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This article describes a method for the modification of paper with single-wall carbon nanohorns (SWCNHs) to form stable suprastructures. The SWCNHs form stable dahlia-like aggregates in solution that are then self-assembled into superior structures if the solvent is evaporated. Dipping paper sections into a dispersion of SWCNHs leads to the formation of a thin film that can be used for microextraction purposes. The coated paper can be easily handled with a simple pipette tip, paving the way for disposable extraction units. As a proof of concept, the extraction of antidepressants from urine and their determination by direct infusion mass spectrometry is studied. Limits of detection (LODs) were 10 ng/L for desipramine, amitriptyline, and mianserin, while the precision, expressed as a relative standard deviation, was 7.2%, 7.3%, and 9.8%, respectively

Post-print RATIONALE: Drug overdose deaths due to fentanyls and other novel psychoactive substances (NPS) are on the rise. The higher potencies of fentanyl analogs compared with morphine require new technologies to identify and quantitate NPS. METHODS: Paper spray tandem mass spectrometry and high-resolution mass spectrometry were used to identify and measure fentanyl analogs as well as common drugs of abuse in urine samples from substance use disorder clinics. Ten-microliter urine samples were deposited directly on paper spray cartridges previously loaded with internal standards, dried, and analyzed with no other sample treatment. Quantitative results were obtained using tandem mass spectrometry (MS/MS). Individual drugs were identified using high resolution accurate mass spectrometry (HRAM), and confirmed by data dependent MS/MS. RESULTS: Calibration curves in urine were linear over a range of 0.5 - 50 ng/mL with R2 of 0.99 or better for eight representative fentanyl analogs. Cartridges preloaded with internal standards demonstrated satisfactory quantitative results compared with LC/MS. Direct identification and confirmation of fentanyl analogs and other common drugs of abuse in urine using high resolution accurate mass and MS/MS fragmentation were demonstrated at low picogram levels. CONCLUSIONS: Paper spray mass spectrometry can reliably identify and quantitate fentanyl analogs and other drugs of abuse in urine. Using paper spray cartridges as collection devices reduces exposure and transportation risks associated with biological fluids. Cartridges pre-loaded with labeled internal standards can be effective for targeted screening of fentanyl analogs and other drugs of abuse. Chris Gill would like to acknowledge the Natural Sciences and Engineering Research Council of Canada (NSERC) for funding under the Discovery Grants program (RGPIN-2016-05380), Vancouver Island University for their ongoing support of my research program, and an anonymous Vancouver Island University Foundation donor who funded the necessary travel to facilitate this timely collaboration. Post-print version This is the peer reviewed version of the following article: Kennedy, J.H., Palaty, J., Gill, C.G., & Wiseman, J.M. (2018). Rapid analysis of fentanyls and other novel psychoactive substances (NPS) in substance use disorder patient urine using paper spray mass spectrometry. Rapid Communications in Mass Spectrometry, 32(15), 1280-1286. DOI: 10.1002/rcm.8164, which has been published in final form at http://dx.doi.org/10.1002/rcm.8164. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. https://viurrspace.ca/bitstream/handle/10613/12782/Gill.RCM2018.pdf?sequence=4

We report a paper-based assay platform for the detection of the kidney disease marker Trefoil Factor 3 (TFF3) in human urine. The sensor is based on a quantitative metalloimmunoassay that can determine TFF3 concentrations via electrochemical detection of environmentally stable silver nanoparticle (AgNP) labels attached to magnetic microbeads via a TFF3 immunosandwich. The paper electroanalytical device incorporates two preconcentration steps that make it possible to detect concentrations of TFF3 in human urine at the low end of the target TFF3 concentration range (0.03-7.0 μg mL(-1)). Importantly, the paper device provides a level of accuracy for TFF3 determination in human urine equivalent to that of a commercial kit. The paper sensor has a dynamic range of ∼2.5 orders of magnitude, only requires a simple, one-step incubation protocol, and is fast, requiring only 10 min to complete. The cost of the materials at the prototypic laboratory scale, excluding reagents, is just US$0.42.

Bisphenol A (BPA) is an endocrine disrupting environmental chemical. Urinary concentrations of BPA were measured in samples collected nationwide from Chinese young adults. The geometric mean urinary concentration of BPA in Chinese young adults was 2.23 ng/mL. The estimated daily exposure dose for BPA was 64.8 ng/kg bw/day. Contributions of various BPA sources to exposure in Chinese young adults were estimated. Dietary intake was the primary exposure pathway. The contribution of dietary intake, indoor dust, paper products and personal care products to BPA intake was 72.5 %, 0.74 %, 0.98 %, 0.22 % of the total exposure dose, respectively. This is the first study on the occurrence of BPA in young adults from most provinces and autonomous regions of China. The results can be used to establish a database for BPA exposure assessment for Chinese general population.

Purpose In thermal paper, Bisphenol S (BPS) is one of the alternatives for bisphenol A (BPA). Due to its structural similarity to BPA, concern has been raised about the safety of BPS. Indeed, handling thermal paper receipts could be a source of occupational exposure to BPS among cashiers, as it was previously described for BPA. In this study, we investigated whether frequent contacts with thermal paper are associated with an increase in urinary BPS levels in cashiers. Method Total (unconjugated and conjugated forms) and free (unconjugated) BPS were measured in urine samples from 17 cashiers and 15 controls, using LC–MS/MS. Spot urine samples, including pre-shift and post-sift samples and first morning void were collected from each volunteer. BPS concentration in thermal paper was determined and the number of receipts handled by cashiers was estimated as well. Results The median urinary total BPS concentration was 0.67 μg/L (0.52 μg/g creatinine) for controls and 2.53 μg/L (2.07 μg/g creatinine) for cashiers. Total BPS concentration was significantly higher in cashiers than in controls. Free BPS was detected in less than 20% of urine samples collected from controls and in less than 50% of urine samples collected from cashiers. Conclusion The detectable levels of BPS in urine of controls suggest an exposure to BPS of the general population. In addition, frequent contact with thermal paper could be responsible for an increase in urinary concentration of total BPS in cashiers.

A prospective, randomised, 6-month feeding trial was performed in 28 adult cats with International Renal Interest Society (IRIS) stage 1 and 2 chronic kidney disease (CKD). All cats were assigned to either a control food: Royal Canin Renal Support A Feline, dry or a test food: Hill's Prescription Diet k/d Feline with chicken, dry. Food intake was recorded daily; body weight weekly; and serum, urine, dual-energy x-ray absorptiometry (DEXA) and body condition assessments were performed at 0, 1, 3 and 6 months. Twenty cats (9 control, 11 test group) completed the study according to protocol. Cats consuming control food had significant loss of body weight (n=14; mean, -13.0 per cent, P

In this work, a disposable paper-plastic hybrid microfluidic lab-on-a-chip (LOC) has been developed and successfully applied for the colorimetric measurement of urine by the smartphone-based optical platform using a "UrineAnalysis" Android app. The developed device was cost-effectively implemented as a stand-alone hybrid LOC by incorporating the paper-based conventional reagent test strip inside the plastic-based LOC microchannel. The LOC device quantitatively investigated the small volume (40 μL) of urine analytes for the colorimetric reaction of glucose, protein, pH, and red blood cell (RBC) in integration with the finger-actuating micropump. On the basis of our experiments, the conventional urine strip showed large deviation as the reaction time goes by, because dipping the strip sensor in a bottle of urine could not control the reaction volume. By integrating the strip sensor in the LOC device for urine analysis, our device significantly improves the time-dependent inconstancy of the conventional dipstick-based urine strip, and the smartphone app used for image analysis enhances the visual assessment of the test strip, which is a major user concern for the colorimetric analysis in point-of-care (POC) applications. As a result, the user-friendly LOC, which is successfully implemented in a disposable format with the smartphone-based optical platform, may be applicable as an effective tool for rapid and qualitative POC urinalysis.

Screening procedures using high resolution (HR)-mass spectrometry (MS) are getting more and more important, e.g., for drug testing or adherence monitoring. Approaches usually include time-consuming sample preparation and compound separation by liquid chromatography (LC). The paper spray ionization (PSI) technique coupled to MS might overcome these steps by direct analysis of complex mixtures without extraction and separation. In recent years, this technology proved its potential for quantification and/or qualitative screening in biofluids. However, so far, PSI-MS was only applied to procedures covering a limited number of targets. Therefore, a PSI-HR-MS/MS approach was developed and successfully validated for comprehensive urine screening. The procedure showed high matrix effects for most drugs but still acceptable limits of identification. Applicability was tested by analyses of three proficiency tests for systematic toxicological analysis and of 103 authentic human urine samples. Its screening power was compared to that of published LC-HR-MS/MS procedures after urine precipitation with conjugate cleavage (UglucP) or dried urine spot workup by conjugate cleavage and liquid extraction (DUSglucE). In the authentic samples, 73% of all 777 drug intakes were detectable with the new approach. The LC-HR-MS/MS screening approaches detected more drugs, particularly in samples with low analyte concentrations, with values of 88% after UglucP or 76% after DUSglucE. In conclusion, the new PSI-HR-MS/MS screening approach was suitable for comprehensive urine screening of different drug classes and might be a promising alternative to conventional procedures. However, limitations should be considered such as detection of drugs in low concentrations and risk of false positive or negative results caused by mixed spectra.

Background Mucopolysaccharidoses (MPSs) are a group of lysosomal storage diseases caused by the deficiency/absence of enzymes which catalyze the degradation of glycosaminoglycans (GAGs). The use of biological samples dried on filter paper has been increasing because it makes it easy to ship them to reference laboratories. Urinary GAGs are the main biomarkers of MPS and, thus, we studied the correlations of determinations to GAGs and creatinine, as well as compared the GAGs' profile on electrophoresis, between urine and dried urine in filter paper (DUFP) samples. We also assessed the GAG stability over time under different storage temperatures. Methods We quantified the GAG concentration in both sample types and compared the results by Pearson correlation. Results The results were very similar, with r = 0.97 for creatinine and with r = 0.94 and r = 0.98 for GAGs for controls and patients, respectively, with similar electrophoretic profiles. The GAG stability in DUFP was up to 30 days at − 20, 4, and 25 °C and up to 21 days at 37 °C. Conclusion Our proposal assessed urinary GAGs in DUFP and concluded that these samples can be used in the investigation of MPS, replacing urine samples in neonatal screening and monitoring of therapies, due to ease of transportation and storage.

The first step in curing a disease is being able to detect the disease effectively. Paper-based microfluidic devices are biodegradable and can make diagnosing diseases cost-effective and easy in almost all environments. We created a three-dimesnional (3D) paper device using wax printing fabrication technique and basic principles of origami. This design allows for a versatile fabrication technique over previously reported patterning of SU-8 photoresist on chromatography paper by employing a readily available wax printer. The design also utilizes multiple colorimetric assays that can accommodate one or more analytes including urine, blood, and saliva. In this case to demonstrate the functionality of the 3D paper-based microfluidic system, a urinalysis of protein and glucose assays is conducted. The amounts of glucose and protein introduced to the device are found to be proportional to the color change of each assay. This color change was quantified by use of Adobe Photoshop. Urine samples from participants with no pre-existing health conditions and one person with diabetes were collected and compared against synthetic urine samples with predetermined glucose and protein levels. Utilizing this method, we were able to confirm that both protein and glucose levels were in fact within healthy ranges for healthy participants. For the participant with diabetes, glucose was found to be above the healthy range while the protein level was in the healthy range.

The aim of the study was to assess occupational exposure to bisphenol A in Finland. Five companies took part in the research project: two paint factories (liquid and powder paints), a composite product factory, a thermal paper factory, and a tractor factory. Exposure was assessed by measuring total bisphenol A excretion (free and conjugated) from urine samples, and its concentrations in the air. The results revealed the specific work tasks in two of five companies in which significant occupational exposure to bisphenol A may occur. In the manufacturing of liquid paint hardener, urine samples collected after the working day showed bisphenol A levels of up to 100-170 µg l-1. Workers in thermal paper manufacturing were also exposed to bisphenol A, especially those working in the manufacture of coating material and operating coating machines. Median concentrations of the post-shift urine samples of coating machine workers were in the range of 130-250 µg l-1. The highest bisphenol A concentrations were in the range of 1000-1500 µg l-1. Recommendations for more effective personal protection resulted in decreased exposure, particularly among coating machine operators. In the rest of the companies, urinary bisphenol A levels were typically in the range of those of the general population. Bisphenol A concentrations in air samples were typically low (

Iodine deficiency is a world-wide health problem. A simple, convenient, and inexpensive method to monitor urine iodine levels would have enormous benefit in determining an individual's recent iodine intake or in identifying populations at risk for iodine deficiency or excess. Current methods used to monitor iodine levels require collection of a large volume of urine and its transport to a testing laboratory, both of which are inconvenient and impractical in parts of the world lacking refrigerated storage and transportation. To circumvent these limitations we developed and validated methods to collect and measure iodine and creatinine in urine dried on filter paper strips. We tested liquid urine and liquid-extracted dried urine for iodine and creatinine in a 96-well format using Sandell–Kolthoff and Jaffe reactions, respectively. Our modified dried urine iodine and creatinine assays correlated well with established liquid urine methods (iodine: R2=0.9483; creatinine: R2=0.9782). Results demonstrate that the dried urine iodine and creatinine assays are ideal for testing the iodine status of individuals and for wide scale application in iodine screening programs.

A rapid and facile analytical method for quantification of ribonucleosides in human urine was developed by the combination of nanocoating cellulose paper based microextraction and nanoelectrospray ionization-tandem mass spectrometry (nESI-MS/MS). Cellulose paper used for microextraction was modified by nano-precision deposition of uniform ultrathin zirconia gel film using a sol-gel process. Due to the large surface area of the cellulose paper and the strong affinity between zirconia and the cis-diol compounds, the target analytes were selectively extracted from the complex matrix. Thus, the detection sensitivity was greatly improved. Typically, the nanocoating cellulose paper was immersed into the diluted urine for selective extraction of target analytes, then the extracted analytes were subjected to nESI-MS/MS detection. The whole analytical procedure could be completed within 10min. The method was evaluated by the determination of ribonucleosides (adenosine, cytidine, uridine, guanosine) in urine sample. The signal intensities of the ribonuclesides extracted by the nanocoating cellulose paper were greatly enhanced by 136-459-folds compared with the one of the unmodified cellulose paper based microextraction. The limits of detection (LODs) and the limits of quantification (LOQs) of the four ribonucleosides were in the range of 0.0136-1.258μgL-1 and 0.0454-4.194μgL-1, respectively. The recoveries of the target nucleosides from spiked human urine were in the range of 75.64-103.49% with the relative standard deviations (RSDs) less than 9.36%. The results demonstrate the potential of the proposed method for rapid and facile determination of endogenous ribonucleosides in urine sample.

Made available in DSpace on 2018-12-11T16:45:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-03-08 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) This paper describes the fabrication of 2D and 3D microfluidic paper-based analytical devices (μPADs) for monitoring glucose, total protein, and nitrite in blood serum and artificial urine. A new method of cutting and sealing filter paper to construct μPADs was demonstrated. Using an inexpensive home cutter printer soft cellulose-based filter paper was easily and precisely cut to produce pattern hydrophilic microchannels. 2D and 3D μPADs were designed with three detection zones each for the colorimetric detection of the analytes. A small volume of samples was added to the μPADs, which was photographed after 15 min using a digital camera. Both μPADs presented an excellent analytical performance for all analytes. The 2D device was applied in artificial urine samples and reached limits of detection (LODs) of 0.54 mM, 5.19 μM, and 2.34 μM for glucose, protein, and nitrite, respectively. The corresponding LODs of the 3D device applied for detecting the same analytes in artificial blood serum were 0.44 mM, 1.26 μM, and 4.35 μM. Departamento de Química Universidade Federal de São Carlos Intituto de Química-UNESP, Av. Francisco Degni s/n Intituto de Química-UNESP, Av. Francisco Degni s/n CNPq: 140484/2013-2 FAPESP: 2015/19890-1 CNPq: 427727/2016-2

Bisphenol A (BPA) is a widely used chemical in the production of many polycarbonate plastics, epoxy resin linings for food and beverage containers and thermal papers. Oral intakes from the contaminated diets were considered as the predominant source of BPA exposure for humans. However, due to the high levels of BPA on thermal receipts and their wide applications in our daily life, the amount of BPA be transferred to the skin after holding thermal paper should not be underestimated, particularly for cashiers. To investigate the contribution of BPA exposure levels via the dermal contact route and the relationship between BPA exposure level and oxidative DNA damage, six male volunteers were recruited and required to simulate the cashiers' work and handle the thermal receipts during the study period. Triclosan (TCS, an antimicrobial compound used widely in personal health and skin care products) was applied as a reference compound. Their urinary BPA, TCS and 8-hydroxy-2′-deoxyguanosine (8-OHdG) concentrations were determined by high performance liquid chromatography/ tandem spectrometer (LC/MS/MS). The results showed that after handling the thermal receipts, the urinary BPA concentrations of volunteers increased 3 times of those before the experimental period. But TCS levels in urine kept stable. There existed a correlation between BPA exposure and 8-OHdG (R2 = 0.237, p

Mucopolysaccharidoses are complex lysosomal storage disorders caused by any of eleven different enzyme deficiencies resulting in the accumulation of substrates, mainly glycosaminoglycans (GAGs), in various tissues and biological fluids.We developed and validated a urine filter paper methodology for the analysis of GAGs using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for mucopolysaccharidoses type I, type II and type VI patients. We focused on 2 objectives: first, its applicability to high-risk screening, and secondly, to facilitate the collection and shipping of samples to reference centers as part of diagnostic investigation, as well as from treated patients needing to be monitored for assessment of the efficacy of treatment. GAGs in urine dried onto filter paper were extracted and subjected to methanolysis to obtain the repeating disaccharides of the molecules. We devised a multiple reaction monitoring method in positive electrospray ionization mode.The use of deuterated internal standards for dermatan sulfate (DS) and heparan sulfate (HS) reduced a troubling matrix effect. The resulting CVs were14%. Linearity assessment showed Pearson correlation coefficients of 0.999 and 0.997, for DS and HS, respectively. The stability on filter paper was good for DS and HS for up to 6 weeks at various temperatures.We devised a robust and efficient LC-MS/MS methodology for GAGS quantification in urine dried on filter paper and subjected to environmental conditions likely to be encountered during collection, storage and shipping of specimens from referring physicians to medical centers.

International audience; Purpose: As an essential component of polycarbonate plastics and epoxy resins, Bisphenol A (BPA) is found in numerous industrial and consumer products. BPA may cause adverse health effects because of its endocrine activity. General population exposure to this compound mainly through diet is well documented. Thermal paper was also identified as a source of BPA through dermal intake. In this study, we investigated whether frequent contact with thermal paper is associated with an increase in urinary BPA excretion.Methods: We evaluated the exposure to BPA in cashiers and in non-occupationally exposed workers from several workplaces. Urinary BPA was quantified in free (unconjugated) and total (unconjugated plus conjugated) forms in 24-h and spot urine samples using LC–MS/MS. BPA concentration in thermal paper was also measured from each workplace. In addition, participants provided information on job, food and drink, tobacco consumption and hands wash during the sampling period through a questionnaire.Results: Urine samples were collected from 90 cashiers and 44 controls. Free and total BPA were detected in all samples. The median urinary total BPA concentration was 3.54 µg/L (2.89 µg/g creatinine) for controls and 8.92 µg/L (6.76 µg/g creatinine) for cashiers. For the free BPA, the median urinary concentration was 0.20 µg/L (0.21 µg/g creatinine) for controls and 0.28 µg/L (0.22 µg/g creatinine) for cashiers. Any correlation was found between the urinary concentration levels and the number of thermal receipts handled. Hand washes frequency, age, job length of service and tobacco consumption had also no effect on the BPA excretions.Conclusion:A significant increase in urinary total BPA concentration was observed for cashiers handling daily thermal paper receipts. However, no significant increase was observed in urinary free BPA concentration. These findings are particularly interesting for risk assessment since all available data on occupational exposure to BPA through thermal paper were obtained from models or from simulated experiments.

A risk assessment study of seven phthalates in paper diapers for newborn babies produced in Japan was performed. The diapers were purchased and the contents of the seven phthalates were determined and estimated amounts of exposure were calculated based on the eluted rate into artificial medium of urine or sweat, average weight of infants, and frequency of use. Di-2-ethylhexyl phthalate and di- n -butyl phthalate were detected in the topsheets and determined to be 0.6 μg/g and 0.2 μg/g, respectively. The daily estimated exposure volume was calculated to be in the range of 1.86 × 10 −10 –2.98 × 10 −6 mg/kg/day as follows: content of seven phthalates in the topsheet (0.1–1 μg/g) × eluted rate of phthalates into artificial sweat (0.0006–2.4%) × weight of the topsheet of a diaper (1.5 g) × the number of diapers used per day (12 sheets) × skin absorption rate (0.005–0.1)/average body weight (2.9 kg). For hazard assessment, we used 0.2–300 mg/kg/day for the seven phthalates based on the data available at international agencies. The margin of exposure to the seven phthalates was 6.71 × 10 4 –1.99 × 10 11 , indicating that the risk of exposure to phthalates from the diapers produced in Japan was negligible.

Dairy cattle barns are a major source of NH3 emissions to the atmosphere. Previous studies have shown that the bedding material used in the barn can influence the magnitude of NH3 emissions, but little is known about which bedding characteristics are important in this respect. The aims of this study were to assess, at a laboratory scale, the relative importance of the chemical [pH, cation exchange capacity (CEC), C:N] and physical (urine absorbance capacity, bulk density) characteristics of 5 bedding materials (chopped wheat straw, sand, pine shavings, chopped newspaper, chopped corn stalks, and recycled manure solids) on NH3 emissions from dairy cattle urine. Recycled manure solids were the most absorbent of the bedding types (4.2 g of urine/g of bedding), and sand was the least (0.3 g of urine/g of bedding). When beddings were soaked in urine to their absorbance capacities, NH3 emissions over 48 h (expressed as a proportion of the urine N absorbed) were not significantly different among bedding types, despite differences in initial bedding pH, CEC, and C:N. When equal volumes of urine were applied to equal depths of dry bedding, NH3 emissions over 48 h were significantly less from sand and pine shavings (23 and 42% of applied urine N, respectively) than from chopped newspaper, chopped corn stalks, and recycled manure solids (62, 68, and 65% of applied urine N, respectively), whereas emissions from chopped wheat straw (55% applied urine N) only differed significantly from that from sand. Differences in the chemical characteristics of the beddings did not explain differences in emission; NH3 emissions increased linearly with CEC contrary to expectations, and there was no significant relationship with initial bedding pH. The physical characteristics of bedding materials were of more importance, as NH3 emissions increased linearly with absorbance capacity and decreased as the bulk density of the packed beddings increased.

Hormonal analysis of urine from free-ranging primates has been limited due to the difficulty of preserving samples under field conditions. Drying urine on filter paper is an alternative for field preservation. This study describes a new laboratory method for eluting steroids from filter paper with methanol, along with a series of experiments used to develop and validate this method. The overall elution recovery of estrone sulfate (ES) from filter paper was 86.4%. Estrone conjugate (E1C) levels in humans and captive orangutans were analyzed by radioimmunoassay (RIA). Values from samples dried on filter paper were significantly correlated with values from matched frozen samples, with elution efficiencies ranging between 97.1% and 102.4%. Creatinine (Cr) measurements from frozen urine compared to urine dried on filter paper were also significantly correlated (r=0.96) with an elution efficiency of 101.7%. After the samples were stored for 2 years, the absolute values of E1C and Cr were significantly lower but were still significantly correlated with frozen urine values. These data demonstrate the effectiveness of filter paper as a medium for preserving urinary steroid samples, and the efficiency of methanol as a solvent for eluting E1C and Cr. This method thus provides a viable alternative to the traditional procedure of freezing urine for field studies, where freezers are not available.

Background The mucopolysaccharidoses (MPS) are inherited metabolic disorders with bone, joint, and visceral abnormalities, leading to multi-organ dysfunction and, sometimes, neurological manifestations. These diseases are caused by storage of glycosaminoglycans (GAGs) and other complex molecules in tissues, among other pathogenic mechanisms. Definitive diagnosis of the affected individual is mainly based on the identification of the specific enzyme deficiency. New therapies are available or are in development for these pathologies, and early diagnosis seems to be important for the therapy outcomes. Almost all MPS patients have increased levels of GAGs in urine being their evaluation usually the first step in the screening of these conditions. Test on urine may be challenging as transportation of liquid urine samples in appropriate conditions for long distances, especially across international borders, could be difficult. Methods With the aim of overcoming the difficulties related to the use of liquid samples, we extended and validated previous studies about colorimetric determination of GAGs in dried-urine filter paper (DUFP) samples. Results In the conditions we described, there are no differences in the concentration of GAGs between urine and DUFP samples. Untreated patients with MPS and normal controls were well discriminated using any of the samples. Conclusions Dried-urine filter paper is a suitable sample for the colorimetric quantitation of GAGs, and that its incorporation as an additional tool for screening of MPS should be considered by reference laboratories.

Creatinine is one of many markers used to investigate kidney function. This paper describes a low-cost enzymatic paper-based analytical device (enz-PAD) for determining urine creatinine. The disposable dead volumes of creatinine enzyme reagents from an automatic analyser cassette were utilised. Whatman No. 3 paper was cut into long rectangular shapes (4×40 mm(2)) on which the enzyme reagents, R1 and R2, were adsorbed in two consecutive regions. The assay was performed by immersing test strips into urine samples contained in microwells to allow creatinine in the sample to react with immobilised active ingredients and, then, traverse via capillary action to the detection area where chromogen products accumulated. The method is based on hydrogen peroxide (H2O2) formation via creatinine conversion using creatininase, creatinase, and sarcosine oxidase. The liberated H2O2 reacts with 4-aminophenazone and 2,4,6-triiodo-3-hydroxybenzoic acid to form quinoneimine with a pink-red colour at the detection zone. The linear range of the creatinine assay was 2.5-25 mg dL(-1) (r(2)=0.983), and the detection limit was 2.0 mg dL(-1). The colorimetric enz-PAD for the creatinine assay was highly correlated with a conventional alkaline picrate method when real urine samples were evaluated (r(2)=0.977; n=40). This simple and nearly zero-cost paper-based device provides a novel alternative method for screening urinary creatinine and will be highly beneficial for developing countries.

Background Rifampicin or rifampin (R) is a common drug used to treat inactive meningitis, cholestatic pruritus and tuberculosis (TB), and it is generally prescribed for long-term administration under regulated dosages. Constant monitoring of rifampicin is important for controlling the side effects and preventing overdose caused by chronic medication. In this study, we present an easy to use, effective and less costly method for detecting residual rifampicin in urine samples using protein (bovine serum albumin, BSA)-stabilized gold nanoclusters (BSA-Au NCs) adsorbed on a paper substrate in which the concentration of rifampicin in urine can be detected via fluorescence quenching. The intensity of the colorimetric assay performed on the paper-based platforms can be easily captured using a digital camera and subsequently analyzed. Results The decreased fluorescence intensity of BSA-Au NCs in the presence of rifampicin allows for the sensitive detection of rifampicin in a range from 0.5 to 823 µg/mL. The detection limit for rifampicin was measured as 70 ng/mL. The BSA-Au NCs were immobilized on a wax-printed paper-based platform and used to conduct real-time monitoring of rifampicin in urine. Conclusion We have developed a robust, cost-effective, and portable point-of-care medical diagnostic platform for the detection of rifampicin in urine based on the ability of rifampicin to quench the fluorescence of immobilized BSA-Au NCs on wax-printed papers. The paper-based assay can be further used for the detection of other specific analytes via surface modification of the BSA in BSA-Au NCs and offers a useful tool for monitoring other diseases. Electronic supplementary material The online version of this article (doi:10.1186/s12951-015-0105-5) contains supplementary material, which is available to authorized users.

Background: quantitative bioassays for determination of urinary creatinine concentrations using liquid urine and dried urine on paper disks (DUPD) specimens were developed and validated. The bioassays were cross-validated by means of measurement of the urinary creatinine concentrations applying DUPD (3 and 5 mm) and liquid urine specimens. Results: LLOQ and LOD for all bioassays were 0.88 and 0.27 mmol/l creatinine, respectively. All bioassays were cross-validated by the analysis of urine samples from healthy voluntary individuals and bioassays performed were statistically not significantly different. Mean observed urinary creatinine concentrations ranged between 0.44 and 25.6, 0.24 and 23.9, 0.13 and 31.26 mmol/l for liquid, DUPD-5mm and DUPD-3mm bioassays, respectively. Conclusion: the advantage of the DUPD bioassays was the relatively small amounts of urine necessary, making these microsampling techniques attractive when limited amounts of urine specimens of, for example, rodents from drug development studies are available.

Simple, efficient, and cost-effective strategies are needed for urine sample preparation in the field diagnosis of infection with Schistosoma haematobium. In this proof-of-concept study, we evaluated inexpensive and widely available paper products (paper towels, school workbook paper, and newspaper) to gravity-filter urine containing 60 eggs/mL of Schistosoma haematobium. Eggs were reliably visualized by light microscopy by using single-ply paper towels as urine filters. This filtration method has broad applicability in clinical and public health settings in resource-constrained environments.

Bisphenol A (BPA) has one of the highest production volumes of all chemicals worldwide. It has been widely studied because of its endocrine modulating activity. In addition to dietary intake, absorption of BPA via the skin from handling thermal papers is believed to be a relevant route of exposure. We studied BPA exposure via thermal paper receipts in simulation experiments performed by three volunteers, and examined urinary excretion of BPA. We also evaluated background BPA excretion among the Finnish working-age population. The geometric mean BPA excretion among non-occupationally exposed working-age Finns (n=121) was 2.6 μg/l, the range being 0.8-18.9 μg/l. The 95th percentile of the non-occupationally exposed people was 8 μg/l, and this was set as the reference limit for the non-occupationally exposed population. In the first simulation experiment, which was conducted under conditions representing the most likely exposure, i.e., the work of a cashier in a supermarket, BPA excretion remained below the reference limit in all participants. In the second simulation experiment, with more intensive, short-time handling of thermal paper (three times 5 min), urinary excretion also remained at or below background levels (highest value being 10.3 μg/l). The calculated maximum BPA excretion per day after handling thermal paper was less than 0.2 μg/kg of body weight, suggesting a total daily intake over 25 times lower than the European Food Safety Authority's (EFSA's) proposal for a temporary tolerable daily intake (temporary TDI) (5 μg/kg/day).

International audience; Bisphenol A (BPA) is an endocrine disrupting environmental contaminant used in a wide variety of products, and BPA metabolites are found in almost everyone's urine, suggesting widespread exposure from multiple sources. Regulatory agencies estimate that virtually all BPA exposure is from food and beverage packaging. However, free BPA is applied to the outer layer of thermal receipt paper present in very high (∼20 mg BPA/g paper) quantities as a print developer. Not taken into account when considering thermal paper as a source of BPA exposure is that some commonly used hand sanitizers, as well as other skin care products, contain mixtures of dermal penetration enhancing chemicals that can increase by up to 100 fold the dermal absorption of lipophilic compounds such as BPA. We found that when men and women held thermal receipt paper immediately after using a hand sanitizer with penetration enhancing chemicals, significant free BPA was transferred to their hands and then to French fries that were eaten, and the combination of dermal and oral BPA absorption led to a rapid and dramatic average maximum increase (Cmax) in unconjugated (bioactive) BPA of ∼7 ng/mL in serum and ∼20 µg total BPA/g creatinine in urine within 90 min. The default method used by regulatory agencies to test for hazards posed by chemicals is intra-gastric gavage. For BPA this approach results in less than 1% of the administered dose being bioavailable in blood. It also ignores dermal absorption as well as sublingual absorption in the mouth that both bypass first-pass liver metabolism. The elevated levels of BPA that we observed due to holding thermal paper after using a product containing dermal penetration enhancing chemicals have been related to an increased risk for a wide range of developmental abnormalities as well as diseases in adults.

The determination of tobacco derived nicotine alkaloids in biofluid samples is of great importance to testing for tobacco use, tobacco cessation treatment, and studies on exposure to secondhand smoke. Paper spray mass spectrometry (MS) has been adapted for direct, quantitative analysis of tobacco alkaloids from biofluid samples, such as blood, urine, and saliva in liquid and dried form. Limits of quantitation as low as several nanograms per milliliter were obtained for nicotine, cotinine, trans-3'-hydroxycotinine, and anabasine. Direct analysis of fresh blood samples has also been achieved with improved sensitivity using print paper substrates of high density. Quantitation of the cotinine in the blood of a rat was performed with both direct analysis using paper spray and a traditional analysis protocol using liquid chromatography MS. Comparable results were obtained and the precision of the two methods was similar. The paper spray MS method is rapid and shows potential for significantly improved analytical efficiency in clinical laboratories as well as for point-of-care tobacco use assessment.

We present a traumatic fatality of a 19-year-old man who had ingested blotter paper containing 25I-NBOMe [2-(4-iodo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)methyl]ethanamine]. Postmortem specimens were analyzed by high performance liquid chromatography with tandem mass spectrometry (HPLC/MS/MS). Toxicology findings for fluids based upon blood or urine calibrators were as follows: peripheral blood, 405 pg/mL; heart blood, 410 pg/mL; urine, 2.86 ng/mL; and vitreous humor, 99 pg/mL. While findings based upon the method of standard additions were: gastric contents, 7.1 μg total; bile, 10.9 ng/g; brain, 2.54 ng/g and liver, 7.2 ng/g. To our knowledge the presented case is the first postmortem case of 25I-NBOMe intoxication documented by toxicological analysis of tissues and body fluids.

Total Sanitation Campaign (TSC) launched by Government of India has been the cornerstone of a decentralized, incentive based approach to achieve rural sanitation objectives. The scheme has developed strategic components to ensure coverage of sanitation facilities through financial and programmatic support for households, and for institutional and community sanitation. Construction of toilets needs to be complemented with mechanisms of waste handling to ensure a safe and hygienic environment in the rural areas. Safe disposal of solid and liquid waste is a major step on the sanitation ladder where communities have begun to successfully tackle. Government at present has specific provision for solid and liquid waste management (SLWM) as an important component under TSC. The Ministry of Drinking Water and Sanitation has been continuously updating technical notes on rural sanitation to reinforce the quality of interventions. The Hand Book on scaling up solid and liquid waste management in rural areas gives a very useful insight on the planning, construction, operation and maintenance of SLWM units in a sustainable way. The effort has been to make a comprehensive document that is user friendly for planning and executing agencies.

Background: The fact that mucopolysaccharidoses (MPSes) are now treatable, and that the earlier treatment is initiated the better, is an indication for neonatal screening. The most efficient approach seems likely to be a multi‐tier procedure in which screening for urinary glycosaminoglycan (GAG) is followed by enzyme determinations in heelprick blood of newborns screening positive. Hitherto the method of choice for the determination of GAG has been the measurement of absorbance by a complex of GAG and 1,9‐dimethylmethylene blue (DMB). Method: We evaluated a DMB method in which absorbance by DMB is measured following its addition to the eluate obtained from paper‐borne newborn urine samples and is normalized relative to urinary creatinine. Calibration is performed with chondroitin‐6‐sulfate (Ch‐6‐S). Results: The limits of detection and quantification of GAG were 1.98 and 5.94 mg/dl, respectively. The within‐run coefficients of variation (CVs) of the GAG/creatinine ratio for 25, 31, and 70 mg/dl solutions of Ch‐6‐S in urine were 21.8, 16.4, and 10.5%, respectively, and the corresponding between‐run CVs were 25.0, 13.5, and 10.1%. Recovery from the urine spiked with 31 mg Ch‐6‐S/dl was 94.8%. Accuracy was also acceptable for all other GAGs except hyaluronic acid. For neonatal screening, the diagnostic threshold was tentatively established as 800 mg GAG/g creatinine, the 95th centile of samples from 903 infants aged 3–28 days, but the value of the GAG/creatinine ratio was negatively correlated with age. Application of the new method to samples from older individuals with and without MPS achieved 100% sensitivity and specificity when used with an age‐dependent threshold taken from the literature on the original DMB method. Conclusion: If used in the first tier of a multi‐tier screening protocol, the proposed method would allow the detection of abnormal levels of all GAGs except hyaluronic acid. J. Clin. Lab. Anal. 24:149–153, 2010. © 2010 Wiley‐Liss, Inc.

We describe the history and current implementation of an inexpensive thin layer chromatography (TLC) method, vertical sandwich‐type continuous/evaporative TLC with fixed mobile phase volume, that is convenient for detecting and identifying reducing sugars of clinical relevance in the paper‐borne blood and urine samples collected in neonatal screening programmes. This method facilitates screening by providing a considerable degree of standardization of chromatographic results. Among some 555,000 newborns to which it has been applied, it has detected 10 cases of classical galactosaemia, 7 cases of galactokinase deficiency, 2 cases of glucosuria, and 3 cases of transitory neonatal diabetes mellitus; the only false negatives we are aware of were two cases of galacto‐4‐epimerase deficiency detected by tandem mass spectrometry. Screening for sugars in urine has allowed the detection of galactosaemia when the accompanying blood sample was invalid because of transfusion or parenteral feeding. The conclusion is that this inexpensive procedure is very useful for the detection of relevant metabolopathies in circumstances where others fail. J. Clin. Lab. Anal. 24:106–112, 2010. © 2010 Wiley‐Liss, Inc.

Aims To evaluate the usefulness of a quantification method using filter paper for analyzing minute voided urine of the mouse. Methods Voided stain on paper (VSOP) method; the correlation between area of stained spot on a filter paper and amount of applied liquid was calculated. Voiding behavior of the mice was analyzed by placing the animal above the same filter paper and recording voided time and area over 2 hr. The usefulness of the VSOP method was tested in analysis of the voiding behavior of five female 7-week-old ddY mice treated with cyclophosphamide (CPM, 150 mg/kg, intraperitoneally) and five control ones, in comparison with the histology of CPM-induced cystitis. Further, the voided volume of male and female ddY mouse ranging from 2 to 13 weeks was assessed. Results There was a linear correlation between liquid volume and stained area on the filter paper (y = 16.472x − 22.411, R2 = 0.9981). Between control mice and those with histologically proven CPM cystitis, there was a significant difference in voided volume (362.7 ± 51.9 and 127.8 ± 100.0 µl

Background: The concentrations of the pyrimidine “de novo” metabolites and their degradation products in urine are useful indicators for the diagnosis of an inborn error of the pyrimidine de novo pathway or a urea-cycle defect. Until now, no procedure was available that allowed the analysis of all of these metabolites in a single analytical run. We describe a rapid, specific method to measure these metabolites by HPLC–tandem mass spectrometry. Methods: Urine or urine-soaked filter-paper strips were used to measure N-carbamyl-aspartate, dihydroorotate, orotate, orotidine, uridine, and uracil. Reversed-phase HPLC was combined with electrospray ionization tandem mass spectrometry, and detection was performed by multiple-reaction monitoring. Stable-isotope-labeled reference compounds were used as internal standards. Results: All pyrimidine de novo metabolites and their degradation products were measured within a single analytical run of 14 min with lower limits of detection of 0.4–3 μmol/L. The intra- and interassay variation for urine with added compounds was 1.2–5% for urines and 2–9% for filter-paper extracts of the urines. Recoveries of the added metabolites were 97–106% for urine samples and 97–115% for filter-paper extracts of the urines. Analysis of urine samples from patients with a urea-cycle defect or pyrimidine degradation defect showed an aberrant metabolic profile when compared with controls. Conclusion: HPLC with electrospray ionization tandem mass spectrometry allows rapid testing for disorders affecting the pyrimidine de novo pathway. The use of filter-paper strips could facilitate collection, transport, and storage of urine samples.

Urea and creatinine were measured by Kodak Ektachem methods and methylmalonic acid by stable-isotope gas chromatography-mass spectrometry in 24 urine samples from patients with methylmalonic acidaemia. For each sample analyses were performed both directly on the liquid urine and on an aliquot which had been blotted onto filter paper and dried. There was good correlation between the methylmalonate/creatinine ratios in liquid and dried urine (r2 = 0.983). Urine dried on filter paper can be used advantageously for monitoring treatment in patients with this disorder.

There is little doubt that mental retardation has been prevented in most babies diagnosed by newborn screening programs for inborn errors and the cost–benefit ratios of these programs have been reported as highly positive. In a previous work we optimised a CE method for quick profiling of organic acidurias, which characterize a large number of inborn errors, so that it permits the separation, detection and even identification in less than 15 min of 22 organic acids in urine samples related to a wide range of metabolic disorders. In the present work we have studied the adequacy of filter paper collection of urine samples to simplify this step, always difficult in babies, when it is not performed by training personnel. The studied parameters were: media and conditions for re-extraction to give the best sensitivity and a more simple procedure when the samples are measured by CE, interferences coming from the diaper, recoveries obtained, possible correction of recoveries with creatinine and stability of the compounds. The whole method we report has the advantages of easy sample collection, easy shipping or delivery, and rapid analysis. Moreover, this method of collection and analysis allows the identification and quantitation of fumaric, methylmalonic, N-acetylaspartic, pyroglutamic and homogentisic acids, as well as glutaric acid for which screening is considered especially advisable.

Propionic acidaemia (PCCD) or deficiency of propionyl-CoA carboxylase (PCC) is one of the most common organic acidaemias. Recent studies have suggested that this disease can cause somatic or cognitive deterioration even in patients without ketosis or metabolic acidosis, or in cases with unusually late onset. This suggests that for this disease a sensitive yet practical screening procedure is required to achieve early treatment. We conducted a pilot study of gas chromatographic-mass spectrometric screening of 12,000 newborns for PCCD using eluates from dried filter-paper urine collected at 4-7 days of age. Methylcitrate (MC) was targeted for PCCD. For bulk screening, 2-hydroxyundecanoate was used as internal standard; for quantification, stable-isotope-labelled MC was used. Urease pretreatment without fractionation allowed satisfactory recovery and reproducibility of the highly polar MC. We detected an asymptomatic male infant with distinctly elevated MC: the creatinine-corrected level relative to 2-hydroxyundecanoate was 4.8 SD above the normal mean. The MC concentration calculated using the stable-isotope-labelled internal standard was 70.6 mmol/mol creatinine 14.7 SD above the normal mean of 3.70. Parallel analysis of the dried blood spot at 4 days of age by tandem MS showed only borderline elevation of propionylcarnitine. The activity of PCC in lymphocytes was 7% of control. Gene analysis revealed that a single missense mutation, TAT to TGT, resulting in Y435C in the beta chain was present in a homozygous form. Dietary treatment including carnitine supplementation decreased this infant's MC level and to date (at 13 months of age), he shows no neurological or somatic abnormalities.

There are several organic acid disorders that require information on alpha-ketoacids, such as maple syrup urine disease or alpha-ketoadipic acidemia. The recovery, stability and diagnostic availability of alpha-ketoacids in dried urine filter paper analyzed by GC-MS with oxime-trimethylsilyl derivatization was studied for organic acidemia screening. The recovery of all nine types of alpha-ketoacids tested, but for phenylpyruvate, 2-ketoadipate, and p-OH-phenylpyruvate, from filter paper samples was acceptable. The stability of pyruvate, branched-chain alpha-ketoacids, alpha-ketoadipate and alpha-ketoglutarate was stable for at least 28 days, although some alpha-ketoacids such as succinylacetone were unstable. It indicated it was difficult to diagnose only tyrosinemia type 1 among nine specimens from organic acidemia patients tested. The method could be applied to global organic acidemia screening.

This review will be concerned primarily with a practical yet comprehensive diagnostic procedure for the diagnosis or even mass screening of a variety of metabolic disorders. This rapid, highly sensitive procedure offers possibilities for clinical chemistry laboratories to extend their diagnostic capacity to new areas of metabolic disorders. The diagnostic procedure consists of the use of urine or filter paper urine, preincubation of urine with urease, stable isotope dilution, and gas chromatography-mass spectrometry. Sample preparation from urine or filter paper urine, creatinine determination, stable isotope-labeled compounds used, and GC-MS measurement conditions are described. Not only organic acids or polar ones but also amino acids, sugars, polyols, purines, pyrimidines and other compounds are simultaneously analyzed and quantified. In this review, a pilot study for screening of 22 target diseases in newborns we are conducting in Japan is described. A neonate with presymptomatic propionic acidemia was detected among 10,000 neonates in the pilot study. The metabolic profiles of patients with ornithine carbamoyl transferase deficiency, fructose-1,6-bisphosphatase deficiency or succinic semialdehyde dehydrogenase deficiency obtained by this method are presented as examples. They were compared to those obtained by the conventional solvent extraction methods or by the tandem mass spectrometric method currently done with dried filter blood spots. The highly sensitive, specific and comprehensive features of our procedure are also demonstrated by its use in establishing the chemical diagnosis of pyrimidine degradation defects in order to prevent side effects of pyrimidine analogs such as 5-flurouracil, and the differential diagnosis of three types of homocystinuria, orotic aciduria, uraciluria and other urea cycle disorders. Evaluation of the effects of liver transplantation or nutritional conditions such as folate deficiency in patients with inborn errors of metabolism is also described.