Interlaboratory trial

实验室间试验
  • 文章类型: Journal Article
    短链氯化石蜡(短链氯化石蜡)被列入《斯德哥尔摩公约》。因此,选择合适的方法对它们的准确量化至关重要。如今,不能保证用作定量标准的商业试剂的质量。作为解决方案,我们采用具有已知同源物组成比的SCCP制剂参考材料作为定量标准,以评估方法的适当性.通过混合SCCP配方和干扰,独立制备分析样品并用作模拟环境样品。短链氯化石蜡的同系物组成概况类似于定量标准和分析样品。质量分数和同系物轮廓,包括碳链长度和氯同系物,14个不同的实验室报告了短链氯化石蜡。对于质量分数,参与者报告的结果是一致的,除了使用低分辨率气相色谱法(GC)的参与者。液相色谱(LC)和GC产生的结果略有不同,尽管定量标准和分析样品之间的同源组成比相似。尽管在整个氯同系物中存在差异,从GC和LC获得的碳链长度分布相似。差异取决于所采用的方法。此外,与低分辨率数据相比,由于高分辨率仪器的质量精度降低了干扰对分析样品的影响,因此高分辨率数据的波动较小。因此,实验室间试验采用相似的同系物组成的定量标准和分析样品证明有价值的方法之间的差异,考虑到设备,分辨率规格,和电离。
    Short-chain chlorinated paraffins (SCCPs) are listed in the Stockholm Convention. Therefore, selecting suitable methods for their accurate quantification is essential. Nowadays, the quality of commercial reagents employed as quantification standards is not guaranteed. As a solution, we adopted an SCCP formulation reference material with known homolog composition ratios as the quantification standard to evaluate the appropriateness of the methods. By mixing the SCCP formulation and interferences, an analytical sample was independently prepared and used as the simulation environmental sample. The homolog compositional profiles of the SCCPs resembled those of the quantification standard and the analytical sample. The mass fractions and the homolog profiles, including the carbon chain length and chlorine homolog profiles, of the SCCPs were reported by 14 different laboratories. For the mass fraction, the results reported by participants were consistent, except for the participants that employed low-resolution gas chromatography (GC). The results generated from liquid chromatography (LC) and GC were slightly different, despite of the similar homolog composition ratios between the quantification standard and the analytical sample. Although there were discreet discrepancies in the overall chlorine homolog profiles, the carbon chain length profiles acquired from GC and LC were similar. The differences depended on the method employed. Additionally, compared with the low-resolution data, the high-resolution data displayed less fluctuation since the effect of the interferences on the analytical sample was reduced because of the mass accuracy of high-resolution instruments. Accordingly, the interlaboratory trial employing the similar homolog compositional profiles of the quantification standard and the analytical sample proved valuable in elucidating the differences among methods, considering equipment, resolution specification, and ionization.
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  • 文章类型: Journal Article
    毛细管区带电泳紫外(CZE-UV)在单克隆抗体的电荷异质性测定中越来越受欢迎,和疫苗。ε-氨基己酸(eACA)CZE-UV方法已用作快速平台方法。然而,在过去的几年里,已经观察到几个问题,例如,电泳分辨率或基线漂移的损失。评估eACA在报告的问题上的作用,各个实验室被要求提供他们常规使用的eACACZE-UV方法,和BGE成分。虽然每个实验室都声称使用He等人。EACACZE-UV法,大多数方法实际上偏离了他的。随后,设计了详细的实验室间研究,其中向每个实验室提供了两种市售的mAb(Waters\'MassCheckStandardmAb(pI7)和NISTmAb(pI9))。以及两个详细的eACACZE-UV协议,高速,和一个长端,高分辨率方法。十个实验室每个都使用自己的仪器参加,和商品,显示出卓越的方法性能(时间校正的主峰面积百分比的RSD从0.2%到1.9%,迁移时间的RSD从0.7%到1.8%(每个实验室n=50),在某些情况下,分析时间短至2.5分钟)。这项研究澄清了eACA不是上述变化的主要原因。本文受版权保护。保留所有权利。
    Capillary zone electrophoresis ultraviolet (CZE-UV) has become increasingly popular for the charge heterogeneity determination of mAbs and vaccines. The ε-aminocaproic acid (eACA) CZE-UV method has been used as a rapid platform method. However, in the last years, several issues have been observed, for example, loss in electrophoretic resolution or baseline drifts. Evaluating the role of eACA on the reported issues, various laboratories were requested to provide their routinely used eACA CZE-UV methods, and background electrolyte compositions. Although every laboratory claimed to use the He et al. eACA CZE-UV method, most methods actually deviate from He\'s. Subsequently, a detailed interlaboratory study was designed wherein two commercially available mAbs (Waters\' Mass Check Standard mAb [pI 7] and NISTmAb [pI 9]) were provided to each laboratory, along with two detailed eACA CZE-UV protocols for a short-end, high-speed, and a long-end, high-resolution method. Ten laboratories participated each using their own instruments, and commodities, showing excellence method performance (relative standard deviations [RSDs] of percent time-corrected main peak areas from 0.2% to 1.9%, and RSDs of migration times from 0.7% to 1.8% [n = 50 per laboratory], analysis times in some cases as short as 2.5 min). This study clarified that eACA is not the main reason for the abovementioned variations.
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  • 文章类型: Journal Article
    核磁共振(NMR)代谢组学目前很受欢迎,可以吸引专门和非专门的NMR小组,其中包括经过分析培训的人员和新来者。包括本科生。由已建立的NMR代谢组学小组进行的最近的实验室间研究表明,最先进的NMR设备和SOP具有很高的可重复性。有,然而,当混合分析专家和新手时,没有评估NMR重现性。使用属于不同实验室的两个NMR仪器对NMR定量再现性进行实验室间评估,该仪器涉及具有不同背景和代谢组学专业知识的几位操作员,目的是评估多用途NMR环境中数据再现性的限制因素。操作员引起的可变性,自动移液器,评估NMR管和NMR仪器以评估定量再现性的限制因素。结果估计在现实生活中的多用途NMR实验室中的预期再现性数据最大为4%的可变性,证明当前的NMR设备和SOP可以补偿一些操作员引起的变异性。
    Nuclear magnetic resonance (NMR) metabolomics is currently popular enough to attract both specialized and non-specialized NMR groups involving both analytical trained personnel and newcomers, including undergraduate students. Recent interlaboratory studies performed by established NMR metabolomics groups demonstrated high reproducibility of the state-of-the-art NMR equipment and SOPs. There is, however, no assessment of NMR reproducibility when mixing both analytical experts and newcomers. An interlaboratory assessment of NMR quantitation reproducibility was performed using two NMR instruments belonging to different laboratories and involving several operators with different backgrounds and metabolomics expertise for the purpose of assessing the limiting factors for data reproducibility in a multipurpose NMR environment. The variability induced by the operator, automatic pipettes, NMR tubes and NMR instruments was evaluated in order to assess the limiting factors for quantitation reproducibility. The results estimated the expected reproducibility data in a real-life multipurpose NMR laboratory to a maximum 4% variability, demonstrating that the current NMR equipment and SOPs may compensate some of the operator-induced variability.
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  • 文章类型: Journal Article
    每个实验室测试都需要通过质量控制进行验证。对于杀菌剂敏感性试验(BST),目前既没有质量控制(QC)菌株,也没有适用于这些菌株的QC范围。作为QC菌株,四个明确定义的实验室参考菌株(金黄色葡萄球菌ATCC®6538、希雷肠球菌ATCC®10541、大肠杆菌ATCC®10536和铜绿假单胞菌ATCC®15442),以前曾用于杀菌剂功效测试,被选中。在有11个参与实验室的实验室间试验中,应针对上述四种菌株和四种杀生物剂苯扎氯铵开发BSTQC范围,氯己定,奥替尼啶和聚己内酯。使用肉汤微量稀释方法探索三个不同批次的胰蛋白酶大豆肉汤的性能,随后使用RangeFinder软件评估数据。因此,为所有参考菌株-杀生物剂组合定义了QC范围,除了铜绿假单胞菌ATCC®15442与两种杀生物剂氯己定和聚己胺。后两个QC范围的发展是不可能的,由于杀生物剂在铜绿假单胞菌ATCC®15442所需的测试范围内的溶解度有限。新开发的QC范围包括三到五个稀释步骤。QC范围的建立将有助于未来BST的验证。
    Every laboratory test needs validation by quality controls. For biocide susceptibility testing (BST), neither quality control (QC) strains nor QC ranges applicable to these strains are currently available. As QC strains, four well-defined laboratory reference strains (Staphylococcus aureus ATCC® 6538, Enterococcus hirae ATCC® 10541, Escherichia coli ATCC® 10536 and Pseudomonas aeruginosa ATCC® 15442), which have been used previously for biocide efficacy testing, were selected. In an interlaboratory trial with eleven participating laboratories, BST QC ranges should be developed for the aforementioned four strains and the four biocides benzalkonium chloride, chlorhexidine, octenidine and polyhexanide. The performance of three different lots of tryptic soy broth was explored using the broth microdilution method and the data were subsequently evaluated using the RangeFinder software. As a result, QC ranges were defined for all reference strain-biocide combinations, except for P. aeruginosa ATCC® 15442 with the two biocides chlorhexidine and polyhexanide. The development of the latter two QC ranges was not possible, due to the limited solubility of the biocides in the test range required for P. aeruginosa ATCC® 15442. The newly developed QC ranges comprise three to five dilution steps. The establishment of QC ranges will contribute to the validation of BST in the future.
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  • 文章类型: Journal Article
    Chlorinated paraffins (CPs) are industrial chemicals that have been primarily used in applications involving metalworking fluids. Among CPs, short-chain chlorinated paraffins (SCCPs) are a well-known environmental pollutant and are listed under Annex A of the Stockholm Convention on Persistent Organic Pollutants. CPs are alkanes substituted with chlorine atoms, and SCCPs are comprised of 10-13 carbon atoms. Reliable quantification of SCCPs is a critical issue because of the large number of SCCP isomers that are in use across multiple industries. Some interlaboratory comparisons of SCCP analyses have been conducted, and the reliability of these results was overwhelmingly determined as inferior to that of comparable PCB and dioxin analyses because of variations in the quality of commercial reagents that were employed as quantification standards. In order to address such inconsistencies, this study endeavored to prepare and evaluate a novel SCCP formulation as a candidate reference material for use as a reliable quantification standard. A subject trial study was hence performed to evaluate methods such as gas- and liquid-chromatography mass spectrometry (GC/MS and LC/MS) on sample matrices (without a clean-up process), and to subsequently elucidate the interpreted specifications for their candidacy as a reliable quantification standard. Results ultimately showed that the SCCP concentrations obtained from GC and LC were comparable. When the homologs reported by a subset of 14 separate laboratories were unified (excluding all results for Cl4 homologs), the carbon chain length profiles obtained from GC and LC were found to be similar; however, the overall chlorine homolog profiles did exhibit slight differences. Moreover, the results from high-resolution MS showed less variation than those from low-resolution MS. Thus, it was overarchingly determined that the deployment of this candidate reference material would serve as an effective mechanism for estimating the comparability of SCCP quantifications/evaluations of standard materials.
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  • 文章类型: Journal Article
    The performance of culture and PCR methods routinely used to diagnose contagious equine metritis (CEM) was evaluated and compared by two interlaboratory trials involving a total of 24 European laboratories, including 22 National Reference Laboratories for CEM. Samples were swab specimens artificially contaminated with bacteria present in the genital tract of Equidae, some with and some without Taylorella equigenitalis, the causative agent of CEM, and T asinigenitalis, responsible for possible misidentification as T equigenitalis Throughout both interlaboratory trials, PCR performed better in terms of specificity and sensitivity than the culture method, supporting the assertion that PCR should be accepted for CEM diagnosis. However, the culture performance during the second interlaboratory trial was better than during the first one, suggesting that the expertise of participants improved. This reveals the advantage of regular interlaboratory trials to constantly improve the expertise of laboratories. It also shows the need to develop new culture media that are more selective and/or better geared to the metabolism of T equigenitalis in order to improve the bacteriological diagnosis of CEM.
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  • 文章类型: Journal Article
    内分泌活性物质会对水生生态系统产生不利影响。特别强调,其中,对雌激素和雌激素模拟化合物的影响。基于效应的筛选方法如体外生物测定是检测和定量已知和未知混合物的内分泌活性的合适工具。本研究描述了Arxula-酵母雌激素筛选(A-YES®)测定的验证,一种基于效应的方法,用于检测水和废水的雌激素潜力。这个报告基因测定,以随时可用的格式提供,是基于人类雌激素受体α的激活。用户友好的A-YES®使没有经验的操作员能够迅速胜任测定。来自四个不同培训水平国家的14个实验室分析了加标和未加标废水和地表水样品中的17β-雌二醇当量浓度(EEQ),在废水进水和加标盐水样品以及三种双酚的混合物中。未处理样品的检测限(LOD)为1.8ng/L的17β-雌二醇(E2)。EEQ高于LOD(平均EEQ值在6.3至20.4ng/L之间)的样品的相对重复性和再现性标准偏差范围为7.5至21.4%和16.6至28.0%,分别。精密度结果与其他常用的雌激素分析方法相当。A-YES®已被证明是准确的,精确和强大的生物测定。结果已包含在ISO草案标准中。该测定法被证明适用于测试典型的废水进水,污水和盐水。其他研究表明,该测定可用于富集样品,这将LOD降低到pg/L范围。A-YES®的验证和相应国际标准的开发是朝着基于效应的水样中雌激素和雌激素样化合物的统一和可靠的生物测定法迈出的一步。
    Endocrine-active substances can adversely impact the aquatic ecosystems. A special emphasis is laid, among others, on the effects of estrogens and estrogen mimicking compounds. Effect-based screening methods like in vitro bioassays are suitable tools to detect and quantify endocrine activities of known and unknown mixtures. This study describes the validation of the Arxula-Yeast Estrogen Screen (A-YES®) assay, an effect-based method for the detection of the estrogenic potential of water and waste water. This reporter gene assay, provided in ready to use format, is based on the activation of the human estrogen receptor alpha. The user-friendly A-YES® enables inexperienced operators to rapidly become competent with the assay. Fourteen laboratories from four countries with different training levels analyzed 17β-estradiol equivalent concentrations (EEQ) in spiked and unspiked waste water effluent and surface water samples, in waste water influent and spiked salt water samples and in a mixture of three bisphenols. The limit of detection (LOD) for untreated samples was 1.8ng/L 17β-estradiol (E2). Relative repeatability and reproducibility standard deviation for samples with EEQ above the LOD (mean EEQ values between 6.3 and 20.4ng/L) ranged from 7.5 to 21.4% and 16.6 to 28.0%, respectively. Precision results are comparable to other frequently used analytical methods for estrogens. The A-YES® has been demonstrated to be an accurate, precise and robust bioassay. The results have been included in the ISO draft standard. The assay was shown to be applicable for testing of typical waste water influent, effluent and saline water. Other studies have shown that the assay can be used with enriched samples, which lower the LOD to the pg/L range. The validation of the A-YES® and the development of a corresponding international standard constitute a step further towards harmonized and reliable bioassays for the effect-based analysis of estrogens and estrogen-like compounds in water samples.
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