Full validation

完全验证
  • 文章类型: Journal Article
    背景:研究组织中的药物处置对于改善给药策略和最大化治疗效果至关重要,然而,由于各种基质之间的差异,开发多组织生物分析方法可能具有挑战性。在这里,我们开发了一种专门用于定量哌拉西林(PIP)的LC-MS/MS方法,头孢唑啉(CFZ),和头孢西丁(CFX)在大鼠血浆和12个组织中,根据FDA和EMA指南,附有每个矩阵的验证数据。
    结果:该方法仅需要少量样品体积(5μL血浆或50-100μL组织匀浆)和相对简单的方案即可同时定量PIP,CFZ,和CFX在不同的生物基质中。流动相A由5mM甲酸铵和0.1%甲酸水溶液组成,流动相B在乙腈中含有0.1%甲酸。将流动相以0.3mL/min的流速泵送通过配备有具有梯度洗脱程序的保护柱的SynergiFusion-RP柱。使用多反应监测以正离子化模式(ESI+)操作质谱仪。
    结论:经过验证的方法已成功用于量化PIP,CFZ,和CFX的血浆和组织样品收集在一个试验大鼠研究,并将进一步用于大型药代动力学研究。据我们所知,这也是第一份长期报告,冻融,和PIP的自动进样器稳定性数据,CFZ,和CFX在大鼠血浆和多个组织中。
    BACKGROUND: The investigation of drug disposition in tissues is critical to improving dosing strategy and maximizing treatment effectiveness, yet developing a multi-tissue bioanalytical method could be challenging due to the differences among various matrices. Herein, we developed an LC-MS/MS method tailored for the quantitation of piperacillin (PIP), cefazolin (CFZ), and cefoxitin (CFX) in rat plasma and 12 tissues, accompanied by validation data for each matrix according to the FDA and EMA guidelines.
    RESULTS: The method required only a small sample volume (5 μL plasma or 50-100 μL tissue homogenates) and a relatively simple protocol for simultaneous quantitation of PIP, CFZ, and CFX within different biological matrices. Mobile phase A was composed of 5 mM ammonium formate and 0.1 % formic acid in water, while mobile phase B contained 0.1 % formic acid in acetonitrile. The mobile phase was pumped through a Synergi Fusion-RP column equipped with a guard column with a gradient elution program at a 0.3 mL/min flow rate. The mass spectrometer was operated in positive ionization mode (ESI+) using multiple reaction monitoring.
    CONCLUSIONS: The validated method has been successfully applied to quantify PIP, CFZ, and CFX from the plasma and tissue samples collected in a pilot rat study and will further be used in a large pharmacokinetic study. To our knowledge, this is also the first report presenting long-term, freeze-thaw, and autosampler stability data for PIP, CFZ, and CFX in rat plasma and multiple tissues.
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  • 文章类型: Journal Article
    背景:新烟碱(NEO)用于薄荷的植物检疫处理。L作物,这种做法要求在非常低的浓度下精确控制这些有害物质。
    目的:本研究的目的是应用一种基于总误差方法学的同时验证和评估测量不确定度的方法。为了准确量化MenthaSpicata中两个近地天体的存在。我用一个快速的,Easy,便宜,有效,Rugged,和安全(QuEChERS)-LC-MS/MS方法。
    方法:采用QuEChERS提取方法对吡虫啉和啶虫脒进行定量,与LC-MS/MS相结合,确保分析方法的准确性,并管理与其常规使用相关的风险。基于β含量的完整而详尽的验证方法,γ-置信公差区间用于不确定度评估,利用广义枢轴量(GPQ)概念和蒙特卡罗模拟,这避免了对额外数据的需要,同时在预定可接受的限度内实现每个浓度水平的中间精度。
    结果:验证程序基于两个近地天体的二次模型的选择,允许在工作浓度范围内通过LC-MS/MS测定法验证啶虫脒和吡虫啉。通过β含量值(66.7、80和90%)和风险值(10和5%)的变化证明了不确定性分布区间的灵活性,保持在20%的可接受性范围内,相对扩展不确定度不超过15%和11%。
    结论:用于分析两个近地天体的QuEChERS-LC-MS/MS方法已使用不确定度曲线策略成功地得到了充分验证。
    结论:实施总体验证策略,其中包括验证和不确定性评估,称为不确定性曲线,用于对MenthaSpicata的两个重要近地天体进行量化。L使用QuEChERS-LC-MS/MS这种定性方法是通过在每个浓度水平的中间精度条件下利用分析验证的数据来计算方法的测量不确定性而进行的,而无需额外的努力。之后,我们证明了该策略用于啶虫脒和吡虫啉的LC-MS/MS定量的灵活性,使用能够选择和修改β含量和γ置信度值的决策工具。
    BACKGROUND: Neonicotinoids (NEOs) are used for the phytosanitary treatment of Mentha Spicata.L crops, and this practice requires precise control of these harmful substances at very low concentrations.
    OBJECTIVE: The objective of this study is to apply an approach allowing simultaneously validation and evaluation of measurement uncertainty based on total error methodology, in order to accurately quantify the presence of two NEOs in Mentha Spicata.L utilizing a Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS)-LC-MS/MS methodology.
    METHODS:  The quantification of imidacloprid and acetamiprid employing a QuEChERS extraction method, coupled with LC-MS/MS, ensuring the accuracy of the analytical method and managing the risks associated with its routine use. A complete and exhaustive validation approach based on the \"β-content, γ-confidence\" tolerance interval was used for the uncertainty assessment, using the generalized pivot quantity (GPQ) concept and Monte Carlo simulation, which avoids the need for additional data while achieving intermediate precision for each concentration level within predetermined acceptable limits.
    RESULTS: The validation procedure is based on the choice of a quadratic model for the two NEOs, allowing the validation of acetamiprid and imidacloprid by LC-MS/MS assay within the range of working concentration. The flexibility of the uncertainty profile intervals was demonstrated with a variation in β-content values (66.7, 80, and 90%) and risk values (10 and 5%), which remained within the acceptability limits of 20%, and the relative expanded uncertainty did not exceed 15 and 11%.
    CONCLUSIONS: A QuEChERS-LC-MS/MS method for the analysis of two NEOs has been successfully fully validated using the uncertainty profile strategy.
    CONCLUSIONS: Implementation of an overall validation strategy, which involves both the validation and uncertainty assessment known as the uncertainty profile, for the quantification of two important NEOs in Mentha Spicata.L using QuEChERS-LC-MS/MS. This qualimetric approach has been conducted by computing the measurement uncertainty of the method utilizing data from analytical validation under conditions of intermediate precision at each level of concentration without additional effort. After that we have demonstrated the flexibility of this strategy for the LC-MS/MS quantification of acetamiprid and imidacloprid, using a decision tool that enables the choice and modification of β-content and γ-confidence values.
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  • 文章类型: Journal Article
    A fully validated bio-analytical method based on Matrix-Assisted-Laser-Desorption/Ionization-Time of Flight Mass Spectrometry was developed for quantitation in human plasma of the anti-tumor peptide CIGB-300. An analog of this peptide acetylated at the N-terminal, was used as internal standard for absolute quantitation. Acid treatment allowed efficient precipitation of plasma proteins as well as high recovery (approximately 80%) of the intact peptide. No other chromatographic step was required for sample processing before MALDI-MS analysis. Spectra were acquired in linear positive ion mode to ensure maximum sensitivity. The lower limit of quantitation was established at 0.5 μg/mL, which is equivalent to 160 fmol peptide. The calibration curve was linear from 0.5 to 7.5 μg/mL, with R(2)>0.98, and permitted quantitation of highly concentrated samples evaluated by dilution integrity testing. All parameters assessed for five validation batches met the FDA guidelines for industry. The method was successfully applied to analysis of clinical samples obtained in a phase I clinical trial following intravenous administration of CIGB-300 at a dose of 1.6 mg/kg body weight. With the exception of Cmax and AUC, pharmacokinetic parameters were similar for ELISA and MALDI-MS methods.
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  • 文章类型: Journal Article
    在精细化学品行业,包括制药和农业化学品,分析测试由生产部门或合同研究机构在产品研发的某个阶段进行。这些外部组织需要保持使用等同于或优于分析方法验证指定的方法执行分析测试的能力。出于这个原因,有必要将分析程序转移到另一个地点。在这次审查中,介绍了分析程序转移与试验验证之间的关系,专注于包括HPLC在内的分析程序。
    In the industry of fine chemicals, including pharmaceutical and agricultural chemicals, analytical tests are performed by production departments or contract research organizations at some stage in the research and development of products. These external organizations are required to maintain the capabilities to perform analytical tests using methods that are equivalent to or better than those specified by analytical method validation. For this reason, transfer of analytical procedures to an alternative site becomes necessary. In this review, the relationship between transfer of analytical procedures and assay validation is introduced, focusing on analytical procedures that include HPLC.
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