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Alpha-pinene is a monoterpene found in the oil of coniferous trees and has a wide variety of applications. Alpha-pinene oxide (APO) is a potential reactive metabolite of alpha-pinene in rodents. The objective of this work is to validate a gas chromatography–mass spectrometry method to quantitate APO in rat and mouse blood and mammary glands in support of studies investigating the toxicity and toxicokinetic behavior of alpha-pinene. The method was validated in male Sprague Dawley rat blood over the concentration range of 5–250 ng/mL. Matrix standard curves were linear (r ≥ 0.99), and accuracy (percent relative error, %RE) was ≤±15% for standards at all levels. Intra- and interday precision (percent relative standard deviation, %RSD) and accuracy (%RE) were evaluated at three concentration levels (10, 50 and 200 ng/mL) and were ≤6.3% and ≤±5.4%, respectively. The limit of detection, determined from the SD of the limit of quantitation (5 ng/mL), was 1.06 ng/mL. Standards as high as 25,000 ng/mL could be accurately quantified after diluting to the validated range (%RE ≤ ±7.1%; %RSD ≤ 5.8%). APO was stable in rat blood for at least 70 days in frozen storage (−80°C). APO could accurately be quantified in male and female Hsd:Sprague Dawley® SD® rat and B6C3F1 mouse blood (mean %RE ≤ ±5.3%; %RSD ≤ 7.8%) and female B6C3F1 and Sprague Dawley rat mammary glands (mean %RE ≤ ±14.6%; %RSD ≤ 8.1%) using a primary matrix standard curve. These results demonstrate that the method is suitable for the analysis of APO in rodent blood and mammary glands generated from toxicokinetic and toxicology studies.

Development and Validation of an Analytical Method for Quantitation of Alpha-Pinene Oxide in Rodent Blood and Mammary Glands by GC–MS