建立了测定大鼠血浆中 N6-羟苄腺苷的超高效液相色谱-四极杆飞行时间质谱（ UPLC-QTOF-MS）分析方法，并应用于 N6-羟苄腺苷药代动力学研究。采用 Agilent ZORBAX SB-C18色谱柱（150 mm×3 mm，1.8μm）对目标物分离，以0.2％（ v／v）甲酸水溶液和乙腈为流动相进行梯度洗脱；以6-苄氨基嘌呤作为内标，在电喷雾离子源正离子模式下进行定量分析。结果表明：N6-羟苄腺苷在0.625～160 ng／mL 的范围内线性关系良好（ r＞0.99）；方法的回收率在88.41％～108.26％之间；检出限达到0.1 ng／mL；日内和日间准确度（以 RE 计，RE＝（测定浓度-加标浓度）／加标浓度×100％）均在±15％之内，精密度（以 RSD 计）均小于6％。该方法选择性好，灵敏度高，重现性好，结果准确，可用于 N6-羟苄腺苷的血药浓度监测及药代动力学研究。
An ultra performance liquid chromatography-quadrupole time of flight mass spec-trometry( UPLC-QTOF-MS)method was developed and validated for the determination of N 6-( 4-hydroxybenzyl ) adenine riboside and its pharmacokinetics in rat plasma. The chromato-graphic conditions were optimized. The separation was performed on an Agilent ZORBAX SB-C18 column(150 mm×3 mm,1. 8 μm)with a gradient elution of 0. 2%(v/v)formic acid aque-ous solution and acetonitrile as the mobile phases at a flow rate of 0. 35 mL/min. The detection was accomplished in positive mode with electrospray ionization( ESI)by UPLC-QTOF-MS,and 6-benzylamino purine was used as the internal standard( IS). The results showed that the linear range of calibration curve was 0. 625-160 ng/mL for N6-(4-hydroxybenzyl)adenine riboside in rat plasma with the correlation coefficient more than 0. 99. The recoveries were 88. 41% -108. 26%. The limit of detection was 0. 1 ng/mL. The intra-day and inter-day precisions( RSDs) were less than 6%,and intra-day and inter-day accuracies( REs,RE=( measured concentration-spiked concentration)/spiked concentration×100%)were less than ±15%. The method is rap-id,sensitive and accurate for the quantitation of N6-(4-hydroxybenzyl)adenine riboside,which can be used for the study of pharmacokinetics of N6-(4-hydroxybenzyl)adenine riboside.
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