Desoxyrhapontigenin (DRG, 4-methoxyresveratrol or trans-3,5-dihydroxy-4′-methoxystilbene) is a naturally occurring resveratrol (RES) derivative with a variety of biological activities. To facilitate its further medicinal exploration, a reliable LC–MS/MS method was developed and validated for the quantification of DRG in rat plasma using heavy isotope labelled RES as an internal standard. The ESI was operated in its negative ion mode while DRG and RES were determined by multiple reaction monitoring (MRM) using precursor-to-product ion transitions of m/z 241.1 → 180.8 and m/z 233.0 → 191.0, respectively. This LC–MS/MS method displayed excellent selectivity, sensitivity (LLOQ = 2.5 ng/ml), accuracy (both intra- and interday mean analytical recovery within 100 ± 15%) and precision (both intra- and interday CV < 15%). The mean matrix factors were all within 1.000 ± 0.150 with CV < 15%. The pharmacokinetic profiles of DRG were subsequently examined in Sprague-Dawley rats. Upon intravenous administration (4 or 10 mg/kg), DRG displayed very rapid clearance (Cl = 338 ± 66 or 275 ± 30 ml/min/kg) and short mean residence time (MRT = 12.9 ± 4.7 or 10.4 ± 0.5 min). After oral administration of DRG fully solubilized by 2-hydroxypropyl-β-cyclodextrin (HP-β-CD), the plasma profiles of DRG were highly erratic with a low absolute bioavailability (F < 9.83 ± 5.31%). When DRG was given at a higher dose (50 mg/kg) in suspension form, the F was increased to 24.1 ± 5.6%. The pharmacokinetic comparison among DRG, RES and some of its hydroxyl analogues stilbenes was performed. The information obtained from this study will facilitate further exploration on DRG as well as other RES derivatives.