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A fast and sensitive high-performance liquid chromatographic (HPLC) method for the determination of vitamin A (all-trans retinol) in 50 microliter human tear fluid is described. After deproteinization with ethanol and extraction with n-hexane, retinol is separated from the extract on a straight-phase HPLC column and detected fluorometrically. Vitamin A can be determined in concentrations as low as 0.4 ng/ml. A single analysis can be completed in 12 min while the analysis of a series of 60 samples takes about 8 h. The within-assay and between-assay coefficients of variation were 3.1 and 4.2% resp. The between-assay analytical recovery of retinol added to tear fluid was 97.4 +/- 6.0% (mean +/- SD). Retinol was determined in tear fluid of nine adult volunteers. The amounts observed ranged from less than 0.4 - 10.6 ng/ml.
A J Speek, and
E J van Agtmaal, and
S Saowakontha, and
W H Schreurs, and
N J van Haeringen
November 2004,
Journal of agricultural and food chemistry,
A J Speek, and
E J van Agtmaal, and
S Saowakontha, and
W H Schreurs, and
N J van Haeringen
July 1991,
Analytical biochemistry,
A J Speek, and
E J van Agtmaal, and
S Saowakontha, and
W H Schreurs, and
N J van Haeringen
February 1984,
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A J Speek, and
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N J van Haeringen
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A J Speek, and
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N J van Haeringen
January 1983,
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A J Speek, and
E J van Agtmaal, and
S Saowakontha, and
W H Schreurs, and
N J van Haeringen
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A J Speek, and
E J van Agtmaal, and
S Saowakontha, and
W H Schreurs, and
N J van Haeringen
July 1980,
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A J Speek, and
E J van Agtmaal, and
S Saowakontha, and
W H Schreurs, and
N J van Haeringen
July 2003,
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A J Speek, and
E J van Agtmaal, and
S Saowakontha, and
W H Schreurs, and
N J van Haeringen
May 1987,
Analytical biochemistry,
A J Speek, and
E J van Agtmaal, and
S Saowakontha, and
W H Schreurs, and
N J van Haeringen
January 1987,
Documenta ophthalmologica. Advances in ophthalmology,
