An improved HPLC method for determination of carotenoids in human serum

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 824; no. 1; pp. 99 - 106
• Main Authors: Rajendran, V., Pu, Y.S., Chen, B.H.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 25.09.2005; Elsevier Science
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; beta Carotene - analogs & derivatives; beta Carotene - blood; beta Carotene - chemistry; Biological and medical sciences; Carotenoids; Carotenoids - blood; Carotenoids - chemistry; Chromatography, High Pressure Liquid - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; HPLC; Human serum; Humans; Lutein - blood; Lutein - chemistry; Medical sciences; Pharmacology. Drug treatments; Reproducibility of Results; Stereoisomerism; Xanthophylls; Zeaxanthins; Human serum; HPLC; Carotenoids; Human; Biological fluid; HPLC chromatography; Serum; Carotenoid; Quantitative analysis
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2005.07.004

An HPLC method was developed to determine the various carotenoids in human serum. A C-30 column and a mobile phase of 100% methanol (A) and 100% methylene chloride (B) with the following gradient elution were used: 90% A and 10% B in the beginning, maintained for 5 min, decreased to 78% A at 15 min, 62% A at 30 min, 52% A at 40 min, 41% A at 50 min, 38% A at 55 min, maintained for 3 min, and returned to 100% A at 65 min. A total of 21 carotenoids, including all- trans forms of lutein, zeaxanthin, α-cryptoxanthin, β-cryptoxanthin, α-carotene, β-carotene and lycopene, as well as their 14 cis-isomers were resolved within 51 min at a flow rate of 1.0 mL/min and detection at 476 nm. all- trans-β-Carotene was found to be present in highest amount (256.3–864.2 ng/mL), followed by all- trans-lycopene (64.4–569.2 ng/mL), all- trans-lutein (137.9–450.3 ng/mL), all- trans-α-cryptoxanthin (55.7–188.2 ng/mL), all- trans-β-cryptoxanthin (43.1–134.5 ng/mL), all- trans-α-carotene (20.0–122.1 ng/mL) and all- trans-zeaxanthin (9.1–21.3 ng/mL). Similar trend was observed for cis-isomers of carotenoids.