Ceylan Burhan, Tekkeli Evrim Kepekci, Önal Cem
Institute of Health Sciences, Department of Pharmacognosy and Natural Products Chemistry, Bezmialem Vakif University, Fatih, Istanbul, Turkey.
Faculty of Pharmacy, Department of Analytical Chemistry, Bezmialem Vakif University, Fatih, Istanbul, Turkey.
J Fluoresc. 2022 Jan;32(1):319-325. doi: 10.1007/s10895-021-02848-4. Epub 2021 Nov 19.
In this study, a new, fast and sensitive HPLC method with fluorometric detection was developed for the determination of mesalazine in human plasma and applied to a pharmacokinetic study. Mesalazine was precolumn derivatized with NBD-Cl and the fluorescent derivative was separated on a C18 (150 × 4.6 mm × 2.6 μm) analytical column at 30 ºC using a mobile phase composed of acetonitrile-0.1% o-phosphoric acid in water (70:30, v/v) by isocratic elution with flow rate of 1.0 mL min. The method was based on the measurement of the derivative using fluorescence detection (λ = 280 nm, λ = 325 nm). The retention time of mesalazine is 3.08 ± 0.06 min. Nortriptiline was used as internal standard. This currently developed method was validated according to ICH criteria by evaluating the specificity, linearity, precision, accuracy and robustness. The method was determined to be linear in a concentration range of 0.25-1.5 μg mL with the correlation coefficient of 0.9997. LOD and LOQ were found to be 0.075 and 0.25 μg mL, respectively. Intraday and interday RSD values were less than 5.92%. The plasma concentration-time profile and pharmacokinetic parameters such as AUC, AUC, C, t, t, were calculated according to the assays. The presented method can certainly be used for bioequivalence and bioavailability investigations and routine analysis of the drug in plasma.
本研究建立了一种新型、快速且灵敏的带荧光检测的高效液相色谱法，用于测定人血浆中的美沙拉嗪，并将其应用于药代动力学研究。美沙拉嗪用NBD-Cl进行柱前衍生化，衍生后的荧光产物在C18（150×4.6 mm×2.6 μm）分析柱上于30℃分离，流动相为乙腈-0.1%磷酸水溶液（70:30，v/v），等度洗脱，流速为1.0 mL/min。该方法基于荧光检测（λ = 280 nm，λ = 325 nm）对衍生物进行测定。美沙拉嗪的保留时间为3.08±0.06 min。去甲替林用作内标。根据ICH标准，通过评估特异性、线性、精密度、准确度和稳健性对当前开发的方法进行了验证。该方法在0.25 - 1.5 μg/mL的浓度范围内呈线性，相关系数为0.9997。检测限和定量限分别为0.075和0.25 μg/mL。日内和日间相对标准偏差值均小于5.92%。根据测定结果计算血浆浓度-时间曲线以及药代动力学参数，如AUC、AUC、Cmax、tmax、t1/2等。所提出的方法肯定可用于生物等效性和生物利用度研究以及血浆中药物的常规分析。
