BIOCHEMICAL VALUES OF BLOOD SERA FROM FARM ANIMALS

The significanceof the main biochemical values (proteins, lipids, carbohydrates, colouring agents, low-molecular nitrogenous and mineral matters) in blood sera (plasm) from farm animals is described in the paper. The change in the content (decrease or increase) of these indicators against the normal level can have different reasons: noncontagious and infectious diseases, imbalance of ration and feeding regime

COMBINING QuEChERS PREPARATION AND MICELLAR ELECTROKINETIC CHROMATOGRAPHY FOR DETERMINATION OF NEONICOTINOID INSECTICIDES IN FRUITS AND VEGETABLES

The possibility of electrophoretic separation and simultaneous determination of 7 neonicotinoids (imidacloprid, acetamiprid, thiamethoxam, thiacloprid, nitenpyram, clothianidin and dinotefuran) by micellar electrokinetic chromatography is shown. We propose a method for extracting and concentrating of neonicotinoid insecticides from vegetables and fruits using the sample preparation QuEChERS. The optimum compositions of the salting-out mixtures for extraction and sorbents for purification of the resulting extracts from vegetables and fruits were selected. Extraction of pesticides from the fruits was performed by acetonitrile using a mixture of sodium citrates (Na3C6H5O7·2H2O, Na2HC6H5O7·1,5Н2О) with the following clean-up of 2 ml of the extract with sorbent PSA (100 mg) and anhydrous magnesium sulfate (300 mg). During the analysis of vegetables ethyl acetat was used as an extractant in the presence of anhydrous magnesium sulfate and sodium chloride with the following clean-up of 2 ml of the extract with sorbent PSA (100 mg) and graphitized carbon black (10 mg). In optimal conditions the recovery of the analytes ranged from 62 to 81 % for fruits and from 38 to 76 % for vegetables. The limits of quantification of neonicotinoids with a 10.0 g weight sample were from 0.25 to 0.65 mg·kg-1 and from 0.04 to 0.13 mg·kg-1 for fruits and vegetables respectively. The relative standard deviation of the test results does not exceed 0.1. Duration of analysis is 1-1.5 hr.Key words: neonicotinoids, micellar electrokinetic chromatography, QuEChERS, vegetables, fruits(Russian)DOI:http://dx.doi.org/10.15826/analitika.2015.19.1.003 D.S. Bol’shakov1, V.G. Amelin1,2, T.B. Nikeshina1 1Federal Centre for Animals Health (ARRIAH), Yur’evets, Vladimir, Russian Federation2Vladimir State University named after Alexander and Nikolay Stoletovs, Vladimir, Russian Federatio

Combining QuEChERS preparation and micellar electrokinetic chromatography for determination of neonicotinoid insecticides in fruits and vegetables

Показана возможность электрофоретического разделения и одновременного определения семи неоникотиноидов (имидаклоприда, ацетамиприда, тиаметоксама, тиаклоприда, нитенпирама, клотианидина и динотефурана) методом мицеллярной электрокинетической хроматографии. Предложен способ извлечения и концентрирования неоникотиноидных инсектицидов из овощей и фруктов с использованием дисперсионной твердофазной экстракции QuEChERS. Выбраны оптимальные составы высаливающих смесей для экстракции и сорбентов для очистки полученных экстрактов. Извлечение пестицидов из фруктов проводили ацетонитрилом с использованием смеси цитратов натрия и очисткой 2 мл полученного экстракта смесью 100 мг сорбента PSA и 300 мг безводного сульфата магния. Для анализа овощей в качестве экстрагента применяли этилацетат в присутствии безводного сульфата магния и хлорида натрия, с последующей очисткой 2 мл экстракта 100 мг сорбента PSA и 10 мг графитированной сажи. В оптимальных условиях степени извлечения пестицидов варьируются от 62 до 81 % для фруктов и от 38 до 76 % для овощей. Нижние границы определяемых содержаний пестицидов при массе пробы 10.0 г с учетом концентрирования составили от 0.25 до 0.65 мг/кг и от 0.04 до 0.13 мг/кг для фруктов и овощей соответственно. Относительное стандартное отклонение результатов анализа не превышает 0.1. Продолжительность анализа - 1-1.5 ч.The possibility of electrophoretic separation and simultaneous determination of 7 neonicotinoids (imidacloprid, acetamiprid, thiamethoxam, thiacloprid, nitenpyram, clothianidin and dinotefuran) by micellar electrokinetic chromatography is shown. We propose a method for extracting and concentrating of neonicotinoid insecticides from vegetables and fruits using the sample preparation QuEChERS. The optimum compositions of the salting-out mixtures for extraction and sorbents for purification of the resulting extracts from vegetables and fruits were selected. Extraction of pesticides from the fruits was performed by acetonitrile using a mixture of sodium citrates (Na₃C₆H₅O₇·2H₂O, Na₂HC₆H₅O₇·1,5Н₂О) with the following clean-up of 2 ml of the extract with sorbent PSA (100 mg) and anhydrous magnesium sulfate (300 mg). During the analysis of vegetables ethyl acetat was used as an extractant in the presence of anhydrous magnesium sulfate and sodium chloride with the following clean-up of 2 ml of the extract with sorbent PSA (100 mg) and graphitized carbon black (10 mg). In optimal conditions the recovery of the analytes ranged from 62 to 81 % for fruits and from 38 to 76 % for vegetables. The limits of quantification of neonicotinoids with a 10.0 g weight sample were from 0.25 to 0.65 mg·kg⁻¹ and from 0.04 to 0.13 mg·kg⁻¹ for fruits and vegetables respectively. The relative standard deviation of the test results does not exceed 0.1. Duration of analysis is 1-1.5 hr

Torovirus infection in animals: a review

Massive digestive disorders of neonatal calves, clinically manifested as diarrhea causing severe dehydration, toxemia, immunodeficiency and metabolic disorders, induce huge economic losses in animal husbandry. Etiopathogenetic lesions of the digestive organs are characterized by significant polymorphism, including a wide range of various (physiological, sanitary and infectious) factors. Massive gastroenteritises in neonatal calves are primarily caused by such infectious agents as viruses, bacteria and protozoa. Massive diarrheas are registered in 70–80% of newborn calves by the end of the first day of life. Diseased newborn calves die on day 5–10 and mortality ranges from 15 to 55%. Rotavirus, coronavirus, pestivirus, parvovirus, enterovirus and kobuvirus, along with bacteria, are most frequently detected in faecal samples collected from neonatal calves with diarrhea. Diagnostic and vaccine products for prevention of these infections have been developed in the Russian Federation. At the end of the 20th – the beginning of the 21st century a large number of cattle were imported to Russia from the countries affected with different contagious diseases (USA, Denmark, France, Slovakia, Austria, Hungary, Germany, the Netherlands, Australia, Finland, etc.). Despite the high activity and field effectiveness of vaccines against rotavirus and coronavirus infections and viral diarrhea, massive neonatal calf diarrheas causing significant economic losses were registered in a number of large-scale livestock farms. Torovirus as well as the above-mentioned pathogens were detected in fecal samples from diseased calves. This report provides data on torovirus infection indicating a wide geographical distribution of animal torovirus in many countries of the world. All this suggests the need to take into account torovirus infection when conducting epizootological investigations in farms affected with massive gastrointestinal diseases of neonatal calves

ИДЕНТИФИКАЦИЯ И ОПРЕДЕЛЕНИЕ ТОКСИКАНТОВ С ИСПОЛЬЗОВАНИЕМ СТАНДАРТНОЙ ДОБАВКИ В ПИЩЕВЫХ ПРОДУКТАХ, ПРОДОВОЛЬСТВЕННОМ СЫРЬЕ И КОРМАХ МЕТОДОМ ВЫСОКОЭФФЕКТИВНОЙ ЖИДКОСТНОЙ ХРОМАТОГРАФИИ / ВРЕМЯПРОЛЕТНОЙ МАСС-СПЕКТРОМЕТРИИ ВЫСОКОГО РАЗРЕШЕНИЯ

The methodology of identification and defining 300 toxicants of different classes in food products and food raw materials out of one sample have been proposed and realized by the TOF mass spectrometry of high resolution combined with high performance liquid chromatography and simple express sample preparation. Analytes included pesticides, mycotoxins and veterinary drugs.  Simple sample preparation variations of milk, meat, fat, eggs, liver, kidney, feed and grain were suggested. Sample preparation involved the extraction with acetonitrile and removal of toxins fat by extraction with hexane. Under the electrospray ionization conditions most analytes develop protonated and deprotonated forms and rare adducts with ammonium, sodium and potassium. Identification of analytes was carried out using the «TargetAnalysis-1.3» program. Retention time, the accuracy of the masses of the ions and matching of the isotopic distribution (mSigma) were used as identification parameters. Low limits of detection of analytes were shown to be 0.0005-50 ng / ml. It was found that, given such low detection limits, the dilution of the extract with water to eliminate the matrix effect is possible. The lower limit of the defined content with the sample preparation and dilution amounted to 1 (500) mg / kg. The degree of extraction of analytes from the analyzed samples ranged from 78 to 110 % depending on the nature of the analyte and the matrix. A scheme for the identification and defining toxins by standard addition was proposed. The advantages of the standard addition method compared to the method of the calibration curve in the determination of toxicants in real samples were demonstrated. The relative standard deviation of the test results does not exceed 0.11. The identification period was 40-60 min, and the definition of identified toxicants was in the range of 2-3 hours.Keywords: high performance liquid chromatography, tandem quadrupole-of-flight mass spectrometry, food, feed, pesticides, mycotoxins, veterinary drugs, food and non-food dyes(Russian)DOI: http://dx.doi.org/10.15826/analitika.2015.19.2.010 V.G. Amelin1,2, A.М. Andoralov2,3, N.M. Volkova1,2, A.I. Korotkov2,3,T.B. Nikeshina1, I.I. Sidorov3, A.A. Timofeev1,21Federal Centre for Animal Health (ARRIAH), Yurievets, Vladimir, Russian Federation 2Vladimir State University, Vladimir, Russian Federation 3Bryansk Interregional Veterinary Laboratory, Bryansk region, p. Suponevo, Russian FederationПредложена методология идентификации и определения из одной навески 300 токсикантов различных классов в пищевых продуктах и продовольственном сырье методом времяпролетной масс-спектрометрии высокого разрешения в сочетании с высокоэффективной жидкостной хроматографией  и простой, экспрессной пробоподготовкой. Аналиты – пестициды, микотоксины, антибиотики, красители в пищевых продуктах и кормах. Предложены простые варианты пробоподготовки молока, мяса, жира, яиц, печени, почек, кормов и зерна. Пробоподготовка включает экстракцию токсикантов ацетонитрилом и удаление жиров экстракцией гексаном. В условиях электрораспылительной ионизации большинство аналитов образуют протонированные и депротонированные формы, и редко встречаются аддукты с аммонием, натрием и калием. Идентификация аналитов проведена с использованием программы «TargetAnalysis-1.3». Идентификационными параметрами служили время удерживания, точность измеренных масс ионов и соответствие изотопному распределению (mSigma). Показаны низкие пределы обнаружения аналитов  (0.0005 - 50 нг/мл). Установлено, что с учетом столь низких пределов определения возможно разбавление экстракта водой до устранения матричного эффекта. Нижняя граница определяемых содержаний с учетом пробоподготовки и разбавления составила 1(500) мкг/кг. Степень извлечения аналитов из анализируемых проб колеблется от 78 до 110 % в зависимости от матрицы и природы аналита. Предложена схема идентификации и определения токсикантов методом стандартной добавки. Показаны преимущества метода стандартной добавки перед методом градуировочного графика при определении токсикантов в реальных пробах. Относительное стандартное отклонение результатов анализа не превышает 0.11. Продолжительность идентификации 40-60 мин и определения обнаруженных токсикантов – 2-3 ч.Ключевые слова: высокоэффективная жидкостная хроматография, тандемная квадруполь-времяпролетная масс-спектрометрия, пищевые продукты, корма, пестициды, микотоксины, ветеринарные препараты, пищевые и непищевые красителиDOI: http://dx.doi.org/10.15826/analitika.2015.19.2.01

The identification and determination of toxicants in food products and raw food materials by HPLC / TOF high resolution mass spectrometry

Предложена методология идентификации и определения из одной навески 300 токсикантов различных классов в пищевых продуктах и продовольственном сырье методом времяпролетной масс-спектрометрии высокого разрешения в сочетании с высокоэффективной жидкостной хроматографией и простой, экспрессной пробоподготовкой. Аналиты - пестициды, микотоксины, антибиотики, красители в пищевых продуктах и кормах. Предложены простые варианты пробоподготовки молока, мяса, жира, яиц, печени, почек, кормов и зерна. Пробоподготовка включает экстракцию токсикантов ацетонитрилом и удаление жиров экстракцией гексаном. В условиях электрораспылительной ионизации большинство аналитов образуют протонированные и депротонированные формы, и редко встречаются аддукты с аммонием, натрием и калием. Идентификация аналитов проведена с использованием программы «TargetAnalysis-1.3». Идентификационными параметрами служили время удерживания, точность измеренных масс ионов и соответствие изотопному распределению (mSigma). Показаны низкие пределы обнаружения аналитов (0.0005 - 50 нг/мл). Установлено, что с учетом столь низких пределов определения возможно разбавление экстракта водой до устранения матричного эффекта. Нижняя граница определяемых содержаний с учетом пробоподготовки и разбавления составила 1(500) мкг/кг. Степень извлечения аналитов из анализируемых проб колеблется от 78 до 110 % в зависимости от матрицы и природы аналита. Предложена схема идентификации и определения токсикантов методом стандартной добавки. Показаны преимущества метода стандартной добавки перед методом градуировочного графика при определении токсикантов в реальных пробах. Относительное стандартное отклонение результатов анализа не превышает 0.11. Продолжительность идентификации 40-60 мин и определения обнаруженных токсикантов - 2-3 ч.The methodology of identification and defining 300 toxicants of different classes in food products and food raw materials out of one sample have been proposed and realized by the TOF mass spectrometry of high resolution combined with high performance liquid chromatography and simple express sample preparation. Analytes included pesticides, mycotoxins and veterinary drugs. Simple sample preparation variations of milk, meat, fat, eggs, liver, kidney, feed and grain were suggested. Sample preparation involved the extraction with acetonitrile and removal of toxins fat by extraction with hexane. Under the electrospray ionization conditions most analytes develop protonated and deprotonated forms and rare adducts with ammonium, sodium and potassium. Identification of analytes was carried out using the «TargetAnalysis-1.3» program. Retention time, the accuracy of the masses of the ions and matching of the isotopic distribution (mSigma) were used as identification parameters. Low limits of detection of analytes were shown to be 0.0005-50 ng / ml. It was found that, given such low detection limits, the dilution of the extract with water to eliminate the matrix effect is possible. The lower limit of the defined content with the sample preparation and dilution amounted to 1 (500) mg / kg. The degree of extraction of analytes from the analyzed samples ranged from 78 to 110 % depending on the nature of the analyte and the matrix. A scheme for the identification and defining toxins by standard addition was proposed. The advantages of the standard addition method compared to the method of the calibration curve in the determination of toxicants in real samples were demonstrated. The relative standard deviation of the test results does not exceed 0.11. The identification period was 40-60 min, and the definition of identified toxicants was in the range of 2-3 hours

Spectrophotometric and enzymatic methods for biochemical analysis of blood sera from farm animals

Conventional and present-day principles of spectrophotometric and enzymatic determination of major biochemical values of animal blood serum (plasma) are described in the paper: total protein, albumin, urea, uric acid, creatinine, bilirubin (total and conjugated bilirubin), glucose, lactic acid, triglycerides, Cholesterine, phospholipids, calcium, magnesium, phosphorus, potassium, sodium, ferrum, chlorides, alkaline phosphatase, lactate dehydrogenase, creatine kinase, a-amylase, aspartate aminotransferase, alanine aminotransferase, Cholinesterase, γ-glutamyltransferase and hemoglobin

Development and validation of fluoroquinolone detection method in chicken eggs using high-performance liquid chromatography

The method for the simultaneous detection of nine fluoroquinolone antibiotics (marbofloxacin, ofloxacin, pefloxacin, norfloxacin, ciprofloxacin, enrofloxacin, danofloxacin, sarafloxacin and difloxacin) in chicken eggs using high-performance liquid chromatography with fluorescence detection was developed. The detected range of fluoroquinolones is from 10 to 100 μg/kg in a 4g sample. Based on the validation studies the combined standard uncertainty and relative expanded uncertainty were calculated. The technique enables to detect antibiotic residues in their admissible levels equal to 100 μg/kg, in compliance with SanPiN 2.3.2.2804-10 and Common sanitary-epidemiological and hygienic requirements to the goods subject to sanitary and epidemiological surveillance (control)

Determination of adulterations in dairy products and vegetable oils based on their fatty acid compositions

Improved method for determination of fatty acid compositions of dairy and plant products that allows detection of adulterations in vegetable oils, milk and dairy products is described in the paper

Capabilities of capillary electrophoresis for analysis of ready-to-use pharmaceutical products with an antibacterial effect

The capability to monitor quality of ready-to-use pharmaceuticals using capillary electrophoresis methods is evaluated in the paper. Optimal conditions for separation and determination of different classes of such antibacterial agents as penicillins, fluoroquinolones, nitrofurans, sulfanilamides, metronidazole and chloramphenicol are chosen. The range of content of active constituents is 1-2500 mg/g for solid samples and 0,001-0,50% for liquid preparations