytes studied and normal levels were determined with a coefficient of variation of 2–3%.

References (19)

S. Vadhanavikit et al.
Anal. Biochem.
(1984)
S. Vadhanavikit et al.
Biochem. Biophys. Res. Commun.
(1984)
D.D. Stump et al.
J. Chromatogr.
(1984)
J.K. Lang et al.
Anal. Biochem.
(1986)
T. Okamoto et al.
J. Chromatogr.
(1985)
P.O. Edlund et al.
J. Pharm. Biomed. Anal.
(1984)
P.O. Edlund
J. Pharm. Biomed. Anal.
(1986)
K. Hirota et al.
J. Chromatogr.
(1984)
G.R. Warnick et al.
J. Lipid Res.
(1978)

There are more references available in the full text version of this article.

Cited by (133)

Coenzyme Q<inf>10</inf> for Patients With Cardiovascular Disease: JACC Focus Seminar
2021, Journal of the American College of Cardiology
Citation Excerpt :
However, under certain circumstances, such as aging (14), statin usage (15,16), and some pathophysiologic states, endogenous biosynthesis is reduced and exogenous CoQ10 may take on a supplementary role. In humans, CoQ10 status may be assessed with measurement of plasma or serum CoQ10 concentration using high-performance liquid chromatography (17,18), although it is unclear whether such measurements reflect actual tissue status (19). In a U.S. population, plasma CoQ10 concentration ranges from 0.50 to 1.91 μmol/l (19), with sex and racial differences observed (20).
Coenzyme Q₁₀ (CoQ₁₀) is a naturally occurring compound that is found in animals and all humans. It has a fundamental role in cellular energy production. Although it is produced in the body, tissue deficiency can occur due to medications such as statins, which inhibit the mevalonate pathway. The clinical syndromes of statin-associated muscle symptoms (SAMS) and some of the features observed in patients with heart failure (HF) may be related to blood and tissue deficiency of CoQ₁₀. Numerous clinical trials of CoQ₁₀ in SAMS have yielded conflicting results. Yet, the weight of evidence as reflected in meta-analyses supports the use of exogenous CoQ₁₀ in SAMS. In patients with HF, large-scale randomized clinical trials are lacking, although one relatively contemporary trial, Q-SYMBIO, suggests an adjunctive role for CoQ₁₀. The possibility that statin-related CoQ₁₀ deficiency may play a role in patients with diastolic HF is an intriguing hypothesis that warrants further exploration.
Kaempferol increases levels of coenzyme Q in kidney cells and serves as a biosynthetic ring precursor
2017, Free Radical Biology and Medicine
Citation Excerpt :
Standards of Q4, Q6, Q9 and Q10 were purchased from Sigma-Aldrich. Dipropoxy-Q10 was synthesized essentially as described by Edlund [17] for diethoxy-Q10, except 1-propanol was substituted for ethanol while maintaining the other reagents and conditions. Mouse kidney proximal tubule epithelial (Tkpts) cells [18], were provided by Dr. Elsa Bello-Reuss (Texas Tech University Health Science Center) and Dr. Judit K. Magyesi (University of Arkansas for Medical Sciences, Little Rock, AR).
Coenzyme Q (Q) is a lipid-soluble antioxidant essential in cellular physiology. Patients with Q deficiencies, with few exceptions, seldom respond to treatment. Current therapies rely on dietary supplementation with Q₁₀, but due to its highly lipophilic nature, Q₁₀ is difficult to absorb by tissues and cells. Plant polyphenols, present in the human diet, are redox active and modulate numerous cellular pathways. In the present study, we tested whether treatment with polyphenols affected the content or biosynthesis of Q. Mouse kidney proximal tubule epithelial (Tkpts) cells and human embryonic kidney cells 293 (HEK 293) were treated with several types of polyphenols, and kaempferol produced the largest increase in Q levels. Experiments with stable isotope ¹³C-labeled kaempferol demonstrated a previously unrecognized role of kaempferol as an aromatic ring precursor in Q biosynthesis. Investigations of the structure-function relationship of related flavonols showed the importance of two hydroxyl groups, located at C3 of the C ring and C4′ of the B ring, both present in kaempferol, as important determinants of kaempferol as a Q biosynthetic precursor. Concurrently, through a mechanism not related to the enhancement of Q biosynthesis, kaempferol also augmented mitochondrial localization of Sirt3. The role of kaempferol as a precursor that increases Q levels, combined with its ability to upregulate Sirt3, identify kaempferol as a potential candidate in the design of interventions aimed on increasing endogenous Q biosynthesis, particularly in kidney.
Simultaneous analysis of circulating 25-hydroxy-vitamin D<inf>3</inf>, 25-hydroxy-vitamin D<inf>2</inf>, retinol, tocopherols, carotenoids, and oxidized and reduced coenzyme Q10 by high performance liquid chromatography with photo diode-array detection using C18 and C30 columns alone or in combination
2013, Journal of Chromatography A
Citation Excerpt :
Using HPLC, the yield was approximately 98%. Et-UN10 and Bu-UN10 standards were synthesized analogous to a previous report [54]. Briefly, 0.1 g UN10 was dissolved in 1 mL hexane followed by addition of 4 mL EtOH or BuOH and 200 μL 1 M KOH.
Circulating lipid-phase micronutrients (LPM) such as 25-hydroxylated D vitamers, retinol, tocopherols, carotenoids including their isomers, and coenzyme Q10 play important roles in health maintenance and disease prevention and can serve as useful biomarkers. We developed fast, affordable, and accurate HPLC assays that simultaneously measured all above LPM in a single run using UV/VIS detection at 265 nm, 295 nm, and 480 nm with (1) a C18 column alone; (2) a C30 column alone; or (3) each of these columns connected in series. The C18 column alone could separate all major LPM of interest in less than 17 min but insufficiently resolved the lycopene isomers, the 25-hydroxylated D vitamers, lutein from zeaxanthin and β- from γ-tocopherol. The C30 column alone separated all LPM of interest including many isomeric analytes but failed to resolve the Q10 compounds, which co-eluted with carotenoids. Connecting the C18 and C30 columns in series with a detector after the C30 column and a pressure resistant detector between the columns resulted in ideal resolution and accurate quantitation of all LPM of interest but required software capable of processing the acquired data from both detectors. Connecting the C18 and C30 columns in series with exclusively one detector after the C30 column resulted in carotenoid-Q10 interferences, however, this was remedied by heart-cutting 2D-LC with a 6-port valve between the columns, which resolved all analytes in 42 min. Faster run times led to some analytes not being resolved. Many variations of these methods are possible to meet the needs of individual requirements while minimizing sample material and turn-around-times.
UbiN, a novel Rhodobacter capsulatus decarboxylative hydroxylase involved in aerobic ubiquinone biosynthesis
2023, FEBS Open Bio
Plasma α-Tocopherol and Ubiquinone and Their Relations to Muscle Function in Healthy Humans and in Cardiac Diseases
2023, Vitamin E in Health and Disease
COVID-19 and the Assessment of Coenzyme Q10
2022, Methods in Molecular Biology

View all citing articles on Scopus

¹: Temporary address till May 1988: Equine Drug Testing and Toxicology, New York State College of Veterinary Medicine, Cornell University, 925 Warren Road, Ithaca, NY 14850, U.S.A.

View full text

Determination of coenzyme Q10, α-tocopherol and cholesterol in biological samples by coupled-column liquid chromatography with coulometric and ultraviolet detection

Abstract

Anal. Biochem.

Biochem. Biophys. Res. Commun.

J. Chromatogr.

Anal. Biochem.

J. Chromatogr.

J. Pharm. Biomed. Anal.

J. Pharm. Biomed. Anal.

J. Chromatogr.

J. Lipid Res.

Determination of coenzyme Q₁₀, α-tocopherol and cholesterol in biological samples by coupled-column liquid chromatography with coulometric and ultraviolet detection