Smart-detection approach for protein residues to evaluate the cleaning efficacy of reusable medical devices.

BACKGROUND: Healthcare-associated infections are challenging problems to eradicate. In practice, infection prevention and control regarding the effective cleaning of reusable medical devices is performed. However, real-time monitoring of potentially contaminated devices is hindered by the complex, costly investigations and the time required. AIM: To develop an inexpensive approach providing high-sensitivity, rapid, and low-cost protein residue determination on medical devices. METHODS: Colourimetric reactions for protein were modified and downscaled to produce a colour product on the moving drop platform and to operate combined with our custom-built smartphone application in order to evaluate and save protein contamination data. The intensity of a product colour was detected by photographing with a smartphone camera and was proportional to the protein concentration in the linear range of 5-20 µg/mL. The protein content was processed on a custom-built smartphone application, namely PromoveCheck. FINDINGS: The proposed procedure was highly sensitive, yielding limits of detection and limits of quantification, at 0.03 and 0.10 µg based on the Bradford reaction. It was applied to reusable medical devices, showing no significant differences from the spectrophotometric method and a correlation with the o-phthaldialdehyde method (P<0.05). CONCLUSION: This approach demonstrates low cost, sensitivity, efficiency and traceability for investigating reusable medical device cleanliness. The PromoveCheck app can report and acquire protein content that can be used for traceability and alerts concerning questionable or discovered contaminant devices, improving the quality management of the Central Sterile Supply Department.

Evaluation and application of a paper-based device for the determination of reactive phosphate in soil solution.

The evaluation and validation of a new low-cost microfluidic paper-based analytical device (µPAD) for the determination of reactive phosphate in soil solution is described. This device allows up to 15 replicate measurements of reactive phosphate on one credit card-sized device and requires only a desktop or hand scanner for signal detection and quantification. The proposed method showed a linear response between 0.1 and 1.0 mg L and between 1.0 and 10.0 mg L P with a limit of detection of 0.05 mg L P. When applied to the analysis of soil solution, there was excellent agreement between results obtained using the µPAD and those obtained by a reference spectrophotometric method, as indicated by the following regression equation: [P] = (0.997 ± 0.005)[P] - (0.020 ± 0.008) ( = 0.997; = 110). It was found that the ambient temperature storage stability of the µPAD could be extended to 15 d by incorporating a removable polymeric interleaving sheet between the adjacent paper layers of the device. The observed sensitivity of the µPADs to sunlight, which was manifested by photoreduction of the chromogenic molybdate reagent used in the assay, was overcome by preparing the µPADs with an ultraviolet-filtering laminating material. The proposed method is rapid, with a reaction time of only 10 min, is easy to perform, and is suitable for application in the field.

Simple lab-on-chip approach with time-based detection.

A simple lab-on-chip approach with time-based detection is proposed. A platform is made from a piece of acrylic differently shaped channels for introducing sample and reagent(s) using flow manipulation. Time-based changes involving migration of the reaction zone are monitored. The changes can be visually monitored by using a stop-watch with naked eyes observation. Some applications for the determination of ascorbic acid, acetic acid and iron in real samples with different chemistries were demonstrated.

Evaluation of on-line preconcentration and flow-injection amperometry for phosphate determination in fresh and marine waters.

Dissolved reactive phosphorus (DRP) was determined as orthophosphate (PO(4)-P) in fresh and saline water samples by flow-injection (FI) amperometry, without and with in-valve column preconcentration. Detection is based on reduction of the product formed from the reaction of DRP with acidic molybdate at a glassy carbon working electrode (GCE) at 220 mV versus the Ag/AgCl reference electrode. A 0.1M potassium chloride solution was used as both supporting electrolyte and eluent in the preconcentration system. For the FI configuration without preconcentration, a detection limit of 3.4 microg Pl(-1) and sample throughput of 70 samples h(-1) were achieved. The relative standard deviations for 50 and 500 microg Pl(-1) orthophosphate standards were 5.2 and 5.9%, respectively. By incorporating an ion exchange preconcentration column, a detection limit of 0.18 microg Pl(-1) was obtained for a 2-min preconcentration time (R.S.D.s for 0.1 and 1 microg Pl(-1) standards were 22 and 1.0%, respectively). Potential interference from silicate, sulfide, organic phosphates and sodium chloride were investigated. Both the systems were applied to the analysis of certified reference materials and water samples.

Determination of trace amounts of bromide by flow injection/stopped-flow detection technique using kinetic-spectrophotometric method.

A simple, sensitive and selective method for the determination of bromide in seawater by using a flow injection/stopped-flow detection technique was examined. The detection system was developed for a new kinetic-spectrophotometric determination of bromide in the presence of chloride matrix without any extraction and/or separation. The detection was based on the kinetic effect of bromide on the oxidation of methylene blue (MB) with hydrogen peroxide in a strongly acidic solution. Large amounts of chloride could enhance the sensitivity of the method as an activator. The decolorisation of the blue color of MB was used for the spectrophotometric determination of bromide at 746nm. A stopped-flow approach was used to improve the sensitivity of the measurement and provide good linearity of the calibration over the range of 0-3.2mugml(-1) of bromide. The relative standard deviation was 0.74% for the determination of 2.4mugml(-1) bromide (n=5). The detection limit (3sigma) was 0.1mugml(-1) with a sampling frequency of 12h(-1). The influence of potential interfering ions was studied. The proposed method was applied to the determination of bromide in seawater samples and provided satisfactory results.

Simple and selective method for determination of iodide in pharmaceutical products by flow injection analysis using the iodine-starch reaction.

This work exploited the well-known iodine-starch reaction for development of a simple flow-injection (FI) method for determination of iodide in pharmaceutical samples. Iodide in an injected zone was oxidized to iodine. A gas diffusion unit enables selective permeation of iodine through a hydrophobic membrane. Detection was made very selective for elemental iodine by employing formation of the I(3)(-)-starch complex. The detection limit (3 S/ N) of the system was 1 mg I L(-1). For a liquid patent medicine used for asthma treatment we suggested modification of the system. Direct injection of this sample, which contains a particularly high concentration level of iodide (ca. 9000 mg I L(-1)), can be achieved by coupling a dialysis unit to the FI system. This has increased the working range to 6000-10,000 mg I L(-1) without employing complicated nanoliter injection.

Cost-effective flow injection spectrophotometric assay of iron content in pharmaceutical preparations using salicylate reagent.

A new flow injection procedure for an assay of Fe(III) by using salicylate obtained from antipyretic powder, which is a cheap and easily available reagent, is proposed. A red complex was continuously monitored by a laboratory-made green LED colorimeter. A linear calibration was obtained in the range of 1-20mgFel(-1) with a detection limit of 0.5mgFel(-1) and R.S.D.s of 1.4-5.4% (n=3, for 1-20mgFel(-1)). The new procedure was applied to assay iron contents in pharmaceutical preparations. The results were in good agreement with those of the USP standard method.

Flow-injection determination of iodide ion in nuclear emergency tablets, using boron-doped diamond thin film electrode.

The electrochemical determination of iodide was studied at boron-doped diamond thin film electrodes (BDD) using cyclic voltammetry (CV) and flow-injection (FI) analysis, with amperometric detection. Cyclic voltammetry of iodide was conducted in a phosphate buffer pH 5. Experiments were performed using glassy carbon (GC) electrode as a comparison. Well-defined oxidation waves of the quasi-reversible cyclic voltammograms were observed at both electrodes. Voltammetric signal-to-background ratios (S/B) were comparable. However, the GC electrode gives much greater in the background current as usual. The potential sweep rate dependence exhibited that the peak current of iodide oxidation at 1mM varied linearly (r(2) = 0.998) with the square root of the scan rate, from 0.01 to 0.30Vs(-1). This result indicates that the reaction is a diffusion-controlled process with negligible adsorption on BDD surface, at this iodide concentration. Results of the flow-injection analysis show a highly reproducible amperometric response. The linear working range was observed up to 200muM (r(2) = 0.999). The detection limit, as low as 0.01muM (3sigma of blank), was obtained. This method was successfully applied for quantification of iodide contents in nuclear emergency tablets.

A simple flow injection system with bead injection for trace iron determination.

A simple and low cost flow injection (FI) system with bead injection (BI) was developed for determination of low concentration (mumol l(-1)) of iron in water samples. Chelex-100 chelating resin beads, trapped in a jet ring cell, were employed. The intensity of red complex of 1,10-phenanthroline with Fe(2+) was monitored using colorimetric detector with a LED green light source. Amount of total Fe (Fe(2+) and Fe(3+)) and Fe(2+) can be evaluated by with and without reduction of Fe(3+) using ascorbic acid. Lowest detectable levels of Fe(2+) were 0.90 and 0.45 mumol l(-1) for sample loading time of 3 and 5 min, respectively. Working range was up to 3.90 mumol l(-1) using 0.3% w/v 1, 10-phenanthroline. Percent recoveries of spiked water samples (0.90-2.33 mumol l(-1) of Fe(2+)) were 100-110%.

Stopped-flow injection simultaneous determination of phosphate and silicate using molybdenum blue.

Kinetic information for the phosphate-molybdate-ascorbic acid reaction can be obtained by making use of a very simple manually operated stopped-flow injection (FI) system. Various parameters (concentrations of reagents, flow rate, mixing coils, and volume of flow cell) were investigated for determination of phosphate. A stopped-FI system should be arranged for low degree of mixing (of reactants) and low dispersion so that good signals of rate changes will be observed. Simultaneous determination of phosphate and silicate by the stopped-FI technique is proposed, using a laboratory-made semi-automatic stopped-FI Analyzer with LED-based photometer. It is based on kinetic separation of phosphate and silicate using molybdenum blue. The proposed procedure has been demonstrated for the application to water samples. The results obtained agree with that of a standard method.

Determination of trace iron in beer using flow injection systems with in-valve column and bead injection.

Three flow injection (FI) systems were investigated for the determination of trace iron in beer: an FI-in-valve column-flame atomic absorption spectrophotometry (FI-FAAS) system, a spectrophotometric FI system with a column placed at the detection point, and an FI-spectrophotometric system with bead injection (FI-BI). Cationic exchange resin Dowex 50W X8 and iminodiacetate chelating resin, Chelex-100, were employed for the FI-spectrophotometric and FI-FAAS systems, respectively. The FI-in-valve column, packed with the resin, enhances the FAAS performance. The spectrophotometric FI system with a column (packed with Chelex-100) placed at the detection point (in a cell holder of a spectrophotometer) is based on the formation of iron (II)-1,10-phenanthroline complex sorbed onto the resin. No eluent has been found to be suitable. The FI-BI for renewable microcolumn has been proven to be an alternative. The FI-FAAS and FI-BI procedures provide online sample preseparation and preconcentration for the determination of iron in beer. Both are simple, rapid, and economical. The procedures also involve sample preparation (decarbonation and suppression of tannin interference by adding ascorbic acid) and standard addition. The results obtained by FI-FAAS and FI-BI agree with those of AOAC spectrophotometric method.

Gravitational field-flow fractionation in combination with flow injection analysis or electrothermal AAS for size based iron speciation of particles.

A simple gravitational field-flow fractionation (GrFFF) system was used for size separation of micron sized silica particles coated with hydrous iron oxide (geothite). The amount of iron on the particles was monitored either on-line by reverse-flow injection analysis (r-FIA) with chemiluminescence detection using luminol or off-line by electrothermal atomic absorption spectrophotometry (ETAAS). The combination of GrFFF with reverse FIA or with ETAAS has been demonstrated to be a cost-effective tool for size based iron speciation of particles.

Continuous and stopped flow injection for catalytic determination of total iodine in urine.

This paper describes the use of flow injection (FI) techniques for the determination of iodine in urine, based on the catalytic effect of iodide in the redox reaction between Ce(IV) and As(III). The proposed procedures minimize errors in the conventional batch method arising from the reading of absorbance at a fixed time after addition of Ce(IV) reagent. Two FI systems, for the continuous and stopped modes of operation were assembled. In the continuous-FI system, a thermostated bath was used to increase the sensitivity. However this is not necessary for the stopped-FI system. The two systems are comparable in terms of sensitivity, sample throughput and detection limit. The continuous-FI and the stopped-FI exhibited detection limits (3 sigma) of 2.3 and 3 micrograms I l-1 respectively. Both systems have equal sample throughputs of 35 samples h-1. Calibration plots for both techniques are linear. The FI procedures provide very short analysis times compared to the batch procedure. Using the linear regression test, there is no significant difference between the results from the four methods, i.e., continuous-FI, stopped-FI, conventional method and ICP-MS. The proposed methods are readily applicable for automation and can be an alternative to the conventional procedure for the survey of the iodine deficiency disorder. A condition for sample digestion is also proposed to reduce the amount of chloric acid required for complete digestion. Kinetic information of the reaction can also be obtained from the stopped flow mode.

Determination of ethanol in liquor by near-infrared spectrophotometry with flow injection.

A simple procedure for the determination of ethanol in a liquor by near-infrared (NIR) spectrophotometry with flow injection (FI) is proposed. A liquor sample is equilibrated off-line with dried chloroform to extract ethanol into the organic phase. The extract is injected into a carrier stream of dried chloroform passing through a home-made flow through cell (1 mm path length) sitting in a NIR spectrophotometer for continuous monitoring of absorbance at 2305 or 2636 nm. The ethanol content can be evaluated from a calibration established by a plot of change in absorbance versus concentration of ethanol standard solutions. Optimization of the system will be discussed. A calibration is linear in the range of 20-50% (v/v) ethanol. A throughput of 240 injections h(-1) can be obtained. The procedure is validated by comparing the results with an analysis using gas chromatography.

Dynamic surface tension analysis of dodecyl sulfate association complexes.

First, a novel calibration method is used to expand the current understanding of spherical drop growth and elongation that occurs during on-line measurements of surface pressure using the dynamic surface tension detector (DSTD). Using a novel surface tension calibration method, the drop radius is calculated as a function of time from experimental drop pressure data and compared to the theoretical drop radius calculated from volumetric flow rate. From this comparison, the drop volume at which the drop shape starts to deviate ( approximately 4 mul) from a spherical shape is readily observed and deviates more significantly by approximately 6 mul drop volume (5% deviation in the ideal spherical drop radius) for the capillary sensing tip employed in the DSTD. From this assessment of drop shape, an experimental method for precise drop detachment referred to as pneumatic drop detachment is employed at a drop volume of 2 mul (two second drops at 60 mul/min) in order to provide rapid dynamic surface tension measurements via the novel on-line calibration methodology. Second, the DSTD is used to observe and study kinetic information for surface-active molecules and association complexes adsorbing to an air-liquid drop interface. Dynamic surface tension measurements are made for sodium dodecyl sulfate (SDS) in the absence and presence of either tetra butyl ammonium (TBA) or chromium (III). Sensitive, indirect detection of chromium and other multiply charged metals at low concentrations is also investigated. The DSTD is utilized in examining the dynamic nature of SDS: cation association at the air-liquid interface of a growing drop. Either TBA or Cr(III) were found to substantially enhance the surface tension lowering of dodecyl sulfate (DS), but the surface tension lowering is accompanied by a considerable kinetic dependence. Essentially, the surface tension lowering of these DS: cation complexes is found to be a fairly slow process in the context of the two second DSTD measurement. The limit of detection for both SDS and chromium (III) is in the 300-400 part-per-billion (by mass) range.

Spectrophotometric determination of copper in alloys using naphthazarin.

Naphthazarin (5,8-dihydroxy-1,4-naphthoquinone; Naph) is proposed as a chromogenic reagent for the spectrophotometric determination of copper(II). The polynuclear complex has a mole ratio of Cu:Naph=4:6 in a 50% v/v ethanol/water medium containing 0.1 M ammonium acetate and 1.5% (w/v) sodium dodecyl sulfate. The copper-naphthazarin complex shows an absorption maximum at 330 nm with a molar absorptivity of 1.84x10(4) l mol(-1) cm(-1). Beer's law is obeyed up to 4.5 ppm of copper(II). The method was applied for copper determination in alloy samples with satisfactory results.

Flow injection dialysis for the determination of anions using ion chromatography.

Flow injection dialysis (FID) coupling with ion chromatography (IC) is proposed for simultaneous determination of some anions (bromide, chloride, fluoride, nitrate, nitrite, phosphate and sulfate). A standard or sample containing the anions is injected into a donor stream of a mixture (0.022 M Na(2)CO(3) and 0.028 M NaHCO(3)) flowing into a dialysis cell. The bolus of the dialysate containing the anions, in the acceptor stream of water, flows to the IC injection valve where a portion of the bolus is injected into the IC and analysed under normal IC conditions, with a conductivity detector. FID provides on-line separation and dilution of the analytes from matrix especially from some species such as proteins, surfactant, particulates which may cause damage to the IC columns. Prolongation of life-time of the IC columns is an additional advantage to others which will be discussed. On-line dialysis-IC was also investigated.

Flow injection spectrophotometric or conductometric determination of ascorbic acid in a vitamin C tablet using permanganate or ammonia.

Two simple flow injection (FI) procedures for the determination of ascorbic acid content in a vitamin C tablet are proposed: spectrophotometric involving injection into a stream of acidic potassium permanganate solution and monitoring its color change due to the redox reaction; FI conductometry based on the neutralization of ascorbic acid injected into a flowing ammonia solution yielding a change in conductivity. The procedures have been applied to the analysis of locally commercial vitamin C tablet samples. A through-put of at least 90 injections h(-1) can be achieved. The relative standard deviation was found to be 2.5% (for a 50 mg vitamin C tablet; n=7) for both. Results obtained by either procedure agree with a standard titrimetric method.

Flow-injection spectrophotometric determination of calcium using murexide as a color agent.

FI spectrophotometric determination of calcium using murexide has been developed. The problem of the color of the dye fading and/or its complex in an alkaline medium in the batch method can be overcome by taking advantage of FIA. A calcium solution is injected into an ethylenediamine-ethylenediamine hydrochloride buffer (1 M, pH 11) which also serves as a masking agent, and is then merged with the aqueous murexide (0.005%, w/v) and continuously monitored. Simple FIA manifolds, including an LED colorimeter detector hooked up to a PC-based data acquisition and evaluation system are described. Optimization of FIA systems has been made. The proposed procedures have been validated by using reference materials and comparing the results with the standard methods, and then applied to ores and drug samples.

Some azo-dye reagents for the spectrophotometric determination of cadmium.

2-[2-(5-Bromopyridyl)azol-4,5-dimethylphenol (BrPDMP) and 2-[2-benzothiazolylazo)-5-dimethylaminophenol (BTADAP) have been synthesised and compared, as reagents for cadmium, with the related dyes BrPADAP and BTDMP. The new dyes both form stable highly coloured 2:1 complexes with cadmium, with molar absorptivities (in o-xylene solution) of 3.8 x 10(4) 1.mole(-1).cm(-1) at 590 nm (BrPDMP) and 4.5 x 10(4) 1.mole(-1).cm(-1) at 600 nm (BTADAP). Cadmium can be determined by extraction under alkaline conditions with a solution of BTADAP in xylene. Beer's law is obeyed up to at least 16 mug of cadmium. A limit of detection of 0.15 mug has been estimated and a coefficient of variation of 3.3% at the 5 mug level was found. The only species which interfere seriously are Co(2+), Ni(2+), and Ca(2+). A 200-fold excess of zinc may be tolerated. The method has been applied to the determination of cadmium in water samples, plant materials and hair. Interferences were overcome by preliminary extraction into Aliquat/carbon tetrachloride. The acid dissociation constant of BTADAP (pK = 9.5) and formation constant of the cadmium-BTADAP complex (log beta = 15.1) have been determined.