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Qualitative methods hold an important place in drug testing, filling central needs in screening and analyses, among others, linked to per se legislation. Nevertheless, the bioanalytical method validation guidelines do not discuss this type of method or describe method validation procedures ill-adapted to qualitative methods. The output of qualitative methods are typically categorical, binary results, such as presence/absence or above cut-off/below cut-off. As the goal of any method validation is to demonstrate fitness for use under production conditions, qualitative validation guidelines should evaluate performance based on discrete, binary results instead of the continuous measurements obtained from the instrument (e.g. area). A tentative validation guideline for threshold qualitative methods was developed by in silico modelling of measurements and derived binary results. This preliminary guideline was applied to a liquid chromatography-tandem mass spectrometry method for 40 analytes, each with a defined threshold concentration. Validation parameters calculated from the analysis of 30 samples spiked above and below the threshold concentration (false negative rate, false positive rate, selectivity rate, sensitivity rate and reliability rate) showed a surprisingly high failure rate. Overall, 13 out of the 40 analytes were not considered validated. A subsequent examination found that this was attributable to an appreciable shift in the standard deviation of the area ratio on a day-to-day basis, a previously undescribed and unaccounted-for behaviour in the qualitative threshold method validation literature. Consequently, the developed guideline was modified and used to validate a qualitative threshold method, based on the binary results for performance evaluation and incorporating measurement uncertainty.

Chronic wounds present a high risk of infection due to delayed and incomplete healing, leading to increased health risks and financial burden to health-care systems. Numerous approaches to promote wound healing have been extensively explored, especially the development of effective wound dressing materials embedded with therapeutic drug molecules. Despite advances made in this area, a remaining challenge to be addressed is the controlled, on-demand release of therapeutic molecules using noncytotoxic stimulus, for example, near-infrared (NIR) excitation. Here, we report a platform that allows for the development of electrospun poly(vinyl alcohol) (PVA) fibrous hybrids embedded with upconverting nanoparticles (UCNPs) and UV-cleavable levofloxacin conjugates for wound dressings. Upon irradiation with NIR light, the excited UCNPs emit UV light around 365 nm, which can cleave the

A study of impaired driving rates in the province of Québec is currently planned following the legalization of recreational cannabis in Canada. Oral fluid (OF) samples are to be collected with a Quantisal® device and sent to the laboratory for analysis. In order to prepare for this project, a qualitative decision point analysis method monitoring for the presence of 97 drugs and metabolites in OF was developed and validated. This high throughput method uses incubation with a precipitation solvent (acetone:acetonitrile 30:70 v:v) to boost drug recovery from the collecting device and improve stability of benzodiazepines (e.g., α-hydroxyalprazolam, clonazepam, 7-aminoclonazepam, flunitrazepam, 7-aminoflunitrazepam, N-desmethylflunitrazepam, nitrazepam). The Quantisal® device has polyglycol in its stabilizing buffer, but timed use of the mass spectrometer waste valve proved sufficient to avoid the glycol interferences for nearly all analytes. Interferences from OF matrices and 140 potentially interfering compounds, carryover, ion ratios, stability, recovery, reproducibility, robustness, false positive rate, false negative rate, selectivity, sensitivity and reliability rates were tested in the validation process. Five of the targeted analytes (olanzapine, oxazepam, 7-aminoclonazepam, flunitrazepam and nitrazepam) did not meet the set validation criteria but will be monitored for identification purposes (no comparison to a cut-off level). Blind internal proficiency testing was performed, where six OF samples were tested and analytes were classified as "negative", "likely positive" or "positive" with success. The final validated OF qualitative decision point method covers 92 analytes, and the presence of 5 additional analytes is screened in this high throughput analysis.

Evaluation of particle dynamics at the nano- and microscale poses a challenge to the development of novel velocimetry techniques. Established optical methods implement external or internal calibrations of the emission profiles by varying the particle velocity and are limited to specific experimental conditions. The proposed multiemission particle velocimetry approach aims to introduce a new concept for a luminescent probe, which guarantees accurate velocity measurements at the microscale, independent of the particle concentration or experimental setup, and without need for calibration. The simplicity of these analyses relies on the intrinsic luminescence dynamics of core-shell upconverting nanoparticles. Upon excitation with a focused near-infrared pulsed laser, the nanoparticle emits photons at different wavelengths. The time interval between emissions from different excited states is independent of the local environment or particle velocity. The velocity of the particles is calculated by measuring the distance between the maxima of two different emissions and dividing it by the known difference in luminescence lifetimes. This method is demonstrated using simple digital imaging of nanoparticles flowing in 75-150 µm diameter capillaries. Using this novel approach typically results in a relative standard deviation of the experimental velocities of 5% or lower without any calibration.

Several substances relevant for forensic toxicology purposes have an endogenous presence in biological matrices: beta-hydroxybutyric acid (BHB), gamma-hydroxybutyric acid (GHB), steroids and human insulin, to name only a few. The presence of significant amounts of these endogenous substances in the biological matrix used to prepare calibration standards and quality control samples (QCs) can compromise validation steps and quantitative analyses. Several approaches to overcome this problem have been suggested, including using an analog matrix or analyte, relying entirely on standard addition analyses for these analytes, or simply ignoring the endogenous contribution provided that it is small enough. Although these approaches side-step the issue of endogenous analyte presence in spiked matrix-matched samples, they create serious problems with regards to the accuracy of the analyses or production capacity. We present here a solution that addresses head-on the problem of endogenous concentrations in matrices used for calibration standards and quality control purposes. The endogenous analyte concentration is estimated via a standard-addition type process. This estimated concentration, plus the spiked concentration are then used as the de facto analyte concentration present in the sample. These de facto concentrations are then used in data analysis software (MultiQuant, Mass Hunter, etc.) as the sample’s concentration. This yields an accurate quantification of the analyte, free from interference of the endogenous contribution. This de facto correction has been applied in a production setting on two BHB quantification methods (GC-MS and LC–MS-MS), allowing the rectification of BHB biases of up to 30 μg/mL. The additional error introduced by this correction procedure is minimal, although the exact amount will be highly method-dependent. The endogenous concentration correction process has been automated with an R script. The final procedure is therefore highly efficient, only adding four mouse clicks to the data analysis operations.

A Tee configuration sheath flow cuvette with square cross-section channels has been produced in PDMS for CE detection. The output of a 1.4 W laser diode operating at 450 nm was focused onto the 300 μm core of a 370 μm od fiber optic whose end was inserted into one arm of the Tee for LIF. The optimal configuration had the fiber optic positioned 500 μm downstream from the intersection and the end of the 35 cm 50 μm id 365 μm od capillary just outside the intersection and in the leg of the Tee, resulting in a 90° configuration. Detection limits of 50 and 3 pM and linear calibrations of at least three orders of magnitude were obtained for Lucifer Yellow and fluorescein, respectively.

The stimulant designer drug 3,4-methylenedioxypyrovalerone (MDPV) was first synthesized by Boehringer Ingelheim in 1969 and introduced on the black market in 2006. Only a small number of fatal intoxication cases have been reported in the literature, all with significant blood MDPV concentrations. In this report, we describe one fatality attributed to an idiosyncratic reaction to MDPV. The victim displayed agitation, violent behavior and delirium followed by cardiac arrest. Hyperthermia was observed at the hospital. The MDPV cardiac and femoral blood concentrations were 6 ng/mL. The presence of excited delirium syndrome and MDPV, a drug with a pharmacology similar to cocaine, leads to the conclusion that the victim suffered a fatal adverse reaction to MDPV. This is the first published case of idiosyncratic reaction to MDPV.

Calculating the confidence interval is a common procedure in data analysis and is readily obtained from normally distributed populations with the familiar [Formula: see text] formula. However, when working with non-normally distributed data, determining the confidence interval is not as obvious. For this type of data, there are fewer references in the literature, and they are much less accessible. We describe, in simple language, the percentile and bias-corrected and accelerated variations of the bootstrap method to calculate confidence intervals. This method can be applied to a wide variety of parameters (mean, median, slope of a calibration curve, etc.) and is appropriate for normal and non-normal data sets. As a worked example, the confidence interval around the median concentration of cocaine in femoral blood is calculated using bootstrap techniques. The median of the non-toxic concentrations was 46.7 ng/mL with a 95% confidence interval of 23.9-85.8 ng/mL in the non-normally distributed set of 45 postmortem cases. This method should be used to lead to more statistically sound and accurate confidence intervals for non-normally distributed populations, such as reference values of therapeutic and toxic drug concentration, as well as situations of truncated concentration values near the limit of quantification or cutoff of a method.

Calibration model selection is required for all quantitative methods in toxicology and more broadly in bioanalysis. This typically involves selecting the equation order (quadratic or linear) and weighting factor correctly modelizing the data. A mis-selection of the calibration model will generate lower quality control (QC) accuracy, with an error up to 154%. Unfortunately, simple tools to perform this selection and tests to validate the resulting model are lacking. We present a stepwise, analyst-independent scheme for selection and validation of calibration models. The success rate of this scheme is on average 40% higher than a traditional "fit and check the QCs accuracy" method of selecting the calibration model. Moreover, the process was completely automated through a script (available in Supplemental Data 3) running in RStudio (free, open-source software). The need for weighting was assessed through an F-test using the variances of the upper limit of quantification and lower limit of quantification replicate measurements. When weighting was required, the choice between 1/x and 1/x2 was determined by calculating which option generated the smallest spread of weighted normalized variances. Finally, model order was selected through a partial F-test. The chosen calibration model was validated through Cramer-von Mises or Kolmogorov-Smirnov normality testing of the standardized residuals. Performance of the different tests was assessed using 50 simulated data sets per possible calibration model (e.g., linear-no weight, quadratic-no weight, linear-1/x, etc.). This first of two papers describes the tests, procedures and outcomes of the developed procedure using real LC-MS-MS results for the quantification of cocaine and naltrexone.

In the first part of this paper (I — Description and application), an automated, stepwise and analyst independent process for the selection and validation of calibration models was put forward and applied to two model analytes. This second part presents the mathematical reasoning and experimental work underlying the selection of the different components of this procedure. Different replicate analysis designs (intra/inter-day and intra/interextraction) were tested and their impact on test results was evaluated. For most methods, the use of intra-day/intra-extraction measurement replicates is recommended due to its decreased variability. This process should be repeated three times during the validation process in order to assess the time stability of the underlying model. Strategies for identification of heteroscedasticity and their potential weaknesses were examined and a unilateral F-test using the lower limit of quantification and upper limit of quantification replicates was chosen. Three different options for model selection were examined and tested: ANOVA lack-of-fit (LOF), partial F-test and significance of the second-order term. Examination of mathematical assumptions for each test and LC-MS/MS experimental results lead to selection of the partial F-test as being the most suitable. The advantages and drawbacks of ANOVA-LOF, examination of the standardized residuals graph and residuals normality testing (Kolmogorov-Smirnov or Cramer-Von Mises) for validation of the calibration model were examined with the last option proving the best in light of its robustness and accuracy. Choosing the correct calibration model improves QC accuracy, and simulations have shown that this automated scheme has a much better performance than a more traditional method of fitting with increasingly complex models until QC accuracies pass below a threshold.

A new liquid chromatography-capillary electrophoresis interface has been developed for multidimensional separations. The interface allows two second dimension separation capillaries to sample the chromatography effluent by servo controlled positioning of the capillary inlets in either the effluent or background electrolyte streams. The use of multiple second dimension capillaries allows higher first dimension sampling rates or multiple second dimension separation modes. Fluorescently labelled serum was separated by gel filtration and sub-micellar capillary array electrophoresis producing a multidimensional separation. Alternatively, using different background electrolytes for each capillary, two multidimensional separations with different selectivities were collected simultaneously. An easily fabricated PDMS multichannel sheath-flow cuvette was used for sensitive laser induced fluorescence detection of the labelled serum.

A centrifugal microanalysis system using spectrophotometric detection has been developed for the determination of nitrite and chromium VI. The system was designed to illustrate the potential of centrifugal microfluidic systems for on-site (field) analysis of waters. The performance was significantly better than manual/visual field test systems and approaches that of bench top systems with detection limits of 0.008 mg L(-1) NO(2)(-)-N and 0.03 mg L(-1) Cr(6+). The system used pre-loaded solid reagents on the disc, providing ease of use, and could process up to 24 samples simultaneously. Precision was enhanced by the use of a multiwavelength ratiometric blank estimation technique.

A simple method for producing a sheath flow cuvette in PDMS suitable for post-column detection in CE is described. Two types of cuvette were investigated. In the first, the sheath flow channel had a round cross-section of approximately 635 microm diameter, whereas the second cuvette had a 300x300 microm(2) square channel. Both cuvettes produced laminar flows that ensheathed the separation capillary's effluent allowing sensitive fluorescence measurements. The elasticity of the PDMS allowed the 300x300 microm(2) square sheath flow channel to expand uniformly and accommodate the larger 330-340 microm od round separation capillary, producing a self-aligning cuvette with robust mechanical properties. With this cuvette, linear calibrations of over five orders of magnitude and 15-30 zmol fluorescein detection limits were obtained for 12 and 50 microm id capillaries.

Objective To establish whether second trimester amniotic fluid protein and/or uric acid concentrations were associated with and predictive of infant birth weight and/or gestational age. Methods Second trimester amniotic fluid samples (n = 230) in mothers undergoing age-related amniocentesis for genetic testing were collected and quantified using capillary zone electrophoresis (CZE) for albumin, IgG, transferrin and uric acid. Maternal characteristics (prepregnancy weight and height, parity, ethnicity, smoking status) and infant birth weight, gestational age and gender were obtained from questionnaires and maternal obstetrical chart review. Results Preterm infants had higher concentrations of second trimester amniotic fluid transferrin than term infants (P = 0.0215). Transferrin was negatively associated with length of gestation, whereas uric acid was positively associated with the gestational age in spontaneous vaginal delivery (SVD) infants. Uric acid was also a significant predictor of the infant birth weight in grams. Conclusion Second trimester amniotic fluid transferrin and uric acid concentrations are related to subsequent birth outcomes and might emerge as biomarkers of early fetal development. Copyright © 2008 John Wiley & Sons, Ltd.

This paper investigates the use of scanning electron microscopy (SEM) and atomic force microscopy (AFM) for the characterization of monoliths used in capillary electrochromatography (CEC) while focusing on the nature of the information available from both techniques. SEM imaging revealed a compact structure of non-porous micrometer sized particles homogeneously agglomerated. With a simple AFM methodology, we found by direct observation that the same material exhibits mesopores in the nanometer range while SEM showed non-porous surfaces. These results obtained by AFM clearly showed that micrometer sized particles shrank and micrometer sized pores increased in the monolith when wetted. Thus, AFM was capable of demonstrating the morphological differences between wet and dried monolithic materials that are not possible by other imaging methods at micrometer resolution.

This review surveys the accomplishments in the separation of peptides and proteins by capillary electrochromatography (CEC) over the last decade. A significant number of research articles have been published on this topic since the last review. Peptide and proteins separations have been carried out in all three formats of CEC, i.e., packed bed, continuous bed and open-tubular (OT) format. In addition to electrophoresis, different chromatographic modes have been successfully exploited with the most prevalent being reversed-phase mode followed by ion-exchange. Although many researchers continue to use model proteins and peptides primarily to evaluate the performance of novel stationary phases some researchers have also applied CEC to the analysis of real-life samples. The potential of CEC to yield complementary information and sometimes a superior separation with respect to established techniques, i.e., microbore HPLC and capillary electrophoresis has been demonstrated. Instrumental modifications in order to facilitate coupling of CEC to mass spectrometry have further upgraded the value of CEC for proteomic analysis. Capillaries are still the separation vehicle of choice for most researchers yet the microfluidic platform is gaining momentum, propelled particularly by its potential for multitasking, e.g., performing different chromatographic modes in series.

Trace metals have been concentrated onto activated charcoal by using oxalic acid, iminodiacetic acid, and 8-hydroxyquinoline. The charcoal was directly analyzed in an ICP by placing several particles in a direct sample insertion (DSI) cup, or fabricating a DSI probe with an activated charcoal cap. Detection limits in the tens to hundreds of parts per trillion were found for Pb, Cd, Zn, and Cu. The recovery for Cu was 2–5% but may be improved by reactivation of the plasma-cleaned charcoal. Cleaning the charcoal, derived from wood, in the plasma prior to adsorption was necessary to remove trace metals.

A sample introduction system for inductively coupled plasmas (ICPs) which utilizes an induction coil for vaporizing analyte has been developed. This device uses HCl(g) in the carrier gas to prevent arcing in the induction furnace. Adding HCl has the additional benefit of acting as a physical carrier which can enhance analyte transport and allow the vaporization of relatively non-volatile elements. Experiments were carried out to investigate some of the performance characteristics of this vaporizer. Absolute detection limits using inductively coupled plasma atomic emission spectrometry (ICP-AES) for a number of elements ranged from 1.5 to 30 ng corresponding to relative detection limits which ranged from 0.04 to 0.7 ng ml −1 . Compared with solution nebulization, improvements in detection limits for these elements was 2–3 orders of magnitude. Analysis of Zn, Mn and Cu in a number of reference samples, including river water, lobster tissue and river sediment, demonstrated the applicability of this vaporizer for a variety of matrices.

An inductively coupled plasma was imaged with a charge coupled device camera through interference filters during direct sample insertion. Images were acquired throughout the insertion and revealed how the analyte behaves in the plasma. Graphite cups, graphite tubes and wire loops were used as sample carrying probes. The diameter of the graphite cup has a dramatic effect on the plasma and calcium emission. Narrower cups disturb the plasma the least and keep the calcium within the center of the plasma. The use of a carrier gas through a graphite tube and hollow stem cup shows that a darker central channel is established. The use of a central gas in combination with Freon-12 yields a performance comparable to that of wire loops. In the case of a deep and narrow graphite cup the analyte appears to emerge into the plasma along the walls of the cup.

Axial viewing generally provides an advantage over lateral viewing of the plasma for direct sample insertion. The signal change is element-specific and has produced detection limit improvements as high as a factor of 10 in the set studied. In the axial viewing mode the incandescent cup does not contribute to an increased background; however, the background and subsequent background noise levels are increased for most elements. Carrier gases were introduced into the cup via a hollow stem resulting in a reduction in the background signal, notably in the longer wavelength region. When 1000 ppm Freon-12 in argon is used as a carrier through the center of the cup, the normally problematic refractory elements are vaporized easily. Modifications and improvements to the design of the direct sample insertion device are also discussed.

Soil lead levels were determined on and around a former battery manufacturing site. Lead concentrations ranging from 120 ppm to 5.1’ were found. The highest concentrations were found close to the factory site. When it was possible to obtain samples over a continuous depth range, it was found that lead concentration decreased with depth and that it increased above underground foundations.

Aim: To identify, using capillary electrophoresis and chemometrics, early biomarkers in human amniotic fluid of large-for-gestational-age (LGA) infants. Materials & methods: Second trimester amniotic fluid samples, obtained from mothers undergoing age-related amniocentesis, were analyzed by capillary electrophoresis. Electropherogram data were aligned using correlation-optimized warping. A genetic algorithm using a Bayesian evaluation function and a leave-one-out cross-validation strategy for two birth outcomes: appropriate-for-gestational-age (AGA) versus LGA infants. Results: LGA (n = 23) was differentiated from AGA (n = 86) with a sensitivity of 100% and a specificity of 98% using only two wavelets. The first wavelet is associated with albumin and the second wavelet with an unknown small molecule. Conclusion: The approach developed herein allows LGA fetuses to be metabolically distinguished from AGA fetuses early in pregnancy and indicates that the birth of a LGA infant is already associated with an altered biochemical profile by the second trimester.

Aim: To detect gestational diabetes mellitus biomarkers in human amniotic fluid collected for age-related genetic testing using capillary electrophoresis and a sophisticated data analysis methodology. Materials & methods: Amniotic fluid samples were separated by capillary electrophoresis. Samples were classified using a genetic algorithm with Bayesian benefit function. The best model maximized the sensitivity and specificity and employed a leave-one-out cross-validation strategy. Results: Gestational diabetes mellitus (GDM; n = 14) was distinguished from non-GDM (n = 95) with 86% sensitivity and 99% specificity using two wavelets. These wavelets were located in the unresolved protein region and on the edge of the maternally derived albumin peak. Conclusion: GDM is a maternal pathology; however, it was shown that it alters the biochemical profile of amniotic fluid. Testing for GDM is normally carried out at 24–28 weeks, but changes can be detected at 15 weeks gestation, suggesting that GDM onset occurs early in gestation.

Cyanide is a powerful chemical asphyxiant found in some forensic cases following voluntary (suicide) or involuntary ingestion (fire, accidental exposure). A quantification method for cyanide that is specifically suited to post-mortem forensic purposes was developed. Determination was performed by headspace gas chromatography coupled to mass spectrometry using a GS-GASPRO column on an HP-6890 gas chromatograph with an HP-5973N mass detector. The biological sample was treated with an internal standard, frozen, glacial acetic acid was added and the sample was then incubated at 60°C for 15 min. The headspace was sampled with a disposable syringe, and analyzed to quantify hydrogen cyanide. Isotopically labeled cyanide ((13)C(15)N) was used as the internal standard to minimize matrix effect and sampling error. The method produced an extended linear dynamic range (0.07-50 μg/mL), and a method detection limit of 0.02 μg/mL. Identical calibration curves were obtained when blood, gastric contents and aqueous solutions were used as the calibration standard matrix. This method was also successful in quantitating cyanide in gastric contents, one of the most variable biological fluids. The method has been validated and is being used for current forensic cases such as fire victims and suicides.

Gestational diabetes mellitus (GDM) is a state of hyperglycaemia and increased oxidative stress with onset during pregnancy. Human serum albumin (HSA) was extracted from 26 GDM and 26 nonGDM amniotic fluid samples collected at 15 weeks gestation and analyzed by mass spectrometry. The majority of all albumin isoforms were oxidized with the cysteinylated HSA as the base peak in the deconvoluted spectrum. The HSA peak areas, from a control sample, had 36% relative standard deviation (RSD) across the six experimental days but using the relative isoform distribution improved the precision to 3−6%. The results show that the relative contribution of permanently oxidized HSA was greater (P = 0.002) and reversibly oxidized HSA was lower (P = 0.006) for GDM compared to nonGDM in the samples measured. This implies that the path toward GDM has been set prior to 15 weeks gestation and results in increased protein oxidation.

Effective protein characterization and identification are demanding and time-consuming operations in proteomics because of long-protein purification/separation procedures, and even longer enzymatic digestions. In this work, polymer-based monolithic enzyme reactors were fabricated in fused-silica capillaries, and performance was characterized through protein digestion and identification by MALDI-MS and ESI-MS. Reactors were prepared by fabricating a porous methacrylate base monolith followed by photografting with glycidyl methacrylate, and immobilization of the enzyme(s) with carbonyldiimidazole. Trypsin and Staphylococcus aureus V-8 protease (Glu-C) were used to produce three types of reactors: trypsin-based, Glu-C-based, and trypsin combined with Glu-C. Protein digestions, performed by perfusing protein solutions through the reactor under pressure, were evaluated based on the peptide map generated when directly coupled to an ESI mass spectrometer. Excellent digestion was observed over flow rates from 0.2 to 1 microL/min, which corresponds to reactor residence times of 0.24-1.4 min. As a proof of principle, chromatographic separation of model proteins followed by the digestion of specific fractions using these proteolytic enzyme reactors and ESI-MS is demonstrated.

This short communication describes features of UV-transparent capillaries employed for CEC and CE. A waveguide effect was observed when using UV-transparent capillaries. Through imaging with SEM, the UV-transparent coating was found to be highly porous unlike polyimide coating, which did not exhibit any porosity at all. Prolonged exposure to several commonly employed solvents with elevated pH caused abrasion of the coating at the capillary tip but no swelling of the UV-transparent coating was observed. Lastly, four different cutting techniques were compared to obtain smooth capillary tips.

The developing fetus begins to swallow amniotic fluid (AF) early in gestation, a process that results in ingestion of numerous growth factors. Our objectives were 2-fold: 1) to assess the concentration and distribution of insulin-like growth factor II (IGF II) and its binding proteins (BP) 1 and 3 in 2nd trimester amniotic fluid using ELISA, and 2) to establish whether concentrations of AF IGF II and its binding proteins IGF BP1 and 3, measured early in pregnancy, were associated with and predictive of infant birth weight. Birth weights were categorized using recently developed birth-weight-for-gestational-age percentiles for fetal growth in which infants10% were classified as SGA (small-for-gestational-age) and those90% as LGA (large-for-gestational-age). AF samples were collected after routine genetic testing (15.1 +/- 0.04 wk, range 12-20 wk) from 543 mother-infant pairs in Montreal, QC, Canada. Maternal and fetal characteristics were obtained from questionnaires and medical chart review. Multivariate regression analysis that controlled for maternal height, prepregnancy weight, smoking behavior, infant gender, gestational age, parity, as well as amniocentesis week showed that higher AF IGF BP1 was associated with lower birth weight (partial r2 = 0.0062). Regression analyses revealed that AF IGF BP3 was positively associated with birth weight within LGA and macrosomia subpopulations (partial r2 = 0.0283 and 0.0404, respectively). These results show that 2nd trimester AF IGF BP1, BP3, and IGF II may emerge as early indicators of fetal growth.

This work presents the separation of model proteins by capillary electrochromatography involving a monolithic stationary phase with C 4 functionality. The monolith was fabricated in UV-transparent capillaries by employing a slight modification of a recently published photopolymerization procedure. With the number of theoretical plates per column ranging between 11 000 and 33 000, the separation efficiency proved to be lower than capillary zone electrophoresis where plate numbers ranged between 18 000 and 66 000. However, higher resolution was obtained due to the additional chromatographic separation mechanism. Inter- and intra-column reproducibility were evaluated, the latter could be significantly improved when using a rinsing procedure that contained 0.05% sodium dodecylsulfate in the mobile phase. Plate heights became nearly independent of mobile phase velocities higher than 0.5 mm/s indicating that high velocities can be applied without sacrificing efficiency. Furthermore, peak heights showed a dependence on injection times. For proteins, an increase in capacity factors was found when increasing the percentage of organic solvent in the mobile phase.

We have developed a simple and rapid immunoassay that uses antibody-coated fluorescent microspheres for high sensitivity detection of antigens. Detection limits in the low pg/ml (10−14M) are obtainable with this method. This assay has been demonstrated in a flow cytometer, a capillary sheath flow cuvette and a microfluidic chip format. Multiple antigens can be simultaneously quantitated within a single sample by judicious choice of bead dye concentration and colour combination.

Using short lengths of fused silica capillary tubing embedded in the disk, a system for valving and filtering samples on centrifugal microfluidic devices has been designed and implemented. Sedimentation of turbid samples and transfer of the clear supernatant was also accomplished. Also demonstrated is the transfer of the liquid through a second capillary valve to the final reservoir. By controlling rotational speed, sedimentation and multiple step valving operations are readily implemented and easily prototyped.

A compact, rapid and cost-efficient system for the determination of total mercury (Hg) in a single human hair strand is presented. The system uses induction heating-electrothermal vaporization and gold amalgamation trapping in tandem with either atomic fluorescence spectrometry or atomic absorption spectrometry with background correction detection. Using these techniques, detection limits of 0.1 ng or 0.2 μg g−1 and 0.08 ng or 0.1 μg g−1 (based on a 0.6 mg sample) of Hg were achieved, respectively.

Recently it was demonstrated that a collimated Gaussian beam can be converted to a flattop beam using a Keplerian beam reshaper consisting of two aspheric lenses. Here, using the same optical system, we demonstrate that this flattop profile can be maintained when used in a confocal detection system that focuses the laser beam into a diameter of only 33 microm. The intensity profile of the reshaped beam was determined by imaging the excitation of a constant stream of fluorescein inside a microfluidic device. The resulting shape had a 6.38% RSD intensity across the flat profile when imaged with a CCD camera. This flat illumination profile was then used in a two bead multiplex immunoassay where the colour combination and/or the intensities can be used to determine the presence of an antigen. Detection limits of TNF-alpha and IL-6 were comparable with standard ELISA immunoassays.