Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, USA; Center for Clinical Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, USA.
Department of Pharmacy & Therapeutics, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, USA.
J Pharm Biomed Anal. 2022 Sep 5;218:114853. doi: 10.1016/j.jpba.2022.114853. Epub 2022 May 23.
Sedative use can result in adverse drug reactions. Intensive care unit patients are especially at risk and pharmacokinetic modeling of drug concentrations is an approach to develop precision dosing strategies. However, limited blood sampling availability in critically ill children and need for multiple assays to quantify a variety of commonly used sedatives creates logistical challenges. The goal of this project was to develop a sensitive and selective assay for the simultaneous quantification of a panel of sedatives comprised of midazolam (MDZ), alpha hydroxymidazolam (1- OH MDZ), dexmedetomidine (DEX), morphine (MOR), morphine-3-glucuronide (M3G), morphine-6-glucuronide (M6G), fentanyl (FEN), norfentanyl (NF), and hydromorphone (HM) in small volume pediatric plasma samples. A sensitive and efficient ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS/MS) method was developed following FDA guidance for bioanalytical validation. Minimal sample preparation consisting of simple protein precipitation extraction using acetonitrile with internal standards was utilized. Analyte separation was achieved using a gradient mixture of (A: 0.15% formic acid in water and B: Acetonitrile) and a Waters Acquity C18, 1.7 µm (2.1 × 100 mm) column. Assays were linear over the clinical concentration ranges: MDZ, MOR, HM: 0.5-125 ng/mL; 1-OH MDZ, M3G, M6G: 5-500 ng/mL; and DEX, FEN, NF: 0.05-7.5 ng/mL (R > 0.99 for all). Assay run time was 10 min and required only 100 μL of plasma. Initial testing of samples from pediatric patients demonstrates adequacy of assay to measure sedatives and metabolites at clinical concentrations confidently in low volumes of plasma. This novel highly-sensitive and specific method to measure a total of nine different analytes (five sedatives, four metabolites) simultaneously enables comprehensive analysis of a panel of sedatives in small volumes such as in pediatric ICU patients.
镇静剂的使用会导致药物不良反应。重症监护病房的患者尤其处于危险之中，药物浓度的药代动力学建模是开发精准给药策略的一种方法。然而，危重症儿童的血样采集有限，需要多种检测方法来定量分析各种常用镇静剂，这给检测带来了挑战。本项目的目标是开发一种灵敏、选择性的测定法，用于同时定量测定一组镇静剂，包括咪达唑仑（MDZ）、α-羟咪达唑仑（1-OH MDZ）、右美托咪定（DEX）、吗啡（MOR）、吗啡-3-葡萄糖醛酸（M3G）、吗啡-6-葡萄糖醛酸（M6G）、芬太尼（FEN）、去甲芬太尼（NF）和氢吗啡酮（HM）在小体积儿科血浆样本中。根据 FDA 对生物分析验证的指南，开发了一种灵敏高效的超高效液相色谱-质谱联用（UPLC-MS/MS）方法。仅使用乙腈进行简单的蛋白沉淀提取，作为最小的样品制备。采用（A：水 0.15%甲酸和 B：乙腈）梯度混合和 Waters Acquity C18，1.7µm（2.1×100mm）柱实现分析物分离。MDZ、MOR、HM 的测定线性范围为 0.5-125ng/mL；1-OH MDZ、M3G、M6G 的测定线性范围为 5-500ng/mL；DEX、FEN、NF 的测定线性范围为 0.05-7.5ng/mL（所有测定均 R > 0.99）。分析时间为 10min，仅需 100μL 血浆。对儿科患者样本的初步测试表明，该测定法足以在低体积血浆中自信地测量临床浓度下的镇静剂和代谢物。该新型高灵敏度和特异性方法能够同时测定总共 9 种不同分析物（5 种镇静剂，4 种代谢物），可在小体积（如儿科 ICU 患者）中对一组镇静剂进行全面分析。
