主题:【第三届原创参赛】已完结Guidance for Industry Bioanalytical Method Validation尝试翻译

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The ability to dilute samples originally above the upper limit of the standard curve should be demonstrated by accuracy and precision parameters in the validation.
原始样品稀释过标准曲线上限的能力应用准确度和精密度参数在验证中证明。
    In consideration of high throughput analyses, including but not limited to multiplexing, multicolumn, and parallel systems, sufficient QC samples should be used to ensure control of the assay.  The number of QC samples to ensure proper control of the assay should be determined based on the run size.  The placement of QC samples should be judiciously considered in the run.
高通量分析的考虑,包括但不限于多通道,多层柱和平行系统,应使用足够的QC样品来确保分析控制。确保适当分析控制的QC样品的数量应根据运行量证明。QC样品的位置,应在运行中明智而谨慎的考虑。
For a bioanalytical method to be considered valid, specific acceptance criteria should be set in advance and achieved for accuracy and precision for the validation of QC samples over the range of the standards.
当谈到生物分析方法已验证时,应预先制订明确的可接受标准并在涵盖标准点范围内的质控样品验证中完美达到准确度和精密度。
myreebok
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这两天实在太忙了,又要自检,又要做验证计划,还要到徐州开会。。。尽量更新
myreebok
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F.  Specific Recommendations for Method Validation 方法验证具体建议
The matrix-based standard curve should consist of a minimum of six standard points, excluding blanks, using single or replicate samples.  The standard curve should cover the entire range of expected concentrations.
除空白外,使用单个或重复(重复指按相同SOP配置)样品,至少由6个标准点组成标准曲线。标准曲线应涵盖整个预期浓度范围。
    Standard curve fitting is determined by applying the simplest model that adequately describes the concentration-response relationship using appropriate weighting and statistical tests for goodness of fit.
应用最简单的恰当地描述浓度-响应相互关系模型,为拟合的良好使用适当的权重和统计学测试来确定标准曲线拟合。
    LLOQ is the lowest concentration of the standard curve that can be measured with acceptable accuracy and precision.  The LLOQ should be established using at least five samples independent of standards and determining the coefficient of variation and/or appropriate confidence interval.  The LLOQ should serve as the lowest concentration on the standard curve and should not be confused with the limit of detection and/or the low QC sample.  The highest standard will define the upper limit of quantification (ULOQ) of an analytical method.
LLOQ是标准曲线可以用适当的准确度和精密度来测定的最低浓度。LLOQ应使用至少5个独立的标准点样品并测定CV值和/或适当的置信区间。LLOQ应作为最低标准曲线浓度并不能与检测限和/或低质控样品相混淆。最高标准将定义分析方法的定量上限(ULOQ)
    For validation of the bioanalytical method, accuracy and precision should be determined using a minimum of five determinations per concentration level (excluding blank samples).
为了生物分析方法验证,准确度和精密度应在没个浓度等级使用至少5个测定值来确定(不包括空白样品)。
    The mean value should be within 15% of the theoretical value, except at LLOQ, where it should not deviate by more than 20%.  The precision around the mean value should not exceed 15% of the CV, except for LLOQ, where it should not exceed 20% of the CV.
除LLOQ的平均值应不偏离理论值的20%外,其他点的平均值应在理论值的15%之内。除LLOQ的平均值周边值精密度不得超过20%外,其他点的平均值周边的精密度CV值不能超过15%。
    Other methods of assessing accuracy and precision that meet these limits may be equally acceptable.
符合这些限制的评估准确度和精密度的其他方法可能也同样适用。
myreebok
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The accuracy and precision with which known concentrations of analyte in biological matrix can be determined should be demonstrated.  This can be accomplished by analysis of replicate sets of analyte samples of known concentrations  QC samples  from an equivalent biological matrix.  At a minimum, three concentrations representing the entire range of the standard curve should be studied: one within 3x the lower limit of quantification (LLOQ) (low QC sample), one near the center (middle QC), and one near the upper boundary of the standard curve (high QC).
应证明已知生物基质内分析物浓度的准确度和精密度可以确定。这可以用重复分析一系列相等的生物基质中已知浓度的QC样品来完成。应研究至少三个浓度来代表整个标准曲线范围:一个在3倍LLOQ之内(低质控),一个在中间值附近(中质控),一个在标准曲线上边界(高质控)。
    Reported method validation data and the determination of accuracy and precision should include all outliers; however, calculations of accuracy and precision excluding values that are statistically determined as outliers can also be reported.
报告分析方法验证数据和准确度和精密度的确定应包括所有异常值,然而,准确度和精密度的计算除统计上确定为异常值之外的值也应报告。
    The stability of the analyte in biological matrix at intended storage temperatures should be established.  The influence of freeze-thaw cycles (a minimum of three cycles at two concentrations in triplicate) should be studied.
应确定生物基质中分析物在预期储存温度的稳定性。应研究冻-融周期的影响(至少在两个浓度重复三个周期并进行三次)
              The stability of the analyte in matrix at ambient temperature should be evaluated over a time period equal to the typical sample preparation, sample handling, and analytical run times.

应评估基质中分析物超过典型样品配制、样品处理和分析运行时间在常温下的稳定性
              Reinjection reproducibility should be evaluated to determine if an analytical run could be reanalyzed in the case of instrument failure.
应评估重复进样的再现性来确定分析运行是否可以在仪器故障时再分析。
              The specificity of the assay methodology should be established using a minimum of six independent sources of the same matrix.  For hyphenated mass spectrometry-based methods, however, testing six independent matrices for interference may not be important.
应使用至少6个不同来源的相同基质确定含量方法学的专属性。然而,对于后接质谱的方法,为干扰测试6个独立的基质可能不很重要。
              In the case of LC-MS and LC-MS-MS-based procedures, matrix effects should be investigated to ensure that precision, selectivity, and sensitivity will not be compromised.
在LC-MS和LC-MS-MS的程序案例中,应调查基质影响来确定不会影响精密度,选择性和灵敏度。
              Method selectivity should be evaluated during method development and throughout method validation and can continue throughout application of the method to actual study samples.
在方法研发和方法验证过程中应评估方法选择性并在方法应用于真实样品分析过程中持续保证。
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Acceptance/rejection criteria for spiked, matrix-based calibration standards and validation QC samples should be based on the nominal (theoretical) concentration of analytes.
加样,含基质校准表和验证QC样品的可接受/不合格标准应按额定(理论)分析物浓度制订。
              Specific criteria can be set up in advance and achieved for accuracy and precision over the range of the standards, if so desired.
如果非常想得到涵盖标准范围的完美的准确度和精密度,可以预先设定具体的标准。
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V.      METHOD DEVELOPMENT:  MICROBIOLOGICAL AND LIGAND-BINDING 方法研发:微生物和配体结合
        ASSAYS 分析
Many of the bioanalytical validation parameters and principles discussed above are also applicable to microbiological and ligand-binding assays.        However, these assays possess some unique characteristics that should be considered during method validation.
在上文中讨论的许多生物分析验证参数和原则也同样适用于微生物和配体结合分析。然而,这些分析具有一些独一无二的在方法严重需考虑的特性。
        A.  Selectivity Issues 选择性制订

        As with chromatographic methods, microbiological and ligand-binding assays should be shown to be selective for the analyte.  The following recommendations for dealing with two selectivity issues should be considered:
正如色谱法,微生物和配体结合分析应体现分析的选择性。应考虑以下处理两个选择性制订的建议:
                1.  Interference From Substances Physiochemically Similar to the Analyte 理化结构与分析物相似的物质的干扰
    Cross-reactivity of metabolites, concomitant medications, or endogenous compounds should be evaluated individually and in combination with the analyte of interest.
应分别评估代谢物,合并用药或内部成分的交叉反应并结合分析物的影响。
    When possible, the immunoassay should be compared with a validated reference method (such as LC-MS) using incurred samples and predetermined criteria for agreement of accuracy of immunoassay and reference method.
如可能,免疫分析应使用加标样品并预先制订免疫测定和参考方法一致性的标准来与经验证的参考方法(如LC-MS)相比较
    The dilutional linearity to the reference standard should be assessed using study (incurred) samples.
应使用研究(加标)样品评估参考标准品(一级标准品)的稀释线性。
闲鹤野云
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楼主真的很辛苦,谢谢分享,学习很多新的药物分析词汇
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    Selectivity may be improved for some analytes by incorporation of separation steps prior to immunoassay.
在免疫测定前加入分离步骤,一些分析物的选择性可能会提高。
        2.  Matrix effects Unrelated to the Analyte 与分析物无关的基质影响

              The standard curve in biological fluids should be compared with standard in buffer to detect matrix effects.
生物流体中的标准曲线应与缓冲溶液中的标准相比较来测定基质影响
              Parallelism of diluted study samples should be evaluated with diluted standards to detect matrix effects.
应使用稀释后标准品评估稀释后研究样品的平行性来测定顶基质影响
              Nonspecific binding should be determined. 应测定非特定键合

B.  Quantification Issues 定量限制订
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Microbiological and imunoassay standard curves are inherently nonlinear and, in general, more concentration points may be recommended to define the fit over the standard curve range than for chemical assays.  In addition to their nonlinear characteristics, the response-error relationship for immunoassay standard curves is a non constant function of the mean response (heteroscadisticity).  For these reasons, a minimum of six non-zero calibrator concentrations, run in duplicate, is recommended.  The concentration-response relationship is most often fitted to a 4- or 5-parameter logistic model, although others may be used with suitable validation.  The use of anchoring points in the asymptotic high- and low-concentration ends of the standard curve may improve the overall curve fit. Generally, these anchoring points will be at concentrations that are below the established LLOQ and above the established ULOQ. Whenever possible, calibrators should be prepared in the same matrix as the study samples or in an alternate matrix of equivalent performance.  Both ULOQ and LLOQ should be defined by acceptable accuracy, precision, or confidence interval criteria based on the study requirements.
微生物和免疫测定标准曲线是天生非线性并且通常,建议使用比化学分析更多的涵盖整个标准曲线范围的浓度点来定义拟合。除其非线性特性之外,免疫测试标准曲线的响应-误相互关系是非常量的平均值函数(异方差性)。由于这些原因,推荐使用,至少6个等份的非零校准浓度测定。浓度-响应的相互关系最常见的是与4或5个逻辑模型参数相匹配,尽管其他的可能在有适当的验证的情况下使用。在渐近线高,低浓度使用固定点(确定)标准曲线的终点可以改进整体上的曲线拟合。通常,这些固定点将在低于LLOQ和高于ULOQ处。每当可能的时候,校准物应用与研究样品一样的基质来制备或用等效的基质替代。ULOQ和LLOQ都应根据研究要求用可接受的准确度,精密度或置信区间标准来定义。
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For all assays the key factor is the accuracy of the reported results. This accuracy can be improved by the use of replicate samples.  In the case where replicate samples should be measured during the validation to improve accuracy, the same procedure should be followed as for unknown samples.
对所有的分析来说关键的因数是报告值的准确度。准确度可以使用重复样品来改良。对在验证中使用测定重复样品来改良准确度的情况来说,对未知样品也应采取相同程序。

The following recommendations apply to quantification issues:
以下建议应用于定量限制订
    If separation is used prior to assay for study samples but not for standards, it is important to establish recovery and use it in determining results.  Possible approaches to assess efficiency and reproducibility of recovery are (1) the use of radiolabeled tracer analyte (quantity too small to affect the assay), (2) the advance establishment of reproducible recovery, (3) the use of an internal standard that is not recognized by the antibody but can be measured by another technique.
如果在分析前对研究样品使用分离技术但不对标准品使用,制订回收率并在测定结果中使用它是非常重要的。评估回收率有效性和再现性的可能方法有:1。使用带放射性示踪的分析物(数量足够小不影响分析)2。预先可重现回收率的制订3。使用不被抗体识别但可以用其他技术测定的内标物。
            Key reagents, such as antibody, tracer, reference standard, and matrix should be characterized appropriately and stored under defined conditions.
关键试剂,如:抗体,示踪物,参考标准品和基质应适当定义并在规条件下储存。
            Assessments of analyte stability should be conducted in true study matrix (e.g., should not use a matrix stripped to remove endogenous interferences).
应在真实的基质中进行分析物稳定性评估(例如,不能使用剥离的基质来去除内源性干扰)
            Acceptance criteria:  At least 67% (4 out of 6) of QC samples should be within 15% of their respective nominal value, 33% of the QC samples (not all replicates at the same concentration) may be outside 15% of nominal value.  In certain situations, wider acceptance criteria may be justified.
可接受标准:至少67%(6个中4个)的质控样品应在其各自标定值15%之内,33%的质控样品(不是在相同浓度下重复)可能超出15%的标定值。在某些情况下,可能证明更宽泛的可接受标准。
            Assay reoptimization or validation may be important when there are changes in key reagents, as follows:
分析再优化或验证在以下关键试剂变更时可能很重要: