Retinol, alpha-tocopherol, lutein/zeaxanthin, beta-cryptoxanthin, lycopene, alpha-carotene, trans-beta-carotene, and four retinyl esters in serum determined simultaneously by reversed-phase HPLC with multiwavelength detection

1994 ◽  
Vol 40 (3) ◽  
pp. 411-416 ◽  
Author(s):  
A L Sowell ◽  
D L Huff ◽  
P R Yeager ◽  
S P Caudill ◽  
E W Gunter
Keyword(s):  
Serum Concentration ◽  
Serum Sample ◽  
Beta Carotene ◽  
Reversed Phase ◽  
Alpha Tocopherol ◽  
Reversed Phase Hplc ◽  
Fasting Serum ◽  
Retinyl Esters ◽  
Carotene Beta

Abstract We describe the use of HPLC with multiwavelength detection to measure retinol, alpha-tocopherol, lutein/zeaxanthin, beta-cryptoxanthin, lycopene, alpha-carotene, trans-beta-carotene, beta-carotene, and the linoleate, oleate, palmitate, and stearate esters of retinol in a single 200-microL serum sample. The method is sensitive enough to detect individual retinyl esters in fasting serum from a nonhyperlipidemic population and requires only 12 min for each sample. Serum concentration ranges and means are reported for retinol, alpha-tocopherol, lutein/zeaxanthin, beta-cryptoxanthin, lycopene, alpha-carotene, trans-beta-carotene, and the sum of the retinyl esters from serum analyses of 3480 participants from several different studies.

Simultaneous determination of vitamins A and E and carotenoids in plasma by reversed-phase HPLC in elderly and younger subjects

1993 ◽  
Vol 39 (11) ◽  
pp. 2229-2234 ◽  
Author(s):  
Z Zaman ◽  
P Fielden ◽  
P G Frost
Keyword(s):  
Simultaneous Determination ◽  
High Performance ◽  
Ammonium Acetate ◽  
Beta Carotene ◽  
High Performance Liquid Chromatographic ◽  
Solvent Mixture ◽  
Reversed Phase ◽  
Alpha Tocopherol ◽  
Elderly Subjects

Abstract A reversed-phase high-performance liquid-chromatographic method for the simultaneous determination of retinol, alpha-tocopherol, alpha-carotene, beta-carotene, cryptoxanthin, lutein/zeaxanthin, and lycopene is described. This method was applied to plasma measurements in healthy young and elderly subjects. The plasma, deproteinized with ethanol, is extracted twice with n-hexane. After evaporation, the residue is dissolved in 50 microL of tetrahydrofuran and made up to 200 microL with ethanol. Samples (50 microL) are injected onto a 250 x 4.6 mm column of 5-microns-particle Spherisorb ODS1 (Phase Separations) that had been equilibrated with solvent mixture A:B (90:10 by vol) [A = 100 mmol/L ammonium acetate in methanol: acetonitrile (80:20 by vol) and B = 100 mmol/L ammonium acetate in water] at 2 mL/min. The analytes are eluted by running a 12-min linear gradient to 100% A; solvent A is then maintained for 10 min. Intrabatch CVs were 2.3%, 3.3%, 2.8%, 3.6%, 3.6%, and 3.0% for retinol, alpha-tocopherol, lutein/zeaxanthin, cryptoxanthin, lycopene, and beta-carotene, respectively. The corresponding interbatch CVs were 4.9%, 5.8%, 12.3%, 6.5%, 8.0%, and 3.4%.

Determination of retinol, alpha-tocopherol, and beta-carotene in serum by liquid chromatography with absorbance and electrochemical detection.

1987 ◽  
Vol 33 (9) ◽  
pp. 1585-1592 ◽  
Author(s):  
W A MacCrehan ◽  
E Schönberger
Keyword(s):  
Electrochemical Detection ◽  
Chromatographic Separation ◽  
Protein Denaturation ◽  
Internal Standard ◽  
Beta Carotene ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Alpha Tocopherol ◽  
Phase Gradient

Abstract We describe a method for the determination of retinol, alpha-tocopherol, and beta-carotene in serum, using a liquid-chromatographic separation with wavelength-programmed ultraviolet/visible absorbance and amperometric electrochemical detection with a glassy carbon electrode. After protein denaturation and addition of an internal standard, tocol, 250-microL samples are twice extracted with hexane. The reversed-phase, gradient-elution chromatographic separation provides baseline resolution of: the all-trans isomer of retinol from the cis isomers, alpha- from gamma-tocopherol, and all-trans-beta-carotene from alpha-carotene and from cis-beta-carotene isomers. The linearity of response and the detection limits for the two detectors for the three analytes are measured. A comparison of the values obtained for serum extracts shows good agreement between the absorbance and electrochemical detectors.

Retinol, alpha-tocopherol, lycopene, and alpha- and beta-carotene simultaneously determined in plasma by isocratic liquid chromatography.

1986 ◽  
Vol 32 (5) ◽  
pp. 874-876 ◽  
Author(s):  
D B Milne ◽  
J Botnen
Keyword(s):  
Liquid Chromatography ◽  
Internal Standard ◽  
Beta Carotene ◽  
Reversed Phase ◽  
Small Sample ◽  
Alpha Tocopherol ◽  
Organic Layer ◽  
Retinyl Acetate ◽  
Mobile Phase Flow Rate ◽  
Short Run

Abstract Retinol, alpha-tocopherol, lycopene, and alpha- and beta-carotene can be simultaneously determined in human plasma by reversed-phase liquid chromatography. Plasma--0.5 mL plus added internal standard, retinyl acetate--is deproteinized with 0.5 mL of ethanol, then extracted with 1.0 mL of petroleum ether. The organic layer is removed and evaporated, the residue is redissolved in 0.25 mL of ethanol, and 8-microL samples are injected into a 60 X 4.6 mm column of Hypersil ODS 3-microns particles at 35 degrees C. An isocratic methanol mobile phase, flow rate 0.9 mL/min, is used for the 9-min run. Retinol and retinyl acetate are monitored at 305 nm, the tocopherols at 292 nm, and the carotenoids at 460 nm. Between-run CVs were 3.1, 6.9, 6.1, and 6.5% for retinol, alpha-tocopherol, lycopene, and beta-carotene, respectively. Small sample requirement, simplicity of extraction, short run time, and good reproducibility make this procedure ideal for clinical or research use.

Separation of beta-carotene and lycopene geometrical isomers in biological samples

1993 ◽  
Vol 39 (5) ◽  
pp. 810-814 ◽  
Author(s):  
W Stahl ◽  
A R Sundquist ◽  
M Hanusch ◽  
W Schwarz ◽  
H Sies
Keyword(s):  
Human Serum ◽  
Biological Samples ◽  
Beta Carotene ◽  
Reversed Phase ◽  
Reversed Phase Hplc ◽  
Trans Isomers ◽  
Geometrical Isomers ◽  
Total Beta

Abstract The analysis of beta-carotene and lycopene, the two predominant carotenoids in human serum and tissues, was extended to the level of geometrical (cis-trans) isomers by using an improved reversed-phase HPLC methodology. We separated five geometrical isomers of beta-carotene and seven of lycopene in human serum and tissues. 13-cis-beta-Carotene was identified as the predominant cisisomer in human serum, contributing about 5% to total beta-carotene. In tissue, however, considerable amounts of 9-cis- and traces of 15-cis-beta-carotene were also detected. In contrast to beta-carotene, the lycopene isomer patterns in human serum and tissues are quite similar.

The Reversed-Phase HPLC Behavior of Retinyl Esters

1989 ◽  
Vol 12 (7) ◽  
pp. 1261-1280 ◽  
Author(s):  
Hubert E. May ◽  
Sung I. Koo
Keyword(s):  
Reversed Phase ◽  
Reversed Phase Hplc ◽  
Retinyl Esters

scholarly journals A Reversed-phase HPLC Study of the Complexation of Benzene Derivatives Guest Molecules with 5,17-bis(N-Tolyliminomethyl)-25,27-dipropoxycalix[4]arene in Acetonitrile–Water Solution

1999 ◽  
Vol 23 (1) ◽  
pp. 60-61
Author(s):  
O. I. Kalchenko ◽  
A. V. Solovyov ◽  
J. Lipkowski ◽  
V. I. Kalchenko
Keyword(s):  
Stability Constants ◽  
Water Solution ◽  
Reversed Phase ◽  
Benzene Derivatives ◽  
Reversed Phase Hplc ◽  
Guest Molecules ◽  
Hplc Study

Stability constants of the host–guest complexes of 5,17-bis( N-tolyliminomethyl)-25,27-dipropoxycalix[4]arene with benzene derivatives were determined by reversed-phase HPLC in acetonitrile–water solution.

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