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Propofol is a widely used intravenous anesthetic agent in sedation and general anesthesia. To improve the safety and maintain the depth of anesthesia, it is important to develop a rapid, sensitive, and reliable method to monitor the concentration of propofol in blood during anesthesia continuously. Here, we present a novel strategy based on paper spray ionization-mass spectrometry (PSI-MS) to detect propofol. Samples (in 10 μL) were mixed with methanol as protein precipitation solvent and 2,6-dimethylphenol as internal standard. Protein micro-precipitation was achieved with methanol by vortexing and centrifuging for 5 s each, and propofol was extracted to the supernatant. PSI-MS was performed in negative ionization mode, and MS signal lasted for 1 min. The analysis of a single sample was completed within 2 min. The area ratios of propofol to internal standard were calculated for quantification. Limit of detection of 5.5 ng mL−1 and limit of quantification of 18.2 ng mL−1 were achieved for propofol in whole blood. Calibration curve was linear in the range of 0.02–10 μg mL−1. The developed method was used successfully in monitoring the propofol concentration in 3 patients’ whole blood during anesthesia, showing its further application in controlling and feeding-back target concentration infusion.

A novel all-in-one paper-based sampling concept for mass spectrometric bottom-up protein analysis is here demonstrated in a chip format integrating instant immunocapture, protein reduction, - alkylation and tryptic digestion all in-device. Conventional laboratory grade filter paper was coated with the polymer 2-hydroxyethyl methacrylate-co-2-vinyl-4,4-dimethyl azlactone (pHEMA-VDM) with a subsequent covalent immobilization of the monoclonal antibody E27 targeting the biomarker human chorionic gonadotropin (hCG). In-device protein reduction and alkylation was optimized with regards to reagent concentration and reaction pH. The sampling concept showed a high degree of performance between 10 and 1000 ng/mL (R2 > 0.99) by a five-point calibration curve sampled with hCG spiked to human serum samples and freshly collected whole blood samples, respectively. LOD (experimentally obtained at 100 pg/mL (2.64 pM/0.9 IU/L)) was demonstrated to be up to ten times lower with more than six times faster sample preparation than what has previously been reported for on-paper analysis of hCG in human serum samples.

In this work, we analyzed the performance of the Electro-Filtering Paper Spray Ionization (EFSI) method for detecting compounds in unprocessed samples. A relatively rigid and electrically conductive copper filter was used as a substrate to insure sufficient and efficient sample-solvent extraction and to increase the conductivity for paper spraying. The method was demonstrated as applicable for indirect high-throughput analysis of large-volume unprocessed samples, which is not possible with conventional nanoESI or direct paper spray methods. The new method can generate different desired ion signals for a wide range of compounds by selecting different extraction solvents. Moreover, key parameters related to extraction efficiency were optimized in detail to obtain the most satisfactory extraction efficiency during antibiotic analysis. Finally, under optimal conditions, the EFSI method was successfully used to detect four antibiotics in animal products of egg, chicken, and chicken liver, exhibiting good reproducibility with calibration curves between 81.6% and 96.3%, and R2 values above 0.99. Recoveries of 75.0%ￚ94.6% were obtained for the four antibiotics. Hence, the proposed EFSI-MS is a successful, economical, rapid, and high-throughput method that is effective for both unknown and targeted extraction of unprocessed samples by mass spectrometric analysis.

This study describes a microfluidic paper-based analytical device (μPAD) for separating plasma from whole blood and measuring glucose concentration. A two-dimensional μPAD was fabricated by wax printing, using chromatographic paper and was functionalized by the incorporation of chitosan into the plasma separation zone. Red blood cells were isolated from whole blood in the plasma separation zone consisting of a chitosan polymer structure and a wax barrier. The separated plasma, with buffer, was injected into the test zone to cause a color change. The colorimetric results were digitized with a scanner, and the concentration was calculated using a calibration curve. This μPAD, which separated the plasma from whole blood without a separation membrane, will be useful for constructing point-of-care testing devices that detect analytes in small sample volumes.

Clinical application of direct sampling electrospray ionization mass spectrometry (ESI-MS) remains limited due to problems associated with very “dirty” sample matrices. Herein we report on a microfluidic platform that allows direct mass spectrometric analysis of serum samples of microliter sizes. The platform integrates in-line paper adsorption-based sample clean-up and voltage assisted liquid desorption ESI-MS/MS (VAL DESI-MS/MS) to detect multiple targeted compounds of clinical interest. Adenosine monophosphate (AMP), adenosine diphosphate (ADP), and adenosine triphosphate (ATP) were selected as model analytes. Simultaneous quantification of these compounds in human serum samples was demonstrated. For all the three compounds, linear calibration curves were obtained in a concentration range from 0.20 to 20.0 μmol/L with r2 values ≥ 0.996. Limits of detection were 0.019, 0.015, and 0.011 μmol/L for AMP, ADP, and ATP, respectively. Recovery was found in the range from 96.5% to 103.5% at spiking concentrations of 0.25 and 2.50 μmol/L. The results indicate that the proposed microfluidic mass spectrometric platform is robust and effective. It may have a potential in clinical analysis.

An optical colorimetric and smartphone-integrated paper device (SIPD) is demonstrated for determination of As (III) in water and soil samples using sucrose modified gold nanoparticles (AuNPs/Suc) as a nanoprobe. The mechanism for determination of As(III) is experimentally validated by performing UV-Vis, transmission electron microscope (TEM), Fourier transforms infra-red spectroscopy (FTIR) and dynamic light scattering (DLS) measurements. The density function theory (DFT) calculations using B3LYP with 6-311G (2d,2p) and LANL2DZ basis sets is used to theoretically prove the mechanism for determination of As(III). In addition, the paper fabricated with AuNPs/SuC was used as a nanoprobe for quantitative determination of As(III) using smartphone and ImageJ software. Calibration plot was linear over 10–800 μgL−1 for colorimetric determination of As(III) with limit of detection (LOD) of 4 μgL−1 acquired when the absorbance ratio obtained at 594 nm/515 nm. The linearity range of 50–3000 μgL−1 with LOD of 20 μgL−1 was determined using smartphone-integrated paper device. AuNPs/Suc is successfully employed for determination of As (III) from contaminated water and soil samples in colorimetry and SIPD.

A fast methodology for the assay of parabens in drug and cosmetic preparations has been presented. The procedure developed is based on paper spray tandem mass spectrometry and isotope dilution approach. For each investigated paraben, the corresponding labeled standard has been used in order to improve the accuracy and reproducibility of the analyses. The MS experiments have been performed under MRM conditions, monitoring the transitions [M-H]- → m/z 92 and [M-H]- → m/z 98, respectively, for each analyte and the corresponding labeled internal standard. The quantitative assay has been performed using a calibration curve built from 2 to 15 mg/L. The method accuracy, in all case near 100%, was evaluated using fortified samples at two concentration levels, which are representative of the lower and the higher portion of calibration curve. The good values of LOQ, LOD, and reproducibility confirm the consistency of the developed approach.

With its low-cost fabrication and ease of modification, paper-based analytical devices have developed rapidly in recent years. Microarrays allow automatic analysis of multiple samples or multiple reactions with minimal sample consumption. While cellulose paper is generally used, its high backgrounds in spectrometry outside of the visible range has limited its application to be mostly colorimetric analysis. In this work, glass-microfiber paper is used as the substrate for a microarray. The glass-microfiber is essentially chemically inert SiOx, and the lower background from this inorganic microfiber can avoid interference from organic analytes in various spectrometers. However, generally used wax printing fails to wet glass microfibers to form hydrophobic barriers. Therefore, to prepare the hydrophobic-hydrophilic pattern, the glass-microfiber paper was first modified with an octadecyltrichlorosilane (OTS) self-assembled monolayer (SAM) to make the paper hydrophobic. A hydrophilic microarray was then prepared using a CO2 laser scriber that selectively removed the OTS layer with a designed pattern. One microliter of aqueous drops of peptides at various concentrations were then dispensed inside the round patterns where OTS SAM was removed while the surrounding area with OTS layer served as a barrier to separate each drop. The resulting specimen of multiple spots was automatically analyzed with a time-of-flight secondary ion mass spectrometer (ToF-SIMS), and all of the secondary ions were collected. Among the various cluster ions that have developed over the past decade, pulsed C60+ was selected as the primary ion because of its high secondary ion intensity in the high mass region, its minimal alteration of the surface when operating within the static-limit and spatial resolution at the ∼μm level. In the resulting spectra, parent ions of various peptides (in the forms [M+H]+ and [M+Na]+) were readily identified for parallel detection of molecules in a mixture. By normalizing the ion intensity of peptides with respect to the glass-microfiber matrix ([SiOH]+), a linear calibration curve for each peptide was generated to quantify these components in a mixture.

In this work, ZnO nanorods (ZnO NRs) with different sizes were hydrothermally grown on the surface of Whatman filter paper for the fabrication of a microfluidic paper-based device (μPAD) for the simultaneous detection of glucose and uric acid. As dual enzymatic reaction was employed for the colorimetric detection in this μPAD, the presence of ZnO NRs promoted the enzyme immobilization thus significantly enhancing the colorimetric signal. The coffee ring effect was effectively conquered by the uniform distribution of ZnO NR as well as a specialized double-layered μPAD design. Meanwhile, two color indicators with distinct colors were used to provide complementary results to better quantify the concentration of the analytes by naked eye. As a result, two linear calibration curves were obtained for the detection of glucose (0.01–10 mmol L−1) and uric acid (0.01–5 mmol L−1), along with a LOD of 3 μmol L−1 for glucose and 4 μmol L−1 for uric acid, respectively. The practical usefulness of the proposed μPAD was further validated by the simultaneous analysis of glucose and uric acid in serum samples and urine samples.

In this study, a paper spray ionization mass spectrometric (PS-MS) method was developed for the rapid in situ screening and simultaneous quantitative analysis of bisphenol A and its analogues, i.e., bisphenol S, bisphenol F, and bisphenol AF, in food packaging products. At the optimal PS-MS conditions, the calibration curves of bisphenols in the range of 1-100 μg/mL were linear. The correlation coefficients were higher than 0.998, and the LODs of the target compounds were 0.1-0.3 μg/mL. After a simple treatment by dichloromethane on the surface, the samples were analyzed by PS-MS in situ for rapid screening without a traditional sample pretreatment procedure, such as powdering, extraction, and enrichment steps. The analytical time of the PS-MS method was less than 1 min. In comparison with conventional HPLC-MS/MS, it was demonstrated that PS-MS was a more effective high-throughput screening and quantitative analysis method.

We demonstrate the possibility of measuring FRET efficiency with a low-cost frequency-domain fluorescence lifetime imaging microscope (FD-FLIM). The system utilizes single-frequency-modulated excitation, which enables the use of cost-effective laser sources and electronics, simplification of data acquisition and analysis, and a dual-channel detection capability. Following calibration with coumarin 6, we measured the apparent donor lifetime in mTFP1-mVenus FRET standards expressed in living cells. We evaluated the system's sensitivity by differentiating the short and long lifetimes of mTFP1 corresponding to the known standards' high and low FRET efficiency, respectively. Furthermore, we show that the lifetime of the vinculin tension sensor, VinTS, at focal adhesions (2.30 ± 0.16 ns) is significantly (p l 10 - 6) longer than the lifetime of the unloaded TSMod probe (2.02 ± 0.16 ns). The pixel dwell time was 6.8 μs for samples expressing the FRET standards, with signal typically an order of magnitude higher than VinTS. The apparent FRET efficiency ( E FRET app ) of the standards, calculated from the measured apparent lifetime, was linearly related to their known FRET efficiency by a factor of 0.92 to 0.99 (R2 = 0.98). This relationship serves as a calibration curve to convert apparent FRET to true FRET and circumvent the need to measure multiexponential lifetime decays. This approach yielded a FRET efficiency of 18% to 19.5%, for VinTS, in agreement with published values. Taken together, our results demonstrate a cost-effective, fast, and sensitive FD-FLIM approach with the potential to facilitate applications of FLIM in mechanobiology and FRET-based biosensing.

Rationale Paper spray mass spectrometry (PS-MS) was used to analyze and quantify ampicillin, a hydrophilic compound and frequently utilized antibiotic. Hydrophilic molecules are difficult to analyze via PS-MS due to their strong binding affinity to paper substrates and low ionization efficiency, among other reasons. Methods Solvent and paper parameters were optimized to increase the extraction of ampicillin from the paper substrate. After optimizing these key parameters, a Resolution IV 1/16 fractional factorial design with two center points was employed to screen eight different design parameters simultaneously. Results Pore size, sample volume, and solvent volume were the most significant factors affecting average peak area under the curve (AUC) and the signal-to-blank (S/B) ratio for the 1 μg/mL ampicillin calibrant. After optimizing the key parameters, a linear calibration curve with a range of 0.2 μg/mL to 100 μg/mL was generated (R2 = 0.98) and the limit of detection (LOD) and lower limit of quantification (LLOQ) were calculated to be 0.07 μg/mL and 0.25 μg/mL, respectively. Conclusions The statistical optimization procedure undertaken here increased the mass spectral signal intensity by more than a factor of 40. This statistical method of screening followed by optimization experiments proved faster and more efficient, and produced more drastic improvements than typical one-factor-at-a-time experiments.

Microfluidic paper-based analytical devices (μPADs) were used to detect the iron ion content in the water of a natural hot spring in order to assess the applicability of this process to the environmental analysis of natural water. The μPADs were fabricated using a wax printer after the addition of hydroxylamine into the detection reservoirs to reduce Fe(3+) to Fe(2+), 1,10-phenanthroline for the forming of a complex, and poly(acrylic acid) for ion-pair formation with an acetate buffer (pH 4.7). The calibration curve of Fe(3+) showed a linearity that ranged from 100 to 1000 ppm in the semi-log plot whereas the color intensity was proportional to the concentration of Fe(3+) and ranged from 40 to 350 ppm. The calibration curve represented the daily fluctuation in successive experiments during four days, which indicated that a calibration curve must be constructed for each day. When freshly prepared μPADs were compared with stored ones, no significant difference was found. The μPADs were applied to the determination of Fe(3+) in a sample of water from a natural hot spring. Both the accuracy and the precision of the μPAD method were evaluated by comparisons with the results obtained via conventional spectrophotometry. The results of the μPADs were in good agreement with, but less precise than, those obtained via conventional spectrophotometry. Consequently, the μPADs offer advantages that include rapid and miniaturized operation, although the precision was poorer than that of conventional spectrophotometry.

The assessment of pesticide residue levels demands fast, low cost and easy-to-use procedures which are not found in conventional methods. In this work, SERS substrates based on the deposition of gold nanoparticles (GNPs) on common office paper were prepared using a wax printer. These substrates combined with Data Driven Soft Independent Modelling of Class Analogies (DD-SIMCA), a one-class classifier algorithm, were used for detection of pesticide residues in water extracts of mango peels. Paper-based substrates made sample collection easier compared with conventional SERS methods, since few microliters of the pesticide aqueous extract from fruit peels needed to be deposited onto the substrate. Moreover, one-class classifiers dismiss the need for quantification or calibration curves. Classification of a fruit with residue levels in accordance to regulatory bodies' limits is based on a mathematical threshold. Just as in an authentication problem, all the possibilities for a given analysed fruit are now restricted to agreeing or not agreeing with current regulations. The performance of the one-class model was demonstrated by detecting thiabendazole (TBZ) residues at various mango samples, with all results being confirmed by HPLC-DAD analysis. The final model could distinguish samples with TBZ levels above the ones allowed by the Brazilian Health Regulatory Agency with 94% of selectivity and 92% of sensitivity, even in the presence of other pesticides.

This paper describes the development of an original micro-solid phase extraction device and its evaluation for the isolation of F2-isoprostanes (F2-IsoPs) from cord and maternal plasma samples. The unit is very simple and consists in a rotating disc (1.8 cm diameter) of oxidized buckypaper (BP), enwrapped in a polypropylene mesh pouch. Even if the selected F2-IsoPs have logP and pKa values that make them suitable candidates for their sorption on BP, several parameters were optimized to maximize recoveries: time of adsorption and desorption; stirring speed; volume, pH and ionic strength of the sample; type, volume, and fractions of the elution solvent; oxidation grade of BP. Among all, the last one was crucial in affecting extraction yields because of the analyte interactions with polar functionalities, introduced by a preliminary oxidative acid treatment. The investigation established the optimal oxidation time and highlighted the pros and cons of the acid activation step. All extracts were analyzed by means of liquid chromatography-tandem mass spectrometry (LC–MS/MS). Validation was performed according to the main FDA guidelines for bioanalytical methods. Depending on the spike level and analyte, recoveries ranged between 30 and 120% with precision and accuracy values lower than 20%. Quantitative analysis was accomplished by matrix-matched calibration curves whose determination coefficients were higher than 0.95. Lower limit of quantitation (LLOQ) spanned the range 2.45–6.77 μg L−1. The validated method was applied to the analysis of eight pairs of mother/child plasma samples, revealing the presence of 8-iso-15-keto-PGF2α and 8-iso-PGE2 at a concentration of about 10 μg L−1 in most cord plasma samples of preterm newborns.

Nowadays, there is great interest in the use of microfluidic paper-based analytical devices (μPADs) for low-cost diagnostics. In most cases, the test equipment is needed. Here we report a new type of device-independent detection method based on timer-paper-based analytical devices, which can be tested by smartphone, and its application for cholesterol detection. The Quick Response code was designed as the timer component of this device. Wax printing method was employed to print the pattern on filter paper. The color and enzyme reagents have been immobilized in the hydrophilic channel to complete the colorimetric detection for cholesterol. Under laminar flow conditions of the cellulose network, the liquid volume of detection zone has been quantified by monitoring the fluid residence time on different area of the timer-paper-based devices. The precise monitoring of detection time can promote the accuracy of colorimetric detection. This is very important for the quantitative detection of paper-based analytical devices. One significant outcome of this report is that simple and accurate timer can be used for detection process self-clocking. The factors of total detection time have been investigated. The linear range of the calibration curve was 3.0 ~ 6.0 mmol L−1, with correlation coefficient of 0.9956. With the characteristic of easy to use, low cost and accurate monitoring of detection time, this kind of timer-paper-based devices can be applied to cholesterol or other substances rapid detection.

Rationale : Saliva as a sample matrix is rapidly gaining interest for disease diagnosis and point-of-care assays because it is easy to collect (non-invasive) and contains many health-related biomarkers. However, saliva poses particular problems relative to more common urine and blood matrices, which includes low analyte concentrations, lack of understanding of biomolecule transportation and inherent viscosity variability in human samples. While several studies have sought to improve assay sensitivity, few have addressed sample viscosity specifically. The goal of this study is to minimize the effect of sample viscosity on paper-based analytical devices (PADs) for the measurement of pH and nitrite in human saliva. Methods : PADs were used to measure salivary pH from 5.0 to 10.0 with a universal indicator consisting of chlorophenol red, phenol red and phenolphthalein. Nitrite determination was performed using the Griess reaction. Artificial saliva with viscosity values between 1.54 and 5.10 mPa∙s was tested on the proposed PAD. To ensure the proposed PADs can be tailored for use in-field analysis, the devices were shipped to Australia and tested with human specimens. Results : Initial experiments showed that viscosity had a significant impact on the calibration curve for nitrite; however, a more consistent curve could be generated when buffer was added after the sample, irrespective of sample viscosity. The linear range for nitrite detection was 0.1 to 2.4 mg/dL using the improved method. The nitrite measurement in artificial saliva also showed a good correlation with the standard spectrophotometry method (p=0.8484, paired sample t-test, n=20). Measured pH values from samples with varying viscosities correlated well with the results from our pH meter. Conclusions : The inherent variation of salivary viscosity that impacts nitrite and pH results can be addressed using a simple washing step on the PAD without the need for complex procedures.

In this research, the thin film microextraction method was applied for the extraction of Ni(II) ion from aqueous matrixes. Chemically modified cellulosic filter paper with phosphorus was used as a thin film extractor. After extraction, the thin film was treated with a solution of dimethylglyoxime. The colored film was captured by flatbed scanner and the absorbance of the images was extracted by some suitable software. Under the optimum conditions and at the pH 7.0, with the sample volume of 100 mL, the stirring rate of 800 rpm, and the extraction time of 50 min, the calibration curve was obtained in the range of 0.05–5 mg/L Ni(II) (R2 = 0.989). Limit and relative standard deviation were achieved to be 18 µg/L and less than 6.7%, respectively. Relative recoveries were obtained in the range of 87%–105%. Finally, the proposed method was found to be simple and cost-effective, with adequate analytical performance for the rapid detection of Ni(II) in river and wastewater samples.

With the emergence of artesunate antimalarial counterfeiting in Southeast Asia and sub-Saharan Africa, we present the production of a rapid, inexpensive and simple colorimetric-based testing kit for the detection of counterfeit artesunate in order to preserve life and prevent the development of multi-drug resistant malaria. The kit works based on paper microfluidics which offer several advantages over conventional microfluidics, and has great potential to generate inexpensive, easy-to-use, rapid and disposable diagnostic devices. Here, we have developed a colorimetric assay that is specific to artesunate and turns yellow upon addition of the sample. The test can be done within minutes, and allows for a semi-quantitative analysis of the artesunate tablets by comparing the developed yellow color on the paper test to a color-coded key chart that comes with the kit. A more accurate and precise analysis is done by utilizing a color analyzer on an iPhone camera that measures the color intensity of the developed color on the paper chip. A digital image of the chip was taken and analyzed by measuring the average gray intensity of the color developed on the paper circle. A plot of the artesunate concentration versus the average gray scale intensity was generated. Results show that the intensity of the yellow color developed on the paper test was consistent and proportional to the amount of artesunate present in the sample. With artesunate concentrations ranging from 0.0 to 20 mg/mL, a linear calibration plot was obtained with a detection limit of 0.98 mg/mL.

In this work, filter paper was used as a low cost substrate for silver nanoparticles in order to perform the detection and quantification of acetylsalicylic acid by SERS in a commercial tablet. The reaction conditions were 150mM of ammonium hydroxide, 50mM of silver nitrate, 500mM of glucose, 12min of the reaction time, 45°C temperature, pretreatment with ammonium hydroxide and quantitative filter paper (1-2μm). The average size of silver nanoparticles deposited on the paper substrate was 180nm. Adsorption time of acetylsalicylic acid on the surface of the silver-coated filter paper was studied and an adsorption time of 80min was used to build the analytical curve. It was possible to obtain a calibration curve with good precision with a coefficient of determination of 0.933. The method proposed in this work was capable to quantify acetylsalicylic acid in commercial tablets, at low concentration levels, with relative error of 2.06% compared to the HPLC. The preparation of filter paper coated with silver nanoparticles using Tollen's reagent presents several advantages such as low cost of synthesis, support and reagents; minimum amount of residuals, which are easily treated, despite the SERS spectroscopy presenting fast analysis, with low sample preparation and low amount of reactants as in HPLC analysis.

To the Editor: Paper spray (PS)1 is a recently described method for the direct mass spectrometry (MS) analysis of blood and other biological samples (1–3). Paper spray is performed by depositing a sample such as whole blood onto a paper substrate contained within a disposable cartridge; extraction and ionization occur directly from the disposable cartridge with an automated MS attachment (Fig. 1). No punching or offline extraction is required. Total analysis time, including sample extraction, is several minutes. Fig. 1. Paper spray for direct analysis of blood spots by MS. Ion chronograms from the lowest calibrator in whole blood, overlaid with representative chronograms from blank blood, are shown for cyclosporine and sirolimus. Quantification was performed with the entire area under the curve for each analyte, normalized by SIL-IS, to obtain calibration curves as shown. Because of its simplicity, paper spray could lower the barrier for implementation of MS-based assays in clinical laboratories. We previously used PS-MS/MS to monitor tacrolimus in clinical samples and showed good correlation with 2 immunoassays and an external HPLC-MS/MS method (4). Here we extend this approach by demonstrating the simultaneous quantification of cyclosporine and sirolimus by PS-MS/MS. Therapeutic drug monitoring of these drugs is important for organ transplant patient care. The method used was similar to our …

Glutathione is an important antioxidant found in body fluids and tissues, which inhibit damage to essential cellular constituents caused by reactive oxygen species. The analysis of glutathione levels in biological systems is important in early clinical diagnosis. A novel, cost-effective synthetic strategy has been developed for the fluorescent probe ethylenediamine passivated carbon dots. Tissue paper was chosen as the carbon source for this "green one pot" synthesis. Glutathione could induce quenching of the fluorescence intensity of ethylenediamine passivated carbon dots through surface interactions, resulting from their aggregation. Based on this, a novel fluorescence sensor was fabricated for the determination of Glutathione in body fluids. A linear calibration graph was obtained in the range of 6.0 × 10-7 to 5.0 × 10-8 M with a detection limit of 1.74 × 10-9 M. The developed sensor was successfully used for the determination of glutathione in artificial saliva samples.

In this work, the establishment of a microdialysis-paper spray ionization-mass spectrometry (MS) system was described. A homemade microdialysis module was employed for sampling, and microdroplets were generated at the outlet of the capillary conducting the dialysate. Online MS analysis of each microdroplet was immediately accomplished, interfacing by paper spray ionization. Analytical performance of the method was investigated and improved through the introduction of thinner capillary tubes and the optimization of spray solvent and paper substrate. For microdroplets with concentrated salt at 50 nL, the limit of detection at 0.8 ppm (or 40 pg absolute) and a highest resolution at about 1.5 s were achieved. The integrated system was applied into the online monitoring of glucose concentration in cell culture mediums. A satisfactory linearity of the calibration curve between the relative MS intensity and the glucose concentration was observed. Furthermore, as a model, hormone regulation of the glucose concentration was investigated. This work demonstrated the potential application of the label-free, online "MS sensor" into studies on cellular metabolism.

Drug levels in patients' bloodstreams vary among individuals and consequently therapeutic drug monitoring (TDM) is fundamental to controlling the effective therapeutic range. For TDM purposes, different analytical approaches have been used, mainly based on immunoassay, liquid chromatography-ultraviolet, liquid chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. More recently a matrix-assisted laser desorption/ionisation method has been proposed for the determination of irinotecan levels in the plasma of subjects under therapy and this method has been cross-validated by comparison with data achieved by LC-MS/MS. However, to reach an effective point-of-care monitoring of plasma drug concentrations, a TDM platform technology for fast, accurate, low-cost assays is required. In this frame, recently the use of paper spray mass spectrometry, which is becoming a popular and widely employed MS method, has been proposed. In this paper we report the results obtained by the development of a paper spray-based method for quantitative analysis in plasma samples of imatinib, a new generation of anticancer drug. Preliminary experiments showed that poor sensitivity, reproducibility and linear response were obtained by the “classical” paper spray set-up. In order to achieve better results, it was thought of interest to operate in presence of a higher and more homogeneous electrical field. For this aim, a stainless steel needle connected with the high voltage power supply was mounted below the paper triangle. Furthermore, in order to obtain valid quantitative data, we analysed the role of the different equilibria participating to the phenomena occurring in paper spray experiments, depending either on instrumental parameters or on the chemical nature of analyte and solvents. A calibration curve was obtained by spiking plasma samples containing different amounts of imatinib (1) with known amounts of deuterated imatinib (1d3) as internal standard, with molar ratios [1] / [1d3] in the range 0.00–2.00. A quite good linearity was obtained ( R2 = 0.975) and some experiments performed on spiked plasma samples with known amounts of 1 confirmed the validity of this method.

In this work, a double-layered three-dimensional (3D) microfluidic paper-based analytical device (μPAD) with high resolution temporally resolved chemiluminescence (CL) emissions were designed for multiplexed CL analysis. The temporally resolved CL emissions were obtained by virtue of the 3D branched microfluidic channel design, which create time delays for luminol transported from one detection zone to another. The peak intensity and peak shape of the temporally resolved CL emissions were quite stable and base-line separated with resolution as high as 21.2–24.4. Then, the fabricated μPAD was applied to multiplexed determination of glucose, lactate, cholesterol, and choline as model analytes. The sample was added to four detection zone modified with CL catalyst cobalt ion and different oxidase by virtue of chitosan. When luminol flowed to μPAD, four temporally resolved CL peaks were successively generated from the cobalt ion catalyzed CL reactions between luminol and generated H2O2 from the specific enzymatic reactions between the oxidase and the analytes. The generated four CL emission peaks in the CL kinetic curve increased in proportion to the concentrations of glucose, lactate, cholesterol, and choline, respectively. Finally, four linear calibration curves were obtained for the detection of glucose (0.01–1.0 mmol/L), lactate (0.02–5.0 mmol/L), cholesterol (0.01–0.4 mmol/L), and choline (0.001–1.0 mmol/L). The detection limits were as low as 8 μmol/L, 15 μmol/L, 6 μmol/L, and 0.07 μmol/L for glucose, lactate, choline, and cholesterol detection, respectively. The present work provides a new strategy for the fabrication of simple and sensitive 3D μPAD with high resolution temporally resolved CL emissions for multiplexed CL analysis, which holds great application potential for point-of-care diagnosis.

In this paper, chemically modified cellulose paper was introduced as a novel extracting phase for thin film microextraction (TFME). Different reagents (Octadecyltrichlorosilane, diphenyldichlorosilane, cyclohexyl isocyanate and phenyl isocyanate) were used to modify the cellulose papers. The modified papers were evaluated as a sorbent for the extraction of some synthetic and natural estrogenic hormones (17α-ethynylestradiol, estriol and estradiol) from aqueous samples. Liquid chromatography–fluorescence detection was used for the quantification of the extracted compounds. The cellulose paper modified with phenyl isocyanate showed the best affinity to the target compounds. TEME parameters such as desorption condition, shaking rate, sample ionic strength and extraction time were investigated and optimized. Limit of detections were between 0.05 and 0.23 μg L −1 and relative standard deviations were less than 11.1% under the optimized condition. The calibration curves were obtained in the range of 0.2–100 μg L −1 with a good linearity ( r 2 > 0.9935). Wastewater, human urine, pool and river water samples were studied as real samples for the evaluation of the method. Relative recoveries were found to be between 75% and 101%.

A paper-based immunodevice was fabricated for point-of-care test (POCT). The array device was simple and easily assembled. It comprised as many as 4×10 detection points on a single paper array. The immunosensor array was prepared by covalently immobilizing capture antibodies on corresponding working zone on a disposable paper array. With a sandwich-type immunoreaction, the CuO nanoparticles (CuO NPs)-labeled secondary antibody (Ab2) bioconjugates were captured in each working zone. The coordination of dithizone (DZ) at the surface of CdTe quantum dots (CdTe QDs) could strongly quench the green emission of CdTe QDs by a fluorescence resonance energy transfer (FRET) mechanism. After the Cu(2+) was released from CuO NPs-Ab2, the fluorescence of CdTe QDs-DZ was "turn-on". The fluorescence intensity would increase with the increasing of analytes. The calibration plot showed a good linear relationship between the fluorescence intensity and the logarithm value of the analytes concentration with the low detection limit. The immunosensor array was performed for cancer screening. The high throughput, low-cost, acceptable stability, reproducibility, sensitivity and accuracy showed good applicability of the proposed multiplex immunoassay in clinical diagnosis. The results indicated that the device could be applied to comprehensive sample and point-of-care detection.

With a great quantity of solid dosage tested by dissolution technology, developing a rapid and sensitive method to access the content of drug within dissolution media is highly desired by analysts and scientists. Traditionally, dissolution media is not compatible with mass spectrometry since the inorganic salts in the media might damage the mass spectrometer. Here, paper spray ionization mass spectrometry (PSI-MS), one of the ambient mass spectrometry technologies, is developed to characterize the content of drugs in dissolution media. The porous structure of paper can effectively retain salts from entering mass spectrometer. This makes the measurement of drug content within dissolution media by mass spectrometer possible. After the experimental parameters were optimized, calibration curves of model drugs - enalapril, quinapril and benazepril were established by using corresponding deuterated internal standards. PSI-MS was then deployed to characterize the content of enalapril from the dissolution testing of enalapril tablets. The results from PSI-MS are comparable to those from HPLC characterization. More importantly, the analysis time of 6 samples is shortened from 90min to 6min. Detection limit of enalapril maleate tablets by PSI-MS is 1/300 of LC. PSI-MS is rapid, sensitive and accurate in analyzing drug content from dissolution tests.

A microfluidic paper-based analytical device (μPAD) fabricated by wax printing was designed to assess occupational exposure to metal-containing aerosols. This method employs rapid digestion of particulate metals using microliters of acid added directly to a punch taken from an air sampling filter. Punches were then placed on a μPAD, and digested metals were transported to detection reservoirs upon addition of water. These reservoirs contained reagents for colorimetric detection of Fe, Cu, and Ni. Dried buffer components were used to set the optimal pH in each detection reservoir, while precomplexation agents were deposited in the channels between the sample and detection zones to minimize interferences from competing metals. Metal concentrations were quantified from color intensity images using a scanner in conjunction with image processing software. Reproducible, log-linear calibration curves were generated for each metal, with method detection limits ranging from 1.0 to 1.5 μg for each metal (i.e., total mass present on the μPAD). Finally, a standard incineration ash sample was aerosolized, collected on filters, and analyzed for the three metals of interest. Analysis of this collected aerosol sample using a μPAD showed good correlation with known amounts of the metals present in the sample. This technology can provide rapid assessment of particulate metal concentrations at or below current regulatory limits and at dramatically reduced cost.

This paper describes the first approach at combining paper microfluidics with electrochemiluminescent (ECL) detection. Inkjet printing is used to produce paper microfluidic substrates which are combined with screen-printed electrodes (SPEs) to create simple, cheap, disposable sensors which can be read without a traditional photodetector. The sensing mechanism is based on the orange luminescence due to the ECL reaction of tris(2,2'-bipyridyl)ruthenium(II) (Ru(bpy)(3)(2+)) with certain analytes. Using a conventional photodetector, 2-(dibutylamino)ethanol (DBAE) and nicotinamide adenine dinucleotide (NADH) could be detected to levels of 0.9 μM and 72 μM, respectively. Significantly, a mobile camera phone can also be used to detect the luminescence from the sensors. By analyzing the red pixel intensity in digital images of the ECL emission, a calibration curve was constructed demonstrating that DBAE could be detected to levels of 250 μM using the phone.

A novel and simple method to detect isothiazolinone-type biocides (2-methyl-3-isothiazolinone (MI), 5-chloro-2-methyl-3-isothiazolinone (CMI), 1,2-benzisothiazolinone (BIT) and 2-octyl-3-isothiazolinone (OIT)) in paper used for food packaging by ultrasonic extraction coupled with UPLC-MS/MS was developed. Parameters affecting process efficiency such as extraction solvents, UPLC mobile phase, gradient elution procedure and MS/MS conditions were studied to optimise the operating conditions. Using the optimised gradient elution procedure, the retention time was less than 6 min. The limits of detection (LODs) were found to be between 0.001 and 0.010 mg kg⁻¹, which was validated using actual concentrations. After diluting the standard solution with a blank matrix, the linear calibration curve ranges were 0.002-1.000 mg kg⁻¹ for BIT and OIT, 0.005-1.000 mg kg⁻¹ for MI, and 0.020-1.000 mg kg⁻¹ for CMI, with correlation coefficients higher than 0.9985 (n = 6). A good level of precision with a mean recovery greater than 81.3% and a relative standard deviation (RSD) less than 6.2% were also obtained. A methodology has been proposed for the analysis of isothiazolinones in paper.

This article describes the use of microfluidic paper-based analytical devices (muPADs) to perform quantitative chemical assays with internal standards. MicroPADs are well-suited for colorimetric biochemical assays; however, errors can be introduced from the background color of the paper due to batch difference and age, and from color measurement devices. To reduce errors from these sources, a series of standard analyte solutions and the sample solution are assayed on a single device with multiple detection zones simultaneously; an analyte concentration calibration curve can thus be established from the standards. Since the muPAD design allows the colorimetric measurements of the standards and the sample to be conducted simultaneously and under the same condition, errors from the above sources can be minimized. The analytical approach reported in this work shows that muPADs can perform quantitative chemical analysis at very low cost.

A novel test sample for the spatially resolved quantification of illicit drugs on the surface of a fingerprint using time-of-flight secondary ion mass spectrometry (ToF-SIMS) and desorption electrospray ionization mass spectrometry (DESI-MS) was demonstrated. Calibration curves relating the signal intensity to the amount of drug deposited on the surface were generated from inkjet-printed arrays of cocaine, methamphetamine, and heroin with a deposited-mass ranging nominally from 10 pg to 50 ng per spot. These curves were used to construct concentration maps that visualized the spatial distribution of the drugs on top of a fingerprint, as well as being able to quantify the amount of drugs in a given area within the map. For the drugs on the fingerprint on silicon, ToF-SIMS showed great success, as it was able to generate concentration maps of all three drugs. On the fingerprint on paper, only the concentration map of cocaine could be constructed using ToF-SIMS and DESI-MS, as the signals of methamphetamine and heroin were completely suppressed by matrix and substrate effects. Spatially resolved quantification of illicit drugs using imaging mass spectrometry is possible, but the choice of substrates could significantly affect the results.

A simple and low-cost fabrication method for paper-based diagnostic devices (PBDDs) is described in this study. Street-available polymer solutions were screen printed onto filter papers to create hydrophobic patterns for fluidic channels. In order to obtain fully functional hydrophobic patterns for fluids, the original polymer solutions were diluted with butyl acetate to yield a suitable viscosity range between 30-200 cP for complete patterning on paper. Typical pH and glucose tests with color indicators were performed on the screen printed PBDDs. Images of the PBDDs were analyzed by computers to obtain calibration curves for pH between 2 and 12 and glucose concentration ranging from 10-1000 mmol dm(-3). Detection of formaldehyde in acetone was also carried out to show the possibility of using this PBBD for analytical detection with organic solvents. An exemplar PBDD with simultaneous pH and glucose detection was also used to demonstrate the feasibility of applying this technique for realistic diagnostic applications.

Residual dried blood spots (DBSs) on filter paper from neonatal screening have been proposed as samples for population survey of lead contamination. We have investigated the EDTA effect on lead release in the eluting solution.Furnace atomic absorption spectrophotometry has been used for lead measurements. Standard, blank and sample solutions contained 2% m/v NH(4)H(2)PO(4), 0.5% v/v Triton X-100 and 0.2% v/v HNO(3) as matrix modifier solution (MMS) with or without EDTA. A calibration curve was established from aqueous standard solutions. Paper discs from DBS and blank, punched near the DBS, were eluted in MM solution and, where required, EDTA at different concentrations. Specimens were leftover DBSs with different storage times, matched samples from 20 adult patients consisting of liquid whole blood (LWB) containing 5 mmol/L EDTA, DBSs eluted in MM solution with 5 mmol/L EDTA or without EDTA.Optimal lead recovery from DBS required 5 mM EDTA in the eluting solution. Mean lead levels of LWB and DBSs eluted with EDTA were similar and higher than DBSs without EDTA (P0.001). Without EDTA, the median value of lead optical density was lower for 6-month-old DBSs than for blanks (P0.001).Residual DBSs can be used for population survey, but 5 mmol/L EDTA in the extracting solution is required to fully recover lead.

Here we report development of a smartphone app (application) that digitizes the colours of a colorimetric sensor array. A conventional colorimetric sensor array consists of multiple paper-based sensors, and reports the detection results in terms of colour change. Evaluation of the colour changes is normally done by the naked eye, which may cause uncertainties due to personal subjectivity and the surrounding conditions. Solutions have been particularly sought in smartphones as they are capable of spectrometric functions. Our report specifically focuses on development of a practical app for immediate point-of-care (POC) multi-analyte sensing without additional devices. First, the individual positions of the sensors are automatically identified by the smartphone; second, the colours measured at each sensor are digitized based on a correction algorithm; and third, the corrected colours are converted to concentration values by pre-loaded calibration curves. All through these sequential processes, the sensor array taken in a smartphone snapshot undergoes laboratory-level spectrometry. The advantages of inexpensive and convenient paper-based colorimetry and the ubiquitous smartphone are tied to achieve a ready-to-go POC diagnosis.

Quantitative determination of Ca, Mg, and Zn in fingernails was performed with laser-induced breakdown spectroscopy. Two different methods of producing solid standards for calibration were explored – preparation of keratin pellets and deposition of aqueous solutions to filter papers. Measurements of the temperature and electron density of the plasma produced on keratin pellets, filter paper, and nails were performed, and it was determined that the standards prepared on filter paper gave plasma temperatures and electron densities closer to those observed on the nails. The ablation rate of the filter paper was also more similar to that of the nails. Using calibration curves produced using these filter paper standards, Ca, Mg, and Zn were determined in fingernails of 11 subjects. For comparison, the same samples were digested and atomic absorption was used to determine the same three elements. The differences in results are discussed in light of sample homogeneity and instrumental precision; the best agreement was obtained for determination of Zn. The work suggests that the filter paper method of standard preparation may be appropriate for LIBS analysis of other samples that give relatively low temperature, low electron density plasmas (i.e., polymers).

Many diagnostic tests in a conventional clinical laboratory are performed on blood plasma because changes in its composition often reflect the current status of pathological processes throughout the body. Recently, a significant research effort has been invested into the development of microfluidic paper-based analytical devices (μPADs) implementing these conventional laboratory tests for point-of-care diagnostics in resource-limited settings. This paper describes the use of red blood cell (RBC) agglutination for separating plasma from finger-prick volumes of whole blood directly in paper, and demonstrates the utility of this approach by integrating plasma separation and a colorimetric assay in a single μPAD. The μPAD was fabricated by printing its pattern onto chromatography paper with a solid ink (wax) printer and melting the ink to create hydrophobic barriers spanning through the entire thickness of the paper substrate. The μPAD was functionalized by spotting agglutinating antibodies onto the plasma separation zone in the center and the reagents of the colorimetric assay onto the test readout zones on the periphery of the device. To operate the μPAD, a drop of whole blood was placed directly onto the plasma separation zone of the device. RBCs in the whole blood sample agglutinated and remained in the central zone, while separated plasma wicked through the paper substrate into the test readout zones where analyte in plasma reacted with the reagents of the colorimetric assay to produce a visible color change. The color change was digitized with a portable scanner and converted to concentration values using a calibration curve. The purity and yield of separated plasma was sufficient for successful operation of the μPAD. This approach to plasma separation based on RBC agglutination will be particularly useful for designing fully integrated μPADs operating directly on small samples of whole blood.

Purpose To compare the quantity of fluorescein delivered to the eye via fluorescein-impregnated paper strips of various sizes and surface areas and via various microliter volumes of fluorescein sodium using an in vitro assay. Methods A commercially available fluorescein-impregnated strip (75 mm2) and three modified strips of reduced fluorescein-impregnated surface areas (10, 7.5, and 5.0 mm2) were used. The amount of fluorescein delivered to the eye for each of the four strips was approximated by applying each strip to a Whatman No. 1 filter paper under conditions simulating application of the strip to the eye, extracting the fluorescein from the filter paper in an aqueous solution, and performing spectrophotometric analysis at 484 nm. Similarly, this filter paper analytical system was calibrated using 1, 2, and 3 microl volumes of 2% w/v fluorescein delivered to the filter paper. Results Using calibration curves, linearity was observed between absorbance and concentration of fluorescein sodium with an R2 value > or = 0.99. Using these calibration curves, the amount of fluorescein delivered to the eye for the four strips and the three fluorescein solution samples was determined. Fluorescein-impregnated strips with surface areas of 75, 10, and 5 mm2 delivered approximately the same quantity of fluorescein to the ocular surface as 3 microl, 1 microl, and 0.5 microl of fluorescein 2% solution, respectively. Conclusions The surface area of the fluorescein-impregnated portion of the strip can be designed to control the amount of fluorescein delivered to the eye.

Diffuse reflectance spectroscopy can be successfully used for the quantitative determination of small amounts of pollutants like metals. The remission function was found to be linearly proportional to the concentration, when we applied the Kubelka-Munk equation. The color reactions of Cu(II), Co(II), and Ni(II) with dithiooxamide, were realised on filter paper. Reaction between Fe(III) and ammonium thiocyanate was realized on filter paper and gelatine matrix. All measurements were accomplished with a laboratory-constructed reflectometer. We have obtained a calibration curve by plotting the optical density of reflectance A(R) vs log of the mol l(-1) concentration. Limits of detection at the 10(-4) M level were estimated for all the compounds. Linear dynamic range extend over one order of magnitude and shows the potential of device for the quantitative analysis of environmental pollutants.

The use of silica-based substrates for room temperature phosphorescence (RTP) was investigated. Contrary to reports in the literature, bare silica was shown to provide higher sensitivity (S/B) for RTP than filter paper when the specific surface areas of the two substrates were similar. The surface heterogeneity of bare silica resulted in nonlinear, discontinuous response curves for a model analyte, phenanthrene. However, analytically useful calibration curves were obtained on silica thin-layer phases in the presence of coadsorbed surfactant. The dynamic range on the silica substrate was 4 times greater and the background phosphorescence was 2 orders of magnitude lower than filter paper. The programming of figures of merit for silica-based substrates via the silica specific surface area is demonstrated.

Four different paper types were evaluated in vitro for their suitability for the collection of gingival crevicular fluid and its subsequent quantification using both the Periotron® 6000 and measurement of the area of paper wetted. Whatman 3MM chromatography paper (Wh.3MM) was found to be the most suitable and strip dimensions of 2x8 mm were found to be the most absorbent whilst remaining compatible with the dimensions of the Periotron electrodes. The 2×8 mm Wh.3MM paper strips were used to derive a quadratic calibration curve, for a range of fluid volumes, which was shown to be a better fit than a straight line. Recovery of human serum proteins applied to these strips exceeded 90% even after compression of the strip between the electrodes of the Periotron during fluid volume measurement.

The present study was undertaken to compare the calibration curve of the periotron* (HAR-6000) with those of two earlier models (HAR-600) instrument, which was found to be appreciably more sensitive in detecting small volumes of fluid than the two older models, is curvilinear in shae while the latter two models display linear calibration characteristics Calibration curves were also obtained for one of the HAR-600 models with the periopaper strip inserted up to and beyond the manufactures recommendations. The results obtained in this case differed from those published by others. Using fluids possessing different dielectric dissipation constants, calibration curves were determined for the HAR-6000 model over a wide range of volumes, The calibration data obtained best fitted quadratic functions and the relative positions of the curves on the axes are in the correct relationship to their dielectric constants Inter-instrument and other variations have once more stressed the importance of individual instrument calibration by uers.