Tamoxifen (TAM) is the choice of a drug approved by the Food and Drug Administration (FDA) for the treatment of estrogen-positive receptor (ER+) breast cancer. Sulphoraphane (SFN), a natural plant antioxidant compound, also acts on estrogen-positive breast cancer receptor. Thus, a combination of TAM with SFN is preferred as it helps to minimize the drug-related toxicity and increases the therapeutic efficacy by providing synergistic anticancer effects of both drugs. In the present study, a new simple, sensitive, precise, and selective UPLC-MS/MS method was developed for the simultaneous quantification of tamoxifen and sulphoraphane using propranolol as an internal standard (IS) in rat plasma. Chromatographic separation was achieved on reverse phase Acquity UPLC BEH C¹⁸ column (50 mm × 2.1 mm, i.d., 1.7 μm) with an isocratic mobile phase composed of solvent A (0.1% formic acid in acetonitrile) and B (0.1% formic acid in water) (80:20, v/v) at a flow-rate of 0.4 mL/min. The detection and quantification of analytes was performed on Waters Zspray^TM Xevo TQD using selected-ion monitoring operated under a positive electrospray ionization mode. The transitions were m/z = 372.0 [M+H]⁺ → 71.92 for tamoxifen, m/z = 177.9 [M+H]⁺ → 113.9 for sulphoraphane and m/z = 260.3 [M+H]⁺ → 116.1 for propranolol. The method was linear over the concentration range of 8–500 ng/mL (r² = 0.9996) for tamoxifen, 30–2000 ng/mL (r² = 0.9998) for sulphoraphane with insignificant matrix effect and high extraction recovery on spiked quality control (QC) samples. The intra- and inter-batch precisions and accuracy were within the acceptable limits, and both the analytes were found to be stable throughout the short term, long term and freeze thaw stability studies. The validated method was successfully applied for the simultaneous estimation of TAM and SFN in an oral pharmacokinetic study in female Wistar rats. This developed UPLC-MS/MS method could be a valuable tool for future pharmacokinetic interaction, therapeutic drug monitoring and pharmacokinetic characterization of novel formulations.

他莫昔芬（TAM）是经美国食品药品监督管理局（FDA）批准用于治疗雌激素阳性受体（ER +）乳腺癌的药物的选择。Sulphoraphane（SFN）是一种天然植物抗氧化剂，也可作用于雌激素阳性乳腺癌受体。因此，TAM与SFN的组合是优选的，因为其通过提供两种药物的协同抗癌作用有助于最小化与药物相关的毒性并提高治疗功效。在本研究中，开发了一种新的简单，灵敏，精确和选择性的UPLC-MS / MS方法，用于在大鼠血浆中使用普萘洛尔作为内标（IS）同时定量他莫昔芬和萝卜硫烷。在反相Acquity UPLC BEH C ¹⁸上完成色谱分离色谱柱（50 mm×2.1 mm，内径，1.7μm），由溶剂A（乙腈中的0.1％甲酸）和B（水中0.1％的甲酸）（80:20，v / v）组成的等度流动相流速为0.4 mL / min。使用在正电喷雾电离模式下运行的选择离子监测，在Waters Zspray ^TM Xevo TQD上进行分析物的检测和定量。他莫昔芬的转变为m / z  = 372.0 [M + H] ⁺ →71.92，对于萝卜硫烷，m / z  = 177.9 [M + H] ⁺ →113.9，m / z  = 260.3 [M + H] ⁺→普萘洛尔为116.1。 他莫昔芬的浓度范围为8–500 ng / mL（r ² = 0.9996），线性范围为30–2000 ng / mL（r ² = 0.9998）对萝卜硫烷具有微不足道的基质效应和对加标质量控制（QC）样品的高提取回收率。批内和批间的精度和准确度均在可接受的范围内，并且在短期，长期和冻融稳定性研究中，两种分析物均稳定。经验证的方法已成功应用于雌性Wistar大鼠的口服药代动力学研究中，同时估算TAM和SFN。这种开发的UPLC-MS / MS方法可能是未来药代动力学相互作用，治疗药物监测和新型制剂药代动力学表征的有价值的工具。