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Tocopherols in human milk: analytical method using high-performance liquid chromatography.

C J Lammi-Keefe

Journal of Pediatric Gastroenterology and Nutrition

|November 1, 1986

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This study details a method for quantifying tocopherols, essential vitamin E compounds, in human milk using high-performance liquid chromatography (HPLC). The validated technique ensures accurate measurement of key tocopherol isomers like alpha- and gamma-tocopherol.

Area of Science:

Nutritional Biochemistry
Analytical Chemistry

Background:

Tocopherols, a group of vitamin E compounds, are crucial antioxidants found in human milk.
Accurate quantification of tocopherols is essential for assessing infant nutrition and milk quality.

Purpose of the Study:

To develop and validate a normal-phase high-performance liquid chromatography (HPLC) method for determining tocopherol isomers in human milk.
To enable the separation and quantification of alpha- and gamma-tocopherol, the primary tocopherol forms in human milk.

Main Methods:

Lipid extraction from human milk followed by direct injection into the HPLC system.
Utilized a Varian Model 5000 Liquid Chromatograph with an Alltech-NH2 column.
Employed n-hexane:2-propanol (98:2) as mobile phase at a flow rate of 2.0 ml/min, with detection at 290 nm.

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Main Results:

Successful chromatographic separation of all four tocopherol isomers (alpha, beta, gamma, delta).
Quantification of alpha- and gamma-tocopherol, the predominant isomers in human milk, was achieved.
The method demonstrated good recovery rates, reproducibility, and precision.

Conclusions:

The described HPLC method provides a reliable and efficient means for analyzing tocopherols in human milk.
This technique facilitates accurate assessment of vitamin E status in breastfed infants.