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Rapid Extraction (rapid + extraction)
Selected AbstractsRapid extraction of high-quality genomic DNA from Porphyra yezoensis (Bangiales, Rhodophyta)PHYCOLOGICAL RESEARCH, Issue 1 2000Maiko Nakajima SUMMARY We developed a simple, rapid and stable method for extraction of high molecular weight DNA from the marine red alga Porphyra yezoensis Ueda using both guanidium treatment and QIAGENÔ kit (Funakoshi, Tokyo, Japan). The method does not require expensive equipment and complex steps. The DNA yield averaged 1.5 ,g 100 mg,1 of Porphyra tissue and the A260/A280 and A230/A260 ratios of the DNA were approximately 1.8 and 0.4, respectively. It was of sufficient quality to be used for not only polymerase chain reactions but also other DNA manipulation techniques such as restriction digestion and construction of genomic libraries. [source] ESI+ MS/MS confirmation of canine ivermectin toxicity,JOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 1 2009A. F. Lehner Abstract Ivermectin is a semisynthetic macrocyclic lactone anthelmintic of the avermectin family derived from Streptomyces fermentation products. Avermectins are used as antiparasitic agents in domestic animals; although considered relatively safe, one must consider animal species, breed, weight, and age in dosage determinations. In January 2006, two canines were presented to the UK Livestock Disease Diagnostic Center after dying from suspected ivermectin overdoses [30,50 mg/kg body weight]. To confirm this clinical diagnosis we developed a rapid, sensitive semiquantitative ElectroSpray Ionization,Mass Spectrometry (ESI/MS) method for ivermectin in canine tissue samples. Pharmaceutical ivermectin contains two ivermectins differing by a single methyl group, and each compound forms interpretation-confounding adducts with tissue Na+ and K+ ions. We now report that ivermectin administration was clearly confirmed by comparison with standard and dosage forms of ivermectin, and simple proportionalities based on mass spectral intensity of respective molecular ions allowed semiquantitative estimates of injection site tissue concentrations of 20 and 40 µg/g tissue (wet weight) in these animals, consistent with the history of ivermectin administration and the clinical signs observed. There is a distinct need for both rapid detection and confirmation of toxic exposures in veterinary diagnostics, whether for interpretation of clinical cases antemortem or for forensic reasons postmortem. It is vital that interpreters of analytical results have appropriate guidance in the scientific literature and elsewhere so as to enable clear-cut answers. The method presented here is suitable for routine diagnostic work in that it allows rapid extraction of ivermectin from tissue samples, avoids the need for high-performance liquid chromatography and allows ready interpretation of the multiple ivermectin species seen by ESI+ MS/MS in samples originating from veterinary dosage forms. Copyright © 2008 John Wiley & Sons, Ltd. [source] Simple and rapid extraction, separation, and detection of alkaloids in beveragesJOURNAL OF SEPARATION SCIENCE, JSS, Issue 21 2008Christine L. Copper Abstract Implementation of an uncomplicated SPE process for the rapid extraction and preconcentration of the alkaloids, colchicine, strychnine, aconitine, and nicotine, from water, apple juice, and nonfat milk samples is presented. When coupled to analysis via micellar EKC (MEKC), the total analysis time per sample was less than 15 min for the water and juice samples and less than 20 min for the milk. The SPE process allowed for anywhere from a three to a fourteen-fold improvement in the LOD for each alkaloid when compared to detecting the alkaloids in a nontreated water sample matrix. Following SPE, the LODs for colchicine, strychnine, and nicotine were sufficient to meet levels from 150 to 5000 times more dilute than the LD50 for a 50 kg individual drinking 12 oz of a contaminated beverage. Aconitine, on the other hand, was detected at approximately the LD50 level. The percent recoveries for the SPE ranged from 37% to as high as 99%. Nicotine attained the highest recovery efficiencies, followed by colchicine, and finally, aconitine and strychnine, which were nearly identical. The greatest recovery efficiencies were achieved from apple juice and water, whereas nonfat milk yielded the lowest. [source] RP-HPTLC densitometric determination and validation of vanillin and related phenolic compounds in accelerated solvent extract of Vanilla planifolia,*JOURNAL OF SEPARATION SCIENCE, JSS, Issue 18 2007Upendra Kumar Sharma Abstract A simple, fast and sensitive RP-HPTLC method is developed for simultaneous quantitative determination of vanillin and related phenolic compounds in ethanolic extracts of Vanilla planifolia pods. In addition to this, the applicability of accelerated solvent extraction (ASE) as an alternative to microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE) and Soxhlet extraction was also explored for the rapid extraction of phenolic compounds in vanilla pods. Good separation was achieved on aluminium plates precoated with silica gel RP-18 F254S in the mobile phase of methanol/water/isopropanol/acetic acid (30:65:2:3, by volume). The method showed good linearity, high precision and good recovery of compounds of interest. ASE showed good extraction efficiency in less time as compared to other techniques for all the phenolic compounds. The present method would be useful for analytical research and for routine analysis of vanilla extracts for their quality control. [source] A new and simple method for rapid extraction and isolation of high-quality RNA from grape (Vitis vinifera) berriesJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 2 2008Francesca Fort Abstract BACKGROUND: A fast and easy method to isolate RNA from grape (Vitis vinifera) berries is proposed. RESULTS: The procedure uses high concentrations of cetyltrimethylammonium bromide (CTAB) and includes boric acid with the purpose of increasing the RNA extraction. Eppendorf tubes can be used since the amount of sample and volumes of reagents needed are small. The method is based on genomic DNA digestion with DNase I, which avoids the disadvantages of using LiCl (e.g. possible amplification of rare transcripts and reduction or even suppression of the activity of RNA-dependent DNA polymerase). The use of DNase reduces the duration of the isolation method to 8 h. The RNA yield (32.9,49.2 µg g,1) and purity indices were satisfactory in all cases. CONCLUSION: Although there is a referenced grape berry method (RNeasy Midi Kit + polyethylene glycol) that is faster than the one proposed, it is 3.7 times more expensive and gives a lower yield. The ability to successfully amplify from the RNA extract was verified by measuring the ,-tubulin and lipoxygenase gene expression. Copyright © 2007 Society of Chemical Industry [source] A short-cut DNA extraction from cod caviarJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 3 2006Futoshi Aranishi Abstract Caviars represent the most consumed form of fish roe products. Due to high demand, ingredient roes of fish are often susceptible to illegal substitution with those of related fish. This study developed a simple and inexpensive protocol enabling the rapid extraction of DNA of acceptable quality and amount to PCR amplification from both cod caviars and their ingredient pollack roes. The protocol was based on extracting total genomic DNA from eggs using urea and a Chelex 100 chelating resin, and could be completed in less than 15 min. Approximately 8 µg of DNA were reproducibly obtained from single eggs of cod caviars and pollack roes in eight individual experiments, and the quality and amount of DNA were sufficient to serve as template for hundreds of PCR reactions of polymorphic DNA markers for phylogenetic analysis. Being applicable to various caviars, this protocol can be useful to detect illegal substitution among ingredient roes of related fishes in PCR-based food inspection. Copyright © 2005 Society of Chemical Industry [source] Application to routine analysis of a method to determine multiclass pesticide residues in fresh vegetables by gas chromatography/tandem mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 11 2002J. L. Martínez Vidal The use of gas chromatography/tandem mass spectrometry (GC/MS/MS) applied to determine multiple pesticide residues in fresh vegetables has been thoroughly studied. A single injection method to detect, confirm and quantify 54 multiclass pesticides has been developed and applied in a routine analysis laboratory. The proposed method consists of a rapid extraction of 15,g of vegetable sample with dichloromethane. An additional clean-up step is not necessary even when injecting 10,µL of extract. Instead the gas chromatograph was fitted with a carbofrit inserted into the glass liner and a guard column. In addition, the detection mode chosen (MS/MS) provides additional selectivity. The method has been validated and applied to 1300 samples in a routine laboratory following specified quality criteria. The recovery efficiencies obtained for all the pesticides ranged between 70.2 and 110.8% at two different fortification levels. The relative standard deviation for quantification (RSD) was lower than 16.7% for all the compounds. Important experimental parameters, such as the conditioning of carbofrit, overload of the analytical column, and cleanliness of the ion trap, were evaluated for their influence on the performance of the method. Copyright © 2002 John Wiley & Sons, Ltd. [source] Application of an efficient strategy based on MAE, HPLC-DAD-MS/MS and HSCCC for the rapid extraction, identification, separation and purification of flavonoids from Fructus Aurantii ImmaturusBIOMEDICAL CHROMATOGRAPHY, Issue 3 2010Chen Wang Abstract This study presents an efficient strategy based on microwave-assisted extraction (MAE), HPLC-DAD-MS/MS and high-speed counter-current chromatography (HSCCC) for the rapid extraction, identification, separation and purification of active components from the traditional Chinese medicine Fructus Aurantii Immaturus. An LC-DAD-MS/MS method was applied for the screening and structural identification of main components in crude extract, and five components were preliminarily identified as neoeriocitrin, narirutin, naringin, hesperidin and neohesperidin according to their UV and mass spectra. An efficient MAE method for the extraction of the three most abundant components (narirutin, naringin and neohesperidin) was optimized by the combination of univariate and multivariate approaches. The crude extract was then separated and purified by HSCCC and a total of 61.6 mg of narirutin, 207.3 mg of naringin and 159.5 mg of neohesperidin at high purities of 98.1, 97.2 and 99.5%, respectively, were obtained from 1.42 g of crude extract. The recoveries of these compounds were 86, 93 and 89%, respectively. Copyright © 2009 John Wiley & Sons, Ltd. [source] Solid-Phase Extraction of Metoprolol onto (Methacrylic acid- ethylene glycol dimethacrylate)-based Molecularly Imprinted Polymer and Its Spectrophotometric DeterminationCHINESE JOURNAL OF CHEMISTRY, Issue 4 2010Mohammad Saber Tehrani Abstract A new adsorbent for molecularly imprinted solid phase extraction (MISPE) of metoprolol was synthesized using methacrylic acid (MAA) as functional monomer and ethylene glycol dimethacrylate (EGDMA) as cross-linking agent causing a non-covalent, bulk, thermal radical-polymerization. Control polymer (non-imprinted polymer) was prepared under well defined conditions without the use of metoprolol. The synthesized polymers were characterized by IR spectroscopy, X-ray diffraction and thermal analysis techniques. This polymer was used for the rapid extraction and preconcentration of metoprolol from real samples prior to spectrophotometric determination. Extraction efficiency and the influence of flow rates of sample and stripping solutions, pH, type of eluent for elution of metoprolol from polymer, break through volume and limit of detection were studied. The detection limit of the proposed method is 0.4 ng·mL,1. The method was applied successfully to the recovery and determination of metoprolol in tablets, human urine and plasma samples. [source] Preconcentration and Determination of Copper(II) Using Octadecyl Silica Membrane Disks Modified by 1,5-Diphenylcarhazide and Flame Atomic Absorption SpectrometryCHINESE JOURNAL OF CHEMISTRY, Issue 11 2007Ali MOGHIMI Abstract A simple and reproducible method for the rapid extraction and determination of trace amounts of copper(II) ions using octadecyl-bonded silica membrane disks modified by 1,5-diphenylcarbazide (DPC) and atomic absorption spectrometry was presented, which was based on complex formation on the surface of the ENVI-18 DISKTM disks followed by stripping of the retained species by minimum amounts of appropriate organic solvents. The elution was efficient and quantitative. The effect of potential interfering ions, pH, ligand amount, stripping solvent, and sample flow rate were also investigated. Under the optimal experimental conditions, the break-through volume was found to be about 1000 mL providing a preconcentration factor of 400. The maximum capacity of the disks was found to be (255±5) µg for Cu2+, and the limit of detection of the proposed method was 5 ng per 1000 mL. The method was applied to the extraction and recovery of copper in different water samples. [source] | ||||||||||