Additional Extraction Step Research Articles

Mutual separation of Am and Cm using ADAAM (Alkyl DiAmide AMine) and reduction of volumes for liquid waste generated via batch-wise multistage extractions

ABSTRACT The mutual separation of Am and Cm is conducted using an alkyl-diamide amine (ADAAM(EH)) extractant. ADAAM(EH) exhibits an extremely high separation factor (SF) with respect to Am and Cm separation (SF = 5.9) in an HNO3-n-dodecane system. The batch-wise multistage extractions are performed using a system containing 0.2 M ADAAM(EH) and 1.5 M HNO3, with their distribution ratios (D) of the system for Am and Cm equal to 2.55 and 0.434, respectively. In this multistage extraction, 3 stages of feed, 9 stages involving scrubbing, and 11 stages involving the use of an organic solvent give 96.5% and 1.06% yields of Am and Cm, respectively. After the mutual separation, the aqueous and the organic phases including actinides increase four times in volume compared to the feed solution. Additional extraction steps are to reduce the volume of the solutions including actinides. We use tetraethyl diglycolamide (TEDGA)/water for Am concentration, tetraoctyl diglycolamide/n-dodecane for the intermediate extraction, and TEDGA/water for stripping in the Cm concentration step. Taking the additional steps, Am and Cm can be recovered in just two or three stages in the aqueous phases.

Isolation of Polyphenols from Two Waste Streams of Clingstone Peach Canneries Utilizing the Cloud Point Extraction Method

This study aimed to assess the feasibility of employing cloud point extraction (CPE) as an efficient way of extracting polyphenols from peach waste (PW). Four distinct food-grade surfactants (Genapol X-080, PEG 8000, Tween 80, and lecithin) were evaluated at concentrations ranging from 2–10% w/v to determine the efficiency of the technique in two separate PW streams [i.e., lye peeling waste stream (LPWS) and total wastewater stream (TWS)]. Low amounts (2% w/v) of surfactants in a single-step CPE were found to result in less than ~61% polyphenol recovery in LPWS and less than ~69% polyphenol recovery in the TWS, necessitating additional extraction steps. In both PW streams, the single-step polyphenol recovery was improved by 25–67% utilizing a higher amount of surfactants (5–10% w/w), leading to a statistically significant figure (p < 0.05). The CPE procedure was conducted under optimal conditions, including a temperature of 65 °C, a sodium chloride concentration of 3% w/v, a pH level of 3.5, and a surfactant concentration of 5% w/v. The polyphenol recovery was efficient when the CPE procedure was conducted twice. Tween 80 proved to be the most efficient surfactant among the four tested surfactants, achieving recoveries above 98% in both PW streams. Under optimum extraction conditions, the total polyphenol content and antiradical activity of PW extracts were evaluated. The results showed statistically significant differences (p < 0.05) between the two PW streams, with the LPWS having approximately 12 times higher polyphenol content and being more potent, achieving ~64% antiradical activity. Using the LPWS instead of the TWS is a more cost-effective and feasible option for the industry. In addition, the considerable volume of the TWS makes it challenging to handle and demands a correspondingly major amount of surfactant. Considering that Tween 80 is a low-toxicity surfactant and that the CPE method is simple, fast, cost-effective, highly accurate, and selective, the extracted polyphenols from two PW streams could be exploited as natural antioxidants to be used directly in the food industry. These findings could have major implications for the manufacturing of sustainable and naturally-derived food additives.

Nanoemulsion-Loaded Capsules for Controlled Delivery of Lipophilic Active Ingredients.

Nanoemulsions have become ideal candidates for loading hydrophobic active ingredients and enhancing their bioavailability in the pharmaceutical, food, and cosmetic industries. However, the lack of versatile carrier platforms for nanoemulsions hinders advanced control over their release behavior. In this work, a method is developed to encapsulate nanoemulsions in alginate capsules for the controlled delivery of lipophilic active ingredients. Functional nanoemulsions loaded with active ingredients and calcium ions are first prepared, followed by encapsulation inside alginate shells. The intrinsically high viscosity of the nanoemulsions ensures the formation of spherical capsules and high encapsulation efficiency during the synthesis. Moreover, a facile approach is developed to measure the nanoemulsion release profile from capsules through UV–vis measurement without an additional extraction step. A quantitative analysis of the release profiles shows that the capsule systems possess a tunable, delayed‐burst release. The encapsulation methodology is generalized to other active ingredients, oil phases, nanodroplet sizes, and chemically crosslinked inner hydrogel cores. Overall, the capsule systems provide promising platforms for various functional nanoemulsion formulations.

The analysis of nitrate explosive vapour samples using Lab-on-a-chip instrumentation

The detection and analysis of explosives and explosive-related compounds is a heightened priority in recent years for homeland security and counter-terrorism applications. This study aimed to evaluate the use of a commercial Lab-On-a-Chip (LOC) instrument for the analysis of explosive vapours, with the long-term goal of developing a portable instrument for passively detecting explosives in air samples. A simple method to collect explosive vapour residues was developed using a glass vial containing varying amounts of the target explosives (1 mg/mL). Standards were diluted to the desired concentration in 150 μL of acetone to facilitate the evaporation. The top of the vial was covered with a circular 0.5 cm diameter filter paper and exposed to a range of temperatures from 22 °C to 80 °C for 15 min. Following evaporation, the filter paper chads were folded and inserted into the LOC wells containing the separation buffer for the analysis, avoiding any further extraction step. After successfully separating and detecting eight explosives via liquid analysis, three explosives were chosen as targets for the vapour analysis experiments. 1,3,5-Trinitrobenzene (TNB), 2,4,6-Trinitrotoluene (TNT), and 2,4,6-Trinitrophenylmethylnitramine (Tetryl) were successfully separated, detected and identified following the vapour extraction of explosive standards onto filter paper chads. Limits of detection for the liquid analysis were demonstrated to be 2.32 ng for TNB, 2.35 ng for Tetryl, and 3.25 ng for TNT. The minimum detectable mass found for the vapour analysis was 6.03 for TNB, 9.99 ng for TNT, and 14.22 ng for Tetryl. The average recovery from the paper chads was 29% for Tetryl, 47% for TNB, and 75% for TNT (n = 4), comparable with findings from previous studies. Results show that a minimum temperature of 40 °C is necessary to vaporize the compounds using acetone, while the best results were achieved when heating the vial to 80 °C. The use of a filter paper to collect the explosives residues, avoiding any additional extraction step, and the ability to analyze these compounds using a LOC instrument, makes this approach a future alternative method for explosive residues detection in the headspace.

Multivariate Analysis of MALDI Imaging Mass Spectrometry Data of Mixtures of Single Pollen Grains.

Mixtures of pollen grains of three different species (Corylus avellana, Alnus cordata, and Pinus sylvestris) were investigated by matrix-assisted laser desorption/ionization time-of-flight imaging mass spectrometry (MALDI-TOF imaging MS). The amount of pollen grains was reduced stepwise from > 10 to single pollen grains. For sample pretreatment, we modified a previously applied approach, where any additional extraction steps were omitted. Our results show that characteristic pollen MALDI mass spectra can be obtained from a single pollen grain, which is the prerequisite for a reliable pollen classification in practical applications. MALDI imaging of laterally resolved pollen grains provides additional information by reducing the complexity of the MS spectra of mixtures, where frequently peak discrimination is observed. Combined with multivariate statistical analyses, such as principal component analysis (PCA), our approach offers the chance for a fast and reliable identification of individual pollen grains by mass spectrometry. Graphical Abstract ᅟ.

Reporter-Embedded SERS Tags from Gold Nanorod Seeds: Selective Immobilization of Reporter Molecules at the Tip of Nanorods.

Reporter-embedded (RE) tags are a new generation of sensitive, stable surface enhanced Raman scattering (SERS) tags with Raman reporters embedded between gold nanoparticle (NP) cores and gold (or silver) shells. Most of the reported RE tags have been designed using Au nanospheres as a seed material. Herein, we investigated the synthesis and SERS properties of AuNR/reporter/Ag tags by using gold nanorod (AuNR) seeds with anisotropic physical and optical features. Several highlighted points were discovered, including the following: (1) The cetyltrimethylammonium bromide (CTAB) layer induced the coexistence of chemically and physically adsorbed Raman reporters on AuNR. Conventional washing of the AuNR-reporter complex with water results in the formation of an "internal-external" mixed tag. To obtain a "pure" RE structure, an additional extraction step involving a CTAB solution was essential. (2) The anisotropic distribution of CTAB on AuNR resulted in the preference of the Raman reporters to adsorb to the hotspot at the AuNR tip, which made it a perfect match for improving the SERS signal of the tag. (3) An anisotropic silver coating occurred with the shell thickness on the AuNR side growing much faster than the shell thickness at the tip. This feature ensured that the tag grew to a suitable size with enough silver for SERS enhancement without shadowing the effective Raman reporters at the tip too much. (4) RE tags showed better in vitro and in vivo signal stabilities compared with their external labeling counterparts. Moreover, a novel pH-sensitive SERS peak test was proposed by using 4-mercaptobenzoic acid as the Raman reporter to verify thin coverage by a silver layer. We believe this tag can be broadly applied for molecular detection and bioimaging, and the proposed preparation and structure verification methods can provide universal guidance in the design of novel RE tags.

OFFGEL fractionation of peptides: Where really is your sample?

Shotgun proteomics of complex samples is generally coupled with at least one peptide fractionation step and, to this effect, peptide isoelectric focusing (IEF) in immobilized pH gradient (IPG) is one of the most used techniques. Fractionation with the OFFGEL 3100 Agilent Technologies apparatus allows the easy recovery of peptides that, after focusing, diffuse into the liquid phase above the gel strip. In this work we investigate the efficiency of peptide diffusion during OFFGEL fractionation and demonstrate that a recovery based only on the spontaneous diffusion process is far from being optimal. We show that a simple additional extraction step with acetonitrile increases of about 40% the amount of material that can be recovered after the focusing. Moreover, we show that the two populations of peptides obtained from the passive elution and from the extraction process are also qualitatively different and only partially overlapping.

Performance of two MALDI-TOF MS systems for the identification of yeasts isolated from bloodstream infections and cerebrospinal fluids using a time-saving direct transfer protocol

The rapid and correct identification of pathogens is of paramount importance for the treatment of patients with invasive infections. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) can speed up the identification of bacteria and fungi and has quickly been embraced by medical microbiology laboratories worldwide. Different MALDI-TOF systems have been compared in studies focussing on identification rates of different pathogens. Another aspect that has not been systematically assessed is the performance in daily routine and handling, which is important especially for microbiology routine laboratories. We compared two widespread commercial systems, Microflex LT Biotyper (Bruker) and VitekMS (bioMérieux), for the identification of 210 relevant clinical yeasts under routine conditions, using a time-saving direct transfer protocol. We assessed the need for an additional extraction step, the threshold for species identification and the duration of measurements with the two systems. The tested yeasts included 34 Candida albicans isolates, 144 non-albicans Candida spp. and 32 yeasts of different genera. The results of the two MS systems were compared with that of biochemical identification and, in case of discrepancies, DNA sequencing of the internal transcribed spacer or the large subunit of ribosomal DNA. Both systems correctly identified 96.2% of isolates [202/210, non-significant (n.s.)]. Misidentifications were observed for VitekMS only (n=5, no major errors, n.s.). VitekMS was the slower system (19.8 vs. 8.0min for 10 samples, p=0.002) but had the advantage of a more effective direct transfer protocol with less need for an additional extraction step.

Assessing vehicle effects on skin absorption of non-volatile compounds using membrane-coated fiber arrays

Context: The membrane-coated fiber (MCF) array technique was previously demonstrated to be a rapid, quantitative assessment of the percutaneous absorption of volatile compounds, capable of studying chemical mixtures and their synergistic effects. In particular, three commercially available fiber coatings (polydimethylsiloxane, polyacrylate and carbowax) were shown to be representative of molecular interactions relevant to skin absorption.Objective: This study evaluated the potential use of these fibers as a high throughput screening approach for topical formulations. More specifically, the MCFs were evaluated for their ability to predict the skin permeability of caffeine (CF), cortisone (CT), mannitol (MN) and salicylic acid (SA) applied in water or ethanol as unsaturated and/or saturated concentrations, using an additional extraction step to obtain analytes appropriate for liquid chromatography.Methods: Compound extraction from a donor solution was carried out by fiber immersion for a set period of time. The compound was then re-extracted into acetonitrile and quantitated by liquid chromatography coupled to a mass spectrometer. Radiolabel equivalents of the compounds were analyzed in a similar manner with the exception of detection method used.Results: Fiber extraction of radiolabeled compounds (CF, MN, SA in water) was proved to be unsuccessful, whereas the extraction of their non-radiolabeled equivalents (CF, CT, SA) revealed poor linearity, and poor between-day and within-day reproducibility. Similar unsatisfactory results were observed regardless of whether a single fiber was used, or whether multiple fibers were used simultaneously. Furthermore, incompatibility between the MCF and solvent (vehicle) was observed, which disputes its potential use as a formulation screening technique.Conclusion: On the basis of these findings, the MCF array is not suitable to describe the vehicle effects on skin absorption of non-volatile compounds.

Biotests for environmental quality assessment of composted sewage sludge

The quality of sewage sludge-based products, such as composts and growth media, is affected by the contamination of sewage sludge with, potentially, hundreds of different substances. Therefore, it is difficult to achieve the reliable environmental quality assessment of sewage sludge-based products solely based on chemical analysis. In the present work, we demonstrate the use of the kinetic luminescent bacteria test (ISO 21338) to evaluate acute toxicity and the Vitotox™ test to monitor genotoxicity of sewage sludge and composted sewages sludge. In addition, endocrine-disrupting and dioxin-like activity was studied using yeast-cell-based assays. The relative contribution of industrial waste water treated at the Waste Water Treatment Plants led to elevated concentrations of polyaromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and polychlorinated dibenzo-p-dioxins and -furans (PCDD/F) in sewage sludge. The effect of elevated amounts of organic contaminants could also be identified with biotests able to demonstrate higher acute toxicity, genotoxicity, and potential for endocrine-disruptive properties. Additional extraction steps in kinetic luminescent bacteria test with DMSO and hexane increased the level of toxicity detected. Composting in a pilot-scale efficiently reduced the amounts of linear alkylbenzensulphonates (LASs), nonylphenols and nonylphenolethoxylates (NPE/NPs) and PAH with relative removal efficiencies of 84%, 61% and 56%. In addition, decrease in acute toxicity, genotoxicity and endocrorine-disrupting and dioxin-like activity during composting could be detected. However, the biotests did have limitations in accessing the ecotoxicity of test media rich with organic matter, such as sewage sludge and compost, and effects of sample characteristics on biotest organisms must be acknowledged. The compost matrix itself, however, which contained a high amount of nutrients, bark, and peat, reduced the sensitivity of the genotoxicity tests and yeast bioreporter assays.

A simple quantitation method for benzoylecgonine from oral fluid, blood, and urine samples used for determining 22 illicit and licit drugs by GC–MS with liquid–liquid extraction

The analytical system used for determination of 22 illicit and licit drugs by gas chromatography–mass spectrometry following liquid–liquid extraction has been extended with an additional extraction step with n-butyl acetate:CH2Cl2 = 1:8 to determine benzoylecgonine from the same oral fluid, blood, or urine samples. The extraction recovery of benzoylecgonine was 39.7% for oral fluid, 14.9% for blood, and 26.7% for urine; the cutoffs were 25, 8.0, and 8.0 ng/ml, respectively. The method was fully validated and proved suitable in further proficiency tests and for analysis of 2738 oral fluid, 197 blood, and 1298 urine samples.

Procedure for tissue sample preparation and metabolite extraction for high-throughput targeted metabolomics

Reproducible quantification of metabolites in tissue samples is of high importance for characterization of animal models and identification of metabolic changes that occur in different tissue types in specific diseases. However, the extraction of metabolites from tissue is often the most labor-intensive and error-prone step in metabolomics studies. Here, we report the development of a standardized high-throughput method for rapid and reproducible extraction of metabolites from multiple tissue samples from different organs of several species. The method involves a bead-based homogenizer in combination with a simple extraction protocol and is compatible with state-of-the-art metabolomics kit technology for quantitative and targeted flow injection tandem mass spectrometry. We analyzed different extraction solvents for both reproducibility as well as suppression effects for a range of different animal tissue types including liver, kidney, muscle, brain, and fat tissue from mouse and bovine. In this study, we show that for most metabolites a simple methanolic extraction is best suited for reliable results. An additional extraction step with phosphate buffer can be used to improve the extraction yields for a few more polar metabolites. We provide a verified tissue extraction setup to be used with different indications. Our results demonstrate that this high-throughput procedure provides a basis for metabolomic assays with a wide spectrum of metabolites. The developed method can be used for tissue extraction setup for different indications like studies of metabolic syndrome, obesity, diabetes or cardiovascular disorders and nutrient transformation in livestock.

Magic angle spinning magnetic resonance: a novel method opening up translational research into NAFLD?

NAFLD (non-alcoholic fatty liver disease) and NASH (non-alcoholic steatohepatitis) are of increasing importance, both in connection with insulin resistance and with the development of liver cirrhosis. Histological samples are still the 'gold standard' for diagnosis; however, because of the risks of a liver biopsy, non-invasive methods are needed. MAS (magic angle spinning) is a special type of NMR which allows characterization of intact excised tissue without need for additional extraction steps. Because clinical MRI (magnetic resonance imaging) and MRS (magnetic resonance spectroscopy) are based on the same physical principle as NMR, translational research is feasible from excised tissue to non-invasive examinations in humans. In the present issue of Clinical Science, Cobbold and co-workers report a study in three animal strains suffering from different degrees of NAFLD showing that MAS results are able to distinguish controls, fatty infiltration and steatohepatitis in cohorts. In vivo MRS methods in humans are not obtainable at the same spectral resolution; however, know-how from MAS studies may help to identify characteristic changes in crowded regions of the magnetic resonance spectrum.

Determination of the fruit content of cherry fruit preparations by gravimetric quantification of hemicellulose

A method recently developed for the determination of the fruit content of strawberry fruit preparations by gravimetric quantification of hemicellulose was extended to cherry fruit preparations. Isolation of the alcohol-insoluble residue (AIR) and sequential fractionation of the cell wall compounds from cherries ( Prunus cerasus L. cv. ‘Oblacinska’) was performed yielding the amount of fresh cherries per gram hemicellulose. Cherry fruit preparations with varying fruit contents (30–40%) were produced using different hydrocolloid systems (pectin, starch, guar gum, xanthan gum, carrageenan, and combinations thereof). After separation of the hydrocolloids by enzymatical digestion and successive extraction, the fruit preparations were subjected to AIR isolation. The AIR was fractionated to yield the hemicellulose fraction, which was quantified gravimetrically for the calculation of the fruit content. Compared to strawberries, modifications including additional extraction steps for the sequential fractionation were required to separate the pectin of the cherries exhaustively. Calculated and initial fruit contents were in good agreement for the single hydrocolloid components pectin and starch as well as for the combinations pectin/xanthan gum and pectin/carrageenan (26.8% vs. 30%, 38.6% vs. 40%, 42.5% vs. 40%, 37.6% vs. 40%, and 41.2% vs. 40%), whereas the preparations produced with more complex hydrocolloid systems (pectin/xanthan gum/guar gum and starch/xanthan/guar gum) showed larger deviations in their contents (46.2% vs. 40%, 49.6% vs. 40%). It is concluded that the novel method is generally suitable for the determination of the fruit content of fruit preparations, but steps of sample preparation need to be individually adapted to the different fruit matrices.

A rapid high resolution chromatographic method for serum barbiturates: Instant thin-layer chromatography

1. Barbiturates were extracted from acidified serum with chloroform and diethyl ether. 2. The various barbiturates in the extract were separated by chromatography on fibre glass sheets impregnated with silica gel. 3. The barbiturates were visualized by being sprayed with Rhodamine B and the fluorescence quenching effect observed under ultraviolet light. 4. The R f values of various barbiturates in three different solvent systems are described. 5. Serum barbiturates could be separated in 5 or 20 min into groups having clinical significance. Urinary barbiturates could also be identified in a similar manner but an additional extraction step was required.

A rapid high resolution chromatographic method for serum barbiturates: Instant thin-layer chromatography

Barbiturates were extracted from acidified serum with chloroform and diethyl ether. The various barbiturates in the extract were separated by chromatography on fibre glass sheets impregnated with silica gel. The barbiturates were visualized by being sprayed with Rhodamine B and the fluorescence quenching effect observed under ultraviolet light. The Rf values of various barbiturates in three different solvent systems are described. Serum barbiturates could be separated in 5 or 20 min into groups having clinical significance. Urinary barbiturates could also be identified in a similar manner but an additional extraction step was required.

A rapid high resolution chromatographic method for serum barbiturates: Instant thin-layer chromatography

Barbiturates were extracted from acidified serum with chloroform and diethyl ether. The various barbiturates in the extract were separated by chromatography on fibre glass sheets impregnated with silica gel. The barbiturates were visualized by being sprayed with Rhodamine B and the fluorescence quenching effect observed under ultraviolet light. The Rf values of various barbiturates in three different solvent systems are described. Serum barbiturates could be separated in 5 or 20 min into groups having clinical significance. Urinary barbiturates could also be identified in a similar manner but an additional extraction step was required.