HPLC quantification of 5‐hydroxyeicosatetraenoic acid in human lung cancer tissues

A simple and cost‐effective HPLC method was established for quantification of 5‐hydroxyeicosatetraenoic acid (5‐HETE) in human lung cancer tissues. 5‐HETE from 27 patients' lung cancer tissues were extracted by solid‐phase extraction and analyzed on a Waters Symmetry C₁₈ column (4.6 × 250 mm, 5 µm) with a mobile phase consisting of methanol, 10 mm ammonium acetate, and 1 m acetic acid (70:30:0.1, v:v:v) at a flow rate of 1.0 mL/min. The UV detection wavelength was set at 240 nm. The calibration curve was linear within the concentration range from 10 to 1000 ng/mL (r² > 0.999, n = 7), the limit of detection was 1.0 ng/mL and the limit of quantitation was 10.0 ng/mL for a 100 µL injection. The relative error (%) for intra‐day accuracy was from 93.14 to 112.50% and the RSD (%) for intra‐day precision was from 0.21 to 2.60% over the concentration range 10–1000 ng/mL. By applying this method, amounts of 5‐HETE were quantitated in human lung cancer tissues from 27 human subjects. The established HPLC method was validated to be a simple, reliable and cost‐effective procedure that can be applied to conduct translational characterization of 5‐HETE in human lung cancer tissues. Copyright © 2009 John Wiley & Sons, Ltd.