A robust and rapid liquid chromatography tandem mass spectrometric method for the quantitative analysis of 5-azacytidine.
A robust and rapid liquid chromatography tandem mass spectrometric method for the quantitative analysis of 5-azacytidine.
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DOI：
10.1002/bmc.3562
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发表时间：
2016-03
期刊：
影响因子：
--
通讯作者：
Rudek MA
中科院分区：
文献类型：
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作者：
Anders NM;Wanjiku TM;He P;Azad NS;Rudek MA
DNA甲基转移酶抑制剂5-氮胞苷正在作为治疗各种实体肿瘤的口服制剂进行临床评估。建立了一种灵敏、可靠的LC-MS/MS定量测定5-氮胞苷的方法，用于药代动力学研究。采用OASIS MCX离子交换固相萃取96孔板从血浆中提取目标药物。色谱柱为YMC J‘SphereM80 C18柱，流动相为甲醇-水-甲酸(15：85：0.1，v/v/v)，总分析时间为7分钟。采用AB Sciex 5500三极四极质谱计，正电喷雾电离模式，对5-氮胞苷进行检测。测定范围为5~500 ng/mL，准确度为97.8~109.1%，精密度(%CV≤为9.8%)。用四氢尿苷稳定血/血浆样品中的5-氮胞苷。随着四氢尿苷的加入，5-氮胞苷在-70°C的长期冰冻血浆稳定性已被测定至少323天。用该方法测定了一名癌症患者每日口服300 mg的5-氮胞苷的血浆总浓度。
The DNA methyltransferase inhibitor 5-azacytidine is being evaluated clinically as an oral formulation to treat various solid tumors. A sensitive, reliable method was developed to quantitate 5-azacytidine using LC-MS/MS to perform detailed pharmacokinetic studies. The drug of interest was extracted from plasma using Oasis MCX ion exchange solid-phase extraction 96-well plates. Chromatographic separation was achieved with an YMC J'sphere M80 C18 column and isocratic elution with a methanol-water-formic acid (15:85:0.1, v/v/v) mobile phase over a 7 minute total analytical run time. An AB Sciex 5500 triple quadrupole mass spectrometer operated in positive electrospray ionization mode was used for the detection of 5-azacytidine. The assay range was 5 to 500 ng/mL and proved to be accurate (97.8-109.1%) and precise (%CV ≤9.8%). Tetrahydrouridine was used to stabilize 5-azacytidine in blood/plasma samples. With the addition of tetrahydrouridine, long-term frozen plasma stability for 5-azacytidine at -70°C has been determined for at least 323 days. The method was applied for the measurement of total plasma concentrations of 5-azacytidine in a cancer patient receiving a 300 mg oral daily dose.