Experimental Comparison of the Different Approaches To Estimate LOD and LOQ of an HPLC Method

Vial, Jérôme; Jardy, Alain

doi:10.1021/ac981179n

Cited by 303 publications

(194 citation statements)

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“…Following the canonical calibration curve, statistical analyses fit a linear regression that relates the concentrations and the observed intensities on the original (i.e., not log-transformed) scale. Many methods for fitting linear regression and estimating the LOB, LOD have been proposed [12,13,14,15], compared [9,16,17,18], and implemented in open-source software such as Quasar [19] and Skyline [20]. Most methods acknowledge that the variance of the observed intensities increases with the strength of the intensity, and specify regression with unequal variance, or use a robust estimation procedure.…”

Section: Introductionmentioning

confidence: 99%

Nonlinear Regression Improves Accuracy of Characterization of Multiplexed Mass Spectrometric Assays

Galitzine

Egertson

Abbatiello

et al. 2018

Molecular & Cellular Proteomics

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AbbreviationsDIA data-independent acquisition LOB limit of blank LOD limit of detection LLOQ lower limit of quantitation/quantification PRM parallel reaction monitoring SRM selected reaction monitoring 2 SummaryThe need for assay characterization is ubiquitous in quantitative mass spectrometry-based proteomics. Among many assay characteristics, the limit of blank (LOB) and limit of detection (LOD) are two particularly useful figures of merit. LOB and LOD are determined by repeatedly quantifying the observed intensities of peptides in samples with known peptide concentrations, and deriving an intensity versus concentration response curve. Most commonly, a weighted linear or logistic curve is fit to the intensity-concentration response, and LOB and LOD are estimated from the fit. Here we argue that these methods inaccurately characterize assays where observed intensities level off at low concentrations, which is a common situation in multiplexed systems. This manuscript illustrates the deficiencies of these methods, and proposes an alternative approach based on non-linear regression that overcomes these inaccuracies. We evaluated the performance of the proposed method using computer simulations, and using eleven experimental datasets acquired in Data-Independent Acquisition (DIA), Parallel Reaction Monitoring (PRM), and Selected Reaction Monitoring (SRM) mode. When the intensity levels off at low concentrations, the non-linear model changes the estimates of LOB/LOD upwards, in some datasets by 20-40%. In absence of a low concentration intensity leveling off, the estimates of LOB/LOD obtained with non-linear statistical modeling were identical to those of weighted linear regression. We implemented the non-linear regression approach in the open-source R-based software MSstats, and advocate its general use for characterization of mass spectrometry-based assays.3

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Section: Introductionmentioning

confidence: 99%

Nonlinear Regression Improves Accuracy of Characterization of Multiplexed Mass Spectrometric Assays

Galitzine

Egertson

Abbatiello

et al. 2018

Molecular & Cellular Proteomics

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“…The limit of detection (LOD) was calculated as described elsewhere [25]. Data from three separate experiments were normalized.…”

Section: Methods Used In Data Analysismentioning

confidence: 99%

An efficient design strategy for a whole-cell biosensor based on engineered ribosome binding sequences

et al. 2011

Anal Bioanal Chem

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In prokaryotes, the ribosome binding sequence (RBS), located in the 5′ untranslated region (5′ UTR) of an mRNA, plays a critical role in enhancing mRNA translation and stability. To evaluate the effect of the RBS on the sensitivity and signal intensity of an environmental whole-cell biosensor, three Escherichia coli-based biosensors that respond to benzene, toluene, ethylbenzene, and the xylenes (BTEX) were constructed; the three biosensors have the same Pu promoter and xylR regulator from the Pseudomonas putida TOL plasmid but differ in the engineered RBS in their reporter genes. The results from time and dose-dependent induction of luminescence activity by 2-chlorotoluene showed that the BTEX-SE and BTEX-SD biosensors with engineered RBS had signal intensities approximately 10-35 times higher than the primary BTEX-W biosensor. The limits of detection (LOD) of the BTEX-SE and BTEX-SD biosensors were also significantly lower than the LOD of the BTEX-W biosensor (20 ± 5 μmol L −1 and 25 ±). Moreover, the BTEX-SE and BTEX-SD biosensors responded three times more rapidly to the analytes. These results suggest that rationally designed RBS in the 5′ UTR of a reporter gene may be a promising strategy for increasing the sensitivity, signal intensity, and response speed of whole-cell biosensors.

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“…The LOD is the smallest concentration of analyte detectable in the sample by an analytical method with a given level of confidence, and the LOQ is the lowest solute concentration that can be determined with an acceptable level of uncertainty. 34,35 LOD and LOQ were determined according to the ICH (International Conference on Harmonization) guideline which suggests calculation based as 3.3 and 10 times the ratio between the standard deviation of the intercept (response), s, and the slope estimated, S, from the calibration curve of the analytes. 36 The precision of the method was evaluated in terms of repeatability and intermediate precision expressed as relative standard deviation (RSD %).…”

Section: Methods Validationmentioning

confidence: 99%

Multiresidue determination of pesticides in carrots using pressurized liquid extraction and gas chromatography with mass spectrometry detector

Abad¹,

Winck²,

Silva³

et al. 2010

J. Braz. Chem. Soc.

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Neste estudo, 14 agrotóxicos (captan, clorotalonil, deltametrina, dicloran, fention, folpete, iprodiona, linurom, malationa, prometrina, procloraz, procimidona, triclorfom e trifluralina) foram usados no desenvolvimento de um método multiresíduos (MRM), utilizando extração de cenouras com líquido pressurizado (PLE). A etapa de clean-up foi realizada por dispersão da matriz em fase sólida (MSPD), sendo testados como material dispersante o Florisil, XAD-4, XAD-7 e C18. Diclorometano, acetato de etila e acetona foram empregados como solventes para PLE, a 75 ºC e 1.500 psi. As recuperações dos analitos para cenouras liofilizadas ficaram na faixa de 70 a 133%. Entretanto, os percentuais de recuperação obtidos para as cenouras úmidas não foram satisfatórios (< 70%). Os coeficientes de determinação das curvas analíticas situaram-se na faixa de 0,9821 a 0,9997; os limites de detecção (LOD) e de quantificação (LOQ) para os vários compostos investigados ficaram entre 0,0024-0,1968 mg kg -1 e entre 0,0072-0,5963 mg kg -1 , respectivamente.In this study, 14 pesticides (captan, chlorothalonil, deltamethrin, dicloran, fenthion, folpet, iprodione, linuron, malathion, prometryn, prochloraz, procymidone, trichlorphon and trifluralin) were employed for developing a multiresidue method (MRM) using pressurized liquid extraction (PLE). In situ clean-up was achieved using matrix solid phase dispersion (MSPD), where Florisil, XAD-4, XAD-7 and C18 were tested as dispersing material for carrot samples. Dichloromethane, ethyl acetate and acetone were used as solvents in a PLE unit at 75 ºC and 1,500 psi. Recovery was in the range of 70 to 133 %. Results obtained with wet carrots were not satisfactory, since most of the recovery values were below 70%. Determination coefficients of the analytical curves presented values between 0.9821 and 0.9970. Limits of detection (LOD) and quantification (LOQ) for the various components varied from 0.0024 to 0.1968 mg kg -1 , and 0.0072 to 0.5963 mg kg -1 , respectively. Keywords: multiresidue method, PLE, MSPD, carrots, GC/MS IntroductionIn the last few decades, pesticides have been used on an increasingly wider scale throughout the world. On account of the large number of active ingredients used in agricultural production, complex analytical procedures are required for the detection of a great number of possible compounds, with few number of extraction and clean-up steps. These compounds are characterized by their diversity, their different physical and chemical properties (polarity, solubility, pKa etc) and their low concentrations in real samples. 1 Some of the analytical methods aim to determine a single component (SRM) and others, more sophisticated, are capable of simultaneously determine more than one residue in a single analysis. These last ones are called pesticide multiresidue methods (MRM) and comprise a higher degree of analytical difficulty. Multiresidue methodology by definition requires determinative steps for separating analytes from one another so each one can be detected and measured ...

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Experimental Comparison of the Different Approaches To Estimate LOD and LOQ of an HPLC Method

Cited by 303 publications

References 10 publications

Nonlinear Regression Improves Accuracy of Characterization of Multiplexed Mass Spectrometric Assays

Nonlinear Regression Improves Accuracy of Characterization of Multiplexed Mass Spectrometric Assays

An efficient design strategy for a whole-cell biosensor based on engineered ribosome binding sequences

Multiresidue determination of pesticides in carrots using pressurized liquid extraction and gas chromatography with mass spectrometry detector

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