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Chromatography is a classical technique for protein separation. However, the chromatography column is filled with tightly packed separation materials and requires an additional pressurizing pump to propel the flow of fluidic samples, largely restraining their applications. Here, we combine heterostructured porous particles with paper strips, realizing spontaneous separation of similarly sized proteins. The interconnected nanofibrous structure and good hydrophility of paper strips enable the spontaneous flow of the liquid sample, and the heterostructured porous particles provide versatile tools for protein separation via electrostatic interaction. The fabricated paper strips are inexpensive, user-friendly, and disposable and exhibit good separation performance. This work may offer a new avenue for fabricating on-site bioseparation tools and purifying various biomacromolecules.

In this study, we developed a point-of-care assay platform with simultaneous detection and self-calibration capabilities for multiple targets based on a microfluidic paper-based analytical device (μPAD). This system is easily manufactured using a wax printing method on chromatographic paper. The design pattern consists of a zone of detection and a calibrant zone for controlled loading using wax barriers with different thicknesses. We showed the utility and applicability of this approach by a proof-of-concept study for two clinically important markers: glucose and lactate. With the naked eye, the results could be fully distinguished and recorded to evaluate the analytical performance with a flatbed scanner. The detection limits of glucose and lactate were 0.3125 mM and 0.2975 mM, respectively, and simultaneous detection was possible from a small sample (0.4 μL) with high sensitivity. Furthermore, this device has a self-calibration function, which minimizes the influence of environmental conditions (i.e., ambient light intensity, temperature, humidity, and pressure). Therefore, the developed multiplex paper-based device is promising for clinical multianalyte point-of-care testing since it is easy to manufacture, cost-effective, user-friendly, and highly sensitive.

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

Biochemical diagnosis of hereditary metabolic diseases requires the detection and simultaneous identification of a large number of compounds, hence the interest in metabolic profiles. Acylcarnitine profile allows the identification and quantification of more than thirty compounds. As part of the accreditation process for medical biology examinations according to standard NF EN ISO 15189, the group from SFEIM recommends an approach to accredit acylcarnitine profile. Validation parameters and recommendations are discussed in this specific framework.

Rheumatoid arthritis (RA) is an inflammatory chronic disease characterized by inflammation, pain, swelling and disability, and radiosynovectomy is one of the disease treatment lines. In this study, the possibility of providing rhenium-186/rhenium-188 chitosan radiopharmaceuticals, optimization of conditions for their production and bio-distribution are reported.In order to build perrhenic acid for labeling, natural rhenium was exposed to radiation. Radionuclidic and radiochemical purities of (186/188Re)-NaReO4 were examined by gamma spectroscopy and paper chromatography methods, respectively.Labeling of chitosan with rhenium was done in different acidic situations. The radiochemical purity 186/188Re-chitosan was applied by radio thin layer chromatography (RTLC). Lastly, the bio-distribution of the radiolabeled chitosan was studied in various organs after intra articular injection of the complex to lab rats. Gamma spectrometry confirmed the high rhenium radionuclidic purity. Chromatography results showed that perrhenic acid was produced with purity greater than 97% and rhenium chitosan labeling was done over 98% in pH = 3. Dissection results showed a high bio-distribution of 186/188Re-chitosan after injection into the joint with no leakage to surrounding organs.According to the results, there is a possibility of labeling rhenium with chitosan in very high radiochemical purity. Regarding the high retention of these radiopharmaceuticals in joints with no leakage to surrounding organs, 186/188Re-chitosan can be applied as new radiosynovectomy drugs for rheumatoid arthritis treatment.

Surface-enhanced Raman scattering (SERS) is a powerful technique for sensitive detection, but it normally has difficulty in multicomponent detection in a complex system, especially for simultaneous analysis of mixture of heavy metal ions. In this work, a simple paper chromatography tandem SERS (PC-SERS) separation/detection platform is proposed by ion-sputtering gold on a filter paper. Based on SEM results, the great electromagnetic field inside nanogaps of Au nanoislands on the paper surface is evaluated with FDTD simulation. It is found that the PC-SERS platform has good uniformity (RSD = 10.12%) and long-time stability. The as-prepared PC-SERS platform was applied to efficiently separate and detect a mixture of pesticides (MG, MB, and CV) in pond water without any pretreatment process, and the limits of detection (LODs) were down to 10 nM. As a crucial application for food safety, several heavy metal ions such as Cd2+, Cu2+, and Ni2+ in grinded rice were successfully detected by the PC-SERS method taking advantage of the sandwich structure based on 4-mercaptobenzoic acid (4-MBA) molecules, which were modified onto sputtering the Au filter paper and gold nanoparticles (Au NPs) to link metal ions and acted as Raman signal molecules. All the LODs for metal ions were down to 1 μM. Due to the easiness of fabrication, good reproducibility, and simple pretreatment step, the PC-SERS platform holds promise in multicomponent detection in a real sample.

Identification of the etiological chemical agent(s) associated with a case(s) of allergic contact dermatitis (ACD) is important for both patient management and public health surveillance. Traditional patch testing can identify chemical allergens to which the patient is allergic. Confirmation of allergen presence in the causative ACD-associated material is presently dependent on labeling information, which may not list the allergenic chemical on the product label or safety data sheet. Dermatologists have expressed concern over the lack of laboratory support for chemical allergen identification and possibly quantification from patients' ACD-associated products. The aim of this review was to provide the clinician a primer to better understand the analytical chemistry of contact allergen confirmation and unknown identification, including types of analyses, required instrumentation, identification levels of confidence decision tree, limitations, and costs.

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

After the cigarette is extinguished, many toxic compounds remain in the environment and accumulate in the air or on surfaces. This exposure is termed thirdhand smoke (THS) and its risks are poorly known. The aim of the study was to evaluate the cellular effects of THS from smokers' homes. Papers were placed in nine smoker's home and three nonsmoker's homes. An area equivalent to the paper size was cleaned with a cotton wipe. A549, Hep-2 and 3T3 cells were exposed to THS for 24 h and cellular functions were assessed by MTT, neutral red (NR) reuptake and trypan blue exclusion assays. High levels of nicotine were found in samples from smokers' homes. Cellular proliferation was similar in almost all samples after THS exposure. Few changes in the cellular functions were observed, mainly higher mitochondrial activity, in paper samples. We are able to detect markers of THS collected from smokers' homes, but a clear evidence of cellular toxicity cannot be demonstrated by the present assays. This is the first study to evaluate the cellular effects of THS samples collected from smokers' homes.

High-performance affinity chromatography is limited by its high cost and high pressure. Paper is made up of porous fiber networks and has the properties of low cost, ease of fabrication, and biodegradable. Due to these advantages, herein, we immobilized beta2-adrenoceptor (β2-AR) onto the surface of the polytetrafluoroethylene membrane, a paper-based material, and constructed a G protein-coupled receptor (GPCR)-in-paper chromatographic platform. This platform was characterized by Fourier transform infrared spectroscopy, fluorescence analysis, X-ray photoelectron spectroscopy, and chromatographic studies. These morphological and elemental analysis showed that β2-AR was successfully immobilized on the paper surface. The specific drugs have good retentions on the GPCR-in-paper chromatographic platform. The association constants of salbutamol, terbutaline and bambuterol to β2-AR were calculated to be 2.02 × 104 M−1, 1.15 × 104 M−1, 1.75 × 104 M−1 by adsorption energy distribution, which were in good line with the values from frontal analysis, zonal elution and previous literatures. We demonstrated that the GPCR-in-paper platform was cost-effective, easy to be modified for protein immobilization, and applicable in the receptor-drug interaction analysis. We believe such a platform sheds new light on paper chromatography for receptor-drug interaction analysis and other applications.

Herein, we report three-dimensional paper chromatography (3D-PC) as a micro-chromatographic platform. The method was based on applying the origami microfluidic device for separation, coupled by colorimetric methods for simultaneous determination. The microfluidic device fabrication was a facile printing approach. Two azo food dyes, Tartrazine (E102) and Indigo carmine (E132), were selected as a model analyte, while carbonate-bicarbonate buffer was used as the mobile phase. Our micro-chromatographic device is associated with two big advantages including needing very small volume of mobile phase ( ~12 µL) and ultrafast separation time (~35 s). Under the optimal conditions, the method provided acceptable linear ranges of 0. 0 g L 1–18.0 g L 1 (R2 = 0.997) for E102 and 0.070 g L 1–10.0 g L 1 for E132 and the limits of detection (3σ/slope) were evaluated as 0.620 and 0.060 g L 1, respectively. The proposed method was successfully applied in the separation and quantification of these dyes in commercial food products such as jelly, candy, and four kinds of drink samples without any sample preparation prior to analysis. The mean recovery values for the real sample analysis were in the range of 100.14%–102.38% for E132 and E102 respectively. The inter-device relative standard deviations were in the ranges of 1.5%–11.8%. In total, our chromatographic μPAD is small (1.0 cm × 1.0 cm × 0.5 cm), portable, inexpensive, no need of specialized user, requires low volumes of sample (0.5 μL), and can perform separation using 12 μL of aqueous mobile phase in very short time.

In this work, we present a facile preparation of a paper-based glucose assay for rapid, sensitive, and quantitative measurement of glucose in blood plasma and urine. Two copper phosphorescent complexes [Cu(2,9-dimethyl-4,7-diphenyl-1,10-phenanthroline)(2,6-dimethylphenylisocyanide)2][B(C6H3(CF3)2)4] (Cu1) and [Cu(2,9-dimethyl-1,10-phenanthroline)(2,6-dimethylphenylisocyanide)2][B(C6H3(CF3)2)4] (Cu2) and a new silver congener [Ag(P3)CNAg(P3)][B(C6H3(CF3)2)4] (Ag3) (P3 = PPh2C6H4-PPh-C6H4PPh2 [bis(o-diphenylphosphinophenyl)phenylphosphine]) have been synthesized and their oxygen sensing abilities were investigated. The dimetallic phosphine-based Ag3 complex, having a high oxygen sensing ability, was employed as an efficient signal transducer in enzymatic reactions to recognize blood plasma glucose and urine glucose, which provided a wide linear response for a concentration range between 1.0 and 35 mM and a rapid response, with a limit of detection (LOD) of 0.09 mM for glucose. In practical application, this Ag3 paper-based device offers great analytical reliability and accuracy upon monitoring glucose concentrations in blood plasma.

Surface-enhanced Raman scattering (SERS) is a powerful analytical tool which enables the detection and identification of analytes adsorbed on nanostructured noble metals. However, SERS analysis of complex mixtures can be challenging due to spectral overlap and interference. In this report, we demonstrate a method to simplify the identification of mixed-analyte samples by coupling SERS detection with chromatographic separation on a nanoplasmonic paper substrate. This "nanopaper" substrate is a silver coated glass microfiber filter paper which possesses large SERS enhancement and can serve as a stationary phase in paper chromatography. Nanopaper is easily synthesized using the silver mirror reaction, making it a highly accessible technology. Nanopaper was successfully used as a combined paper chromatography-SERS (PC-SERS) substrate in the separation and identification of mixed organic dyes. It was further employed to separate and identify lycopene and β-carotene in commercial food products, demonstrating the versatility and utility of nanopaper in the identification of complex mixtures.

The design and fabrication of reconfigurable, modular paperfluidics driven by a prefabricated reusable block library, asynchronous modular paperfluidic linear instrument-free (Ampli) block, are reported. The blocks are inspired by the plug-and-play modularity of electronic breadboards that lower prototyping barriers in circuit design. The resulting biochemical breadboard is a paperfluidic construction set that can be functionalized with chemical, biological, and electrical elements. Ampli blocks can form standard paperfluidic devices without any external instrumentation. Furthermore, their modular nature enhances fluidics in ways that fixed devices cannot. The blocks' ability to start, stop, modify, and reverse reaction flows, reagents, and rates in real time is demonstrated. These enhancements allow users to increase colorimetric signals, fine tune reaction times, and counter check multiplexed diagnostics for false positives or negatives. The modular construction demonstrates that field-ready, distributed fabrication of paper analytical systems can be standardized without requiring the “black box” of craft and technique inherent in paper-based systems. Ampli assembly and point-of-care redesign extends the usability of paper analytical systems and invites user-driven prototyping beyond the lab setting demonstrating “Design for Hack” in diagnostics.

Orientador: Celso Dario Ramos Dissertação (mestrado profissional) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: O uso do radionuclídeo tecnécio-99m ligado a diversos fármacos é de suma importância na medicina nuclear, devido às suas características físico-químicas e radiológicas. Dentro dos serviços, é necessário um controle de qualidade de marcação eficaz, confiável e reprodutível visando eficácia do procedimento para que não exista concentração indesejável do material radioativo em outros órgãos além daqueles de interesse, devido às falhas da marcação. Um dos controles empregados nos serviços de medicina nuclear brasileiros é conhecido como determinação da pureza radioquímica (DPR), feito pela técnica de cromatografia em papel (coluna ascendente) e as aferições das atividades presentes nas amostras são feitas no medidor de atividades conhecido, também, como calibrador de doses. Este equipamento é eficiente sob o ponto de vista de aferir as atividades das alíquotas de radiofármacos administradas aos pacientes, entretanto, para avaliar a DPR são utilizadas atividades da ordem de milionésimo das administradas aos pacientes e, nessa faixa de atividade, supõe-se que o equipamento necessita de caracterizações e condições específicas para garantir a eficácia deste controle. O método padrão ouro para a aferição da DPR utiliza o contador gama tipo poço, entretanto, no Brasil o número de serviços de medicina nuclear com este tipo de equipamento é pequeno. Assim, o trabalho visou comparar os valores de eficiência obtidos nestes dois equipamentos. Na comparação, utilizou-se do teste não paramétrico de Wilcoxon com valor de significância (p-value) de 0,05 e não se encontrou diferenças significativas entre as medidas feitas com os equipamentos. Foram também determinadas às condições confiáveis para a utilização do calibrador de doses nas faixas de atividade empregas na DPR e as condições para que a radiação de fundo (background) do equipamento seja tal que não interfira na quantificação do teste cromatográfico Abstract: The use of Technetium-99m radionuclide isotope combined with various drugs (labeling) has huge importance in nuclear medicine, due to its physico-chemical and radiological characteristics. Within the nuclear medicine services, an effective and reproducible quality control for these radiotracers is required for the security of the procedure so that there is no concentration undesirable of the radioactive material in organs other than those of interest due to marking failures. One of the controls employed in the brazilian nuclear medicine service is known as radiochemical purity (RP), made by paper chromatography technique (ascending column) and the measurements of the activities present in the samples are made in the doses calibrator. This equipment is efficient from the point of view of checking the activities generally administered to the patients, however, in order to evaluate the RP, activities of the order of millionth of the ones administered to the patients are used and, in this range, the equipment needs specific characterizations and conditions to guarantee effectiveness of this control. The gold standard method for the RP measurement is the gamma counter, however, in Brazil the number of nuclear medicine services with this type of equipment is small. Thus, the work aimed at comparing the efficiency values obtained in these two equipments. In the comparison, the nonparametric test, Wilcoxon, with significance value (p-value) of 0.05 was used and no significant differences between the measurements made with the equipment were found. It was also determined the reliable conditions for the use of the dose calibrator in the activity ranges used in the RP and the conditions that the equipment background radiation be useful in order to not interfere in the paper chromatographic test quantification Mestrado Oncologia Mestra em Ciências

Knowledge of the radiochemical purity of radiopharmaceuticals is mandatory and can be evaluated by several methods and techniques. Planar chromatography is the technique normally employed in nuclear medicine since it is simple, rapid and usually of low cost. There is no standard system for the chromatographic technique, but price, separation efficiency and short time for execution must be considered. We have studied an alternative system using common chromatographic stationary phase and alcohol or alcohol:chloroform mixtures as the mobile phase, using the lipophilic radiopharmaceutical [(99m)Tc(MIBI)₆]⁺ as a model. Whatman 1 modified phase paper and absolute ethanol, Whatman 1 paper and methanol:chloroform (25:75), Whatman 3MM paper and ethanol:chloroform (25:75), and the more expensive ITLC-SG and 1-propanol:chloroform (10:90) were suitable systems for the direct determination of radiochemical purity of [(99m)Tc(MIBI)₆]⁺ since impurities such as (99m)Tc-reduced-hydrolyzed (RH), (99m)TcO(4)(-) and [(99m)Tc(cysteine)₂]⁻ complex were completely separated from the radiopharmaceutical, which moved toward the front of chromatographic systems while impurities were retained at the origin. The time required for analysis was 4 to 15 min, which is appropriate for nuclear medicine routines.

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

� Background and Aims During evolution, plants have acquired and/or lost diverse sugar residues as cell-wall constituents. Of particular interest are primordial cell-wall features that existed, and in some cases abruptly changed, during the momentous step whereby land-plants arose from charophytic algal ancestors. � Methods Polysaccharides were extracted from four charophyte orders [Chlorokybales (Chlorokybus atmophyticus), Klebsormidiales (Klebsormidium fluitans, K. subtile), Charales (Chara vulgaris, Nitella flexilis), Coleochaetales (Coleochaete scutata)] and an early-diverging land-plant (Anthoceros agrestis). ‘Pectins’ and ‘hemicelluloses’, operationally defined as extractable in oxalate (100 � C) and 6 M NaOH (37 � C), respectively, were acid- or Driselase-hydrolysed, and the monosaccharides analysed chromatographically. One unusual monosaccharide, ‘U’, was characterized by 1 H/ 13 C-nuclear magnetic resonance spectroscopy and also enzymically. � Key Results ‘U’ was identified as 3-O-methyl-D-galactose (3-MeGal). All pectins, except in Klebsormidium ,c ontained acid- and Driselase-releasable galacturonate, suggesting homogalacturonan. All pectins, without exception, released rhamnose and galactose on acid hydrolysis; however, only in ‘higher’ charophytes (Charales, Coleochaetales) and Anthoceros were these sugars also efficiently released by Driselase, suggesting rhamnogalacturonan-I. Pectins of ‘higher’ charophytes, especially Chara, contained little arabinose, instead possessing 3-MeGal. Anthoceros hemicelluloses were rich in glucose, xylose, galactose and arabinose (suggesting xyloglucan and arabinoxylan), none of which was consistently present in charophyte hemicelluloses. � Conclusions Homogalacturonan is an ancient streptophyte feature, albeit secondarily lost in Klebsormidium. When conquering the land, the first embryophytes already possessed rhamnogalacturonan-I. In contrast, charophyte and land-plant hemicelluloses differ substantially, indicating major changes during terrestrialization. The presence of 3-MeGal in charophytes and lycophytes but not in the ‘intervening’ bryophytes confirms that cell-wall chemistry changed drastically between major phylogenetic grades.

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

The ability to rapidly identify and quantitate, over a wide range of concentrations, anthocyanins in food and therapeutic products is important to ensuring their presence at medicinally significant levels. Sensitive, yet mild, analysis conditions are required given their susceptibility to degradation and transformation. Paper spray ionization has been used to detect and quantify the levels of anthocyanin levels in extracts of fresh and dried elderberries, and elderberry stems, as well as 3 commercially available nutraceutical formulations. The component cyanidin glucosides, including cyanidin-3-sambubioside, cyanidin-3-glucoside, cyanidin-3,5-diglucoside, cyanidin-3-sambubioside-5-glucoside, and the aglycone cyanidin, were readily detected in a range of sources. Quantitation was achieved by establishing a calibration plot from dilutions of a stock solution of cyanidin-3,5-diglucoside containing malvidin-3,5-diglucoside as an internal standard at a fixed concentration. The same standard was used to quantify the anthocyanin content in the fruit and nutraceutical formulations. Wide 5-fold variations in anthocyanin concentration were detected in the nutraceutical formulations from different suppliers ranging from 1050 to 5430 mg/100 g. These concentrations compared with 500 to 2370 mg/100 g measured in the dried stems and fruit, respectively.

Distinguishing a specific biomarker from a biofluid sample containing a large variety of proteins often requires the selective preconcentration of that particular biomarker to a detectable level for analysis. Low-cost, paper-based device is an emerging opportunity in diagnostics. In the present study, we report a novel Zinc oxide nanorods functionalized paper platform for the preconcentration of Myoglobin, a cardiac biomarker. Zinc oxide nanorods were grown on a Whatman filter paper no. 1 via the standard hydrothermal route. The growth of Zinc oxide nanorods on paper was confirmed by a combination of techniques consisting of X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS,) scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDX) analysis. The Zinc oxide nanorods modified Whatman filter paper (ZnO-NRs/WFP) was further tested for use as a protein preconcentrator. Paper-based ELISA was performed for determination of pre-concentration of cardiac marker protein Myoglobin using the new ZnO-NRs/WFP platform. The ZnO-NRs/WFP could efficiently capture the biomarker even from a very dilute solution (Myoglobin

The development of a new tracer based on the cyclic sulfonamides (sultams) was investigated.3-(Methoxy-phenyl-methyl)-1,6-dimethyl-1H benzo[c][1,2] thiazine 2,2-dioxide (benzo-δ-sultam) was synthesized and characterized by elemental analysis, FT-IR spectroscopy and single crystal X-ray structure determination. The prepared cyclic sulfonamide was labeled with non-commercial (62)Zn radioisotope for fast in vivo targeting and Coincidence imaging purposes (radiochemical purity 97 % ITLC, 96 % HPLC, specific activity 20-23 GBq/mmol). In vivo biodistribution of the final complex was investigated in Sprague Dawley(®) rats bearing fibro sarcoma tumor after 2, 4 and 8 h post injection and compared with free Zn(+2) cation.Using instant paper chromatography method, the physicochemical properties of labeled compounds were found sufficiently stable in organic phases, e.g. a human serum, to be reliably used in bioapplications.The complex exhibited a rapid as well as high tumor uptake (tumor to blood ratio 4.38 and tumor to muscle ratio 9.63) resulting in an efficient tumor targeting agent.

Vermicompost was produced from flower waste inoculated with biofertilizers using the earthworm Eisenia fetida. Principal component analysis (PCA) and cluster analysis (CA) were carried out on the basis of physicochemical parameters of vermicomposted samples. From the results of the PCA and CA, it was possible to classify two different groups of vermicompost samples in the following categories: E2 and E5; and E1, E3, E4, and control. Scanning electron microscopy and biodynamic circular paper chromatography analysis were used to investigate the changes in surface morphology and functional groups in the control and vermicompost products. SEM analysis of E1-E5 shows more fragment and pores than the control. Chromatographic analysis of vermicompost indicated the mature condition of the compost materials.

Four marine-derived fungal isolates were screened for the production of inulinase enzyme from low cost substrates under solid state fermentation (SSF), one of them identified as Aspergillus terreus showed the highest inulinase activity using artichoke leaves as a solid substrate. Sequential optimization strategy, based on statistical experimental designs was employed to optimize the composition of the medium, including Plackett–Burman and Taguchi's (L9 3^4) orthogonal array designs. Under the optimized conditions, inulinase activity (21.058 U/gds) reached the predicted maximum activity derived from the taguchi methodology, which increased about 4.79-folds the initial production medium. Fructose was produced, as an end product of inulin hydrolysis proving that the enzyme produced was exoinulinase. The marine-derived A. terreus is suggested as a new potential candidate for industrial enzymatic production of fructose from low cost substrate containing inulin as an economic source.

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.

A simple and inexpensive paper chromatographic method was developed for the isolation of a small peptide from Selaginella bryopteris fronds. The content of peptides is low in most plants and the isolation and purification procedure is troublesome. This method may be used as a first step for the detection of small peptides in the plant extracts. De novo sequence determination by tandem mass spectrometry indicated that the peptide is a tetrapeptide.

Reports of low-quality pharmaceuticals have been on the rise in the past decade, with the greatest prevalence of substandard medicines in developing countries, where lapses in manufacturing quality control or breaches in the supply chain allow substandard medicines to reach the marketplace. Here, we describe inexpensive test cards for fast field screening of pharmaceutical dosage forms containing beta lactam antibiotics or combinations of the four first-line antituberculosis (TB) drugs. The devices detect the active pharmaceutical ingredients (APIs) ampicillin, amoxicillin, rifampicin, isoniazid, ethambutol, and pyrazinamide and also screen for substitute pharmaceuticals, such as acetaminophen and chloroquine that may be found in counterfeit pharmaceuticals. The tests can detect binders and fillers such as chalk, talc, and starch not revealed by traditional chromatographic methods. These paper devices contain 12 lanes, separated by hydrophobic barriers, with different reagents deposited in the lanes. The user rubs some of the solid pharmaceutical across the lanes and dips the edge of the paper into water. As water climbs up the lanes by capillary action, it triggers a library of different chemical tests and a timer to indicate when the tests are completed. The reactions in each lane generate colors to form a "color bar code" which can be analyzed visually by comparison with standard outcomes. Although quantification of the APIs is poor compared with conventional analytical methods, the sensitivity and selectivity for the analytes is high enough to pick out suspicious formulations containing no API or a substitute API as well as formulations containing APIs that have been "cut" with inactive ingredients.

A paper-based solid-phase assay is presented for transduction of nucleic acid hybridization using immobilized quantum dots (QDs) as donors in fluorescence resonance energy transfer (FRET). The surface of paper was modified with imidazole groups to immobilize QD-probe oligonucleotide conjugates that were assembled in solution. Green-emitting QDs (gQDs) were FRET-paired with Cy3 acceptor. Hybridization of Cy3-labeled oligonucleotide targets provided the proximity required for FRET-sensitized emission from Cy3, which served as an analytical signal. The assay exhibited rapid transduction of nucleic acid hybridization within minutes. Without any amplification steps, the limit of detection of the assay was found to be 300 fmol with the upper limit of the dynamic range at 5 pmol. The implementation of glutathione-coated QDs for the development of nucleic acid hybridization assay integrated on a paper-based platform exhibited excellent resistance to nonspecific adsorption of oligonucleotides and showed no reduction in the performance of the assay in the presence of large quantities of noncomplementary DNA. The selectivity of nucleic acid hybridization was demonstrated by single-nucleotide polymorphism (SNP) detection at a contrast ratio of 19 to 1. The reuse of paper over multiple cycles of hybridization and dehybridization was possible, with less than 20% reduction in the performance of the assay in five cycles. This work provides an important framework for the development of paper-based solid-phase QD-FRET nucleic acid hybridization assays that make use of a ratiometric approach for detection and analysis.

Background: This review aims to present the relevant background information and current research status in concentration of polyphenols using membrane technologies. The potential implementation of membrane separation to bioactive compounds like soluble phenolics from aqueous and organic solvent solutions is gaining increasing interest in the recent years. This review does not pretend to cover the abundant published literature on the subject, but to be representative for the observed tendencies in membrane processes applications for concentration of polyphenols derived from natural products. The first part of the article includes general information regarding the polyphenols and the traditional methods for their separation (such as: thin layer chromatography; paper chromatography; gas chromatography; high performance liquid chromatography; capillary electrophoresis), while the second part presents a review of different membrane processes applied for concentration of polyphenols. Three main sources for such implementations are discussed: (1) aqueous or organic solvent extracts from plant material, (2) fruits, and (3) recovery of polyphenols from industrial waste liquids. A diversity of membrane processes are considered in a large scope of implementations ranging from lab-scale studies to pilot and semiindustrial scale operations. Conclusion: Conclusion: Membrane technology is an excellent candidate to make a paradigm shift in biological active compounds fractionation/separation processes. Presented results clearly demonstrate that membrane processes are of great advantages over traditionally used methods; however, characterization of separated polyphenols has to be improved. Most of citied authors concentrated their investigation only on the total amount of polyphenols determination. Exhaustive studies including: antioxidant activities, retention index, total soluble solids, or volume reduction factor, have been only carried out by a few authors.

This article presents the results of testing of phenolic compounds (flavonoids, isoflavonoids, xanthones, phenolcarboxylic acids, tannins, coumarins, etc.) in the rhizomes of four Iris species (Iris sibirica L., Iris pseudacorus L., Iris imbricatа Lindl., Iris hungarica Waldst. et Kit.). With the use of paper and thin-layer chromatography, fifteen phenolic compounds were identified: gallic, coumaric, cinnamic, chlorogenic, neochlorogenic, ferulic, caffeic acids; kaempferol, quercetin, hispidulin, daidzein, genistein, formononetin, mangiferin and isomangiferin. Quantitative contents of flavonoids (1.2-3.7%), hydroxycinnamic acids (0.6-6.5%), γ-pyrones (0.01-0.8%), tannins (6-14%), isoflavonoids (1-2%), polyphenolic compounds (up to 3%) in the rhizomes of the Iris species were determined. Chosen plants belong to natural flora and have been often cultivated. However, this phytochemical analysis for the main groups of the biologically active substances shows a perspective use of the Iris species in medicine.Iris species Iridaceae phenolic compounds chromatography qualitative analysis quantitative contents.

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

The aim of the present work was to validate a paper chromatography system as an alternative way to determine the radiochemical purity of Na18F. Methods: The evaluated parameters were specificity, limit of quantification, measurement interval, linearity, precision, accuracy, and robustness. Results: The proposed method proved to be linear (P > 0.05; r2 = 1.000), precise (relative SD, 8.6%), accurate (mean recovery, 95.9%; relative SD, 1.5%–1.8%), and robust under different conditions since no influence of the operative variables on the chromatographic performance was observed. Conclusion: This system can be used as a reliable alternative method to determine the radiochemical purity of Na18F samples that can be easily performed in PET radiopharmacies at low cost.

Sodium monofluoroacetate was first identified in Dichapetalum cymosum, a South African plant that can cause livestock poisoning and death. After, several other plants also showed to contain this toxin, which leads to the “sudden death”. Mascagnia rigida, a well identified poisonous plant, commonly found in northeast of Brazil also cause sudden death in cattle, which shows clinical signs similar to those produced by the ingestion of plants that contain monofluoroacetate. Our aim was to identify the toxic compound present in the aqueous extract of M. rigida. For this purpose, the dried and milled plant was extracted; the extract was lyophilized and submitted to successive chromatographic process, until the desired purity of the active compound was achieved. The study of this material by planar chromatography and by infrared spectrometry indicated that the toxin can be a mixture of mono, di and trifluoroacetate.

Objective To evaluate alternative methods for radiochemical purity testing of indium (In)-111 capromab pendetide after the discontinuation of instant thin-layer chromatography silica gel (ITLC-SG) strips. Design Descriptive experimental study. Setting United States in 2009. Participants Not applicable. Intervention Paper chromatography using Whatman 3MM strips, paper chromatography using Whatman 31ET strips, and mini-column chromatography using silica Sep-Pak cartridges were evaluated in the radiochemical purity testing of 13 consecutive vials of In-111 capromab pendetide prepared for clinical patient imaging procedures. These methods also were evaluated by testing seven aliquots of In-111 capromab pendetide that had been spiked with 5% to 15% In-111 pentetate (diethylene triamine pentaacetic acid). Main outcome measures Correlation coefficients, radiochemical purity values for each method compared with ITLC-SG, and procedure times. Results Correlation coefficients of 0.988, 0.996, and 0.979 were found for ITLC-SG compared with Whatman 3MM, Whatman 31ET, and silica Sep-Pak, respectively. Compared with ITLC-SG, mean (±SD) radiochemical purity values differed by −0.8 ± (range −2.5 to 0.4) for Whatman 3MM, −0.8 ± (–2.3 to 0) for Whatman 31ET, and −0.5 ± (–3.6 to 0.7) for silica Sep-Pak. The approximate time required to develop the chromatography strip or elute the mini-column was 3, 14, 5, and 6 minutes for ITLC-SG, Whatman 3MM, Whatman 31ET, and silica Sep-Pak, respectively. Conclusion All three methods evaluated were acceptable alternatives to ITLC-SG for radiochemical purity testing of In-111 capromab pendetide. Based on its slightly higher correlation to ITLC-SG, slightly tighter SD and range, and slightly shorter development time, Whatman 31ET is preferred in our facility.

A novel saccharide host containing four boronic acid recognition units on a single DNA duplex terminus was constructed. This construct allowed boronic acid sugar recognition in the context of double stranded DNA to be established while highlighting the benefits of multivalency. Following the solid-phase synthesis of a bis-boronic acid tag, two end-functionalized oligonucleotides with complementary sequences were functionalized through amide ligation. By annealing the boronic acid-DNA conjugates, a tetra-boronic acid DNA duplex was assembled. The saccharide binding ability of this tetra-boronic acid host was revealed through cellulose paper chromatography in the absence and presence of various saccharides. While no appreciable saccharide binding was seen in the case of a bis-boronic DNA conjugate, the increased migration of the tetra-boronic acid host relative to the control sequences in the presence of selected monosaccharides underscored the importance of multivalent effects. We thus identified a requirement for multiple recognition sites in these conjugate systems and expect the results to facilitate future efforts toward applying synthetic recognition systems to the realm of macromolecules.

This paper described a convenient semiquantitative method for colorimetric detection of protein with self-calibration integrated on the test strip. Hydrophilic paper was employed as microfluidic device for running colorimetric assay, tree-shaped design was developed to ensure uniform microfluidic flow for multiple branches. The approach was validated with bovine serum albumin (BSA) colorimetric detection, and colorimetric results observed by naked eyes were consistent with that from apparatus. The device could be coupled with digital transmission of images for remote monitoring system for diagnosis, food control, and environmental analysis.

Prototypes of microfluidic paper-based separation devices with amperometric detection were developed and evaluated. Photolithography was used to make a gold electrochemical microcell on polyester and that microcell was coupled to a strip of paper where a chromatographic separation occurs. The device performance was demonstrated with the separation and quantification of uric and ascorbic acid in mixtures. The method provides an analytical alternative for the determination of compounds where low cost and simplicity are essential.

A reagentless bioactive paper-based solid-phase biosensor was developed for detection of acetylcholinesterase (AChE) inhibitors, including organophosphate pesticides. The assay strip is composed of a paper support (1 x 10 cm), onto which AChE and a chromogenic substrate, indophenyl acetate (IPA), were entrapped using biocompatible sol-gel derived silica inks in two different zones (e.g., sensing and substrate zones). The assay protocol involves first introducing the sample to the sensing zone via lateral flow of a pesticide-containing solution. Following an incubation period, the opposite end of the paper support is placed into distilled deionized water (ddH(2)O) to allow lateral flow in the opposite direction to move paper-bound IPA to the sensing area to initiate enzyme catalyzed hydrolysis of the substrate, causing a yellow-to-blue color change. The modified sensor is able to detect pesticides without the use of any external reagents with excellent detection limits (bendiocarb approximately 1 nM; carbaryl approximately 10 nM; paraoxon approximately 1 nM; malathion approximately 10 nM) and rapid response times (approximately 5 min). The sensor strip showed negligible matrix effects in detection of pesticides in spiked milk and apple juice samples. Bioactive paper-based assays on pesticide residues collected from food samples showed good agreement with a conventional mass spectrometric assay method. The bioactive paper assay should, therefore, be suitable for rapid screening of trace levels of organophosphate and carbamate pesticides in environmental and food samples.

The purpose of this study was to develop a simple screening technique for estimation of salivary flow and to test the usefulness of the method for determining decreased salivary flow.A novel assay system comprising 3 spots containing 30 microg starch and 49.6 microg potassium iodide per spot on filter paper and a coloring reagent, based on the color reaction of iodine-starch and theory of paper chromatography, was designed. We investigated the relationship between resting whole salivary rates and the number of colored spots on the filter produced by 41 hospitalized subjects.A significant negative correlation was observed between the number of colored spots and the resting salivary flow rate (n = 41; r = -0.803; P.01). For all complaints of decreased salivary flow (n = 9) having cutoff values100 microL/min for the salivary flow rate, 3 colored spots appeared on the paper, whereas for healthy subjects there wasor =1 colored spot.This novel assay system might be effective for estimation of salivary flow not only in healthy but also in bedridden and disabled elderly people.

Evidence that extracts of intestinal contents and intestinal mucosa from germfree rats fed a defined diet, inhibit the bactericidal activity of rat serum on E.coli is presented and a partial chemical characterization of the active factor, FIB, is given. FIB was present in an aqueous extract of intestinal mucosa and fœees and was non-dialysable. Extraction with chloroform—methanol (4:1, by vol.) followed by acetone did not reduce the activity of FIB, indicating that lipids were not necessary for effect. The inhibiting factor resisted 100° C for 30 min. FIB was also resistant to proteolysis (pronase) for 8 days, and was recovered in the resulting non-dialysable hydrolysate, whereas the dialysable fraction was inactive. Thiol-blocking agents had no effect on FIB activity, indicating that the factor was not dependent upon intact thiol groups. FIB activity was enhanced by weak acid hydrolysis, causing liberation of sialic acid and fucose. These observations suggest that FIB is either of polysaccharide or glycoprotein nature. A general inhibition of the bactericidal activity of serum exerted by polar high molecular substances was unlikely, since this effect could neither be demonstrated for the plant polysaccharides nor for the bovine glycoprotein fraction tested.

Lipopolysaccharides (LPS) were isolated by phenol-water extraction from 34 strains of Veillonella, and examined by paper chromatography and colorimetric methods for the presence of neutral sugars, amino sugars and 2-keto-3-deoxy-octonate (KDO). Several preparations were also examined for neutral sugars by gas liquid chromatography. The LPS had in common glucosamine, galactosamine, L-glycero-D-manno-heptose glucose and KDO. Most LPS contained galactose, and a few rhamnose. D-glycero-D-manno-heptose was found in LPS from one of the strains. Based on the sugar composition of the LPS, the Veillonella strains could be classified into four chemotypes.

Sera from patients with myasthenia gravis contain antibodies to a citric acid extractable antigen present in heart and skeletal muscle. This antigen is located in the sarcolemmal/subsarcolemmal region of the muscle fibres. The antigen is resistant to pH changes between 2 and 10, but is inactivated by treatment at 56° C for 60 min or at 70° C for 30 min. The antigenic determinant is of protein nature.

The production of phospholipase C (haemolysin) of Acinetobacter calcoaceticus in a chemically defined medium is described. A molecular weight of the enzyme of 25—35, 000, and an isoelectric point of 6.75 were estimated by gel-filtration and isoelectric focusing experiments.

The wall teichoic acid of Staphylococcus hyicus has been isolated and characterized. The teichoic acid is a glycerol phosphate polymer with glycosidically linked N-acetylglucosamine. Interaction with concanavalin A and susceptibility to alpha- but not to beta-N-acetylglucosaminidase showed that the sugar is in the alpha-configuration.