Purpose

To develop and validate a one-step, rapid and simple reversed-phase high-performance liquid chromatography (HPLC)-based protocol for the simultaneous measurement of voriconazole (VCZ), posaconazole (POSA), itraconazole (ITC) in serum/plasma.

Methods

Calibration standards (CS) and quality control samples were prepared in drug-free serum by spiking with the triazoles at different concentrations. HPLC was performed with C₁₈ column, isocratic mobile phase after extraction with cold acetonitrile. The standardized method was tested in 2693 patients’ serum/plasma samples.

Results

Linearity of CS for ITC, VCZ and POSA was proportional to the nominal concentration (correlation coefficient > 0.999). Limit of detection (mg/L) for ITC, VCZ and POSA was 0.25, 0.25 and 0.125, respectively. The lower limit of quantification (mg/L) for ITC, VCZ and POSA was 0.5, 0.5 and 0.25, respectively. Precision and accuracy were in acceptable range with 100% average percentage recovery. No interferences from endogenous substances and other antimicrobial compounds were noted. In clinical samples, the therapeutic range achieved for VCZ was 39.9%. Whereas, 61.1% and 44% of samples with ITC and POSA, respectively, were in the sub-therapeutic range.

Conclusion

We developed a rapid and simple HPLC method to quantify common triazoles in a single chromatographic run allowing simultaneous measurement of different antifungals in a small volume of serum/plasma. Thus, therapeutic drug monitoring requests can be processed in one run without changing the protocol parameters, column or column conditioning thereby improving turnaround time.

中文翻译：

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快速简单的反相高效液相色谱 (RP-HPLC) 方法同时定量人血清中的三唑类抗真菌药

目的

开发和验证基于反相高效液相色谱( HPLC) 的一步式、快速和简单的方案，用于同时测量血清/血浆中的伏立康唑 (VCZ)、泊沙康唑 (POSA) 和伊曲康唑 (ITC)。

方法

通过添加不同浓度的三唑，在无药物血清中制备校准标准品 (CS) 和质量控制样品。HPLC 使用 C ₁₈柱，等度流动相，用冷乙腈萃取后进行。标准化方法在 2693 名患者的血清/血浆样本中进行了测试。

结果

ITC、VCZ 和 POSA 的 CS 线性与标称浓度成正比（相关系数 > 0.999）。ITC、VCZ 和 POSA 的检测限 (mg/L) 分别为 0.25、0.25 和 0.125。ITC、VCZ 和 POSA 的定量下限 (mg/L) 分别为 0.5、0.5 和 0.25。精密度和准确度在可接受的范围内，平均回收率为 100%。没有发现来自内源性物质和其他抗菌化合物的干扰。在临床样本中，VCZ 的治疗范围为 39.9%。然而，分别有 61.1% 和 44% 的 ITC 和 POSA 样本处于亚治疗范围。

结论

我们开发了一种快速且简单的 HPLC 方法，可在单次色谱运行中对常见的三唑进行定量，从而可以同时测量少量血清/血浆中的不同抗真菌剂。因此，可以在一次运行中处理治疗药物监测请求，而无需更改协议参数、色谱柱或色谱柱调节，从而缩短周转时间。