快速简便反相高效液相色谱法（RP-HPLC）同时测定人血清中三唑类抗真菌药物的含量。

Rapid and Simple Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) Method for Simultaneous Quantifications of Triazole Antifungals in Human Serum.

机构信息

Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.

Department of Microbial Biotechnology, Panjab University, Chandigarh, India.

出版信息

Mycopathologia. 2021 Mar;186(1):27-39. doi: 10.1007/s11046-020-00514-0. Epub 2021 Jan 3.

DOI:10.1007/s11046-020-00514-0

PMID:33389486

Abstract

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 C 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.

摘要

目的

开发并验证一种一步法、快速且简单的反相高效液相色谱（HPLC）方法，用于同时测定血清/血浆中的伏立康唑（VCZ）、泊沙康唑（POSA）和伊曲康唑（ITC）。

方法

通过向无药物血清中加入不同浓度的三唑类药物来制备校准标准品（CS）和质控样品。采用 C 柱，冷乙腈提取后等度洗脱进行 HPLC 分析。该标准化方法在 2693 例患者的血清/血浆样本中进行了测试。

结果

CS 中 ITC、VCZ 和 POSA 的线性与名义浓度成正比（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，定量下限（mg/L）分别为 0.5、0.5 和 0.25。精密度和准确度均在可接受范围内，平均回收率为 100%。未发现内源性物质和其他抗菌化合物的干扰。在临床样本中，VCZ 的治疗范围达到 39.9%。而 ITC 和 POSA 的样本中，分别有 61.1%和 44%处于亚治疗范围。