Clinical Chemistry, Vol 34, 377-381, Copyright © 1988 by American Association for Clinical Chemistry

Concurrent liquid-chromatographic assay of retinol, alpha-tocopherol, beta-carotene, alpha-carotene, lycopene, and beta-cryptoxanthin in plasma, with tocopherol acetate as internal standard

DI Thurnham, E Smith and PS Flora
Nutrition Section, Dudley Road Hospital, Birmingham, U.K.

A method is described for simultaneously determining retinol, alpha- tocopherol, beta-carotene, alpha-carotene, lycopene, and beta- cryptoxanthin in 0.25 mL of plasma. Plasma mixed with sodium dodecyl sulfate is deproteinized with ethanol containing tocopherol acetate, then extracted with heptane. The evaporated organic layer is reconstituted with mobile phase (methanol/acetonitrile/chloroform, 47/47/6 by vol) and injected onto a 100 x 4.6 mm 3-micron column of Spherisorb ODS-2 (LKB) at 1.5 mL/min. The alpha- and beta-carotenes are well resolved during the 6.5-min run. Retinol is monitored at 325 nm, the tocopherols at 292 nm, and the carotenoids at 450 nm. Extraction of concentrations as great as 135 mumol/L is complete. Intrabatch CVs were 1.7%, 2.3%, 4.1%, 10.4%, 6.4%, and 3.6% for retinol, alpha-tocopherol, beta-carotene, alpha-carotene, lycopene, and beta-cryptoxanthin, respectively. Interbatch CVs for measurements on 30 occasions over 11 weeks were about 10% for all components except alpha-tocopherol (5.3%). Results agree well with those for retinol, alpha-tocopherol, and beta- carotene in quality-control samples.

The following articles in journals at HighWire Press have cited this article:

				P. R. Afolabi, F. Jahoor, A. A. Jackson, J. Stubbs, A. M. Johnstone, P. Faber, G. Lobley, E. Gibney, and M. Elia The effect of total starvation and very low energy diet in lean men on kinetics of whole body protein and five hepatic secretory proteins Am J Physiol Endocrinol Metab, December 1, 2007; 293(6): E1580 - E1589. [Abstract] [Full Text] [PDF]

				R. S. Richardson, A. J. Donato, A. Uberoi, D. W. Wray, L. Lawrenson, S. Nishiyama, and D. M. Bailey Exercise-induced brachial artery vasodilation: role of free radicals Am J Physiol Heart Circ Physiol, March 1, 2007; 292(3): H1516 - H1522. [Abstract] [Full Text] [PDF]

				C. I. Gill, S. Haldar, L. A Boyd, R. Bennett, J. Whiteford, M. Butler, J. R Pearson, I. Bradbury, and I. R Rowland Watercress supplementation in diet reduces lymphocyte DNA damage and alters blood antioxidant status in healthy adults Am. J. Clinical Nutrition, February 1, 2007; 85(2): 504 - 510. [Abstract] [Full Text] [PDF]

				J. J. Carrero, J. Fonolla, J. L. Marti, J. Jimenez, J. J. Boza, and E. Lopez-Huertas Intake of Fish Oil, Oleic Acid, Folic Acid, and Vitamins B-6 and E for 1 Year Decreases Plasma C-Reactive Protein and Reduces Coronary Heart Disease Risk Factors in Male Patients in a Cardiac Rehabilitation Program J. Nutr., February 1, 2007; 137(2): 384 - 390. [Abstract] [Full Text] [PDF]

				E. Paterson, M. H. Gordon, C. Niwat, T. W. George, L. Parr, S. Waroonphan, and J. A. Lovegrove Supplementation with Fruit and Vegetable Soups and Beverages Increases Plasma Carotenoid Concentrations but Does Not Alter Markers of Oxidative Stress or Cardiovascular Risk Factors J. Nutr., November 1, 2006; 136(11): 2849 - 2855. [Abstract] [Full Text] [PDF]

				M. A. Atkin, A. Gasper, R. Ullegaddi, and H. J. Powers Oxidative Susceptibility of Unfractionated Serum or Plasma: Response to Antioxidants in Vitro and to Antioxidant Supplementation Clin. Chem., November 1, 2005; 51(11): 2138 - 2144. [Abstract] [Full Text] [PDF]

				N. L. A. Misso, J. Brooks-Wildhaber, S. Ray, H. Vally, and P. J. Thompson Plasma concentrations of dietary and nondietary antioxidants are low in severe asthma Eur. Respir. J., August 1, 2005; 26(2): 257 - 264. [Abstract] [Full Text] [PDF]

				R. Rej Clinical Chemistry through Clinical Chemistry: A Journal Timeline Clin. Chem., December 1, 2004; 50(12): 2415 - 2458. [Abstract] [Full Text] [PDF]

				D. M. Bailey, I. S. Young, J. McEneny, L. Lawrenson, J. Kim, J. Barden, and R. S. Richardson Regulation of free radical outflow from an isolated muscle bed in exercising humans Am J Physiol Heart Circ Physiol, October 1, 2004; 287(4): H1689 - H1699. [Abstract] [Full Text] [PDF]

				H. E Theobald, P. J Chowienczyk, R. Whittall, S. E Humphries, and T. A. Sanders LDL cholesterol-raising effect of low-dose docosahexaenoic acid in middle-aged men and women Am. J. Clinical Nutrition, April 1, 2004; 79(4): 558 - 563. [Abstract] [Full Text] [PDF]

				L. E. Rhodes, H. Shahbakhti, R. M. Azurdia, R. M.W. Moison, M.-J. S.T. Steenwinkel, M. I. Homburg, M. P. Dean, F. McArdle, G. M.J. Beijersbergen van Henegouwen, B. Epe, et al. Effect of eicosapentaenoic acid, an omega-3 polyunsaturated fatty acid, on UVR-related cancer risk in humans. An assessment of early genotoxic markers Carcinogenesis, May 1, 2003; 24(5): 919 - 925. [Abstract] [Full Text] [PDF]

				Y. Kelly, A. Sacker, and M. Marmot Nutrition and respiratory health in adults: findings from the Health Survey for Scotland Eur. Respir. J., April 1, 2003; 21(4): 664 - 671. [Abstract] [Full Text] [PDF]

				F. T Wieringa, M. A Dijkhuizen, C. E West, D. I Thurnham, Muhilal, and J. W. Van der Meer Redistribution of vitamin A after iron supplementation in Indonesian infants Am. J. Clinical Nutrition, March 1, 2003; 77(3): 651 - 657. [Abstract] [Full Text] [PDF]

				M. van Lieshout, C. E West, and R. B van Breemen Isotopic tracer techniques for studying the bioavailability and bioefficacy of dietary carotenoids, particularly {beta}-carotene, in humans: a review Am. J. Clinical Nutrition, January 1, 2003; 77(1): 12 - 28. [Abstract] [Full Text] [PDF]

				T. H Rissanen, S. Voutilainen, K. Nyyssonen, R. Salonen, G. A Kaplan, and J. T Salonen Serum lycopene concentrations and carotid atherosclerosis: the Kuopio Ischaemic Heart Disease Risk Factor Study Am. J. Clinical Nutrition, January 1, 2003; 77(1): 133 - 138. [Abstract] [Full Text] [PDF]

				T. Rissanen, S. Voutilainen, K. Nyyssonen, and J. T. Salonen Lycopene, Atherosclerosis, and Coronary Heart Disease Experimental Biology and Medicine, November 1, 2002; 227(10): 900 - 907. [Abstract] [Full Text] [PDF]

				F. T. Wieringa, M. A. Dijkhuizen, C. E. West, C. A. Northrop-Clewes, and Muhilal Estimation of the Effect of the Acute Phase Response on Indicators of Micronutrient Status in Indonesian Infants J. Nutr., October 1, 2002; 132(10): 3061 - 3066. [Abstract] [Full Text] [PDF]

				M. P. Valero, A. E. Fletcher, B. L. De Stavola, J. Vioque, and V. C. Alepuz Vitamin C Is Associated with Reduced Risk of Cataract in a Mediterranean Population J. Nutr., June 1, 2002; 132(6): 1299 - 1306. [Abstract] [Full Text] [PDF]

				A. J. Edwards, C. H. Nguyen, C.-S. You, J. E. Swanson, C. Emenhiser, and R. S. Parker {alpha}- and {beta}-Carotene from a Commercial Carrot Puree Are More Bioavailable to Humans than from Boiled-Mashed Carrots, as Determined Using an Extrinsic Stable Isotope Reference Method J. Nutr., February 1, 2002; 132(2): 159 - 167. [Abstract] [Full Text] [PDF]

				S. Ching, D. Ingram, R. Hahnel, J. Beilby, and E. Rossi Serum Levels of Micronutrients, Antioxidants and Total Antioxidant Status Predict Risk of Breast Cancer in a Case Control Study J. Nutr., February 1, 2002; 132(2): 303 - 306. [Abstract] [Full Text] [PDF]

				S. E Moore, A. C Collinson, and A. M Prentice Immune function in rural Gambian children is not related to season of birth, birth size, or maternal supplementation status Am. J. Clinical Nutrition, December 1, 2001; 74(6): 840 - 847. [Abstract] [Full Text]

				L. Mabile, A. Piolot, L. Boulet, L.-J. Fortin, N. Doyle, C. Rodriguez, J. Davignon, D. Blache, and S. Lussier-Cacan Moderate intake of n-3 fatty acids is associated with stable erythrocyte resistance to oxidative stress in hypertriglyceridemic subjects Am. J. Clinical Nutrition, October 1, 2001; 74(4): 449 - 456. [Abstract] [Full Text] [PDF]

				A. J Edwards, C.-S. You, J. E Swanson, and R. S Parker A novel extrinsic reference method for assessing the vitamin A value of plant foods Am. J. Clinical Nutrition, September 1, 2001; 74(3): 348 - 355. [Abstract] [Full Text]

				C. R Gale, H. E Ashurst, H. J Powers, and C. N Martyn Antioxidant vitamin status and carotid atherosclerosis in the elderly Am. J. Clinical Nutrition, September 1, 2001; 74(3): 402 - 408. [Abstract] [Full Text]

				M. van Lieshout, C. E West, Muhilal, D. Permaesih, Y. Wang, X. Xu, R. B van Breemen, A. F. Creemers, M. A Verhoeven, and J. Lugtenburg Bioefficacy of {beta}-carotene dissolved in oil studied in children in Indonesia Am. J. Clinical Nutrition, May 1, 2001; 73(5): 949 - 958. [Abstract] [Full Text] [PDF]

				P. Wallstrom, E. Wirfalt, P. H Lahmann, B. Gullberg, L. Janzon, and G. Berglund Serum concentrations of {beta}-carotene and {{alpha}}-tocopherol are associated with diet, smoking, and general and central adiposity Am. J. Clinical Nutrition, April 1, 2001; 73(4): 777 - 785. [Abstract] [Full Text] [PDF]

				M. Chopra, M. E. O'Neill, N. Keogh, G. Wortley, S. Southon, and D. I. Thurnham Influence of Increased Fruit and Vegetable Intake on Plasma and Lipoprotein Carotenoids and LDL Oxidation in Smokers and Nonsmokers Clin. Chem., November 1, 2000; 46(11): 1818 - 1829. [Abstract] [Full Text] [PDF]

				H. Wiseman, J. D O'Reilly, H. Adlercreutz, A. I Mallet, E. A Bowey, I. R Rowland, and T. A. Sanders Isoflavone phytoestrogens consumed in soy decrease F2-isoprostane concentrations and increase resistance of low-density lipoprotein to oxidation in humans Am. J. Clinical Nutrition, August 1, 2000; 72(2): 395 - 400. [Abstract] [Full Text] [PDF]

				M. Chopra, P. E.E. Fitzsimons, J. J. Strain, D. I. Thurnham, and A. N. Howard Nonalcoholic Red Wine Extract and Quercetin Inhibit LDL Oxidation without Affecting Plasma Antioxidant Vitamin and Carotenoid Concentrations Clin. Chem., August 1, 2000; 46(8): 1162 - 1170. [Abstract] [Full Text] [PDF]

				L T McGrath, P Mallon, L Dowey, B Silke, E McClean, M McDonnell, A Devine, S Copeland, and S Elborn Oxidative stress during acute respiratory exacerbations in cystic fibrosis Thorax, June 1, 1999; 54(6): 518 - 523. [Abstract] [Full Text]

				J C Baker, W S Tunnicliffe, R C Duncanson, and J G Ayres Dietary antioxidants and magnesium in type 1 brittle asthma: a case control study Thorax, February 1, 1999; 54(2): 115 - 118. [Abstract] [Full Text]

				C.J. Foy, A.P. Passmore, M.D. Vahidassr, I.S. Young, and J.T. Lawson Plasma chain-breaking antioxidants in Alzheimer's disease, vascular dementia and Parkinson's disease QJM, January 1, 1999; 92(1): 39 - 45. [Abstract] [Full Text] [PDF]

				M. M. Van Duijn, J. Van der Zee, J. VanSteveninck, and P. J. A. Van den Broek Ascorbate Stimulates Ferricyanide Reduction in HL-60 Cells through a Mechanism Distinct from the NADH-dependent Plasma Membrane Reductase J. Biol. Chem., May 29, 1998; 273(22): 13415 - 13420. [Abstract] [Full Text] [PDF]

				K M Silvers, A T Gibson, J M Russell, and H J Powers Antioxidant activity, packed cell transfusions, and outcome in premature infants Arch. Dis. Child. Fetal Neonatal Ed., May 1, 1998; 78(3): 214F - 219. [Abstract] [Full Text]

				S. Zino, M. Skeaff, S. Williams, and J. Mann Randomised controlled trial of effect of fruit and vegetable consumption on plasma concentrations of lipids and antioxidants BMJ, June 21, 1997; 314(7097): 1787 - 1787. [Abstract] [Full Text]

				Y. I. Zhu, W.-C. Hsieh, R. S. Parker, L. A. Herraiz, J. D. Haas, J. E. Swanson, and D. A. Roe Evidence of a Role for Fat-Free Body Mass in Modulation of Plasma Carotenoid Concentrations in Older Men: Studies with Hydrodensitometry J. Nutr., February 1, 1997; 127(2): 321 - 326. [Abstract] [Full Text]

				H A W Neil, L Roe, R J P Godlee, J W Moore, G M G Clark, J Brown, M Thorogood, I M Stratton, T Lancaster, D Mant, et al. Randomised trial of lipid lowering dietary advice in general practice: the effects on serum lipids, lipoproteins, and antioxidants BMJ, March 4, 1995; 310(6979): 569 - 573. [Abstract] [Full Text]