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 C(18) column (4.6 x 250 mm, 5 microm) 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 (r(2) > 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 microL 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.
Georgetown University, Lombardi Comprehensive Cancer Center, Washington, DC 20057, USA.
Mikell
M
Paige
Mary S
MS
Saprito
Dorothy A
DA
Bunyan
Y Michael
YM
Shim
D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000379:methods*; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000032:analysis* / Q000302:isolation & purification; D016014:Linear Models; D008168:Lung / Q000737:chemistry* / Q000473:pathology; D008175:Lung Neoplasms / Q000737:chemistry*; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000191:economics / Q000379:methods*
D006893:Hydroxyeicosatetraenoic Acids; C022022:5-hydroxy-6,8,11,14-eicosatetraenoic acid