Louw Vanessa, Brownfoot Fiona, Cluver Catherine, Decloedt Eric, Kellermann Tracy
Division of Clinical Pharmacology, Department of Medicine, Stellenbosch University, Cape Town, South Africa.
Department of Obstetrics and Gynaecology, University of Melbourne, Victoria, Australia; Mercy Perinatal, Mercy Hospital Heidelberg, Victoria, Australia.
J Pharm Biomed Anal. 2023 Oct 25;235:115633. doi: 10.1016/j.jpba.2023.115633. Epub 2023 Aug 14.
Sulfasalazine has been identified as a candidate molecule to be investigated as an intervention to treat preterm pre-eclampsia during pregnancy. However, placental exposure of sulfasalazine and its systemically absorbed metabolite, sulfapyridine, is unknown. A robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated to simultaneously quantitate these analytes in human placenta with an application to a pilot clinical trial. The placental tissue was homogenised using a water:methanol (1:1, v/v) mixture, followed by sample extraction using both protein precipitation and solid phase extraction. Sulfasalazine-d4 and sulfapyridine-d4 were used as internal standards. An Agilent Poroshell EC-C18 (3.0 ×100 mm, 2.7 µm) column was used for chromatographic separation, with gradient elution employed at a flow rate of 0.450 mL/min over a total run time of seven minutes. The mobile phases consisted of water with 0.1% formic acid (mobile phase A) and acetonitrile:methanol (90:10, v/v) with 0.1% formic acid (mobile phase B). A Shimadzu-8040 mass spectrometer was operated in multiple reaction monitoring (MRM) mode using positive electrospray ionisation (ESI). For both analytes, the assay was validated over the range 30-30,000 ng/mL, or 150-150,000 ng/g. During inter-day validations (n = 18), the average accuracies of quality controls ranged from 101.6% to 112.7% with corresponding precisions of 4.4-6.7% for sulfasalazine, and from 97.4% to 108.4%, with corresponding precisions of 3.7-10.0% for sulfapyridine. No significant matrix effects were observed, and the method proved to be sensitive and specific for both analytes. This study presents the first validated analytical method for quantifying sulfasalazine and sulfapyridine in human placenta as part of a pilot clinical trial to generate preliminary data on its pharmacokinetics and efficacy as in intervention for preterm pre-eclampsia.
柳氮磺胺吡啶已被确定为一种有待研究的候选分子，作为孕期治疗早发型子痫前期的干预措施。然而，柳氮磺胺吡啶及其全身吸收的代谢产物磺胺吡啶在胎盘中的暴露情况尚不清楚。我们开发并验证了一种强大的液相色谱 - 串联质谱（LC-MS/MS）方法，用于同时定量人胎盘中的这些分析物，并应用于一项先导性临床试验。胎盘组织用体积比为1:1的水：甲醇混合物匀浆，然后通过蛋白质沉淀和固相萃取进行样品提取。柳氮磺胺吡啶 - d4和磺胺吡啶 - d4用作内标。使用安捷伦Poroshell EC-C18（3.0×100 mm，2.7 µm）色谱柱进行色谱分离，采用梯度洗脱，流速为0.450 mL/min，总运行时间为7分钟。流动相由含0.1%甲酸的水（流动相A）和含0.1%甲酸的乙腈：甲醇（90:10，v/v）（流动相B）组成。岛津 - 8040质谱仪在多反应监测（MRM）模式下运行，采用正电喷雾电离（ESI）。对于这两种分析物，该测定法在30 - 30,000 ng/mL或150 - 150,000 ng/g范围内得到验证。在日间验证（n = 18）期间，质量控制的平均准确度对于柳氮磺胺吡啶为101.6%至112.7%，相应精密度为4.4 - 6.7%，对于磺胺吡啶为97.4%至108.4%，相应精密度为3.7 - 10.0%。未观察到明显的基质效应，该方法对两种分析物均具有灵敏性和特异性。本研究提出了首个经过验证的用于定量人胎盘中柳氮磺胺吡啶和磺胺吡啶的分析方法，作为一项先导性临床试验的一部分，以生成关于其作为早发型子痫前期干预措施的药代动力学和疗效的初步数据。
