Principais técnicas de preparo de amostra para a determinação de resíduos de agrotóxicos em água por cromatografia líquida com detecção por arranjo de diodos e por espectrometria de massas

The determination of pesticide residues in water samples by Liquid Chromatography require sample preparation for extraction and enrichment of the analytes with the minimization of interferences to achieve adequate detection limits. The Solid Phase Extraction (SPE), Solid Phase Microextraction (SPME), Stir Bar Sorptive Extraction (SBSE) and Dispersive Liquid-Liquid Microextraction (DLLME) techniques have been widely used for extraction of pesticides in water. In this review, the principles of these sample preparation techniques associated with the analysis by Liquid Chromatography with Diode Array Detection (LC-DAD) or Mass Spectrometry (LC-MS) are described and an overview of several applications were presented and discussed.

Sensitive method for determination of piplartine, an alkaloid amide from Piper species, in rat plasma samples by liquid chromatography-tandem mass spectrometry

Piplartine (PPTN) is an alkaloid amide found in Piper species that presents different activities. PPTN determination in rat plasma is necessary to better understand its biological effects. The aim of this study was to develop a sensitive LC-MS/MS method for the determination of PPTN in rat plasma. The performance criteria for linearity, sensitivity, precision, accuracy, recovery, and stability have been assessed and were within the recommended guidelines. The validated method proved to be suitable in a pilot study of PPTN kinetic disposition in rat plasma after a single intraperitoneal dose, and represents an appropriate tool to further pharmacokinetic studies.

Análises quali e quantitativas de micotoxinas em águas da cadeia produtiva do arroz por CCD e CCDAE

This study validated a simple and applied method for determining mycotoxins aflatoxin B1, aflatoxin B2, ochratoxin A, zearalenone and deoxynivalenol, in water from the rice production chain. Five solvent combinations for extraction were tested, with quantification performed by TLC/HPTLC and confirmation by LC-MS/MS. Mycotoxins in water from field and rice industries were evaluated. Mycotoxin recovery levels were around 90%. Two samples from rice parboiling waste were contaminated (deoxynivalenol/aflatoxin B1, 110/9 ng mL-1; and deoxynivalenol, 100 ng mL-1). Zearalenone, deoxynivalenol and ochratoxin A (36, 30 and 28%) were carried to soaking water during parboiling.

Determination of fipronil in bovine plasma by solid-phase extraction and liquid chromatography with ultraviolet detection

A fast and efficient method has been developed and validated for the determination of fipronil in bovine plasma. Samples were subjected to solid-phase extraction (SPE) followed by reversed phase liquid chromatography (LC) separation, using acetonitrile/water (60:40 v/v) as the mobile phase with a flow rate of 1.0 mL/min and ultraviolet (UV) detection at 210 nm. Ethiprole was used as the internal standard (IS). The method was found to be linear over the range 5-500 ng/mL (r = 0.999). The limit of quantitation (LOQ) was validated at 5 ng/mL. The method was successfully applied to monitor plasma concentrations following subcutaneous administration of fipronil in cattle.

Occurrence, biological activities and 13C NMR data of amides from Piper (Piperaceae)

This manuscript describes an update review with up to 285 references concerning the occurrence of amides from a variety of species of the genus Piper (Piperaceae). Besides addressing occurrence, this review also describes the biological activities attributed to extracts and pure compounds, a compiled 13C NMR data set, the main correlations between structural and NMR spectroscopic data of these compounds, and employment of hyphened techniques such as LC-MS, GC-MS and NMR for analysis of amides from biological samples and crude Piper extracts.

Atividade de extratos de Arrabidaea chica (Humb. & Bonpl.) Verlot obtidos por processos biotecnológicos sobre a proliferação de fibroblastos e células tumorais humanas

Arrabidaea chica (H&B) Verlot is a plant popularly known as Pariri and this species is a known source of anthocyanins, flavonoids and tannins. This report describes an approach involving enzymatic treatment prior to extraction procedures to enhance A chica crude extract anticancer activity. Anticancer activity in human cancer cell lines in vitro using a 48 h SRB cell viability assay was performed to determine growth inhibition and cytotoxic properties. The final extraction yield without enzyme treatment was higher (24.28%) compared to the enzyme-treated material (19.03%), with an enhanced aglycones anthocyanin ratio as determined by HPLC- DAD and LC-MS with direct infusion.

Aspectos analíticos e regulatórios na determinação de resíduos de macrolídeos em alimentos de origem animal por cromatografia líquida associada à espectrometria de massas

This is an overview of LC-MS techniques applied for macrolide determination in food, including sample preparation and method validation, as well as the policies adopted by international agencies regarding their presence in food. Techniques for the analysis of macrolides in food normally include solid phase or liquid-liquid extraction followed by HPLC. UHPLC presents advantages in running time, detectability and solvent consumption. Triple-quadrupoles are the most common analyzers in instruments used for the determination of contaminants in food, but time-of-flight and ion-trap spectrometers have been successfully applied for analyses focusing on the investigation of structural formula or the presence of degradation products.

Resíduos de medicamentos veterinários em leite e ovos

The use of veterinary drugs in food producing animals may result in the presence of residues in foods, including milk and eggs. Immunoassay or microbiological tests are used to screen for residues, but chromatographic methods are needed to confirm positive results. In most methods, the sample is extracted with acetonitrile, submitted to clean up or directly analyzed by LC-MS/MS. Results of the Brazilian governmental monitoring programs from 2006 to 2011 have shown that the antiparasitic ivermectin was the drug most frequently found in milk. Residues in eggs are only monitored by one of the programs, and few studies have reported the incidence of veterinary drugs in this matrix in Brazil.

Actividad insecticida sobre Spodoptera frugiperda (Lepidóptera: Noctuidae) de los compuestos aislados de la parte aérea de Piper septuplinervium (Miq.) c. dc. y las inflorescencias de Piper subtomentosum Trel. & Yunck. (Piperaceae)

The bioassay-guided purification of ethanolic extracts from inflorescences of Piper subtomentosum Trel. & Yunck and aerial part of Piper septuplinervium (Miq. ) C. DC. led to isolation of five flavonoids, uvangoletin (1), galangin (2), chrysin (5), 5-hydroxy-4',7-dimethoxy-flavone (6), pinostrobin (7); one amide, N-p-coumaroil-tyramine (4); one acylglycerol, monopalmitin (3); one derivative of acid, protocatechuic acid (8); and glycosydated sterol, daucosterol (9). Their structures were elucidated on the basis of spectroscopy and spectrometry data and by comparison with data reported in the literature. The isolated compounds were tested against Spodoptera frugiperda. The results showed galangin and protocatechuic acid to be the most active (LC 50 13.63 and 17.16 ppm, respectively).

Análise dos produtos de degradação do esfenvalerato por SBSE/CLAE-UV/DAD utilizando planejamento fatorial fracionário

A simple procedure based on stir bar sorptive extraction and high-performance liquid chromatography-ultraviolet/photodiode array detection (SBSE/LC-UV/PAD) to determine intermediates and by-products of esfenvalerate is described. The influence of organic modifier, ionic strength, extraction time, temperature and pH were simultaneously evaluated by using a factorial experimental design. The utilization of different organic solvents and desorption times were also investigated to establish the optimal conditions for SBSE liquid desorption. Among the ten different peaks (intermediates and by-products) detected after degradation of esfenvalerate, eight (including 3-phenoxybenzoic acid and 3-phenoxybenzaldehyde) were successfully extracted by SBSE under the optimized conditions.

Desenvolvimento de procedimento de extração para análise de sedativos e B-bloqueadores em rim suíno

A procedure was developed for determination of 5 sedatives and 14 β-blockers in swine kidney and subsequent analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Three different procedures for extraction were tested, evaluated through recovery studies. The procedure using acetonitrile for extraction and cleanup with freezing at low temperature and dispersive solid phase extraction using 500 mg celite® 545 before the concentration step presented the better results. The dried samples were redissolved with methanol and analyzed using a LC-MS/MS system with electrospray ionization (ESI) operating in positive MRM mode. The recovery values for this procedure were in the 75-88% range. The robustness of the method was tested against small variations. The method was used to analyze carazolol, azaperone and azaperol in collaborative assay, obtaining results close to designed value.

COMPARAÇÃO DE MÉTODOS POR CROMATOGRAFIA LÍQUIDA NA DETERMINAÇÃO DE MULTIRRESÍDUOS DE AGROTÓXICOS EM MORANGOS

Liquid chromatography is often used for the determination of pesticide multiresidues in foods. In Brazil, the strawberry crop is an example of a food with high levels of irregularities because of the application of pesticides. This is a major concern from the perspective of food safety, environmental protection, and certification for food export. The purpose of this study is to evaluate and compare chromatographic separation and detection methods in relation to a newly developed and validated method using ultra high performance liquid chromatography (LC) coupled with tandem mass spectrometry (MS/MS) for the analytical determination of pesticides in strawberries. The comparisons were based on evaluations of the analysis time, consumption of the solvent in the mobile phase, injection volume, detectability, matrix effect, and recovery. The results showed that the LC–MS/MS and UHPLC–MS/MS techniques were both extremely efficient at analyzing pesticide residues with different physico-chemical parameters that were present at low concentrations in a complex matrix. The UHPLC separation method provided better chromatographic performance and productivity, which contributed favorably to routine analytical determinations. Detection by MS/MS had better detectability and selectivity compared with the diode array detector.

Cryptococcus laurentii aplicado em pós-colheita reduz podridões em maçãs

As podridões pós-colheita podem ocasionar perdas substanciais em maçãs (Malus domestica). O mofo azul (Penicillium expansum), a podridão amarga (Glomerella cingulata) e a podridão olho-de-boi (Pezicula malicorticis) estão entre as mais comuns. Grande atenção tem sido dada ao uso de alternativas de controle às doenças pós-colheita. A aplicação pós-colheita de leveduras, como o Cryptococcus laurentii, é uma das opções. Neste estudo testou-se a eficiência de C. laurentii (isolado 36) para o controle de podridões em maçãs 'Fuji' e 'Gala'. Após aplicação dos produtos, através de imersão, os frutos foram armazenados em laboratório (15-20 ºC / 60-70% UR) ou em câmara fria (1 ºC / 90-95% UR). Os patógenos foram inoculados na concentração de 10² conídios ml-1, a levedura a 10(7) células ml-1 e os fungicidas a 150 mg l-1. Em testes de laboratório, a aplicação de C. laurentii reduziu as podridões (G. cingulata, P. expansum e P. malicorticis) da maçã tanto quanto os fungicidas testados (thiabendazol e iprodione). Nos testes efetuados em câmara fria, constatou-se que, após o armazenamento, o tratamento de maçãs com C. laurentii foi tão eficiente quanto os tratamentos com fungicidas (thiabendazol, iprodione, digluconato de clorohexidina, dicloro-s-triazinatriona sódica, dicloroisocianurato de sódio e hipoclorito de sódio) na redução da podridão causada por P. expansum. Aplicação de C. laurentii não alterou a firmeza de polpa nem o teor de sólidos solúveis totais (°Brix) dos frutos.

Polyclonal antibodies to the coat protein of Apple stem grooving virus expressed in Escherichia coli: production and use in immunodiagnosis

The coat protein gene of Apple stem grooving virus (ASGV) was amplified by RT-PCR, cloned, sequenced and subcloned in the expression vector pMal-c2. This plasmid was used to transform Escherichia coli BL21c+ competent cells. The ASGV coat protein (cp) was expressed as a fusion protein containing a fragment of E. coli maltose binding protein (MBP). Bacterial cells were disrupted by sonication and the ASGVcp/MBP fusion protein was purified by amylose resin affinity chromatography. Polyclonal antibodies from rabbits immunized with the fusion protein gave specific reactions to ASGV from infected apple (Malus domestica) cv. Fuji Irradiada and Chenopodium quinoa at dilutions of up to 1:1,000 and 1:2,000, respectively, in plate trapped ELISA. The ASGVcp/MBP fusion protein reacted to a commercial antiserum against ASGV in immunoblotting assay. The IgG against ASGVcp/MBP performed favorably in specificity and sensitivity to the virus. This method represents an additional tool for the efficient ASGV-indexing of apple propagative and mother stock materials, and for use in support of biological and molecular techniques.

Tasasähkönjakeluverkon vaihtosuuntaaja galvaanisella erotuksella

Suomessa sähkönjakeluverkko koostuu pääasiassa 20 kV ja 400 V jännitetasoista. Tällöin sähkö viedään lähelle kuluttajia suuremmalla jännitetasolla ja muunnetaan alhaisemmaksi lähellä asiakkaita. Haittapuolena on se, että haja-asutusalueilla jakelumuuntajien määrä kasvaa suureksi, koska ne täytyy sijoittaa lähelle kuluttajaa. Vaihtoehtona on toteuttaa osa sähkönjakelusta tasajännitteellä, jolloin tehollinen jännite olisi suurempi. Tällöin sähköä voitaisiin siirtää pidempiä matkoja ilman, että asiakaskohtaisia tai muutaman asiakkaan kattavia 20 kV siirtolinjoja tarvitsisi käyttää. Tämä taasen edellyttää asiakaskohtaisien vaihtosuuntaajien käyttöä. Tässä työssä esiteltävällä 1 kVA vaihtosuuntaajalla muodostetaan tasasähköjakeluverkosta saatavasta 750 V tasasähköstä yksivaiheista (230 VRMS, 50 Hz) verkkojännitettä. Laite on suunniteltu toteuttamaan galvaaninen erotus mahdollisimman hyvän hyötysuhteen puitteissa. Vaihtosuuntaaja on toteutettu käyttämällä resonanssikonvertteria, joka vaihtosuuntaa jakelujännitteen korkeataajuiseksi vaihtojännitteeksi. Tämän jälkeen toteutetaan galvaaninen erotus käyttäen suurtaajuusmuuntajaa. Tätä seuraa syklokonvertteri, joka pulssintiheysmodulaatiota soveltaen muodostaa lähtöjännitteen. Tämä suodatetaan lopuksi LC- alipäästösuotimella säröltään standardin mukaiseksi. Laite on jaettu työssä kolmeen osaan, joista jokaisen toiminta on selitetty ja simuloitu itsenäisesti. Lopussa koko järjestelmä on simuloitu yhtenä kokonaisuutena. Hyötysuhteeksi arvioitiin karkeasti 94 % ja lopullisista tuloksista voidaan päätellä, että laitteen toteuttaminen prototyypiksi voisi olla kannattavaa.