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Attogram detection limit for aqueous dye samples by laser-induced fluorescence.

A modified flow cytometer has been used to detect attogram quantities of aqueous rhodamine 6G by laser-induced fluorescence analysis. A detection limit of 28 attograms (35,000 molecules) was obtained, nearly two orders of magnitude better than earlier measurements. The detection limit in concentration units was 1.4 x 10(-13) mole per liter. During the 1-second measurement period, the total volume sampled was 0.42 microliter. On average, only half a rhodamine 6G molecule was present in the 6-picoliter probed volume.

Flow Cytometry↗

Detection limit of methods to assess fluid status changes in dialysis patients.

Technical systems for an accurate and practicable fluid management of dialysis patients are urgently needed, since current clinical methods are partially subjective, imprecise, and time consuming. Such new systems should not only allow the determination of the target normohydration weight, but also must be able to detect clinically relevant changes in fluid volume ( approximately 1 l). This study focuses on the systematic analysis of the detection limit of several candidate methods for fluid management. In a cohort of 16 new dialysis patients, several candidate methods were applied in parallel during each treatment of the initial weight reduction phase: the measurement of vena cava diameter (VCD), vena cava collapsibility index (CI), the blood volume drop during an ultrafiltration (UF) bolus (Deltarelative blood volume (RBV)-), the rebound after the UF bolus (DeltaRBV+), and the extracellular fluid volume determined with whole body bioimpedance spectroscopy (BIS). A clinical reference method was used to manage the fluid status of patients. All methods showed significant correlations with predialysis weight. The detection limits W(lim) of candidate methods for changes in fluid status were assessed as W(lim)=0.87 kg+/-0.64 kg (BIS), 1.74 kg+/-1.56 kg (VCD), 2.3 kg+/-1.0 kg (DeltaRBV-), 7.4 kg+/-8.5 kg (CI), 40 kg+/-108 kg (DeltaRBV+). Only BIS shows a satisfactorily low detection limit W(lim), whereas W(lim) was rated as critical for the VCD and DeltaRBV- methods, and as unacceptable for the CI and DeltaRBV+ methods. Bioimpedance spectroscopy appears to be the most promising method for a practical fluid management system in dialysis.

Adult↗

A first and straightforward approach to detection limits for narrow-bore HPLC and CE.

Two methodologies have been proposed for a simple statistical estimation of the detection limits in microbore-HPLC and CE techniques. Due to the impossibility to measure the blank, by using the extrapolated or approximated expressions it is possible to obtain a straightforward approach of the detection limits from the study of the residuals of the calibration data set, avoiding tedious treatment of the signals. The utility of these expressions has been checked on the data from an experimental microbore HPLC system.

Calibration↗

Comparison of detection limits in environmental analysis--is it possible? An approach on quality assurance in the lower working range by verification.

Detection limit, reporting limit and limit of quantitation are analytical parameters which describe the power of analytical methods. These parameters are used for internal quality assurance and externally for competing, especially in the case of trace analysis in environmental compartments. The wide variety of possibilities for computing or obtaining these measures in literature and in legislative rules makes any comparison difficult. Additionally, a host of terms have been used within the analytical community to describe detection and quantitation capabilities. Without trying to create an order for the variety of terms, this paper is aimed at providing a practical proposal for answering the main questions for the analysts concerning quality measures above. These main questions and related parameters were explained and graphically demonstrated. Estimation and verification of these parameters are the two steps to get real measures. A rule for a practical verification is given in a table, where the analyst can read out what to measure, what to estimate and which criteria have to be fulfilled. In this manner verified parameters detection limit, reporting limit and limit of quantitation now are comparable and the analyst himself is responsible to the unambiguity and reliability of these measures.

Journal Article↗

A new system to detect residual subgingival calculus: in vitro detection limits.

AIM: We recently introduced an experimental surface detection system based on a conventional dental ultrasonic scaler. This device automatically discriminates cementum and dental calculus, which is the prerequisite for complete and thorough calculus removal. In the present study, the detection limits of this device were tested in vitro. MATERIAL AND METHODS: From 50 extracted teeth, subgingival calculus was gradually removed using a Gracey curette. During this stepwise procedure, detection properties of the surface detection system were continuously monitored and systematically verified until the system stopped discriminating calculus from the root surface. By measuring the diameter, circumference and area of the smallest, yet recognizable deposit, and of the no longer recognizable deposit, the cut-off point of the discriminative capability of the detection device was determined. RESULTS: The cut-off points for the correct classification of residual deposits averaged on a diameter of 219 microm, an area of 21,600 microm2, and a circumference of 748 microm. This means a sensitivity of 73% and a specificity of 80% in this critical area. CONCLUSIONS: This calculus detection system was able to detect small deposits. In clinical practice, this device may support dentists in deciding whether to stop or to continue the debridement.

Dental Calculus↗

A polypyrrole-based solid-contact Pb(2+)-selective PVC-membrane electrode with a nanomolar detection limit.

Ion-selective electrodes (ISE) based on conventional plasticized PVC membranes with solid inner contact (SC) have so far had unsatisfactory lower detection limits. Here it is shown that electropolymerization of pyrrole in the presence of potassium hexacyanoferrate(II)/(III) on Pt results in adequate inner contact for SC-ISE. The nanomolar lower detection limit achieved with the Pb(2+)-selective PVC membranes investigated is comparable with values obtained with optimized internal solutions of liquid-contact ISE.

Electrodes↗

Air-cooled cold trap channel integrated in a microfluidic device for monitoring airborne BTEX with an improved detection limit.

We integrated an air-cooled cold trap (CT) channel in a microfluidic device for monitoring airborne benzene, toluene, ethylbenzene, and xylene (BTEX) gases and demonstrated its effect on improving the detection limit of the microfluidic device. The device consists of concentration and detection cells formed of 3 x 1 cm Pyrex plates. We first introduced a sample gas into the concentration cell, and the gas was adsorbed onto an adsorbent in the channel. We then raised the temperature using a thin-film heater and introduced the desorbed gas into the detection cell. To prevent dilution of the gas before detection, we propose an improvement to the concentration cell structure that involves the integration of the CT channel. We examined the CT effect by comparing three types of concentration cell with different channel structures. We found that we could detect a gas concentration about 2 orders of magnitude lower than in our previous work by optimizing the channel structure and integrating a CT channel. As an example of BTEX detection,we obtained a 0.05 ppm detection limit for toluene gas with a sampling time of 30 min.

Adsorption↗

A mixture model with detection limits for regression analyses of antibody response to vaccine.

Antibody concentration values as determined by quantitative assays often are left-censored due to detection limits or limits established for purpose of specificity. Standard analyses which assume the data arise from a single lognormal response distribution may not be appropriate, when more observations are censored than would be expected under such a model. Interference from maternal antibodies due to vaccination at an early age, for example, could result in a high proportion of nonresponders to vaccine. A mixture model consisting of a censored lognormal distribution and a point distribution located below the detection limit is proposed for such situations. Antibody data from a study of measles vaccine are used to illustrate the utility of this approach and the interpretation of the model parameters.

Antibodies, Viral↗

Bayesian decision threshold, detection limit and confidence limits in ionising-radiation measurement.

Based on Bayesian statistics and the Bayesian theory of measurement uncertainty, characteristic limits such as the decision threshold, detection limit and limits of a confidence interval can be calculated taking into account all sources of uncertainty. This approach consists of the complete evaluation of a measurement according to the ISO Guide to the Expression of Uncertainty in Measurement (GUM) and the successive determination of the characteristic limits by using the standard uncertainty obtained from the evaluation. This procedure is elaborated here for several particular models of evaluation. It is, however, so general that it allows for a large variety of applications to similar measurements. It is proposed for the revision of those parts of DIN 25482 and ISO 11929 that are still based on conventional statistics and, therefore, do not allow to take completely into account all the components of measurement uncertainty in the calculation of the characteristic limits.

Algorithms↗

Energy-filtering TEM at high magnification: spatial resolution and detection limits.

Energy-filtering TEM (EFTEM) has turned out to be a very efficient and rapid tool for the chemical characterization of a specimen on a nanometer and even subnanometer length scale. Especially, the detection and measurement of very thin layers has become a great application of this technique in many materials science fields, e.g. semiconductors and hard disk technology. There, the reliability of compositional profiles is an important issue. However, the experimentally obtainable spatial resolution strongly influences the appearance of a thin layer in an EFTEM image, when dimensions reach subnanometer levels, which mainly leads to a broadening of the layer in the image. This fact has to be taken into account, when measuring the thickness of such a thin layer. Additionally, the convolution decreases contrast which makes the layer less visible in the image and finally determines the detection limit. In this work we present a systematic study on specifically designed Mn/PdMn multilayer test specimens to explore the practical aspects of spatial resolution and detection limits in EFTEM. Although specific to the ionization edges used, we will present general conclusions about the practical limitations in terms of EFTEM spatial resolution. Additionally, work will be shown about low energy-loss imaging of thin oxide layers, where delocalization is the main factor responsible for broadening.

Journal Article↗

Extending the detection limit of solid-phase electrochemical enzyme immunoassay to the attomole level.

Electrochemical enzyme immunoassay methodology has been developed to take advantage of the selectivity of antibody reactions, the amplification feature of an enzyme-based assay, and the ease with which small amounts of the enzyme-generated product can be detected electrochemically. A heterogeneous sandwich enzyme immunoassay was used in this work as the model assay. In this type of assay, the antigen is sandwiched between the enzyme conjugate and a primary antibody that is adsorbed to the solid phase. Alkaline phosphatase is a suitable enzyme for electrochemical assays since it catalyzes the conversion of electroinactive phenyl phosphate to electroactive phenol. The product, phenol, is then quantitated by liquid chromatography with electrochemical detection in a thin-layer flow cell with a carbon paste electrode at 0.895 V vs Ag/AgCl. The current produced by the oxidation of phenol is directly proportional to the analyte (antigen) concentration. The problem associated with these types of solid-phase immunoassays is that the adsorption of the primary antibody is desired while the adsorption of other assay proteins is not. The detection limits are generally defined by the ability to control this nonspecific adsorption. The detection limit of a previous electrochemical assay for rabbit IgG was 100 pg/ml and was limited by a large background current observed in the absence of antigen. In the present study, each step of the assay was examined in order to determine the sources of this background current, and it was found that the major contribution was from the nonspecific adsorption of the enzyme conjugate. Using combinations of Tween 20 and bovine serum albumin as blocking agents, the level of nonspecific adsorption was reduced by 96%.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The dependence of radioimmunoassay detection limits on antibody affinity.

Competitive radioimmunoassay standard curves for progesterone were established with tritiated progesterone and six different anti-progesterone monoclonal antibodies ranging in affinity from 1.23 x 10(8) to 2.82 x 10(10) l/mol. The detection limits for these curves ranged from 133 to 6,670 pmol/l final assay concentration (16.7 to 839.1 pg/tube). Separately, a mass-action mathematical model was used to predict lower detection limits for assays run under equivalent conditions with the six antibodies. When compared to the experimental data the predictions were reliable for the higher affinity antibodies but became less accurate as the affinity decreased.

Animals↗

Detection limits and receiver operating characteristic curve analysis in the evaluation of specific IgE assays.

One of the major sources of uncertainty and controversy in allergy testing is the definition and clinical significance of a positive test result in the low assay range. Use of analytical detection limit theory can guide the diagnostic allergy laboratory director in setting a lower assay cutoff and in the evaluation of the recommended cutoff by the manufacturer of the assay. Several methods for calculating lower limit of detection are applicable to specific IgE assay methods. Receiver operating characteristic (ROC) curves illustrate the relationship between statistical sensitivity and specificity as cutoffs are adjusted, and statistical analysis of ROC curves is an invaluable aspect of the comparative performance evaluation of two or more assays relative to an independent standard such as skin testing.

False Negative Reactions↗

Determination of the instrumental detection limits of commercial nonylphenol ethoxylates with a wide range of molecular masses using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Recent interest in the environmental fate and impact of nonylphenol ethoxylates (NPEs) has necessitated the need for the development of specific, and yet efficient, analytical methodologies for their detection. Using seventeen commercial NPEs, all having the general formula 4-(C(9)H(19))-C(6)H(4)-(OCH(2)CH(2))(n)-OH, we have established minimum concentrations at which matrix-assisted laser desorption ionization (MALDI)/time-of-flight (TOF)/mass spectrometry (MS) can be used in their detection. The NPEs were dissolved in an acetonitrile/tetrahydrofuran (3:1) solvent system prior to mixing with the matrix (alpha-cyano-4-hydroxycinnamic acid, dissolved in acetonitrile/tetrahydrofuran) solution, and the resulting MALDI-TOF mass spectra produced mostly sodiated molecules [M + Na](+).For the NPEs in which the major components contain three to ten ethoxy units, the sodiated molecules were detected at a minimum analyte concentration of 10 &mgr;g/L, whereas those in which the major components contain eight to fifteen ethoxy units were detected at a minimum concentration of 30 &mgr;g/L. Those NPEs containing oligomers in excess of twenty ethoxy units were only detectable at much higher concentration. For examples, Surfonic(R) N-300, N-400, N550, N-700, N-800, and N-1000 had detection limits of (&mgr;g/L): 1000, 1600, 2000, 2600, 3500 and 4500, respectively. These variations in instrumental detection limits must be considered when analyzing environmental samples for the presence of nonylphenol ethoxylates. Copyright 1999 John Wiley & Sons, Ltd.

Journal Article↗

Detection limit, cut-off and specificity of an improved rapid assay for cardiac troponin T.

The performance of an improved version of the troponin T rapid test TROPT Sensitive was investigated in a multicentre evaluation at twelve centres. The detection limit and the cut-off were determined in a method comparison with Elecsys Troponin T using a total of 365 samples from patients with suspected acute coronary syndromes and 91 samples from healthy blood donors or non-cardiological patients. The analytical specificity was determined by measuring 1271 blood samples from blood donors without any myocardial injury. The test cut-off (90% of results positive) is 0.08 microg/L, and the detection limit is about 0.05 microg/L. The analytical specificity of the test is between 99.7 and 99.9%. With its small area of undefined significance between positive and negative results and its high sensitivity and specificity, TROPT Sensitive is very well suited to the reliable detection of troponin T positive patients with acute coronary syndromes.

Biomarkers↗

On the estimate of capillary electrophoresis blanks: application to detection limits evaluation as recommended by IUPAC.

A new method to obtain the signal associated with an electrophoretic blank is presented to be included in the calibration procedures. The signal assigned to the blank is obtained by direct integration of the background noise using extrapolated values of the base-peak width at different concentrations in order to obtain the zero concentration. Detection limits that are better adjusted to a statistical evaluation are thus implemented, as recommended by the International Union of Pure and Applied Chemistry (IUPAC). The detection limit has been examined for a series of linear alkyl benzenesulfonates (C10-C13) and total linear alkyl benzenesulfonates that are widely used in many different commercial products.

Benzenesulfonates↗

Evaluation of probe chemistries and platforms to improve the detection limit of real-time PCR.

A validated PCR-based Salmonella method targeting a 94-bp sequence of the ttr gene was used as a model to compare six different combinations of reporter and quencher dyes of a TaqMan probe, on three different instruments, to improve the detection limit in a real-time PCR assay with the aim of a same-day analysis. The use of locked nucleic acids (LNA) and Scorpion probes were also tested. The combination FAM-BHQ1 or Cy5-BHQ3, both dark quenchers, gave the best results (Cycle threshold (Ct) of 25.42+/-0.65 and 24.47+/-0.18 at 10(3) DNA copies). When comparing different probe technologies, the LNA probe (FAM-BHQ1) was the most sensitive with the strongest fluorescence signal (dR last 48066), resulting in 0.6 to 1.1 lower Ct values than a DNA TaqMan probe, and 1.9 to 4.0 lower Ct than the Scorpion system (FAM-BHQ1). The RotorGene real-time PCR instrument gave 0.4-1.0 lower Ct values (more sensitive) than the Mx3005p, and 1.5-3.0 lower than the ABI 7700. Using the LNA in a RotorGene instrument, we detected the following Salmonella DNA copies in 1-ml pre-enriched samples: fishmeal (100 copies), chicken rinse (100 copies) and pig feces (10 copies). The detection probability of the final assay on inoculated fecal samples was 100% at 2x10(4) copies per ml. In conclusion, the LNA probe with annealing temperature of 65 degrees C could be useful for more sensitive detection limits.

Animals↗

Use of coincidence techniques to improve the detection limits of electron spectroscopy in STEM.

A method of improving the detection limits of microanalysis using electron energy losses due to inner-shell excitation is proposed. This is based on coincidence detection of the energy-loss electrons with another signal which also results from inner-shell excitations, i.e., Auger electrons or characteristic X-rays. It is concluded that there will be a significant improvement in the detection of monolayers on a homogeneous monatomic substrate by using coincidence of energy-loss electrons with Auger electrons.

Electron Probe Microanalysis↗