Toxicology Unit, Department of Clinical Biochemistry, King's College Hospital NHS Foundation Trust, London, United Kingdom.
Ther Drug Monit. 2011 Dec;33(6):735-41. doi: 10.1097/FTD.0b013e3182381bb1.
Itraconazole and posaconazole are used in the prevention and treatment of invasive fungal infections. However, the oral bioavailability of both compounds varies widely, and dose-serum concentration relationships are poorly defined for these analytes. The aim of this work was to develop and validate a simple assay that could be implemented in most laboratories for the purpose of therapeutic drug monitoring.
Calibrators (n = 7) and internal quality control solutions (n = 3) were prepared in pooled human serum. Sample (100 μL), internal standard solution (25 μL), Tris solution (2 mol/L; pH 10.6), and extraction solvent (methyl tert-butyl ether, 600 μL) were vortex mixed and centrifuged. The solvent layer was removed and evaporated to dryness and the residue reconstituted in water:methanol (1 + 3, 50 μL). A portion (5 μL) of the reconstituted extract was analyzed using a 3-μm Gemini C6 phenyl column with fluorescence detection (excitation 260 nm, emission 350 nm). The method was used to measure itraconazole and hydroxyitraconazole, or posaconazole, in serum samples taken 1-2 hours before the next dose, from patients forming part of a study into management and diagnostic strategies for invasive aspergillosis.
Response was linear over the calibration ranges. Accuracy and imprecision were 92-111.4% and 3.2-13.4% (relative standard deviation), respectively. No interferences were noted. There was a good agreement with nominal values of each analyte in an external quality assessment scheme. In patients prescribed either 400 mg/d of itraconazole (n = 46) or 600-800 mg/d of posaconazole (n = 28) only 24% and 7% of samples, respectively, had serum itraconazole or posaconazole concentrations above the target threshold suggested in published guidelines.
A simple, sensitive high-performance liquid chromatographic method has been developed for the analysis of itraconazole, hydroxyitraconazole, and posaconazole in serum/plasma. Few of the samples measured from patients participating in the clinical study attained concentrations of the drug/metabolite in serum that have been recommended for effective antifungal therapy.
伊曲康唑和泊沙康唑用于预防和治疗侵袭性真菌感染。然而，这两种化合物的口服生物利用度差异很大，并且这些分析物的剂量-血清浓度关系也未得到很好的定义。本研究的目的是开发和验证一种简单的测定方法，以便在大多数实验室中用于治疗药物监测。
校准品（n=7）和内部质量控制溶液（n=3）用人混合血清制备。取样品（100μL）、内标溶液（25μL）、三（羟甲基）氨基甲烷溶液（2mol/L，pH 10.6）和萃取溶剂（甲基叔丁基醚，600μL），涡旋混合并离心。去除溶剂层并蒸发至干燥，残留物用 50μL 水：甲醇（1+3）重新溶解。用 3μm Gemini C6 苯基柱和荧光检测（激发波长 260nm，发射波长 350nm）分析 5μL 重新溶解的提取物的一部分。该方法用于测量来自正在进行侵袭性曲霉菌病管理和诊断策略研究的患者的下一次给药前 1-2 小时采集的血清样本中的伊曲康唑和羟基伊曲康唑或泊沙康唑。
校准范围内的响应呈线性。准确度和精密度分别为 92-111.4%和 3.2-13.4%（相对标准偏差）。未观察到干扰。在外部质量评估计划中，每个分析物的标称值与实际值吻合良好。在服用 400mg/d 伊曲康唑（n=46）或 600-800mg/d 泊沙康唑（n=28）的患者中，分别只有 24%和 7%的样本的血清伊曲康唑或泊沙康唑浓度高于已发表指南建议的目标阈值。
已开发出一种简单、灵敏的高效液相色谱法，用于分析血清/血浆中的伊曲康唑、羟基伊曲康唑和泊沙康唑。从参与临床研究的患者中测量的少数样本达到了有效抗真菌治疗所需的药物/代谢物血清浓度。
