Holman Miranda Kay, Brown Stacy D, Frempong Dorcas, Puri Ashana, Dinh Steven
Department of Biological Sciences, College of Arts and Sciences, East Tennessee State University, TN 37604, Johnson, USA.
Department of Pharmaceutical Sciences, Bill Gatton College of Pharmacy, East Tennessee State University, TN 37604, Johnson, USA.
J Anal Methods Chem. 2022 Apr 21;2022:7437905. doi: 10.1155/2022/7437905. eCollection 2022.
Cromolyn sodium (CS) is a mast cell stabilizer administered to treat allergic diseases. A topical system would sustain its delivery and may be designed for treatment of atopic dermatitis. Established HPLC protocols for detection of CS are time consuming and intensive, indicating the need for a more streamlined method. This study aimed at developing and validating a sensitive and selective LC-MS method for quantifying CS in skin permeation studies that was less time and resource demanding. The optimized method involved an isocratic mobile phase (10 mM NHHCO, pH 8.0, 90% and ACN, 10%) at a flow rate of 0.25 mL/min. Detection involved direct MS/MS channels with m/z 467.0255 (precursor) and / 379.0517 (fragment) using argon as the collision gas. CS calibrants were prepared in PBS, pH 7.4, and methanol for validation (0.1-2.5 g/mL). To ensure no skin interference, dermatomed porcine skin was mounted on Franz diffusion cells that were analyzed after 24 h. The skin layers were also separated, extracted in methanol, and analyzed using the developed method. Retention time was 1.9 min and 4.1 min in methanol and buffer, respectively. No interfering peaks were observed from the receptor and skin extracts, and linearity was established between 0.1 and 2.5 g/mL. Interday and intraday accuracy and precision were within the acceptable limit of ±20% at the LLOQ and ±15% at other concentrations. Overall, the simplified, validated method showed sensitivity in detecting CS in skin without interference and was applied to demonstrate quantification of drug in skin following 4% cromolyn sodium gel exposure.
色甘酸钠（CS）是一种用于治疗过敏性疾病的肥大细胞稳定剂。一种局部给药系统将持续释放该药物，并且可以设计用于治疗特应性皮炎。已有的用于检测CS的高效液相色谱（HPLC）方法耗时且繁琐，这表明需要一种更简化的方法。本研究旨在开发并验证一种灵敏且具选择性的液相色谱-质谱（LC-MS）方法，用于在皮肤渗透研究中定量CS，该方法对时间和资源的需求较少。优化后的方法采用等度流动相（10 mM NH₄HCO₃，pH 8.0，90%水和10%乙腈），流速为0.25 mL/min。检测采用直接质谱/质谱通道，以氩气作为碰撞气体，母离子m/z为467.0255，子离子m/z为379.0517。CS校准品在pH 7.4的磷酸盐缓冲液（PBS）和甲醇中制备用于验证（0.1 - 2.5 μg/mL）。为确保无皮肤干扰，将猪皮切片安装在Franz扩散池中，24小时后进行分析。还分离了皮肤层，用甲醇提取，并使用所开发的方法进行分析。在甲醇和缓冲液中的保留时间分别为1.9分钟和4.1分钟。在受体和皮肤提取物中未观察到干扰峰，在0.1至2.5 μg/mL之间建立了线性关系。在LLOQ时，日间和日内准确度和精密度在±20%的可接受限度内，在其他浓度下为±15%。总体而言，该简化且经过验证的方法在检测皮肤中的CS时具有灵敏度且无干扰，并被用于证明4%色甘酸钠凝胶暴露后皮肤中药物的定量。
