Saba Alessandro, Raffaelli Andrea, Cupisti Adamasco, Petri Antonella, Marcocci Claudio, Salvadori Piero
Dipartimento di Chimica e Chimica Industriale, Università di Pisa, Via Risorgimento 35, 56126 Pisa, Italy.
J Mass Spectrom. 2009 Apr;44(4):541-8. doi: 10.1002/jms.1530.
A previously reported method for the assessment of the ratio of tetrahydrocortisol (THF) + allo-tetrahydrocortisol (A-THF) to tetrahydrocortisone (THE) by HPLC-MS-MS has been significantly improved, in order to increase either ruggedness and reliability. That was achieved by the introduction of an on-line sample cleanup stage, which made use of a perfusion column as a solid phase microextraction (SPE) cartridge. The set of analytes was expanded, by introducing cortisol and cortisone, whose ratio supply additional diagnostic information. The response factors of both THF and A-THF has been checked, resulting almost identical, as well as the influence of the matrix on the calibration curves which, although different for water and urine, had similar effect on the ratios of interest. As a consequence, the calibration solutions can be prepared in pure water. The influence of several different storage procedures has also been tested, resulting in no substantial effect on the final result. Finally, the improved method has been used to run real samples from healthy volunteers, with satisfactory results.
一种先前报道的通过高效液相色谱-串联质谱法（HPLC-MS-MS）评估四氢皮质醇（THF）+表四氢皮质醇（A-THF）与四氢可的松（THE）比值的方法已得到显著改进，以提高耐用性和可靠性。这是通过引入在线样品净化步骤实现的，该步骤使用灌注柱作为固相微萃取（SPE）柱。通过引入皮质醇和可的松扩大了分析物的范围，它们的比值提供了额外的诊断信息。已检查了THF和A-THF的响应因子，结果几乎相同，还检查了基质对校准曲线的影响，虽然水和尿液的影响不同，但对感兴趣的比值有类似影响。因此，可以在纯水中制备校准溶液。还测试了几种不同储存程序的影响，结果对最终结果没有实质性影响。最后，使用改进后的方法对健康志愿者的实际样本进行检测，结果令人满意。
