Clinical Pharmacology, Sunovion Pharmaceuticals, Marlborough, MA.
Bioanalytical Services, Frontage Laboratories, Exton, PA.
J Appl Lab Med. 2020 Jan 1;5(1):41-53. doi: 10.1373/jalm.2019.029397.
The development of more efficient drug delivery devices for ciclesonide inhalation products requires an ultrasensitive bioanalytical method to measure systematic exposure of ciclesonide (CIC) and its active metabolite desisobutyryl-ciclesonide (des-CIC) in humans.
Serum sample was extracted with 1-chlorobutane. A reversed-phase liquid chromatography coupled with atmospheric pressure photoionization-tandem mass spectrometry (LC-APPI-MS/MS) method was used for quantification of 1-500 pg/mL for both analytes in a 0.500-mL serum. The analysis time was 4.7 min/injection. CIC-d11 and des-CIC-d11 were used as the internal standards.
Calibration curves showed good linearity (r2 > 0.99) for both analytes. This novel method was precise and accurate with interassay precision and accuracy of all within 9.6% CV and ± 4.0% bias for regular QC samples. Extraction recovery was approximately 85% for both analytes. Serum samples are stable for 3 freeze-thaw cycles, 24 h at bench top, and up to 706 days at both -20 °C and -70 °C. This method was successfully used to support a pharmacokinetic (PK) comparison between the inhalation suspensions and an inhalation aerosol of ciclesonide in healthy participants. The method robustness was also supported by the good incurred sample reanalysis reproducibility.
APPI, a highly selective and sensitive ionization source, made possible for quantifying CIC and des-CIC with a lower limit of quantification (LLOQ) of 1 pg/mL in human serum by LC-MS/MS. A 10-fold sensitivity improvement from the most sensitive reported method (LLOQ, 10 pg/mL) is essential to fully characterize the PK profiles of CIC and des-CIC in support of the clinical development of the ciclesonide-related medications for patients.
为了开发更有效的环索奈德吸入产品的药物输送装置，需要一种超灵敏的生物分析方法来测量人体中环索奈德（CIC）及其活性代谢物去异丁酰环索奈德（des-CIC）的系统暴露。
血清样品用 1-氯丁烷提取。采用反相液相色谱-大气压光电离串联质谱（LC-APPI-MS/MS）法，在 0.500 mL 血清中定量分析 1-500 pg/mL 的两种分析物。分析时间为 4.7 min/进样。CIC-d11 和 des-CIC-d11 用作内标。
校准曲线对两种分析物均表现出良好的线性（r2 > 0.99）。该新方法具有良好的精密度和准确性，所有常规 QC 样品的日内和日间精密度和准确度均在 9.6%CV 内和 ± 4.0%偏差内。两种分析物的提取回收率均约为 85%。血清样品在 3 次冻融循环、室温下 24 小时以及-20°C 和-70°C 下长达 706 天内稳定。该方法成功用于比较健康参与者吸入混悬液和吸入气雾剂中环索奈德的药代动力学（PK）。良好的进样重复分析重现性也支持该方法的稳健性。
APPI 是一种高度选择性和灵敏的离子源，通过 LC-MS/MS 实现了人血清中环索奈德和去异丁酰环索奈德的定量检测，其定量下限（LLOQ）为 1 pg/mL。与最灵敏的报道方法（LLOQ，10 pg/mL）相比，灵敏度提高了 10 倍，这对于充分描述 CIC 和 des-CIC 的 PK 特征以支持环索奈德相关药物的临床开发至关重要。
