A three-dimensional origami microfluidic device for paper chromatography: Application to quantification of Tartrazine and Indigo carmine in food samples.

Herein, we report three-dimensional paper chromatography (3D-PC) as a micro-chromatographic platform. The method was based on applying the origami microfluidic device for separation, coupled by colorimetric methods for simultaneous determination. The microfluidic device fabrication was a facile printing approach. Two azo food dyes, Tartrazine (E102) and Indigo carmine (E132), were selected as a model analyte, while carbonate-bicarbonate buffer was used as the mobile phase. Our micro-chromatographic device is associated with two big advantages including needing very small volume of mobile phase ( ~12 µL) and ultrafast separation time (~35 s). Under the optimal conditions, the method provided acceptable linear ranges of 0. 0 g L1-18.0 g L1 (R2 = 0.997) for E102 and 0.070 g L1-10.0 g L1 for E132 and the limits of detection (3σ/slope) were evaluated as 0.620 and 0.060 g L1, respectively. The proposed method was successfully applied in the separation and quantification of these dyes in commercial food products such as jelly, candy, and four kinds of drink samples without any sample preparation prior to analysis. The mean recovery values for the real sample analysis were in the range of 100.14%-102.38% for E132 and E102 respectively. The inter-device relative standard deviations were in the ranges of 1.5%-11.8%. In total, our chromatographic µPAD is small (1.0 cm × 1.0 cm × 0.5 cm), portable, inexpensive, no need of specialized user, requires low volumes of sample (0.5 µL), and can perform separation using 12 µL of aqueous mobile phase in very short time.

A paper-based analytical device coupled with electrochemical detection for the determination of dexamethasone and prednisolone in adulterated traditional medicines.

The adulteration of herbal medicines by dexamethasone or prednisolone is regarded as a serious problem in many communities. Herein, a novel platform for the separation and quantification of both target steroids in herbal medicines based on electrochemical paper-based analytical devices (ePADs) has been created. The ePAD was composed of Whatman SG81 chromatography paper, 3D-printed devices and a commercial screen-printed electrode. Whatman SG81 silica-coated paper was used for the separation of dexamethasone and prednisolone based on the difference in their partition coefficients during the flow of the mobile phase. The optimal mobile phase was composed of 60% ethyl acetate in cyclohexane and required 7 min for separation. The separated steroids on the paper were then quantified by electrochemical detection using differential pulse voltammetry, in which the 3D-printed devices facilitated the measurement. Analytical detection ranges of 10-500 µg mL-1 were obtained for both dexamethasone and prednisolone (r2 = 0.988 and 0.994, respectively). The limits of detection for dexamethasone and prednisolone were 3.59 and 11.98 µg mL-1, respectively, whereas the limits of quantification were 6.00 and 20.02 µg mL-1, respectively. The amounts of both target steroids derived from real herbal medicine samples determined by the proposed method were comparable to those obtained with assays using standard high-performance liquid chromatography. In addition, a simple evaporation step can be used to increase the concentration of the samples before analysis. These ePADs are simple, low-cost, rapid, and very promising for on-site quantitative detection.

Rapid multiplex nucleic acid amplification test developed using paper chromatography chip and azobenzene-modified oligonucleotides.

Although nucleic acid amplification test (NAT) is widely used for pathogen detection, rapid NAT systems that do not require special and expensive instruments must be developed in order to enable point of care (POC)-NATs, which contribute to early initiation of treatment. As a POC-NAT system, Kaneka DNA chromatography chip (KDCC), developed using DNA tag-bound primer through modified substance, was shown to be suitable for POC testing, due to the rapid detection time, simple procedures, and low manufacturing costs. However, owing to some modifications in primer, the detection performance and amplification speed were shown to be reduced when using KDCC, counteracting the increased speed of detection. To solve these issues and improve the speed of this NAT system, we investigated a better modification substance for KDCC. Here, azobenzene-modified primers were shown to have the highest amplification speed and detection performance in KDCC, of all modifications tested in this study, showing 10-100-fold lower detection limit but maintaining the same reaction time. Additionally, rapid herpes simplex virus detection system with azobenzene modified primers was developed. We believed that this breakthrough will contribute toward enabling greater utilization of POC-NATs for medical care, especially in developing countries and clinics.

Impedance Analysis of Colloidal Gold Nanoparticles in Chromatography Paper for Quantitation of an Immunochromatographic Assay.

A detection method of gold nanoparticles in chromatography paper has been developed for a simple, cost-effective and reliable quantitation of immunochromatographic strip test. The time courses of the solution resistance in chromatography paper with the gold nanoparticles solution are electrochemically measured by chrono-impedimetry. The dependence of the solution resistance on the concentration of gold nanoparticles has been successfully observed. The main factor to increase the solution resistance may be obstruction of the ion transport due to the presence of gold nanoparticles. The existence of gold nanoparticles with 1.92 × 10(9) particles/mL in an indistinctly-colored chromatography paper is also identified by a solution resistance measurement. This indicates that the solution resistance assay has the potential to lower the detection limit of the conventional qualitative assay.

Ambient ionization based on mesoporous graphene coated paper for therapeutic drug monitoring.

Paper spray (PS), as a new ambient ionization method, has been applied for direct qualitative and quantitative analysis. The high sensitivity and minimum internal energy (low spray voltage) with optimized paper spray conditions is a significant request for real application in POCT. In this study, a simple and efficient ambient ionization method is developed by spraying from a mesoporous graphene foams (MGFs)-modified paper surface. The good electrical conductivity of MGFs results in obvious spray voltage decrease. Meanwhile, the MGFs-paper substrate has a well improvement in separation and elution efficiency ascribing to ultrahigh specific surface area and π-π electrostatic stacking property of graphene. In combination a commercial triple quadrupole mass spectrometer, the paper spray is successfully used for analysis of amphetamine in saliva. The linear dynamic ranges expand 10 fold in comparison with unmodified chromatography papers and the low limit of quantitation (LOQ) is as low as 1 pg/mL. A small sample volume (0.5 µL) could be analyzed immediately after spotting, without any pretreatment. The performance of this method was demonstrated for application in fast point-of-care mass spectrometry.

Development of a test strip for rapid detection of lactoperoxidase in raw milk.

Traditional methods for detecting lactoperoxidase (LP) are complex and time-consuming, so a test strip was made based on the enzymatic reaction principle to enable quick and convenient detection of LP in raw milk. In this study 0.1 mol/L citric acid (CA)/0.2 mol/L disodium hydrogen phosphate (NaP) buffer solution (pH 5.0), 22 mmol/L 3,3',5,5'-tetramethylbenzidine (TMB), 0.6 mmol/L hydrogen peroxide (H2O2), and 0.5% Tween-20 or 0.3% cetyltrimethyl ammonium bromide (CTAB) were optimal for preparing a quick, sensitive, and accurate LP test strip. The coefficient of variation (CV) of the estimated LP concentrations ranged from 2.47% to 6.72% and the minimum LP concentration detected by the test strip was 1-2 mg/L. Estimates of active LP in sixteen raw milk samples obtained using the test strip or the TMB method showed a good correlation (r=0.9776). So the test strip provides a quick, convenient, and accurate method for detecting the LP concentration of raw milk.

A simple method for patterning poly(dimethylsiloxane) barriers in paper using contact-printing with low-cost rubber stamps.

This paper presents a simple and low-cost method for patterning poly(dimethylsiloxane) (PDMS) barriers in porous support such as paper for the construction of flexible microfluidic paper-based analytical devices (µPADs). The fabrication method consisted of contact-printing a solution of PDMS and hexane (10:1.5 w/w) onto chromatographic paper using custom-designed rubber stamps containing the patterns of µPADs. After penetrating the paper (∼30 s), the PDMS is cured to form hydrophobic barriers. Under optimized conditions, hydrophobic barriers and hydrophilic channels with dimensions down to 949±88 µm and 771±90 µm (n=5), respectively, were obtained. This resolution is well-suitable for most applications in analytical chemistry. Chemical compatibility studies revealed that the PDMS barriers were able to contain some organic solvents, including acetonitrile and methanol, and aqueous solutions of some surfactants. This find is particularly interesting given that acetonitrile and methanol are the most used solvents in chromatographic separations, non-aqueous capillary electrophoresis and electroanalysis, as well as aqueous solutions of surfactants are suitable mediums for cell lyses assays. The utility of the technique was evaluated in the fabrication of paper-based electrochemical devices (PEDs) with pencil-drawn electrodes for experiments in static cyclic voltammetry and flow injection analysis (FIA) with amperometric detection, in both aqueous and organic mediums.

Evanescent field: a potential light-tool for theranostics application.

A noninvasive or minimally invasive optical approach for theranostics, which would reinforce diagnosis, treatment, and preferably guidance simultaneously, is considered to be major challenge in biomedical instrument design. In the present work, we have developed an evanescent field-based fiber optic strategy for the potential theranostics application in hyperbilirubinemia, an increased concentration of bilirubin in the blood and is a potential cause of permanent brain damage or even death in newborn babies. Potential problem of bilirubin deposition on the hydroxylated fiber surface at physiological pH (7.4), that masks the sensing efficacy and extraction of information of the pigment level, has also been addressed. Removal of bilirubin in a blood-phantom (hemoglobin and human serum albumin) solution from an enhanced level of 77 µM/l (human jaundice >50 µM/l) to ~30 µM/l (normal level ~25 µM/l in human) using our strategy has been successfully demonstrated. In a model experiment using chromatography paper as a mimic of biological membrane, we have shown efficient degradation of the bilirubin under continuous monitoring for guidance of immediate/future course of action.

Assessment of the within- and between-lot variability of Whatman™ FTA(®) DMPK and 903(®) DBS papers and their suitability for the quantitative bioanalysis of small molecules.

BACKGROUND: To ensure that PK data generated from DBS samples are of the highest quality, it is important that the paper substrate is uniform and does not unduly contribute to variability. This study investigated any within and between lot variations for four cellulose paper types: Whatman™ FTA(®) DMPK-A, -B and -C, and 903(®) (GE Healthcare, Buckinghamshire, UK). The substrates were tested to demonstrate manufacturing reproducibility (thickness, weight, chemical coating concentration) and its effect on the size of the DBS produced, and the quantitative data derived from the bioanalysis of human DBS samples containing six compounds of varying physicochemical properties. RESULTS & DISCUSSION: Within and between lot variations in paper thickness, mass and chemical coating concentration were within acceptable manufacturing limits. No variation in the spot size or bioanalytical data was observed. CONCLUSION: Bioanalytical results obtained for DBS samples containing a number of analytes spanning a range of chemical space are not affected by the lot used or by the location within a lot.

Chromatographic separation and detection of target analytes from complex samples using inkjet printed SERS substrates.

In principle, surface enhanced Raman spectroscopy (SERS) is thought to provide unique identification of a target analyte, even in complex samples or in the presence of multiple analytes. In practice, however, this is not always true for real-world samples due to various forms of interference. In this report, we build upon our previous work on inkjet-printed SERS substrates by using paper and polymer membranes to integrate sample cleanup and analyte separation with SERS detection. Inkjet-printed paper SERS substrates provide a highly sensitive chemical detection platform of unprecedented cost and simplicity. In addition, paper inherently provides unique capabilities, such as capillary-actuated fluid transport and selective molecular retention. Utilizing these properties, we demonstrate two-dimensional chromatographic separation and SERS detection on inkjet-printed paper SERS substrates. Then, we leverage the separation properties of paper and polymer membranes for real applications that feature complex sample matrices, including the detection of down to 5 ppm melamine in infant formula, as well as the quantification of nanograms of heroin in samples contaminated with a highly fluorescent background. The results presented here demonstrate that inkjet-printed paper SERS devices not only provide advantages in terms of sensitivity and cost, but the paper provides inherently integrated sample cleanup capabilities that are not available in traditional SERS substrates and microfluidic SERS devices. These unique capabilities of paper SERS devices enable the identification of targeted analytes even in complex real-world samples.

Validation of stationary phases in (111)In-pentetreotide planar chromatography.

INTRODUCTION: Since Pall-German stopped manufacturing ITLC-SG, it has become necessary to validate alternative stationary phases. OBJECTIVE: To validate different stationary phases versus ITLC-SG Pall-Gelman in the determination of the radiochemical purity (RCP) of (111)In-pentetreotide ((111)In-Octreoscan) by planar chromatography. MATERIAL AND METHODS: We conducted a case-control study, which included 66 (111)In-pentetreotide preparations. We determined the RCP by planar chromatography, using a freshly prepared solution of 0,1M sodium citrate (pH 5) and the following stationary phases: ITLC-SG (Pall-Gelman) (reference method), iTLC-SG (Varian), HPTLC silica gel 60 (Merck), Whatman 1, Whatman 3MM and Whatman 17. For each of the methods, we calculated: PRQ, relative front values (RF) of the radiopharmaceutical and free (111)In, chromatographic development time, resolution between peaks. We compared the results obtained with the reference method. The statistical analysis was performed using the SPSS program. The p value was calculated for the study of statistical significance. RESULTS: The highest resolution is obtained with HPTLC silica gel 60 (Merck). However, the chromatographic development time is too long (mean=33.62minutes). Greater resolution is obtained with iTLC-SG (Varian) than with the reference method, with lower chromatographic development time (mean=3.61minutes). Very low resolutions are obtained with Whatman paper, essentially with Whatman 1 and 3MM. Therefore, we do not recommend their use. CONCLUSIONS: Although iTLC-SG (Varian) and HPTLC silica gel 60 (Merck) are suitable alternatives to ITLC-SG (Pall-Gelman) in determining the RCP of (111)In-pentetreotide, iTLC-SG (Varian) is the method of choice due to its lower chromatographic development time.

A screening test for capsaicin-stimulated salivary flow using filter paper: a study for diagnosis of hyposalivation with a complaint of dry mouth.

OBJECTIVE: The purpose of this study was to develop a simple screening technique for diagnosis of hyposalivation with dry mouth by estimation of capsaicin-stimulated salivary flow using filter paper. STUDY DESIGN: An assay system comprising 5 spots containing starch and potassium iodide on filter paper incorporating or without capsaicin and a coloring reagent was designed. We investigated whether the number of colored spots using the filter paper incorporating capsaicin could distinguish between healthy subjects and subjects with hyposalivation and dry mouth. RESULTS: In the healthy group (>200 µL/min; n = 33), the capsaicin-stimulated salivary flow significantly increased as compared with the resting salivary flow, from 1.2 ± 1.4 to 2.9 ± 1.3 colored spots (P < .05). In contrast, the hyposalivation group with dry mouth (<100 µL/min; n = 32) hardly changed (4.4 ± 1.0 vs 4.9 ± 0.2), except for 3 subjects who had considerable elevated secretion on capsaicin stimulation. CONCLUSION: By measuring resting and stimulated salivary flows, this method should be useful for evaluating retained functional ability of salivary glands and screening of hyposalivation with dry mouth.

Inkjet-printed paperfluidic immuno-chemical sensing device.

This paper reports on an inkjet printing method for the fabrication of lateral flow immunochromatographic devices made from a single piece of filter paper by patterning microfluidic channels and dispensing immunosensing inks, requiring only a single printing apparatus. This "paperfluidic" immunosensing device allows for a less time-consuming and more low-cost fabrication compared with the conventional immunochromatographic strips requiring multiple pads, plastic or nylon backing, and a plastic case. A sandwich immunoreaction was performed on the patterned immunosensing paper device, and the sensitivity of the device was optimized with an IgG model analyte. Inkjet-printed antibodies on the test line and the control line were immobilized by physical adsorption, resulting in a very simple fabrication method applicable for pure cellulose surfaces. The color intensity in the test line and the control line was determined both by naked eye and by means of a color scanner in combination with a simple computer program. With the resulting paperfluidic immunosensing device, human IgG concentrations at least down to 10 µg/l could be detected within 20 min. Additionally, in order to demonstrate the feasibility of a total multianalyte sensing system, a combined immuno-chemical sensing device was also fabricated by patterning an additional microfluidic channel for a chemical assay onto the same paper substrate. This low-cost multianalyte paperfluidic sensing device thus demonstrates the feasibility of simple, portable, and disposable tools for pathogen detection in the field of medical, environmental, and food analyses, possibly resulting in useful devices in remote settings and less-industrialized countries.

A pilot study of a simple screening technique for estimation of salivary flow.

OBJECTIVES: The purpose of this study was to develop a simple screening technique for estimation of salivary flow and to test the usefulness of the method for determining decreased salivary flow. STUDY DESIGN: A novel assay system comprising 3 spots containing 30 microg starch and 49.6 microg potassium iodide per spot on filter paper and a coloring reagent, based on the color reaction of iodine-starch and theory of paper chromatography, was designed. We investigated the relationship between resting whole salivary rates and the number of colored spots on the filter produced by 41 hospitalized subjects. RESULTS: A significant negative correlation was observed between the number of colored spots and the resting salivary flow rate (n = 41; r = -0.803; P < .01). For all complaints of decreased salivary flow (n = 9) having cutoff values <100 microL/min for the salivary flow rate, 3 colored spots appeared on the paper, whereas for healthy subjects there was < or =1 colored spot. CONCLUSION: This novel assay system might be effective for estimation of salivary flow not only in healthy but also in bedridden and disabled elderly people.

Capillary blotting of glycosaminoglycans on nitrocellulose membranes after agarose-gel electrophoresis separation.

A method for the blotting and immobilizing of several nonsulfated and sulfated complex polysaccharides on membranes made hydrophilic and positively charged by cationic detergent after their separation by conventional agarose gel electrophoresis is illustrated. This new approach to the study of glycosaminoglycans (GAGs) utilizes the capacity of agarose gel electrophoresis to separate single species of polysaccharides from mixtures and the membrane technology for further preparative and analytical uses.Nitrocellulose membranes are derivatized with the cationic detergent cetylpyridinium chloride and mixtures of GAGs are capillary blotted after their separation in agarose gel electrophoresis. Single purified species of variously sulfated polysaccharides are transferred on derivatized membranes with an efficiency of 100% and stained with alcian blue (irreversible staining) and toluidine blue (reversible staining). This enables a lower amount limit of detection of 0.1 microg. Nonsulfated polyanions, for example hyaluronic acid, may also be transferred to membranes with a limit of detection of approximately 0.1-0.5 microg after irreversible or reversible staining. The membranes may be stained with reversible staining and the same lanes are used for immunological detection or other applications.

[Comparative study of immunochromatographic sets for rapid diagnosis of syphilis].

The paper presents the results of the comparative study of 9 commercial kits made in foreign countries for the rapid diagnosis of syphilis, performed at the Central Research Institute of Dermatovenereological Infections, Russian Agency for Health Care, within the framework of the International Sexual Transmitted Diseases Diagnostic Initiative Program under the aegis of the World Health Organization during 2002-2005. The above diagnostic kits were assessed in the context of the availability of and statement simplicity in the instruction for use; the technological complexity in performing a study procedure and the simplicity in interpreting the findings; the necessity of employing additional laboratory equipment. The parameters of the clinical efficiency (sensitivity and specificity) were studied; a passive sensitized red blood cell agglutination test was used as a reference method. The comprehensive study showed that the "Determine Syphilis TP test system ("Abbot Lab", USA) yielded the best results with a total of 10 scores at 100% sensitivity and 100% sensitivity) while the "Bioline Syphilis anti-TP Test Card" ("Pacific Biotech Co.", Thailand) provided a total of 9 scores at 96% sensitivity and 98% specificity.

Chromatography paper strip sampling of enteric adenoviruses type 40 and 41 positive stool specimens.

BACKGROUND: The enteric subgroup F adenoviruses type 40 (Ad40) and 41 (Ad41) are the second most important cause of acute infantile gastroenteritis after rotaviruses. Repeated community outbreaks have been associated with antigenic changes among the Ad40 and Ad41 strains due to host immune pressure. Therefore large field epidemiological surveys and studies on the genetic variations in different isolates of Ad40 and Ad41 are important for disease control programs, the design of efficient diagnostic kits and vaccines against subgroup F adenoviruses. A novel method using sodium dodecyl sulphate SDS/EDTA-pretreated chromatography paper strips was evaluated for the collection, storage and shipping of Ad40/41 contaminated stool samples. RESULTS: This study shows that adenoviral DNA can be successfully detected in the filter strips by PCR after four months storage at -20 degrees C, 4 degrees C, room temperature (20-25 degrees C) and 37 degrees C. Furthermore no adenoviral infectivity was observed upon contact with the SDS/EDTA-pretreated strips. CONCLUSIONS: Collecting, storing and transporting adenovirus type 40 and 41 positive stool samples on SDS/EDTA-pretreated chromatography filter strips is a convenient, biosafe and cost effective method for studying new genome variants and monitoring spread of enteric adenovirus strains during outbreaks.

[Physical and technical aspects of research on mechanical blood emission]. / Fiziko-tekhnicheskie printsipy issledovaniia mekhanoémissii krovi.

Mechanic emission is an effect observed in dissipation of mechanical energy due to mechanochemical reactions and electrophysic processes at tension, deformation or friction of blood samples dried on the chromatographic paper. The effect is accompanied by electron emission and electromagnetic radiation. The physical principles of mechanic emission were technically realized while developing and designing a microcomputer approximate analyzer (MPA) for spectral analysis of blood mechanic emission when it was recorded in the optical, electric, and radio channels of information conversion. The arrangement of MPA and analytical methods for its technical parameters are given in the paper. MPA may be useful in supplementary non-invasive differential diagnosis of cancer and inflammatory diseases, as well as in the solution of some diagnostic goals of radiology and cardiology.