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Rapid and Simple Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) Method for Simultaneous Quantifications of Triazole Antifungals in Human Serum
Mycopathologia ( IF 3.6 ) Pub Date : 2021-01-03 , DOI: 10.1007/s11046-020-00514-0
Hansraj Choudhary 1 , Shreya Singh 1 , Rachna Singh 2 , Ritesh Agarwal 3 , Harsimran Kaur 1 , Anup Ghosh 1 , Arunaloke Chakrabarti 1 , Shivaprakash M Rudramurthy 1
Affiliation  

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:

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。

更新日期:2021-01-03
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