Modified Digestion Method Research Articles

Determination of concentrations of Sr and Ba in coal and coal combustion by-products: A comparison between results by ICP-MS and XRF techniques

Concentrations of trace elements in coal and coal combustion products are commonly analyzed by inductively coupled plasma-mass spectrometry (ICP-MS) due to large number of elements detected and the relatively the low detection limits of this technique. Like other geological samples, complete dissolution of coal and coal combustion products is also essential for accurate ICP-MS results. In this study, Sr and Ba in coal and coal combustion products (fly and bottom ashes) from two coal-combustion power plants from North China analyzed by XRF and ICP-MS were comparatively studied. The concentrations of Sr and Ba analyzed by ICP-MS, when a mixture of acids (2 ml HF + 5 ml HNO3 for each 50 mg coal sample and 5 ml HF + 2 ml HNO3 for each 50 mg ash sample) was used for microwave-assisted digestion, do not fit well with the their relative XRF results. This is most probably due to the formation of the fluorides during microwave digestion, and this assumption is supported by the presence of various fluoride compounds, NaMgAl(F, OH)6·H2O, NH4MgAlF6, AlF3, and K2SiF6, in the residues of all the coal and ash samples in our sequential extraction experiment. Cations of Sr and Ba were probably trapped into the divalent cation sites of the fluorides. Concentrations of Sr and Ba analyzed by ICP-MS using increased HF and HF/HNO3 ratio (7 ml HF and 2 ml HNO3 for each 50 mg coal/ash sample) are in better agreement with the XRF results. Our results indicate that excess amount of HF probably has led to the suppression of these elements due to fluoride precipitation. The results indicate that the modified digestion method is capable of achieving complete digestion of coal samples and results in a reliable analysis of Sr and Ba concentrations in coal samples and most fly ashes by ICP-MS. However, the formation of insoluble fluorides is probably not completely suppressed for some bottom ash samples, which can result in underestimation of Sr and Ba concentrations. Nevertheless, XRF analysis can serve as a reliable cross-check method to assist in the evaluation of the accuracy of ICP-MS results.

Optimization and Identification of Single Mutation in Hemoglobin Variants with 2,2,2 Trifluoroethanol Modified Digestion Method and Nano-LC Coupled MALDI MS/MS.

Background: Hemoglobin (Hb) variants arise due to point mutations in globin chains and their pathological treatments rely heavily on the identification of the nature and location of the mutation in the globin chains. Traditional methods for diagnosis such as HPLC and electrophoresis have their own limitations. Therefore, the present study aims to develop and optimize a specific method of sample processing that could lead to improved sequence coverage and analysis of Hb variants by nano LC−MALDI MS/MS. Methods: In our study, we primarily standardized various sample processing methods such as conventional digestion with trypsin followed by 10% acetonitrile treatment, digestion with multiple proteases like trypsin, Glu−C, Lys−C, and trypsin digestion subsequent to 2,2,2 trifluoroethanol (TFE) treatment. Finally, the peptides were identified by LC−MALDI MS/MS. All of these sample processing steps were primarily tested with recombinant Hb samples. After initial optimization, we found that the TFE method was the most suitable one and the efficiency of this method was applied in Hb variant identification based on high sequence coverage. Results: We developed and optimized a method using an organic solvent TFE and heat denaturation prior to digestion, resulting in 100% sequence coverage in the β−chains and 95% sequence coverage in the α−chains, which further helped in the identification of Hb mutations. A Hb variant protein sequence database was created to specify the search and reduce the search time. Conclusion: All of the mutations were identified using a bottom−up non−target approach. Therefore, a sensitive, robust and reproducible method was developed to identify single substitution mutations in the Hb variants from the sequence of the entire globin chains. Biological Significance: Over 330,000 infants are born annually with hemoglobinopathies and it is the major cause of morbidity and mortality in early childhood. Hb variants generally arise due to point mutation in the globin chains. There is high sequence homology between normal Hb and Hb variant chains. Due to this high homology between the two forms, identification of variants by mass spectrometry is very difficult and requires the full sequence coverage of α− and β−chains. As such, there is a need for a suitable method that provides 100% sequence coverage of globin chains for variant analysis by mass spectrometry. Our study provides a simple, robust, and reproducible method that is suitable for LC−MALDI and provides nearly complete sequence coverage in the globin chains. This method may be used in the near future in routine diagnosis for Hb variant analysis.

Pectic Arabinans in Quinoa Seeds (Chenopodium quinoa Willd.) Are Acylated with p‐Coumaric Acid

A modified digestion method was used to hydrolyze large amounts of quinoa meal to study polysaccharide‐bound phenolic acids. Besides the well‐characterized oligosaccharide O‐(2‐O‐trans‐feruloyl‐α‐l‐arabinofuranosyl)‐(1→5)‐l‐arabinofuranose, O‐(2‐O‐trans‐coumaroyl‐α‐l‐arabinofuranosyl)‐(1→5)‐l‐arabinofuranose was isolated from the hydrolysate and identified by LC‐MS, GC‐MS, and NMR experiments. This study demonstrates for the first time that p‐coumaric acid can be linked to pectic side chains of dicotyledonous plants.

Collaborative study for quantification of Anisakis larvae in spiked salmon fillets (Salmo salar) by a modified Codex digestion method

A collaborative study for quantification of Anisakis larvae in spiked farmed salmon fillets was conducted by 12 laboratories in 2012, using a modified Codex method (CODEX STAN 244). N = 6 samples with a different number of Anisakis larvae including two blind duplicates were analyzed by participants. All laboratories applied the same modified digestion method with stirring speed of 500–600 rpm, using a triangular stirring bar and a maximum digestion temperature of 35 °C in order to reduce fragmentation of larvae. All participants found correctly no larvae in the control samples while the mean recovery rate of all spiked larvae was 86.6 %. The among-laboratory reproducibility was 16.5 %. Differences were noticed in the interpretation of Anisakis fragments, which occurred as a result of prolonged frozen storage of the samples.

An improved digestion technique for determination of platinum group elements in geological samples

This paper reports a modified digestion method for the determination of platinum group elements (PGEs) in geological samples. Samples are first digested by HF in a custom made 120 ml PTFE beaker on a hot plate to remove silicates. The dried residue is then digested with HF + HNO3 at 190 °C for about 48 h after the beaker is sealed in a stainless steel pressure bomb. Although only HF + HNO3 are used for sample digestion at relatively low temperature, ultramafic rocks with refractory minerals can be effectively decomposed by this method. Analytical results for CCRMP (CANMET, Ottawa, Canada) certified reference materials WGB-1 (gabbro), UMT-1 (ultramafic ore tailings), WPR-1 (peridotite) and WMS-1(Massive Sulphide), agree well with the certified values. The proposed method significantly simplifies the sample digestion procedures for PGE analysis compared to those used previously.

Sensitivity and Optimization of Artificial Digestion in the Inspection of Meat for Trichinella spiralis

In many countries, the method of choice in inspecting meat for Trichinella spiralis infection is artificial digestion. We conducted a study of the sensitivity of the artificial digestion method recommended by the International Commission on Trichinellosis for detecting T. spiralis larvae in meat and of the effect of modifications of some procedures used in the method on its sensitivity. As part of this, we evaluated the effects on larval recovery of the vessels used for larval settling, sieve sizes, and temperatures at which larvae passed through the sieves, using larvae from T. spiralis-infected mice. We observed the effects on larval recovery of digestion duration and of modified artificial digestion by using 10-g samples of infected mouse muscle alone or mixed with uninfected pork. The percentages of larvae recovered with the respective use of separatory funnels and conical cylinders were 51.20% and 98.70%. The rates of recovery of T. spiralis larvae at 4 degrees C after passage through sieves of 425-microm mesh (No. 40), 250-microm mesh (No. 60), and 180-microm mesh (No. 80) were 98.42%, 90.59%, and 81.63%, which exceeded the 97.79%, 85.10%, and 61.12% rates of recovery of motile larvae at 40 degrees C and the 95.12%, 78.60%, and 44.16% rates of recovery of dead larvae at 90 degrees C. The larval recovery rate after digestion for 2 hours (96.18%) was greater than that after 0.5 hours (88.00%). We then examined a modified digestion method in which 10-g samples of pork mixed with 300 mL of digestive solution were digested for 2 hours at 43 degrees C followed by chilling of digest solution to 4 degrees C before passing it through a 425-microm mesh (No. 40) sieve and allowing it to settle in a 1-L conical cylinder. With this procedure, the modified method detected T. spiralis in samples of pork meat weighing 10 g and containing either 1 larva per gram or 0.1 larva per gram. Further validation of digestion method incorporating these modifications is required with the use of larger samples of infected muscle from species such as swine, which are routinely tested for T. spiralis for the purpose of food safety.

An Improved Method for Recovery of Muscle-Stage Larvae From Mice Infected with Toxocara canis

We report a modified digestion method that improves the recovery of Toxocara canis larvae from skeletal muscle. Minced muscle tissue from infected mice was incubated in artificial gastric juice for 48 hr at 37 C, and ethanol was added for the second 24 hr. This procedure allowed the larvae to be identified and counted more quickly than with the standard digestion method. This method allows measurement of the total number of larvae present in muscle tissue following oral intubation of embryonated eggs, although it does not permit counting of live larvae.

A Simple Digestion Method for the Quantitative Analysis of Metal in Some Organometallic Compounds

Abstract A new, modified digestion method for the quantitative analysis of metal in various organometallic compounds containing silicon, tin and titanium has been developed by the use of ammonium nitrate as a new oxidizing reagent. It has been shown that this method can be carried out more rapidly and more accurately than the other known methods.