Rapid Sample Preparation Method Research Articles

A simple, rapid and eco-friendly approach for the analysis of aromatic amines in environmental water using single-drop microextraction-gas chromatography.

Single-drop microextraction (SDME) coupled with gas chromatography-flame ionization detector (GC-FID) was developed for the extraction and determination of aromatic amines (AAs) in environmental water samples. A silicon tube was introduced for the SDME procedure by inserting it into the needle of the micro-injector. In this manner, a large volume of extractant is allowed to be suspended for the extraction, leading to the enhancement of method sensitivity and reproducibility. Extraction parameters which control the performance of SDME such as the type of microextraction solvent and volume, sample pH, ionic strength and extraction time were investigated and optimized. Under the optimized conditions, the SDME-GC method exhibited good linearity from 0.5 to 50 μg mL(-1) for aniline and 4-methylaniline and 0.1 to 50 μg mL(-1) for N-methylaniline and N, N-diethylaniline. The enrichment factors were calculated to be 42-509. The SDME-GC method was performed for the determination of AAs in environmental water samples including drinking, lake and sea water, and excellent recoveries and relative standard deviations (RSD values) ranging from 79.5 to 122.7% and 3.2 to 13.3%, respectively, were obtained. The results demonstrated that SDME-GC is a rapid, simple and effective sample preparation method and could be successfully applied for the determination of AAs in environmental water samples.

Dispersive liquid-liquid microextraction for simultaneous determination of six parabens in aqueous cosmetics

A simple and rapid sample preparation method of dispersive liquid-liquid microextraction(DLLME) was applied in the simultaneous determination of six parabens in the aqueous cosmetics. The analysis was performed on gas chromatography coupled with a flame ionization detection(GC-FID). The mixed solution containing 30 μL of chloroform(extraction solvent) and 300 μL of tetrahydrofuran(dispersive solvent) was rapidly injected into the sample solution for the purpose of microextraction. After that, the solution mentioned above was centrifuged at 4000 r/min for 10 min, and then the organic sediment phase was detected by GC-FID. The effects of experimental parameters, such as the extraction solvent and the volume of it, and the dispersive solvent and the volume of it, on the yield of the extraction were studied in detail. Under the optimum conditions, the enrichment factors of the target analytes range from 87 to 214. Linearity ranges are 0.05–10.0 μg/mL for methylparaben and 0.025–5.0 μg/mL for the other five parabens. The relative standard deviations(RSDs) are lower than 8.2%(n=6). The proposed method was applied to the analysis of six parabens in eleven aqueous cosmetics. The recoveries of the target analytes in the spiked real samples are in the range of 81.0%–103%.

Development of a new microextraction method based on elevated temperature dispersive liquid–liquid microextraction for determination of triazole pesticides residues in honey by gas chromatography-nitrogen phosphorus detection

In the present study, a rapid, highly efficient, and reliable sample preparation method named “elevated temperature dispersive liquid–liquid microextraction” followed by gas chromatography-nitrogen-phosphorus detection was developed for the extraction, preconcentration, and determination of five triazole pesticides (penconazole, hexaconazole, diniconazole, tebuconazole, and difenoconazole) in honey samples. In this method the temperature of high-volume aqueous phase was adjusted at an elevated temperature and then a disperser solvent containing an extraction solvent was rapidly injected into the aqueous phase. After cooling to room temperature, the phase separation was accelerated by centrifugation. Various parameters affecting the extraction efficiency such as type and volume of the extraction and disperser solvents, temperature, salt addition, and pH were evaluated. Under the optimum extraction conditions, the method resulted in low limits of detection and quantification within the range 0.05–0.21ngg−1 in honey (15–70ngL−1 in solution) and 0.15–1.1ngg−1 in honey (45–210ngL−1 in solution), respectively. Enrichment factors and extraction recoveries were in the ranges of 1943–1994 and 97–100%, respectively. The method precision was evaluated at 1.5ngg−1 of each analyte, and the relative standard deviations were found to be less than 4% for intra-day (n=6) and less than 6% for inter-days. The method was successfully applied to the analysis of honey samples and difenoconazole was determined at ngg−1 levels.

Determination of micronutrient minerals in coconut milk by ICP OES after ultrasound-assisted extraction procedure

In this paper, multivariate optimisation was applied to the development of an ultrasound-assisted multi-elemental extraction procedure to measure micronutrient minerals in coconut milk samples by inductively coupled plasma optical emission spectrometry (ICP OES). The factorial and Doehlert designs were applied to optimise the process variables. Using this approach, the optimal experimental condition for 1.0g of coconut milk was determined: 10min sonication time; 20.0mL of HNO3 (1.0molL−1); and 5min centrifugation time at 2000rpm. The extraction percentage was calculated using the analyte concentration obtained by a total digestion procedure as a reference. Accuracy was assessed by analysing a certified reference material (NIST 1549). The proposed procedure was applied to the analysis of commercial samples of coconut milk, and the resulting concentration ranges, in μgg−1, were as follows: 5.8–131 (Ca), <0.27–1.56 (Cu), 0.76–5.52 (Fe), 212–1781 (K), 14.9–201 (Mg), <0.11–3.88 (Mn), 274–625 (Na), 26–341 (P) and <0.74–3.2 (Zn). The method described offers a rapid, easy and efficient sample preparation method for direct determination of micronutrient minerals in coconut milk by ICP OES.

Optimisation of a sample preparation method for the determination of fumonisin B1 in rice

A simple, rapid and cost-effective sample preparation method for the determination of fumonisin B1 in rice was optimised using a strategy of sequential experimental designs. Initially, a Plackett–Burman design was applied to select the statistically significant variables for the determination of fumonisin B1, and then, a central composite rotatable design was used to define the optimal conditions of these variables. The method involves extraction with a 50% acetonitrile aqueous solution and glacial acetic acid, liquid–liquid partitioning with addition of anhydrous sodium sulphate and sodium chloride, followed by dispersive SPE clean-up with diatomaceous earth. The final extract was analysed by HPLC-FLD after precolumn derivatisation with ortho-phthaldialdehyde. The optimised method was validated for selectivity, linearity, matrix effect, limits of detection and quantification, trueness, and precision, and then applied to commercial samples of polished rice. This is the first report of the occurrence of fumonisin B1 in commercial samples of polished rice from the Southeast region of Brazil.

Comparison between conventional solid phase extraction and its simplified method for HPLC determination of five flavonoids in orange, tangerine, and lime juice samples

Summary Although conventional solid phase extraction (SPE) is an applicable routine method for extraction of different analytes from various matrices, it requires more time and volume of solvent and sample rather than other routine laboratory extraction methods. In this study, a rapid and simplified sample preparation method based on SPE was studied by eliminating some steps, such as conditioning and washing, for extraction of diosmin, eriocitrin, narirutin, naringin, and hesperidin in orange, tangerine, and lime juice samples. The separation of these flavonoids was achieved by using a C8 column with a mobile phase comprised of water-acetonitrile-acetic acid (78:21:1, v/v/v) at a flow rate of 0.85 mL min−1 and UV detection at 280 nm. To examine the applicability of this method, effective parameters such as type of adsorbent, type and volume of elution solvent, ionic strength, and pH of the sample were studied and applied to the comparison between the conventional SPE and the simplified method. The best re...

Rapid Sample Preparation for Molecular Biological Food Analysis Based on Magnesium Chloride

The prerequisites of rapid and reliable direct quantification of foodborne pathogens deem prior sample preparation a necessity. This study describes a new system for the modular and rapid (3 h) sample preparation method “Matrix-Lysis” for the quantification of Listeria monocytogenes and Salmonella typhimurium from food. In this study a new chemical concept is introduced for Matrix-Lysis, based on solubilization of proteins through the preferential interaction with MgCl2. The results of this study show that the new system has many advantages compared to the previously described Matrix-Lysis systems, both regarding the performance of the whole method as well as the chemistry employed for solubilization of the foodstuffs. Artificial contamination experiments showed average recovery rates of 82 % for S. typhimurium and 73 % for L. monocytogenes from several foodstuffs with detection limits <10 CFU/g. Examination of naturally contaminated acid curd cheese samples, from a recent L. monocytogenes outbreak in Austria and Germany, with the new buffer system resulted in 100 % relative accuracy, 100 % relative specificity, and 100 % relative sensitivity compared to the ISO 11290-1 standard method. The results demonstrate the excellent applicability of Matrix-Lysis as sample preparation for the direct quantification of foodborne pathogens with molecular biological, as well as microbiological methods and the new chemical concept described enables the widespread use of this method for routine applications.

Supercritical fluid extraction of 1,1-dimethylhydrazine from peaty soils

Supercritical fluid extraction of a highly toxic rocket-fuel component (1,1-dimethylhydrazine, UDMH) from peaty soils with carbon dioxide is proposed as a rapid method of sample preparation for ion-chromatographic analysis. Optimum conditions of extraction have been determined. The efficiency of the UDMH extraction can be improved by basic treatments of the soil sample and by the use of proper additives to SC CO2 (cosolvents).

Solvent Bar Microextraction with HPLC for Determination and Protein-Binding Characteristics of Oleanolic Acid and Ursolic Acid

An improved solvent bar microextraction (SBME) combined with HPLC was applied to rapidly determine oleanolic acid (OA) and ursolic acid (UA) in Chinese medicinal herbs (CMHs), and to investigate drug–protein binding. Variables influencing SBME were investigated and optimized. Under optimal conditions, the linear ranges of 3.6–1,820 ng mL−1 for OA and 4.2–2,080 ng mL−1 for UA were obtained with square correlation coefficients of 0.9996 and 0.9997, respectively. The detection limits were 1.3 ng mL−1 for OA and 1.5 ng mL−1 for UA. The relative standard deviations were lower than 9.4 %. The protein-binding rates, the numbers of binding sites, and the binding constants of OA and UA were also obtained via the method. It has been demonstrated that SBME can not only be a simple, rapid and efficient sample preparation method for determination of active components from CMHs but also be a potential research protocol for protein-binding interactions.

Application of Rapid Sample Preparation Method and Monitoring for Cholesterol Content in Chicken Egg and Egg powder

The aim of this study was to develop sample preparation method and evaluate the exact cholesterol content in egg and egg powder purchased from Korean markets, and to determine whether significant differences exist among various egg products, since a variety of products are available in Korean markets and there are no recent databases for cholesterol. To evaluate the cholesterol content in chicken egg sold in Korean local market, a simple method using non-heated saponification to determine cholesterol for emulsified foods was applied. The results of recovery for egg and egg powder were in a range of 92.4-105.0%, with a relative standard deviation between 1.1% and 2.8% by using gas chromatography-flame ionization detector. Therefore, the total cholesterol content in whole egg was estimated between 160.8 and 226.3 mg/egg (AV(average) <TEX>$186.8{\pm}3.5$</TEX>), which is similar or lower than previously reported levels. The value for cholesterol in egg powder was estimated between 2.94 and 3.49 mg/g (AV <TEX>$3.23{\pm}0.15$</TEX>). We suggested method that can be applicable to chicken egg and egg powder matrix as providing rapid and accurate determination of cholesterol in egg and egg powder. This information will be helpful for processed food producers for deciding food labels of cholesterol content.

Simple fabrication of hydrophobic surface target for increased sensitivity and homogeneity in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of peptides, phosphopeptides, carbohydrates and proteins

To enhance sample signals and improve homogeneity in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) analysis, a simple, rapid, and efficient sample preparation method was developed in this study. Polydimethylsiloxane (PDMS) was coated on a stainless steel MALDI plate, forming a transparent, hydrophobic surface that enhanced sample signals without producing observable background signals. Compared to the use of an unmodified commercial metal MALDI plate, peptide signals were enhanced by ~7.1–11.0-fold due to the reduced sample spot area of the PDMS-coated plate, and showed improved peptide mass fingerprinting (PMF) and MS/MS peptide sequencing results. In the analysis of phosphopeptides and carbohydrates with a 2,5-dihydroxybenzoic acid (DHB) matrix, the PDMS-coated plate showed improved sample homogeneity and signal enhancements of ~5.2–8.2-fold and ~2.8–3.2-fold, respectively. Improved sensitivity in the observation of more unique fragment ions by MS/MS analysis, to successfully distinguish isomeric carbohydrates, was also illustrated. In protein analysis with a sinapinic acid (SA) matrix, a ~3.4-fold signal enhancement was observed. The PDMS film coating was easily removed and refabricated to avoid sample carryover, and was applicable to diverse commercial MALDI plates. The PDMS-coated approach is a simple, practical, and attractive method for enhancing analyte signals and homogeneity.

UPLC를 이용한 염모제품에 함유된 유효성분에 대한 품질검사

To identify oxidative hair dyes in hair-coloring products, the thin-layer chromatography (TLC) screening method was used in accordance with Korean Quasi-drug Codex. However, the TLC method is not reliable when there are very small amount of materials to be tested or when values of several components are similar. In this study, Ultra Performance Liquid Chromatography (UPLC) with a rapid sample preparation method was developed for the reliable and sensitive identification of active components contained in oxidative hair-coloring products. Hexane-distilled water was used for the extraction of active components contained in the products prior to UPLC analysis. The limit of detection of active components was 6.7-77.9 , and the limit of quantitation was 22.3-259.7 . Except for -naphthol, the range of recovery ratio was 96.2-101.5%. From this study, we demonstrated that oxidative active hair-coloring components can easily be analyzed by rapid extraction method followed by UPLC analysis.

In Vivo Solid-Phase Microextraction with in Vitro Calibration: Determination of Off-Flavor Components in Live Fish

The presence of off-flavor compounds in fish represents a significant economic problem encountered in aquaculture production. The off-flavor compounds are due to the absorption of substances produced by a range of microorganisms. Currently, a number of strategies have been used to prevent or limit the growth of these microorganisms. Therefore, it is important to evaluate the effectiveness of strategies via monitoring the concentrations of off-flavor compounds in the recirculating aquaculture system. In vivo solid-phase microextraction (SPME), a rapid and simple sample preparation method, will allow monitoring the concentration of off-flavor compounds in live fish. In this research, geosmin and 2-methylisoborneol (2-MIB) produced by cyanobacteria and actinomycetes, which are the major sources for "earthy" and "muddy" flavors in fish, were selected as representatives. In order to accurately quantify these compounds in fish muscle, two kinetic calibration methods, on-fiber standardization and measurement using predetermined sampling rate, were used as quantification methods, which were both validated by traditional methods. The detection limit of in vivo SPME in fish muscle was 0.12 ng/g for geosmin and 0.21 ng/g for 2-MIB, which are both below the human sensory thresholds.

Homogeneous duplex polymerase chain reaction assay using switchable lanthanide fluorescence probes

We have developed a duplex polymerase chain reaction (PCR) assay based on switchable lanthanide chelate complementation probes. In the complementation probe technology, two nonfluorescent oligonucleotide probes, one labeled with a lanthanide ion carrier chelate and another with a light absorbing antenna ligand, form a fluorescent complex by self-assembly of the reporter molecules when the two probes are hybridized in adjacent positions to the target DNA. Here we report the synthesis of a new terbium(III) (TbIII) ion carrier chelate and a new light-absorbing antenna ligand for TbIII and the development of a duplex Chlamydia trachomatis (Ct) PCR assay. For the detection of Ct in urine samples, a specific sequence in Ct cryptic plasmid was amplified and detected using europium(III) (EuIII) complementation probes. An internal amplification control was amplified in each reaction and detected using TbIII complementation probes to verify the Ct negative results. Ct bacteria were concentrated from urine samples with a rapid and simple centrifugation-based sample preparation method. Good diagnostic accuracy (99–100%) was achieved, and also Ct positive reactions yielded a very high EuIII signal-to-background ratio (maximum of 244). High performance of the complementation probes is advantageous when sample may contain impurities after a simple sample preparation.

What Is the Opposite of Pandora’s Box? Direct Analysis, Ambient Ionization, and a New Generation of Atmospheric Pressure Ion Sources

The introduction of DART and DESI sources approximately seven years ago led to the development of a new series of atmospheric pressure ion sources referred to as "ambient ionization" sources. These fall into two major categories: spray techniques like DESI or plasma techniques like DART. The selectivity of "direct ionization," meaning analysis without chromatography and with little or no sample preparation, depends on the mass spectrometer selectivity. Although high resolution and tandem mass spectrometry are valuable tools, rapid and simple sample preparation methods can improve the utility of ambient ionization methods. The concept of ambient ionization has led to the realization that there are many more ways to form ions than might be expected. An interesting example is the use of a flint-and-steel spark source to generate ions from compounds such as phenolphthalein and Gramicidin S.

Evaluation of HemogloBind™ treatment for preparation of samples for cholinesterase analysis.

Acetylcholine is an essential neurotransmitter found throughout the nervous system. Its action on postsynaptic receptors is regulated through hydrolysis by various carboxylesterases, especially cholinesterases (ChEs). The acute toxicity of organophosphate (OP) compounds is directly linked to their action as inhibitors of ChE. One widely used assay for evaluating ChE activity is a spectrophotometric method developed by Ellman et al. When the enzyme source is from tissues or, in particular, blood, hemoglobin displays a spectrophotometric peak at the same wavelength used to analyze cholinergic activity. This creates a substantial background that interferes with the Ellman’s assay and must be overcome in order to accurately monitor cholinesterase activity. Herein, we directly compare blood processing methods: classical method (1.67 ± 0.30 U/mL) and HemogloBind™ treatment (1.51 ± 0.17 U/mL), and clearly demonstrate that pretreatment of blood samples with Hemoglobind™ is both a sufficient and rapid sample preparation method for the assessment of ChE activity using the Ellman’s method.

Chemometrics assisted dispersive liquid-liquid microextraction for quantification of seven UV filters in urine samples by HPLC-DAD

In the present study, dispersive liquid-liquid microextraction followed by high performance liquid chromatography-diode array detection has been developed as simple, rapid, accurate, and efficient sample preparation method for simultaneous determination of seven organic UV filters in urine samples. The influence of the main effects as well as their interactions was studied through a 2((6-2)) fractional factorial design. The candidate parameters were: type and volume of dispersant and extraction solvents, sample pH, and salt concentration. Under final optimal conditions, the analytes were extracted from 5 mL of samples by addition of 0.5 mL of acetonitrile (dispersing solvent) containing 70 μL of carbon tetrachloride (extraction solvent), without modifying the pH of the solution and applying the (+1) level of salt concentration (10% w/v NaCl). The assay was linear (R(2) > 0.997), relative recoveries ranged from 86.9 up to 97.3% and the LOQs between 3 and 45 ng mL(-1) were obtained. The intra- and interday RSDs were lower than 5 and 8% at the middle point of the linear range, respectively. The proposed method was successfully applied to different volunteer urine samples and it was shown that the extraction efficiency was not affected by the type of urine samples.

A Novel Sample Preparation Method Using Rapid Nonheated Saponification Method for the Determination of Cholesterol in Emulsified Foods

In this study, nonheated saponification was employed as a novel, rapid, and easy sample preparation method for the determination of cholesterol in emulsified foods. Cholesterol content was analyzed using gas chromatography with a flame ionization detector (GC-FID). The cholesterol extraction method was optimized for maximum recovery from baby food and infant formula. Under these conditions, the optimum extraction solvent was 10 mL ethyl ether per 1 to 2 g sample, and the saponification solution was 0.2 mL KOH in methanol. The cholesterol content in the products was determined to be within the certified range of certified reference materials (CRMs), NIST SRM 1544 and SRM 1849. The results of the recovery test performed using spiked materials were in the range of 98.24% to 99.45% with an relative standard devitation (RSD) between 0.83% and 1.61%. This method could be used to reduce sample pretreatment time and is expected to provide an accurate determination of cholesterol in emulsified food matrices such as infant formula and baby food. A novel, rapid, and easy sample preparation method using nonheated saponification was developed for cholesterol detection in emulsified foods. Recovery tests of CRMs were satisfactory, and the recoveries of spiked materials were accurate and precise. This method was effective and decreased the time required for analysis by 5-fold compared to the official method.

Integration of shot-gun proteomics and bioinformatics analysis to explore plant hormone responses

BackgroundMultidimensional protein identification technology (MudPIT)-based shot-gun proteomics has been proven to be an effective platform for functional proteomics. In particular, the various sample preparation methods and bioinformatics tools can be integrated to improve the proteomics platform for applications like target organelle proteomics. We have recently integrated a rapid sample preparation method and bioinformatics classification system for comparative analysis of plant responses to two plant hormones, zeatin and brassinosteroid (BR). These hormones belong to two distinct classes of plant growth regulators, yet both can promote cell elongation and growth. An understanding of the differences and the cross-talk between the two types of hormone responses will allow us to better understand the molecular mechanisms and to identify new candidate genes for plant engineering.ResultsAs compared to traditional organelle proteomics, the organelle-enrichment method both simplifies the sample preparation and increases the number of proteins identified in the targeted organelle as well as the entire sample. Both zeatin and BR induce dramatic changes in signaling and metabolism. Their shared-regulated protein components indicate that both hormones may down-regulate some key components in auxin responses. However, they have shown distinct induction and suppression of metabolic pathways in mitochondria and chloroplast. For zeatin, the metabolic pathways in sucrose and starch biosynthesis and utilization were significantly changed, yet the lipid biosynthesis remained unchanged. For BR, lipid biosynthesis and β-oxidation were both down-regulated, yet the changes in sucrose and starch metabolism were minor.ConclusionsWe present a rapid sample preparation method and bioinformatics classification for effective proteomics analysis of plant hormone responses. The study highlighted the largely differing response to zeatin and brassinosteroid by the metabolic pathways in chloroplast and mitochondria.

Rapid determination of cholesterol in milk containing emulsified foods

In this study, a rapid and easy sample preparation method that involved no-heating saponification and dSPE (dispersive solid phase extraction) clean-up was developed to determine the level of cholesterol in milk containing emulsified foods (infant formula, baby food, cheese). The developed method utilised high performance liquid chromatography with an ultraviolet detector (HPLC-UVD) as a separation instrument. The optimum extraction conditions were determined as 10mL isopropyl alcohol with 8.0g (NH4)2SO4 per 1g sample, and saponification was achieved using 25mg KOH, 1.6g NaCl and 100mg of a silica based NH2. Cholesterol levels determined using CRMs (NIST SRM 1849 and 1544) were in the range of the certificated value and the recovery test using spiked materials ranged from 94.34% to 102.34% with a RSD of 0.63–4.10%. This method enables the accurate determination of cholesterol with reduced sample preparation time.