The development of lateral flow devices for urinary biomarkers to assess kidney health.

Serum creatinine levels are insensitive to real-time changes in kidney function or injury. There is a growing interest in assessing kidney injury by measuring biomarkers in body fluid. From our previous studies, we identified and reported three urinary biomarkers namely Uromodulin (UMOD), Osteopontin (OPN), and Interleukin-9 (IL-9) to be associated with kidney health. The availability of a rapid point-of-care test for these urinary biomarkers will potentially accelerate its applicability and accessibility. In this study, we aimed to develop novel lateral flow device (LFD) for UMOD, OPN and IL-9. We tested paired antibodies using Enzyme Linked Immunosorbent Assay wherein we observed functionality only for UMOD and OPN and not for IL-9. A conjugation buffer pH of 7.8 and 8.5 was found suitable at a detection antibody concentration of 15 µg/mL for LFD development. The developed LFDs were found to quantitatively measure UMOD standard (LLOD of 80,000 pg/mL) and OPN standard (LLOD of 8600 pg/mL) respectively. The LFD was also able to measure human urinary UMOD and OPN with a percent CV of 12.12 and 5.23 respectively.

Correction to "Synthesis of Lysoglycerophosphocholines from Crude Soybean Lecithins as Sustainable and Non-toxic Antifouling Agents against the Golden Mussel Limnoperna fortunei".

[This corrects the article DOI: 10.1021/acsomega.2c05645.].

Synthesis of Lysoglycerophosphocholines from Crude Soybean Lecithins as Sustainable and Non-toxic Antifouling Agents against the Golden Mussel Limnoperna fortunei.

This research aimed to produce, on a multigram scale, a new class of non-toxic, halogen- and metal-free antifouling agents from the abundant lecithin byproducts of industrial soybean oil extraction. Three glycerophospholipid analogues were prepared by a facile methanolysis of crude soybean lecithins and a subsequent solvent-free O-alkylation: lysoglycerophosphocholines (LGPCs) and its ether derivatives O-alkyl lysoglycerophosphocholines (ALPCs). As efficient antiproliferative agents, LGPCs and ALPCs are an eco-friendly alternative to current commercial antifoulants which possess significant toxicity to aquatic life. In situ immersion tests of coated stainless-steel nets with previously incorporated automotive paint products, LGPCs and ALPCs (1-O-octadecyl-2-O-acyl-sn-glycero-3-phosphocholine, ALPC18, and 1-O-hexadecyl-2-O-acyl-sn-glycero-3-phosphocholine, ALPC16), in an aquaculture reservoir in SP-Brazil revealed significant growth inhibition against macrofouling species, especially the epibiotic golden mussel (Limnoperna fortunei), when compared with the control. These results promise a more sustainable and ecologically innocuous approach to combating the biofouling phenomenon and the deeply concerning dissemination of the golden mussel which has provoked an economic crisis in the energy and aquaculture sectors.

Anti-Leishmania amazonensis activity of the terpenoid fraction from Eugenia pruniformis leaves.

Leishmaniasis is caused by protozoan parasites belonging to the genus Leishmania and includes cutaneous, mucocutaneous and visceral clinical forms. Drugs currently available for leishmaniasis treatment present high toxicity, and development of parasite resistance. Plants constitute an important source of compounds with leishmanicidal potential. This study aimed to evaluate the anti-Leishmania amazonensis activity of the terpenoid fraction of Eugenia pruniformis leaves (TF-EpL). TF-EpL was active against the promastigote and intracellular amastigote forms of L. amazonensis with IC50(24h) value of 43.60µg/mL and 44.77µg/mL, respectively. TF-EpL altered the cell cycle of the parasite, increasing 2.32-fold the cells in the Sub-G0/G1 phase. TF-EpL also changed the ΔΨm and increased ROS and the number of annexin-V-PI positive promastigotes, which suggests incidental death. ß-sitosterol, ursolic acid, corosolic acid and asiatic acid were isolated from TF-EpL. The results showed the antileishmanial activity of E. pruniformis terpenoids and its potential for further studies as a source of new drugs for leishmaniasis.

Lipoprotein(a) particle number assay without error from apolipoprotein(a) size isoforms.

BACKGROUND: Lipoprotein(a) [Lp(a)] is an important cardiovascular risk factor, but clinical immunoassays are flawed. Apolipoprotein(a) [apo(a)], the characteristic protein of Lp(a), contains a variable number of kringle repeats (size isoforms) that make accurate measurement of Lp(a) difficult. We developed a sandwich enzyme immunoassay that uses a murine monoclonal anti-apo(a) antibody for capture and a polyclonal anti-apolipoprotein B (apo B) for detection. Because Lp(a) contains one molecule each of apo(a) and apo B, the assay measures the number of Lp(a) particles [Lp(a)-P] in the circulation without bias due to apo(a) size isoforms. METHODS: After developing and choosing the best anti-apo(a) clone for Lp(a) capture, we identified suitable reagents and ELISA conditions, and validated assay performance (precision, linearity, limit of detection, interferences, and apo(a) size isoform bias). RESULTS: The Lp(a)-P assay was precise with within-run precision of 5.5% to 7.2% and total imprecision of 6.9% to 12.1%. The assay had a limit of detection of 13 nmol/l and was linear from 2 to 499 nmol/l. There was no interference from plasminogen or apolipoprotein B up to 80 and 200 mg/dl, respectively, and bias plot showed no bias related to apo(a) size (kringle 4 type 2 repeats). CONCLUSIONS: Lp(a)-P assay is sensitive, precise and linear over a wide analytical range and is a suitable alternative for laboratories concerned about inaccuracy due to apo(a) size polymorphism and the poor performance of immunoturbidimetric assays.

Synthesis of natural 1-O-alkylglycerols: a study on the chemoselective opening of the epoxide ring by onium quaternary salts (N and P) and ionic liquids.

A chemoselective route for the synthesis of 1-O-alkylglycerols chimyl (1), batyl (2), and selachyl (3) is reported. These compounds can be naturally isolated from shark liver oil and the skin of animals such as stingrays and chimeras and exhibit potential anti-fouling activity. The synthetic approach developed in this work included two distinct methods of preparation. The first was based on solvent-free reactions catalyzed by onium quaternary salts (N and P) and ionic liquids; the second methodology was based on a series of one-pot reactions.

Importância de uma regulamentação específica com as definições e classificações dos produtos comercializados como suplementos alimentares, alimentos funcionais e nutracêuticos / Importance of a specific legislation with definitions and classifications of products marketed as food supplements, functional foods and nutraceuticals

Este artigo trata da importância de a legislação brasileira regulamentar o termo "suplemento alimentar", a fim de se adequar às legislações internacionais e ao cenário desse mercado, marcado por constantes inovações tecnológicas e, consequentemente, uma ampla variedade de produtos. São descritas as leis e resoluções de diretoria colegiada publicadas pela Agência Nacional de Vigilância Sanitária para o registro desses produtos, que são classificados legalmente como alimentos. O artigo apresenta, ainda, as classificações de legislações internacionais e os casos de adulterações.

This study aims at showing how important is that the Brazilian legislation regulate the term "food supplement", in order to comply with international legislation and the market scenario characterized by constant technological innovations and, consequently, a wide variety of products. Laws and resolutions of the collegiate board are described, which have been published by the Brazilian Health Regulatory Agency for the registration of these products, legally classified as foods. This paper also describes the classifications of international laws and cases of adulteration.

Evaluation of pharmacokinetics and efficacy of ivermectin following oral administration in dogs against experimental infection of Ctenocephalides felis felis and Rhipicephalus sanguineus.

With the increasing number of pets in home the human-animal relationship is increasingly close and care about control disease growing. Ivermectin (IVM) is frequently used because its proven safety. IVM is recommended for the treatment of demodectic scabies and prevention of heartworm in dogs, but informally is extremely used to control of Ctenocephalides felis felis and Rhipicephalus sanguineus. The aim of this study is evaluate the use of IVM in dogs, by the oral route at 0.6µg/kg dose, against experimental infection of these parasites using the construction of the plasma concentration curve and efficacy study. A IVM quantification method in canine plasma using HPLC-FL was developed and validated based on RE n°899/03 ANVISA. The samples collected during the efficacy test was analyzed by this validated method and prove Cmax of 350ng/mL at 4h (tmax) and AUC of 8411ng/h/mL. Spite of formulation have shown good absorption, the highest efficiency values found for Rhipcephalus sanguineus and Ctenocephalides felis felis were very low, 35% and 67% respectively, demonstrating this not be the most appropriate treatment for the control of these parasites.

Determinação simultânea de Rifampicina, Isoniazida, Pirazinamida e Etambutol em comprimidos com doses fixas combinadas / Method for simultaneously determining the Rifampin, Isoniazid, Pyrazinamide and Ethambutol in the combined fixed-dose tablets

Os comprimidos utilizados no tratamento da tuberculose possuem quatro fármacos associados, isoniazida, pirazinamida, etambutol e rifampicina, e são distribuídos gratuitamente pelo Sistema Único de Saúde. Os métodos analíticos oficiais para analisar este medicamento estão especificados na Farmacopeia Americana 36a edição e na Farmacopeia Internacional 4a edição. Porém, estes compêndios oficiais não possuem monografias para análise simultânea dos quatro fármacos. O objetivo deste estudo foi desenvolver uma metodologia para determinar simultaneamente os princípios ativos em comprimidos dose fixa combinada, utilizando-se cromatografia a líquido de alta eficiência com detector de ultravioleta-visível, pois é de grande importância para o controle da qualidade do medicamento. O método desenvolvido utilizou coluna cromatográfica C18 (250 x 4,6) mm e 5 μm, fase móvel constituída de fase aquosa (85 % tampão formiato de amônio 0,3 mol/L pH 5, 15 % metanol e 0,005 mol/L de Cu2+ ou 250 mg/L de CuSO4.5H2O) e fase orgânica (metanol, 0,1 % de trietilamina e 0,2 % de ácido fórmico). O fluxo foi de 1,0 mL/min e comprimento de onda de 265 nm para isoniazida, pirazinamida e o etambutol e de 335 nm para rifampicina. Este método apresentou desvio padrão relativo inferior a 2,0 % na precisão e linearidade para os quatro fármacos estudados.

Tablets containing isoniazid, pyrazinamide, ethambutol and rifampicin are used for tuberculosis treatment, and theyare freely distributed by the Brazilian National Health System. The official analytical methods for testing those substances in fixed-dose combined tablet are described in the United States Pharmacopeia and theInternational Pharmacopoeia. None of these official compendiums refers to the methodologies forconducting the simultaneous analysis of these four drugs. This study aimedatdeveloping an analytical methodology to determine simultaneously allof four drugs in tablets for tuberculosis treatment, bymeans ofhigh performance liquid chromatography with ultraviolet-visible detector. The developed method used a chromatographic column with octadecylsilane stationary phase (250 mm x 4.6 mm x 5 μm particle size). The mobile phase was aqueous (85 % ammonium formate buffer pH 5, 15 % methanol and 250 mg CuSO4.5H2O), and organic phase (methanol, 0.1 % triethylamine and 0.2 % formic acid). The flow was 1.0 mL/min, at a wavelength of 265 nm or, when the equipment allowed, a wavelength of 265 nm for isoniazid, pyrazinamide and ethambutol and 335 nm for rifampicin. The developed methodology showed satisfactory results regarding the precision parameter, with relative standard deviation lower than 2.0 % for the studied drugs.

QuickTox™ Kit for QuickScan DON3 (Vomitoxin) Method Modification.

Lateral flow technology and a reader-based system are used for quantitative determination of deoxynivalenol (DON), also known as vomitoxin, residues in cereal grain commodities by the QuickTox Kit for QuickScan DON (Vomitoxin). The assay has been modified, and a study was conducted in support of a Performance Tested MethodSM (PTM) Modification. The modified assay employs identical biologic reagents as used previously (PTM No. 121202). Compared to the PTM certified product, the new assay uses modified device architecture. Multiple kits and catalog numbers were required in the original kit reflecting the necessity for matrix specific calibration curves affixed to assay strips. A single calibration curve and kit are utilized in the new product; extraction volumes used in sample preparation are varied to accommodate multiple sample types. Extracts are clarified by filtration or settling depending on the sample type. Filtration was used for matrixes examined in these studies. With the original product, the extract was mixed 1:1 with DB1 buffer followed by the addition of the strip which was developed for 10 min. The new product dilutes extracts five-fold offline in DB6 buffer; an aliquot of the dilution is moved to a reaction vial followed by strip development time for 3 min. The new assay performance was evaluated for linearity, robustness, selectivity (inclusivity), lot-to-lot consistency, and both internal and third party matrix studies. All DON positive samples yielded results within previously defined acceptable ranges with dose-dependent correlation values of R2 greater than 0.97 in linearity and internal and external matrix studies. Inclusivity data indicated detection of DON along with acetyl derivatives, glucoside-conjugate, and Nivalenol. Robustness studies showed within range results upon co-variation of multiple user interface parameters, and lot-to-lot consistency challenges demonstrated acceptable results across five manufactured lots.

Immunoprecipitation of apolipoprotein B-containing lipoproteins for isolation of HDL particles.

BACKGROUND: Immunoprecipitation (IP) of non-HDL particles with antisera provides the simplest and most specific method available for the separation of HDL. We compared the LipoSep™ IP reagent with the dextran sulfate/MgCl2 precipitation method (DS). METHODS: The IP reagent (200 µl) was added to an equal volume of serum, vortexed, incubated for 10 min at room temperature, and microcentrifuged at 12,000 rpm for 10 min. RESULTS: Equal volumes of a sample and the IP reagent precipitated apoB to 3.0 g/l without the coprecipitation of HDL. HDL-C measured in the supernatant after IP (Y) gave excellent agreement to DS precipitation (X) with a slope of 1.01, an intercept of 0.070 mmol/l (2.7 mg/dl), and a correlation of 0.99 (n=118; apoB 0.16-2.11 g/l). However, DS failed in most samples with moderate to elevated triglycerides. At triglyceride concentrations from 2.86 to 23.63 mmol/l (253-2091 mg/dl) the initial success rate was 65.4% for IP, while DS successfully precipitated only 5.8%. Success rate on repeat with additional reagent and/or sample dilution gave a success rate of 86.5% for IP and 40.4% for DS. CONCLUSION: The IP reagent and protocol is a simple, effective and highly specific tool for isolating HDL particles in human serum and is effective with high triglyceride samples.

QuickTox Kit for QuickScan DON (Vomitoxin).

The QuickTox Kit for QuickScan DON (Vomitoxin) uses lateral flow technology and a reader-based system for the quantification of deoxynivalenol (DON, vomitoxin) residues in cereal grain commodities. The present work was performed to obtain AOAC Research Institute Performance Tested MethodsM certification for testing wheat, maize, wheat bran, wheat flour, and barley samples with DON contamination levels as high as 5 ppm. Assay performance was examined using naturally contaminated and spiked samples in internal and independent laboratory evaluations and was compared to previously established acceptance criteria. Performance was evaluated with direct regard to linearity, matrix, selectivity, robustness, and stability. All data points in all studies were within the ranges defined in the acceptance criteria. The assay exhibited a linear dose response over the range tested, 0-5.0 ppm, with R2 values exceeding 0.97. RSD of repeatability, or RSDr, values for results ranged from 3.12 to 16.01% across all tested commodities and levels. Assay results were not affected by the presence of other common mycotoxins: aflatoxin B1, fumonisin B1, ochratoxin A, or zearalenone. Selectivity for modified DON was examined, specifically 15-acetyl DON, 3-acetyl DON, DON-3-glucoside, and another tricothecene, nivalenol; all were detected. The assay produced acceptable results in robustness studies when assay timing, temperature, and volume were covaried. The QuickTox Kit for QuickScan DON (Vomitoxin) assay is a convenient and reliable method for determination of DON in grain commodities.

Estudo das propriedades físico-químicas da glibenclamida e excipientes que influem sobre resultados do ensaio de dissolução para medicamento similar e genérico / Effect of the physicochemical properties of glyburide on the results of dissolution test for generic and similar drugs

Alguns insumos farmacêuticos ativos (IFA) possuem como característica, a possibilidade de apresentarem o polimorfismo, que pode se desenvolver em alguma das etapas do processo de produção na indústria. No caso deste não ser caracterizado e especificado, um diferente polimorfo poderá ser utilizado equivocadamente durante o processo de fabricação. A ocorrência de polimorfismo pode originar importantes variações nas propriedades físico-químicas dos IFAs, principalmente quanto à solubilidade. Alguns medicamentos de glibenclamida (GLIB) apresentaram denúncias de ineficácia terapêutica e a presença de polimorfos pode ser uma das possíveis causas. Neste trabalho foram analisados cinco medicamentos e cinco IFAS de diferentes fornecedores. Para os medicamentos foram feitos testes característicos de verificação de equivalência farmacêutica. Nos IFAS, as diversas técnicas empregadas não evidenciaram presença de polimorfos ou alterações importantes nas propriedades físico-químicas e na velocidade de dissolução intrínseca. Entretanto, os perfis de dissolução dos medicamentos, principalmente, entre os dois similares A e B demonstraram diferenças apontadas pelos valores do fator f2, respectivamente, de 20 e 42, os quais indicaram associação destes valores com a presença de distintos excipientes, como por exemplo o manitol e diferentes processos de produção industrial.

Some active pharmaceutical ingredients (API) might present polymorphism at any stage of the industryproduction process. In caseit is not characterized and specified, a different polymorph might beerroneously used during the manufacturing procedure. Polymorphisms cause some variations in thephysicochemical properties of APIs, especially in solubility. Therapeutic inefficacy was detected in someglyburide drug products, and the occurrence of polymorphs might be one of the possible reasons. Thisstudy analyzed five drug products and five APIs., The characteristic pharmaceutical equivalence testswere used for analyzing the drug products. The techniques employed to evaluate the APIs showed nodifferences in polymorphism, no significant changes in the physicochemical properties or in the intrinsicdissolution rate. However, the dissolution profiles of the drug products, mainly between two similarproducts A and B, showed significant differences in the f2 factor values, being 20 and 42, respectively,indicating that these values were related to the occurrence of different excipients, such as mannitol.

Development of a new analytical method for determination of acetylsalicylic and salicylic acids in tablets by reversed phase liquid chromatography / Desenvolvimento de um novo método analítico para determinação de ácidos salicílico e acetilsalicílico em comprimidos por cromatografia líquida de fase invertida

Acetylsalicylic acid (AAS) is a drug utilized as analgesic, anti-inflammatory, and antipyretic medication, available worldwide and commonly used in Brazil. Salicylic acid (AS) is a precursor in AAS synthesis and is also produced during its degradation. The official United States Pharmacopoeia (USP) suggests the determination of these drugs by high performance liquid chromatography (HPLC), with ultraviolet detection, but this method has neither a high sensitivity (S AAS=0.12 mAbs/(μg/mL) and S AS=0.48 mAbs/(μg/mL)) nor resolution (Rs=1.61). The purpose of this study was to develop a new more adequate, accurate method by liquid phase chromatography than the current official methodology, and to use this new method in the determination of the tenors of acetylsalicylic, as of salicylic acids in tablets. The parameters of the chromatographic system for both the AAS and AS were satisfactory. Selectivity was verified by absorption spectra comparison in the ultraviolet (UV) range, during and after substance retention time. The linear range for AAS was 0.21 to 0.39 mg/mL, and that for AS was 6.3 to 11.7 μg/mL. The correlation coefficients (r) of the analytical curves of AAS and AS were 0.9995 and 0.9988, respectively; and the detection and quantification limits for the AS were 0.23 and 0.69 μg/mL. The sensitivity (S AAS=1.88 mAbs/(μg/mL) and S AS=1.84 mAbs/(μg/mL)) and the resolution (Rs =5.06) show the improvement obtained using this method over that described by the USP.

O ácido acetilsalicílico (AAS) é um fármaco utilizado como analgésico, antiinflamatório, antipirético, sendo amplamente comercializado e consumido no Brasil e no mundo. Como precursor de sua síntese utiliza-se o ácido salicílico (AS) que também é produzido através de sua degradação. A metodologia oficial da Farmacopéia Americana (USP) preconiza a determinação destes fármacos por cromatografia líquida de alta eficiência (CLAE) com detecção por ultravioleta, mas este método não possui boa sensibilidade (S AAS=0,12 mAbs/(μg/mL) e S AS=0,48 mAbs/(μg/mL)) e resolução (Rs =1,61). O objetivo deste trabalho, visando a melhor adequação do sistema cromatográfico em relação à metodologia oficial, foi o desenvolvimento e otimização de um novo método por cromatografia em fase líquida para determinar os teores tanto do ácido acetilsalicílico quanto do salicílico em comprimidos. Os parâmetros da adequação do sistema cromatográfico para o AAS e para o AS foram satisfatórios. A seletividade foi verificada por comparações dos espectros de absorção no ultravioleta (UV) antes, durante e depois do tempo de retenção da substância. A faixa linear de trabalho para o AAS foi de 0,21 a 0,39 mg/mL e a do AS foi de 6,3 a 11,7 μg/mL. Os coeficientes de correlação (r) das curvas analíticas do AAS e do AS foram de 0,9995 e 0,9988, respectivamente e os limites de detecção e quantificação para o AS foram 0,23 e 0,69 μg/mL. A sensibilidade (S AAS=1,88 mAbs/(μg/mL) e S AS=1,84 mAbs/(μg/mL)) e a resolução (Rs=5,06) atestam a melhoria em relação ao método descrito na USP.