Simultaneous assay of serum vitamin A and vitamin E by high performance liquid chromatography using time‐switched UV and fluorimetric detectors

Abstract

SummaryAn HPLC method utilizing a UV and a fluorimetricdetector linked in series is described. By use of a simple integrator-controlled time-switched relay, analysis of serum vitamins A and E is accomplished on the same chromatogram and at optimum sensitivity for each detector. A single internal standard (retinyl acetate) monitored only by the UV detector permits measurement of both vitamins over a wide linear range. Precision of the assays is satisfactory, both on a within-day and on a day-to-day basis. Recoveries of both vitamins are virtually 1000/0 whilst sensitivity is 2 pg/L (retinol) and 0.05 mg/L (a-tocopherol).

A prospective study of the relationship between serum vitamins A and E and risk of breast cancer

“…During With the availability of a series of some 5,000 serum samples from a new prospective breast cancer study on women resident on the island of Guernsey, it seemed appropriate to re-investigate the problem using a newly developed high performance liquid chromatography (HPLC) method which included butylated hydroxytoluene (BHT) as an antioxidant (Russell et al, 1986). In addition, serum retinol binding protein (RPB), which is a good indicator of nutritional status (Gofferje, 1978;Tyler et al, 1984) Serum retinol and a-tocopherol were measured by HPLC (Russell et al, 1986). Several workers (Chow et al, 1983;Driskell et al, 1985) have shown that the addition of antioxidant at the extraction stage of the analysis prevents the loss of vitamin A, even in frozen stored serum samples.…”

“…Several workers (Chow et al, 1983;Driskell et al, 1985) have shown that the addition of antioxidant at the extraction stage of the analysis prevents the loss of vitamin A, even in frozen stored serum samples. We have also found that with the addition of BHT, both vitamins A and E are stable, in that there is no significant correlation between time in storage and titre (Russell et al, 1986).The RBP was assayed by the Behring LC-Partigen Immunodiffusion Plate method from Hoechst Pharmaceuticals Ltd., Hounslow, UK. After addition of the serum to each well on the assay plates, they were left for 48 h at room temperature before measurement of the diffusion area.…”

Vitamin A, vitamin E and the risk of cervical intraepithelial neoplasia

“…Vitamin E has been little studied in relation to cervix cancer. An inverse relation has been reported in one study (Knekt, 1988) but not another (Heinonen et al, 1987).We have examined serum levels of vitamin A and vitamin E in young women aged Sera were analysed blindly for vitamin A and vitamin E according to the procedure of Russell et al (1986). Antioxidant (BHT) was added prior to extraction to ensure that both vitamins A and E were stable after prolonged storage (Russell et al, 1986).…”

“…We have examined serum levels of vitamin A and vitamin E in young women aged Sera were analysed blindly for vitamin A and vitamin E according to the procedure of Russell et al (1986). Antioxidant (BHT) was added prior to extraction to ensure that both vitamins A and E were stable after prolonged storage (Russell et al, 1986).…”

“…Antioxidant (BHT) was added prior to extraction to ensure that both vitamins A and E were stable after prolonged storage (Russell et al, 1986). A trend in levels across the three groups was examined by the Wilcoxon test for trend (Cuzick, 1985).…”