Skagg's School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, San Diego, CA 92093-0831, USA.
Ther Drug Monit. 2013 Apr;35(2):203-8. doi: 10.1097/FTD.0b013e31827fb72b.
Previously published methods for determination of efavirenz (EFV) in human dried blood spots (DBS) use costly and complex liquid chromatography/mass spectrometry. We describe the validation and evaluation of a simple and inexpensive high-performance liquid chromatography method for EFV quantification in human DBS and dried plasma spots (DPS), using ultraviolet detection appropriate for resource-limited settings.
One hundred microliters of heparinized whole blood or plasma were spotted onto blood collection cards, dried, punched, and eluted. Eluates are injected onto a C-18 reversed phase high-performance liquid chromatography column. EFV is separated isocratically using a potassium phosphate and acetonitrile mobile phase. Ultraviolet detection is at 245 nm. Quantitation is by use of external calibration standards. Following validation, the method was evaluated using whole blood and plasma from HIV-positive patients undergoing EFV therapy.
Mean recovery of drug from DBS is 91.5%. The method is linear over the validated concentration range of 0.3125-20.0 μg/mL. A good correlation (Spearman r = 0.96) between paired plasma and DBS EFV concentrations from the clinical samples was observed, and hematocrit level was not found to be a significant determinant of the EFV DBS level. The mean observed C DBS/C plasma ratio was 0.68. A good correlation (Spearman r = 0.96) between paired plasma and DPS EFV concentrations from the clinical samples was observed. The mean percent deviation of DPS samples from plasma samples is 1.68%.
Dried whole blood spot or dried plasma spot sampling is well suited for monitoring EFV therapy in resource-limited settings, particularly when high sensitivity is not essential.
先前发表的用于测定人干血斑（DBS）中依非韦伦（EFV）的方法使用昂贵且复杂的液相色谱/质谱法。我们描述了一种简单且廉价的高效液相色谱法，用于在资源有限的环境中进行 EFV 定量检测，该方法使用紫外检测适用于 DBS 和干血浆斑（DPS）中的 EFV 定量。
将 100μL 肝素化全血或血浆点样到采血卡上，干燥，打孔，洗脱。洗脱液注入 C-18 反相高效液相色谱柱。EFV 采用磷酸盐和乙腈流动相进行等度洗脱。紫外检测波长为 245nm。定量采用外标校准。验证后，该方法用于接受 EFV 治疗的 HIV 阳性患者的全血和血浆进行评估。
从 DBS 中药物的平均回收率为 91.5%。该方法在 0.3125-20.0μg/mL 的验证浓度范围内呈线性关系。从临床样本中观察到配对血浆和 DBS EFV 浓度之间具有良好的相关性（Spearman r=0.96），并且未发现红细胞压积水平对 EFV DBS 水平有显著影响。观察到的平均 C DBS/C 血浆比为 0.68。从临床样本中观察到配对血浆和 DPS EFV 浓度之间具有良好的相关性（Spearman r=0.96）。DPS 样本相对于血浆样本的平均偏差百分比为 1.68%。
干全血斑或干血浆斑采样非常适合资源有限的环境中监测 EFV 治疗，尤其是在不需要高灵敏度的情况下。
