Álvaro-Alonso Elena Alba, Lorenzo Mª Paz, Escobar-Rodríguez Ismael, Aguilar-Ros Antonio
Pharmacy Department, Infanta Leonor University Hospital, Av. Gran Vía del Este, 80, 28031, Madrid, Spain.
Center for Metabolomics and Bioanalysis (CEMBIO), Facultad de Farmacia, Universidad CEU San Pablo, CEU Universities, Urbanización Montepríncipe, 28660, Boadilla del Monte, Spain.
BMC Chem. 2022 May 14;16(1):32. doi: 10.1186/s13065-022-00827-9.
The Pharmacy Service of the Infanta Leonor University Hospital acquires, compounds, distributes and dispenses more than 3000 L of methadone oral solution to Drug Addiction Patients Centers per year. Our purpose is to develop and validate an improved high performance liquid chromatography (HPLC) method to quantify methadone hydrochloride in a new oral solution with methylhydroxybenzoate (methylparaben) and propylhydroxybenzoate (propylparaben) to be implemented in physicochemical stability studies that allow to provide more information and even to increase the beyond-use date.
A HPLC-Agilent 1100 equipment, comprising a quaternary pump and an ultraviolet diode-array-detector (DAD) was used. An analytical method development and validation was completed. The curve was constructed from methadone working concentrations of 75-125% (7.5, 9.0, 10.0, 11.0 and 12.5 mg/mL) to assess the linear relationship between the concentration of the analyte and the obtained areas. Precision and accuracy were calculated. Detection and quantification limit (LD, LQ) were estimated using the EURACHEM method. Forced-degradation studies were also performed.
Chromatographic conditions were: flow rate 1.6 mL/min; mobile phase 55% acetonitrile and 45% sodium phosphate 25 mM (pH = 10); injection volume was 5 µL. The column was a Waters-XTerra™ RP18, maintained at 40 °C. DAD was λ = 254 nm. Retention times for methadone, methylparaben and propylparaben were 4.34, 0.70 and 0.88 min respectively. The method was linear (y = 284.3x - 97.8, r = 0.996). Instrumental precision was 0.33% for standards (n = 10); intra-assay precision 0.53% (n = 6) and inter-assay precision 1.95% (n = 12). The relative standard deviation percentage for accuracy was 1.28%. The recovery percentage was 101.5 ± 1.5%. LQ and LD were 2.18 µg/mL and 2.0 µg/mL respectively. The most destabilizing conditions were oxidizing and alkaline. The chromatograms confirmed no interference with the methadone signal.
The HPLC method has proved to be valid and reproducible for methadone quantification in a new oral solution with methylparaben and propylparaben. This assay is a rapid, simple and reliable technique that can be used in daily analysis and physicochemical stability studies.
莱昂诺尔公主大学医院药房每年向药物成瘾患者中心采购、配制、分发和调配超过3000升美沙酮口服溶液。我们的目的是开发并验证一种改进的高效液相色谱（HPLC）方法，用于定量测定一种含有甲基羟基苯甲酸酯（对羟基苯甲酸甲酯）和丙基羟基苯甲酸酯（对羟基苯甲酸丙酯）的新口服溶液中的盐酸美沙酮，以用于物理化学稳定性研究，从而能够提供更多信息，甚至延长有效期。
使用一台配备四元泵和紫外二极管阵列检测器（DAD）的HPLC - Agilent 1100设备。完成了一种分析方法的开发与验证。通过75 - 125%（7.5、9.0、10.0、11.0和12.5毫克/毫升）的美沙酮工作浓度构建曲线，以评估分析物浓度与所得峰面积之间的线性关系。计算了精密度和准确度。采用EURACHEM方法估算检测限和定量限（LD、LQ）。还进行了强制降解研究。
色谱条件为：流速1.6毫升/分钟；流动相为55%乙腈和45% 25毫摩尔/升磷酸钠（pH = 10）；进样体积为5微升。色谱柱为Waters - XTerra™ RP18，保持在40℃。DAD检测波长为λ = 254纳米。美沙酮、对羟基苯甲酸甲酯和对羟基苯甲酸丙酯的保留时间分别为4.34、0.70和0.88分钟。该方法呈线性（y = 284.3x - 97.8，r = 0.996）。标准品的仪器精密度为0.33%（n = 10）；批内精密度为0.53%（n = 6），批间精密度为1.95%（n = 12）。准确度的相对标准偏差百分比为1.28%。回收率为101.5 ± 1.5%。LQ和LD分别为2.18微克/毫升和2.0微克/毫升。最不稳定的条件是氧化和碱性条件。色谱图证实对美沙酮信号无干扰。
对于含有对羟基苯甲酸甲酯和对羟基苯甲酸丙酯的新口服溶液中的美沙酮定量，HPLC方法已证明是有效且可重现的。该测定法是一种快速、简单且可靠的技术，可用于日常分析和物理化学稳定性研究。
