Determination of aluminum in plant tissue digests using a catechol violet colorimetric method

Abstract

A procedure is described for the colorimetric determination of Al in acid digests of plant tissue using catechol violet as the chromogenic reagent. Components of plant tissue digests were examined for interference in the determination of Al using the described procedure. For highest accuracy, Al standards should contain macroelements, microelements, and acid at concentrations generally typical of those present in acid digests of the plant tissue being analyzed. Using the described procedure, recovery of Al added prior to digestion was 99.1% and 97.1% for National Bureau of Standards SRM 1571 (orchard leaves) and SRM 1572 (citrus leaves), respectively.     

A simple and precise method for total chlorophyll analysis in tropical foliage plants 1

Methods currently in use for total chlorophyll analysis of plant tissue were reviewed. Storage of leaf tissue before analysis, filtration methods during analysis and extract longevity were investigated. The final procedure includes the use of leaf area cores and extraction with 100% acetone with results reported by area and oven dry weight. A simple and precise composite method was adopted.     

Thin layer chromatographic screening method for sulfadiazine residues in calf tissues, plasma, and urine

Because of the lack of specificity of the Bratton-Marshall procedure for assaying sulfonamides, a sensitive, specific tissue residue assay for sulfadiazine (SDZ) was developed. The methodology has been extended to provide a highly sensitive screen for sulfonamide residues, which employs 2-dimensional thin layer chromatography in conjunction with fluorescamine derivatization. The procedure described, which has been developed for SDZ in calf tissues, involves direct ethyl acetate extraction of tissue homogenates. Following evaporation of the organic phase, a portion of the residue is spotted on a 20 X 20 cm silica gel 60 plate, which is then developed in 2 dimensions with solvent systems devised to separate SDZ from endogenous substances as well as from 12 other sulfonamides that might be present in calf tissues. The presence of SDZ at a concentration of 0.1 ppm or its absence is easily demonstrated in calf kidney, liver, muscle, plasma, and urine. The basic method can be modified for a particular sulfonamide in a target tissue and can be used as a quantitative assay for sulfonamide residues.     

Determination of albendazole and its major metabolites in the muscle tissues of Atlantic salmon,tilapia, and rainbow trout by high performance liquid chromatography with fluorometric detection

A liquid chromatographic procedure for the determination of albendazole ([5-(propylthio)-1H-benzimidazol-2yl]carbamic acid methyl ester) and its major metabolites, albendazole sulfoxide, albendazole sulfone, and albendazole-2- aminosulfone in rainbow trout, tilapia, and salmon muscle with adhering skin tissue is described. The muscle tissue samples are made alkaline with potassium carbonate and extracted with ethyl acetate. The extracts are further subjected to cleanup by utilizing a number of liquid-liquid extraction steps. After solvent evaporation, the residue is reconstituted in mobile phase and chromatographed. The chromatography is carried out on a reversed phase Luna C(18) column, using acetonitrile/methanol/buffer as a mobile phase and a fluorescence detector. The average recoveries from the fortified muscle tissue of the three fish species for albendazole (25-100 ppb), albendazole sulfoxide (15.5-62 ppb), albendazole sulfone (1-10 ppb), and albendazole-2- aminosulfone (10-100 ppb) were 94, 77, 82, and 67%, respectively. The average CV for each compound was < or =10%. The procedure was validated and then applied to the determination of albendazole and its three major metabolites in the muscle tissue of the three fish species obtained after orally dosing with albendazole.     

Sensitive streptavidin-biotin enzyme-linked immunosorbent assay for rapid screening of chloramphenicol residues in swine muscle tissue

A sensitive, competitive enzyme-linked immunosorbent assay (ELISA) for chloramphenicol (CAP) in swine muscle tissue has been developed. The ELISA is based on an earlier procedure. To improve sensitivity, different optimization procedures were investigated. The introduction of a streptavidin-biotin system and the use of a coating antigen with a lower CAP incorporation resulted in the most sensitive ELISA: the CAP concentration giving 50% inhibition decreased from 125 ng/mL to 3.0 ng/mL. This ELISA procedure was applied for a rapid screening of CAP residues in swine muscle tissue. The tissues were extracted with demineralized water. A concentrated phosphate-buffered saline solution was added to the filtered aqueous extract and this sample solution was directly submitted to the ELISA procedure. The results were compared to values obtained by analysis of a corresponding blank. This blank was prepared by treating a part of the aqueous sample solution with an immobilized monoclonal antibody preparation. This treatment was necessary because aqueous extracts of different swine muscle tissues showed a high variation in dose-response curves, probably caused by the complexity and variability of the matrix. In spiked tissues, the presence of CAP at concentrations of 10 micrograms/kg and higher can be easily demonstrated.     

An automated procedure for the analysis of boron in plant tissue

Abstract

Boron analysis of plant tissue by the quinallzarin procedure has been adapted to the Technicon AutoAnalyzer system. Results obtained are comparable to those by the curcumin method for evaluating the boron status of the plant. Details of the mechanical assembly are presented and precautions to be observed are discussed. The suggested procedure permits the analysis of 30 samples of plant ash extract per hour.     

Development of an enzyme-linked immunosorbent assay for the determination of maduramicin in broiler chicken tissues.

Maduramicin is one of the most widely used coccidiostats in the world. A rapid and accurate analytical method for this drug should provide producers and users with an effective management tool. The current chromatographic methods are sensitive but labor-intensive. This paper reports the development of an enzyme-linked immunosorbent assay (ELISA) based on an immunoaffinity chromatography cleanup procedure for the analysis of maduramicin in broiler chicken tissues (including muscle, liver, and fat). Recoveries from fortified tissue homogenates at levels of 30.0-120.0 microg kg(-)(1) ranged from 76.4 to 107.5% with coefficients of variation of 3.8-16.4%. The limits of detection were 1.0 ng g(-)(1) in muscle, 2.8 ng g(-)(1) in liver, and 1.5 ng g(-)(1) in fat. The ELISA results from the analysis of incurred residue in tissue samples showed the cleanup procedure is viable.     

Simple automated wet digestion of animal tissues for determination of seven elements by atomic absorption spectroscopy

A simple, automated wet digestion procedure was developed for the quantitative determination by atomic absorption spectroscopy (AAS) of arsenic, cadmium, copper, mercury, lead, selenium, and zinc in animal tissue. A commercial digestion block system with automated temperature programming was used. Recoveries of all elements from spiked bovine liver and kidney samples exceed 95%. The analytical results obtained for samples of NBS Bovine Liver (No. 1577a) agree well with certified values. The procedure is safe and requires minimum analyst time.     

Test method for determination of virucidal efficacy of liquid surface disinfectants.

A test method was developed incorporating AOAC use-dilution stainless steel Penicylinders and H.Ep.-2 tissue culture cells for the evaluation of disinfectants as virucides. The subsequent collaborative study using Herpes simplex and poliovirus type 1 in evaluation against a quaternary ammonium compound, phenolic, and iodiphor disinfectants indicated that the basic test procedure is sound. However, an improved virus concentration technique and a modified method for drying virus-contaminated cylinders are recommended. In addition, other types of disinfectants and test viruses should be included in order to demonstrate that the procedure is versatile as well as reproducible.     

Comparison of Five Digestion Procedures for Recovery of Nutrients and Trace Elements in Plant Tissue

Comparison of five digestion procedures for recovery of nutrients and trace elements in plant tissue were examined. The objectives of this study were to compare five digestion procedures: AOAC (dry ashing), two nitric acid, (NA1, commonly used and NA2, our modification), and two aqua regia (AR1, commonly used and AR2, our modification) procedures for recovery of 15 plant nutrients and trace elements in corn, swiss chard, thorn apple, and barley flour (certified material). Generally, the recovery of various elements depended on the digestion procedure used and the plant species. In most instances, NA2 recovered more phosphorus (P), potassium (K), sulfur (S), and manganese (Mn) than the AOAC or the two AR procedures. Also, overall NA2 recovered more K and magnesium (Mg) than NA1. Extractants AR1 and AR2 recovered more iron (Fe) than the two NA procedures, and the same as the AOAC. The AR2 recovered very similar concentrations of nutrients as the AOAC; and in corn tissue, AR2 recovered more zinc (Zn), chromium (Cr), nickel (Ni), copper (Cu), and boron (B) than the AOAC. The AOAC recovered less S and B than the other procedures tested. For instance, in barley flour, AOAC recovered eight to nine times less S, while in corn tissue AOAC recovered around three times less S than the NA procedures. Overall, there are three important findings: (1) NA procedures may be used for recovery of most elements in plant tissue; (2) AR2 procedure is comparable to the official AOAC method for recovery of nutrients and trace elements in plant tissue, and (3) the official AOAC method used in Canadian plant testing laboratories may underestimate the concentration of S and B in plant tissue.     

A phenol‐hypochlorite manual determination of ammonium‐nitrogen in Kjeldahl digests of plant tissue

Abstract

A phenol‐hypochlorite (Berthelot) procedure for determining NH₄‐N in acid digests of plant tissue is described. EDTA suppresses Cu interference and precipitation of hydroxides and phosphates. The oxidizing reagent includes an alkaline buffer to account for variations in acidity between digestion solutions, the main source of error in an unbuffered procedure. The reagent combinations and addition sequence employed allow for delays of up to at least 30 minutes between reagent additions without affecting the final colour intensity. The proposed procedure is as sensitive as similar methods used for determining NH₄‐N in natural waters and yields results which agree well with those obtained by steam distillation.     

Preparative method for gas chromatographic/mass spectral analysis of trace quantities of pesticides in fish tissue.

A large capacity, low efficiency pesticide cleanup method is combined with a small capacity, high efficiency method for gas chromatographic/mass spectral analysis of fish tissue. Trace chemicals are extracted from the tissue, removed from the bulk of the co-extracted lipids through selective desorption from Micro Cel-E, and isolated from the remaining lipids by gel permeation chromatography. The procedure is capable of cleaning up the extract from several hundred grams of fish, minimizes sample contamination, and permits identification of xenobiotic chemicals present at ng/g concentrations.     

Simple assay for analyzing five microcystins and nodularin in fish muscle tissue: hot water extraction followed by liquid chromatography-tandem mass spectrometry

A simple, specific, and sensitive procedure for determining six cyanotoxins, that is, microcystins RR, LR, YR, LA, and LW and nodularin, in fish muscle tissue is presented. This method is based on the matrix solid-phase dispersion technique with heated water as extractant followed by liquid chromatography (LC)-tandem mass spectrometry (MS) equipped with an electrospray ion source. Target compounds were extracted from tissue by 4 mL of water acidified to pH 2 and heated at 80 degrees C. After acidification and filtration, 0.2 mL of the aqueous extract was injected in the LC column. MS data acquisition was performed in the multireaction monitoring mode, with at least two precursor ion > product ion transitions selected for each target compound. Analyte recovery ranged between 61 and 82% and was not substantially affected by either the analyte concentrations or the type of fish. The nonexcellent recovery of some of the microcystins was traced to binding of these compounds to protein phosphatases in fish tissue occurring during sample treatment. The existence of covalently bound microcystins in fish has been evidenced by several studies. Compared to an older sample preparation procedure, this one extracted larger amounts of the analytes in a simpler and much more rapid way. On the basis of a signal-to-noise ratio of 10, limits of quantification were estimated to range between 1.6 and 4.0 ng/g. The effects of temperature and volume of the extractant on the analyte recovery were studied.     

Liquid chromatographic determination of lasalocid sodium in chicken skin: interlaboratory study

The U.S. Food and Drug Administration (FDA) sponsored an interlaboratory study of a liquid chromatographic determinative procedure for lasalocid sodium in chicken skin with adhering fat. Four laboratories analyzed 35 dosed tissue samples and 82 fortified tissue samples containing lasalocid at levels ranging from 0.1 to 0.6 ppm. Samples were homogenized with acetonitrile, washed with hexane, and partitioned into the mobile phase prior to analysis liquid chromatography with fluorescence detection. The results of the interlaboratory study showed good reproducibility for fortified samples. Fortification levels, average recoveries, and interlaboratory percent coefficients of variation were as follows: 0.6 ppm, 0.57 ppm, and 9.7; 0.3 ppm, 0.25 ppm, and 9.1; and 0.15 ppm, 0.14 ppm, and 7.0, respectively. Data for analysis of the dosed tissue also showed good agreement among the laboratories.     

Rapid perchloric acid digest methods for analysis of major elements in plant tissue

Abstract

The nutrient element composition of tissue from a plant species is not fixed. Composition in the plant varies among different plant parts at the same or different physiological stages of maturity, as soil type changes, from day to day, even hour to hour on the same day due to past and current environmental conditions. These observations seem to undermine the general trend in plant tissue analysis to develop quantitative procedures that are time consuming and, therefore, foster insufficient nutrient status monitoring within and/or between different physiological stages of maturity of like or different plant parts. Therefore, until tissue sampling becomes more definitive, a digest procedure with 90–100% recovery should be more than adequate for major elemental analysis. The HClO₄ digestion procedures described here meet that requirement and take only 15 to 30 min to complete. N recovery was greater than 100% with the H₂O₂‐HClO₄ digestion procedure. However, when HNO₃ was combined with H₂O₂ and HClO₄, N was lost, although even then N volatilization occured only when digestion was extended past clearing.     

Plant Alkaline Fusion Technique Followed by Colorimetric Procedure for the Detection and Quantification of Total Silicon in Ornamental Plants

Silicon (Si)–determination procedures currently available require expensive equipment and must be modified to assess Si concentrations in plant tissues. A Si-determination procedure referred to as the plant alkaline fusion technique, or PAFT, was developed to assess total Si concentrations, specifically in plant tissue. The procedure consists of dry-ashing the plant material, alkaline fusion, solubilization of the fusion cake, dilution, and colorimetric determination to quantify total Si content. The fusion technique for solubilizing plant Si can be performed using sodium hydroxide (NaOH), nickel crucibles, and Bunsen burners. Silicon concentrations of two National Institute of Standards and Technology (NIST) standard reference materials (SRM) [pine needles (SRM 1575) and peach leaves (SRM 1547)], a rice straw standard (LSU#5), an NIST soil standard (SRM 2709), and five ornamental plant species were all assessed using the PAFT procedure, and then these values were compared to two different inductively coupled plasma–optical emission spectroscopy (ICP-OES) procedures conducted by independent laboratories. The PAFT Si determination procedure typically reported the greatest value or was within 15% of the two ICP-OES procedures while costing significantly less than both ICP-OES procedures.     

Detection of traces of tetracyclines from fish with a bioluminescent sensor strain incorporating bacterial luciferase reporter genes

Bioluminescent Escherichia coli K-12 strain for the specific detection of the tetracycline family of antimicrobial agents was optimized to work with fish samples. The biosensing strain contains a plasmid incorporating the bacterial luciferase operon of Photorhabdus luminescens under the control of the tetracycline responsive element from transposon Tn10 (Korpela et al. Anal. Chem. 1998, 70, 4457-4462). The extraction procedure of oxytetracycline from rainbow trout (Oncorhynchus mykiss) tissue was optimized. There was neither need for centrifugation of homogenized tissue nor use of organic solvents. The lowest levels of detection of tetracycline and oxytetracycline from spiked fish tissue were 20 and 50 microg/kg, respectively, in a 2-h assay. The optimized assay protocol was tested with fish that were given a single oral dose of high and low concentrations of oxytetracycline. The assay was able to detect oxytetracycline residues below the European Union maximum residue limits, and the results correlated well with those obtained by conventional HPLC (R = 0.81).     

Validation study of gas chromatographic determination of pentachlorophenol in animal liver

A validation study was conducted of a gas chromatographic procedure for the determination of pentachlorophenol (PCP) in chicken, pork, and beef liver. Five analysts representing 5 laboratories analyzed randomly numbered blind duplicates at 3 fortified tissue concentrations and one incurred tissue on 2 consecutive days. The PCP concentrations ranged from approximately 40 to 400 parts per billion (ppb). All data were reported to 3 significant figures in ppb. The coefficients of variation for repeatability were between 2.8 and 8.5%, except for the beef liver, at a mean value of 80 ppb PCP, where the CV was 11.3%. The CVs for reproducibility were in the range of 9.7-16.5% with little significant difference by species. The CV asymptotically approached 10% as the PCP level increased.     

Glucosinolate content and isothiocyanate evolution--two measures of the biofumigation potential of plants.

A total of 570 lyophilised Brassica root and shoot tissue samples were hydrolyzed, and the liberated isothiocyanates (ITCs) were analyzed by gas chromatography-flame photometric detection (GC-FPD). Glucosinolates (GSLs) were extracted from samples of the same tissues and analyzed by high-performance liquid chromatography (HPLC). The concentrations of six GSLs/ITCs (2-propenyl, 3-butyl, 4-pentenyl, benzyl, 4-methylthiobutyl, and 2-phenylethyl) as determined by the two techniques were compared. In 79% of the samples, the concentration of GSLs in the tissues was greater than that of the ITCs released on hydrolysis. Several possible reasons for the difference are proposed, including the effect of tissue storage time, hydrolysis of GSLs may be less efficient than the GSL extraction procedure, or some of the ITCs formed reacted with plant proteins and amino acids in the sample and were therefore not detected in the extract. GSL concentration in plant tissues is used to estimate the biofumigation potential of the plant tissue, whereas the actual biofumigation effect is thought to be due to the ITCs formed by hydrolysis of the plant-based GSLs. The variation between ITC and GSL values therefore has implications for the assessment of the biofumigation potential of the plant tissue.     

Determination of molybdenum in plant tissue by graphite furnace atomic absorption spectrophotometry (GFAAS)

Abstract

The analysis of molybdenum in plant tissue using graphite furnace atomic absorption spectrophotometry (GFAAS) is described. The method involved wet digestion using a mixture of sulphuric, nitric and perchloric acids followed by a solvent extraction procedure. Molybdenum was extracted into di‐iso butyl ketone (DIBK) as the iron‐thiocyanate complex. The extract was then analysed for molybdenum by GFAAS.

The results of analyses of the reference plant materials (orchard leaves and citrus leaves) of the National Bureau of Standards (NBS) compared very well with the certified values. Other types of plant tissue were also analysed and the results correlated well with those obtained by an alternative method.