Sample Preparation System Research Articles

Development of a Novel AMS-14C Sample Preparation System

Development of a Novel AMS-14C Sample Preparation System

LithoSpace: An Idea for an Automated System for in situ Petrographic Thin Section Preparation on Mars and Other Extraterrestrial Rocky Bodies

Optical microscopy analyses using thin sections is one of the most standard techniques in geology. It permits identification of most rock-forming minerals and it is essential for micropaleontology since observation in transmitted light is the only way to observe individual, or colonies of, microfossils in their mineralogical and textural context. Thin sections are also used for other techniques such as Raman spectroscopy. In the framework of exploration of the Martian surface and of the search for potential relics of microbial life, the possibility to observe petrographic thin sections in situ would be revolutionary. Nevertheless, although optical microscopes can be readily designed for space exploration, thin section preparation is not easy to do in situ on extraterrestrial rocky bodies due to the generally harsh environmental conditions and to the difficulty to automate a protocol that normally requires several human interventions. The aim of the LithoSpace project, supported by the French Space Agency (CNES) since 2014, is to work on the development of an automated system permitting preparation of petrographic thin sections on extraterrestrial bodies, such as Mars, the Moon or asteroids. In this paper, we describe the approaches used to study rocks in situ during space missions and compare them to those used on Earth. Then, the relevance of making thin sections during space exploration is highlighted from the description of the data that could be collected from the analysis of these specific samples, using new instruments and instruments already sent to Mars. The standard preparation protocol is described as well as previously envisioned or existing in situ sample preparation systems. We then propose a protocol for automation of the preparation. Particular attention is paid to demonstrating the feasibility of making thin sections in the absence of liquid water and with energy consumption compatible with automated space probes. Tests are then carried out to control the quality of the prepared samples. On the basis of the demonstrated feasibility, an automated system is proposed as a conceptual all-in-one system. Finally, a “proof-of-concept” model developed with the help of students at different educational levels is presented.

Ultrasound-assisted electromembrane extraction with supported semi-liquid membrane

Electromembrane extraction (EME), involving the migration of charged analytes across a supported liquid membrane (SLM) with an external power supply, is a promising sample preparation method in analytical chemistry. However, the presence of boundary double layers at the SLM/solution interfaces often restricts extraction efficiency. To avoid this, the current work proposed an ultrasound-assisted EME (UA-EME) method based on a novel type of supported semi-liquid membrane (SsLM). The characterizations showed that the SsLM was stable under ultrasound conditions. Ultrasound was found to reduce the boundary double layers and thus increase the mass transfer. Major operational parameters in UA-EME including ultrasound power density, temperature, applied voltage and extraction time were optimized with haloperidol, fluoxetine, and sertraline as model analytes. Under the optimal conditions, extraction recoveries of model analytes in water samples were in the range of 66.8%–91.6%. When this UA-EME method was coupled with LC-MS/MS for detection of the target analytes in human urine samples, the linear range of the analytical method was 10–1000 ng mL−1, with R2 > 0.997 for all analytes. The limits of detection (LOD) and limits of quantification (LOQ) were in the range of 1.7–2.1 ng mL−1 and 5.7–6.7 ng mL−1, respectively. The UA-EME expands the application field of ultrasound chemistry and will be very important in development of stable and fast sample preparation systems in the future.

Halocarbon emissions by selected tropical seaweeds exposed to different temperatures

Four tropical seaweeds, Gracilaria manilaensis Yamamoto & Trono, Ulva reticulata Forsskål, Kappaphycus alvarezii (Doty) L.M.Liao and Turbinaria conoides (J.Agardh) Kützing, collected from various habitats throughout Malaysia, were subjected to temperatures of 40, 35, 30, 25 and 20 °C in the laboratory. An exposure range of 21–38 °C is reported for Malaysian waters. The effect of the temperature exposures on the halocarbon emissions of the seaweeds were determined 4 and 28 h after treatment. The emission rates for a suite of six halocarbons commonly emitted by seaweeds, bromoform (CHBr3), dibromomethane (CH2Br2), diiodomethane (CH2I2), iodomethane (CH3I), dibromochloromethane (CHBr2Cl) and dichlorobromomethane (CHBrCl2), were measured using a cryogenic purge-and-trap sample preparation system coupled to a gas chromatography–mass spectrometry. The emission rate of CHBr3 was the highest of the six halocarbons for all the seaweeds under all the temperatures tested, followed by CH2Br2, and CH2I2. The emission rates were affected by temperature change and exposure duration, but overall responses were unique to each seaweed species. Larger decreases in the emissions of CHBr3, CH2Br2, CH2I2 and CHBr2Cl were found for K. alvarezii and T. conoides after 4 h at 40 °C. In both cases there was a >90% (p < 0.05) reduction in the Fv/Fm value suggesting that photosynthetic actitivity was severely compromised. After a 28 h exposure period, strong negative correlations (r = −0.69 to −0.95; p < 0.01) were observed between temperature and the emission of CHBr3, CH2Br2 and CH2I2 for U. reticulata, K. alvarezii and T. conoides. This suggests a potential decrease in the halocarbon emissions from these tropical seaweeds, especially where the temperature increase is a prolonged event. Strong correlations were also seen between seaweed chlorophyll and carotenoid pigment contents and the emission rates for CHBr3, CH2Br2 and CH2I2 (r = 0.48 to 0.96 and −0.49 to −0.96; p < 0.05). These results suggest that the regulation of halocarbon production versus reactive oxygen species production in seaweeds is an area worthy of further exploration.

Abstract 600: Liquid biopsy for neuroendocrine differentiation: Validation of a circulating tumor cell assay for synaptophysin

Abstract Protein expression of synaptophysin (SYP) is characteristic of neuroendocrine subtype tumors. In prostate cancer, neuroendocrine differentiation is correlated with disease progression, poor prognosis, and treatment resistance. Analysis of circulating tumor cells (CTCs) by multiparameter immunofluorescence (IF) microscopy allows non-invasive characterization of cancer cell biomarker expression in real time. This information can be helpful in prognosis, treatment selection, and patient stratification. Here we describe the validation of a biomarker IF assay for enumeration of CTCs and characterization of their SYP expression. The 0920-VB SYP CTC assay workflow includes processing blood samples to slides (AccuCyte® Sample Preparation System), staining slides with a panel of fluorescent markers (RarePlex® Staining Kit and Ventana® DISCOVERY® ULTRA immunostaining system), and multiparameter imaging and analysis (CyteFinder® Instrument). The panel consists of a nuclear dye, and antibodies against cytokeratins and EpCAM (to identify epithelial CTCs), CD45 (to exclude white blood cells) and SYP. Both clinical and spike-in samples were used for assay validation. The model CTCs used to generate spike-in samples included 22Rv1 (prostate carcinoma, SYP positive) and BT-474 (breast carcinoma, SYP negative). Performance metrics for the assay included accuracy, sensitivity, specificity, repeatability, and intermediate precision of SYP detection and CTC enumeration. Single-cell SYP mean fluorescence intensities (MFI) were analyzed to determine protein expression levels. An MFI threshold for SYP positivity was established by maximizing the classification accuracy of the positive and negative cell lines. Using this threshold, 98.7% of 22Rv1 cells were correctly classified as SYP-positive, and 99.7% of BT474 cells were correctly classified as SYP-negative with an inter-run coefficient of variation of 13.9% for the 22Rv1 cell line. In clinical prostate, breast and colon cancer patient samples, subsets of CTCs were found to be SYP positive with the expected cytoplasmic localization of the marker. In summary, this liquid biopsy assay provides an analytically sensitive and specific method for CTC enumeration and SYP biomarker expression analysis, allowing non-invasive detection of neuroendocrine differentiation. Citation Format: Jennifer Chow, A Anders Tevis, Edward Lo, Alisa Clein, Daniel E. Sabath, Arturo B. Ramirez, Eric P. Kaldjian, Tad George. Liquid biopsy for neuroendocrine differentiation: Validation of a circulating tumor cell assay for synaptophysin [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 600.

A New Highly Sensitive Realtime PCR Assay with Faster Turnaround Time for Detecting IDH1 and IDH2 mutations in Acute Myeloid Leukemia (AML) Patients

Introduction:Genetic profiling of AML reveals distinct molecular subgroups that inform disease classification and prognostic stratification (Tayyab M et al., J. Cancer Sci. Ther. 2014). Studies have demonstrated, that the Isocitrate Dehydrogenase Isoenzymes (IDH1/2), regulating cellular metabolism co-operate to result in AML (Papaemmanuil E et al., N. Engl. J. Med. 2016). Both IDH1/2 mutations produce neomorphic activity resulting in the overproduction of an onco-metabolite 2-hydroxyglutarate (2HG) (Gross S et al., J Exp Med. 2010). IDH1 mutations affect codon 132 and IDH2 mutations affect codon 140 or 172. IDH1 mutations have been reported to occur in 6-9% of adult and 1% of pediatric AML cases respectively, while IDH2 mutations have been reported to occur in 8-12% of adult and 1-2% of pediatric AML cases respectively (Abbas, S, et al., Blood. 2010; Andersson AK et al., Leukemia. 2011). The objective of this study was to conduct workflow analysis of the Abbott RealTime IDH1 and IDH2 assays in comparison to the Sanger based sequencing assays for IDH1/2 in blood/bone marrow specimens from subjects with advanced hematological malignancies. Next Generation Sequencing (NGS) was utilized to resolve samples with discordant results.Methods:This study was approved by Medical College of Wisconsin Institutional Review Board. The clinical specimens used were blinded during the study. Blood or bone marrow aspirates from 100 AML subjects sent to the BloodCenter of Wisconsin (BCW) were tested for IDH1/2 with four different assays (Abbott RealTime IDH1, Abbott RealTime IDH2 assays, the BCW's Polymerase Chain Reaction (PCR) based Sanger sequencing assay and NGS HemeOnc panel (contains 30 genes including IDH1/2). Abbott RealTime IDH1 and IDH2 assays are qualitative in vitro PCR tests wherein, the IDH1 assay detects five IDH1 mutations (R132H, R132C, R132L, R132G and R132S) and the IDH2 assay detects nine IDH2 mutations (R140Q, R140L, R140G, R140W, R172K, R172M, R172G, R172S and R172W).The BCW's PCR based Sanger sequencing assay and NGS HemeOnc panel detect mutations in exon 4 of IDH1/2 with a sensitivity limit of 20% and 10% respectively. In this study NGS data was analyzed for only IDH1 and IDH2 .DNA was extracted from 100 clinical specimens either by the Abbott m Sample preparation system or by the Qiagen DNA extraction method and tested using the Abbott RealTime IDH1 and IDH2 assays. Additionally, a set of commercially available controls (IDH1 R132H/ IDH2 R172K, IDH1 R132C/ IDH2 R140Q) with different mutation levels (20%, 5%, 1% and 0.25%) were tested using the above mentioned four assaysResults:The Abbott RealTime IDH1 and IDH2 assays demonstrated 100% sensitivity and 95% specificity compared to the BCW Sanger sequencing assay. Five out of 100 samples with low level IDH2 mutations were missed by BCW Sanger sequencing assay but were detected as mutation positive by Abbott RealTime IDH2 assay. These five discordant samples with low level IDH2 mutations were subsequently tested with the BCW's NGS assay and two of the five samples were detected as positive by the NGS assay (Table 1). Additionally, turnaround time (TAT) analysis demonstrated that the results from the Abbott RealTime IDH1/2 assays were available in 2 business days as compared to 10 days for the BCW Sanger sequencing assay and 21 days for the BCW NGS HemOnc panel.Testing of the mixture of IDH1 and IDH2 mutations with differing mutation levels (20%, 5%, 1% and 0.25%) resulted in the Abbott RealTime IDH1 and IDH2 assay reliably detecting the mutations down to the 1% level (with some variability at 0.25%) (Table2).Conclusions:This study demonstrated that the Abbott RealTime IDH1 and IDH2 assays detected IDH1 and IDH2 mutations at 1% mutation level in AML patients. The Abbott assays were highly sensitive (1%) compared to the PCR based Sanger sequencing and NGS HemeOnc panel, which have sensitivity limits of 20% and 10% respectively. The higher sensitivity and relatively fast TAT of the Abbott assays may potentially improve the ability to diagnose new and relapsed patients in settings of lower disease burden. The relatively fast TAT could facilitate clinicians in making a quicker decision about therapeutic treatment recommendations for afflicted patients. [Display omitted] DisclosuresDash:Abbott Molecular Inc.: Other: I do not have financial relationship, however wanted to mention here is that this study was sponsored by Abbott Molecular Inc which was performed in BloodCenter of Wisconsin.

Effective Failure Analysis for Packaged Semiconductor Lasers with a Simple Sample Preparation and Home-Made PEM System

As the application requirements of semiconductor lasers continue to increase, severe challenges are brought to the reliability of semiconductor lasers. In order to promote the study of laser failure, this paper proposes an effective failure analysis method for packaged semiconductor lasers with a simple sample preparation and home-made photon emission microscopy (PEM) system. The new simple sample preparation process for failure analysis is presented and the necessary polishing fixture is designed so that sample can be obtained without expensive and complex micro-/nano-processing. Two types of home-made PEM experimental systems were established for observing the failure from the front facet and active region of semiconductor lasers. Experimental results showed that, with the proposed sample preparation flow, the home-made PEM experimental system effectively observed the leakage defects from the front facet and dark spot defects (DSDs) in the active region of semiconductor lasers. The method can help researchers and laser manufactures to perform effective failure analysis of packaged semiconductor lasers.

Recent applications of microextraction sample preparation techniques in biological samples analysis

Analysis of biological samples is affected by interfering substances with chemical properties similar to those of the target analytes, such as drugs. Biological samples such as whole blood, plasma, serum, urine and saliva must be properly processed for separation, purification, enrichment and chemical modification to meet the requirements of the analytical instruments. This causes the sample preparation stage to be of undeniable importance in the analysis of such samples through methods such as microextraction techniques. The scope of this review will cover a comprehensive summary of available literature data on microextraction techniques playing a key role for analytical purposes, methods of their implementation in common biological samples, and finally, the most recent examples of application of microextraction techniques in preconcentration of analytes from urine, blood and saliva samples. The objectives and merits of each microextration technique are carefully described in detail with respect to the nature of the biological samples. This review presents the most recent and innovative work published on microextraction application in common biological samples, mostly focused on original studies reported from 2017 to date. The main sections of this review comprise an introduction to the microextraction techniques supported by recent application studies involving quantitative and qualitative results and summaries of the most significant, recently published applications of microextracion methods in biological samples. This article considers recent applications of several microextraction techniques in the field of sample preparation for biological samples including urine, blood and saliva, with consideration for extraction techniques, sample preparation and instrumental detection systems.

Сhromatographic methods for the study of organic extractants of transformer oil

THE PURPOSE. In order to control the technical condition of oil-filled electrical equipment, a number of new analytical methods have recently been created to determine the ultra-low concentrations of impurity compounds in transformer oil. An analysis of the literature data showed that at this stage of the analytical procedure significant errors may occur that worsen the quality of control of the technical condition of oil-filled electrical equipment, and in some cases make its results meaningless. The published literature practically does not discuss the problem of sample preparation of transformer oil, which negatively affects the quality of analytical control, that is, the reliability of the results and, accordingly, the diagnosis of oil-filled electrical equipment. METHODS. The paper discusses a sample preparation system for transformer oil, which is based on various methods for extracting target components from it. The adsorption process is used, the preparation of chemical derivatives, various types of extraction, liquid, gas, solid phase and fluid, as well as using low temperatures, extraction in microwave, magnetic, electromagnetic and centrifugal fields. It is shown that liquid extraction with an organic solvent is widely used from all sample preparation methods, with the help of which furan compounds resulting from the destruction of paper insulation are extracted from transformer oil. The disadvantages of sample preparation using liquid-phase extraction and the possibility of using solid-phase extraction on various adsorbents for this purpose are discussed. RESULTS. The results of an experimental study of the sorption properties of various organic solvents with respect to porous materials differing in the structure of their surface are presented. We used zeolite-bearing rocks of the Tatar-Shatrashansky deposit, synthetic zeolites NaX-13A. Using the ascending variant of liquid column chromatography, the absolute retention values of standard extractants were determined and their dependence on the length of the sorption layer of the porous material was found. CONCLUSIONS. It was shown that the highest retention times for all studied adsorbents are observed for organochlorine extractants, chloroform, carbon tetrachloride, dichloroethane, trichlorethylene. For N. Hexane is a relatively small interaction. Histograms are given of the effect of the retention time of standard sorbates (extractants) on their physicochemical nature.

Capillary flow-based sample preparation system for metabolomic analysis of mammalian cells in suspension.

Sample preparation methodology is critical to obtaining reliable data for studying endogenous metabolites. Dependable preparation techniques require separation of cells from culture media, quenching of enzymatic activity, and extraction of metabolites from the cells. Presented here is a simple, rapid, semi-automated metabolomic sample preparation technique for 20μL samples of RAW 264.7 cells suspended in culture media. This method uses online filter-assisted electroporation-based cell lysis and chilled organic solvent extraction to prepare metabolomic samples from cells in suspension in 2min. Experiments using an isotopically labeled adenosine triphosphate internal standard were carried out to ensure enzymatic quenching by monitoring the ratio of labeled adenosine diphosphate to adenosine triphosphate. Cells were metabolically labeled with 13C-glucose concurrent with sampling aliquots of the cell suspension over the course of 24h. Incorporation of 13C into organic acid metabolites such as itaconate Cell lysates was analyzed by nano-reverse-phase liquid chromatography-mass spectrometry (nano-RP-LC-MS), showing incorporation of 13C into organic acid metabolites such as itaconate.

Estimation of time spent on studies of toxic elements in food in an accredited laboratory testing center

Reducing the time spent on preparing a sample for analysis by 19.5 times, as well as cost items for raw materials and basic materials by 2.6 times, is a weighty argument in favor of introducing the PLP-01M microwave laboratory system. When the estimated price for a unit of research is established, the competitiveness in the market for the services provided significantly increases, the setting of the market price allows increasing revenue by 1.2 times, which will amount to 883 thousand rubles. additional income. Upgrading the equipment used from Kvant-AFA to Kvant-2AT with the simultaneous implementation of the PLP-01M microwave laboratory system will not only improve the used measuring instrument, but also the entire sample preparation system as a whole. The optimal choice of measuring instruments for updating the laboratory base is carried out based on the volume of analyzed samples, the productivity of existing and considered for implementation instruments, the cost of equipment and the solvency of the TLC, as well as the possibility of implementing a new method in the laboratory.

The effectiveness of the introduction of new equipment in the test laboratory center

Improving the quality of the services provided by updating the laboratory base is considered as one of the most important ways to increase competitiveness, achieve competitive advantages, which, in turn, helps to increase the economic efficiency of the testing laboratory center. The introduction of new equipment at the TLC will contribute to an increase in the possible number of studies being carried out, a decrease in random errors, and the achievement of the best indicators of internal laboratory control. The introduction of the PLP-01M microwave laboratory system will significantly reduce the time for preparing a sample for analysis by reducing the decomposition time of the sample by 19.5 times. Upgrading the equipment used from Kvant-AFA to Kvant-2AT with the simultaneous introduction of the PLP-01M microwave laboratory system will not only improve the used measuring instrument, but also the entire sample preparation system as a whole.

Integrated proteomic sample preparation with combination of on-line high-abundance protein depletion, denaturation, reduction, desalting and digestion to achieve high throughput plasma proteome quantification

In this study, we developed an integrated plasma proteome sample preparation system, by which high-abundance proteins from human plasma were first depleted by immunoaffinity column, followed by on-line middle and low-abundance proteins denaturation, reduction, desalting and tryptic digestion. To evaluate the performance of such a system, 20 μL plasma was processed automatically, followed by 1-h gradient liquid chromatography-mass spectrometry analysis (LC-MS). Compared to conventional in-solution protocols, not only the sample preparation time could be shortened from 20 h to 20 min, but also the number of identified proteins were greatly increased by 1.4–2.0 times. Such an integrated system allows us to process 36 human plasma samples per day, with more than 300 proteins and 52 FDA approved disease markers per sample being identified. With combination of such an integrated sample preparation system with label-free single-shot LC-MS/MS, the human plasma proteins could be quantified across more than 6 orders of magnitude of abundance range with high reproducibility (Pearson R = 0.99, n = 9). In addition, the relative quantification of human plasma samples from diabetic retinopathy patients and diabetic patients demonstrated the feasibility of our developed workflow for clinic plasma proteome profiling. All these results demonstrated that our developed integrated plasma proteome sample preparation system would provide a new tool for high throughput biomarker discovery.

Miniaturized Blood Sampling System with Integrated Sample Preparation

Blood sampling as well as sample preparation are time consuming and requires a strict procedure, which is generally performed by medical trained personal. Not carrying out the procedure correctly could result in an infection of the patient or contamination of the sample itself. These limitations should be especially considered in case of pandemic outbreaks. In order to handle such a high number of patients a novel sample preparation system paired with modern blood sampling procedure is necessary. For this reason, a new device for blood sampling and preparation is designed containing an integrated microfluidic system. The fabrication is carried out by utilizing micro moulding of PDMS as well as micro milling. A first set of initial experiments as part of a first-generation study shows promising results. However, further steps of optimisation considering flow time and preparation cycle are part of a second-generation study.

Method Development on Automated Sample Preparation System for Cu and Assay Testing in a Product with Multiple Apis and New Strategy for Quick Development and Application

Automation of analytical sample preparation and testing is always attractive due to its high efficiency and continuous operation, but it can also be very challenging due to the operational complexity and method development. Studies focused on method development in automated systems are extremely rare and little has been published on the subject. In this paper, we investigate several unique and advanced aspects of this subject. The example product is selected containing two APIs to be tested by a single method. The product strengths of the two APIs are significantly different, making development of the single method much more challenging. In addition, we apply the additional challenge of using a “green chemistry” approach, avoiding any organic solvent in the dissolving media in order to achieve an environmentally friendly solution. A thorough study on the selection and optimization of several key operational parameters are then discussed. The successful method is achieved using a new strategy called “Automated Online Standard” methodology. This novel approach is compared to the conventional procedure in several scenarios, demonstrating its high effectiveness with significantly improved recovery. The proposed approach can substantially reduce the method development time, generally considered the biggest obstacle to practical deployment of automated systems. The automated system applied in this paper is most suitable for content uniformity testing, but is also applicable for blend uniformity testing, potency assays and degradation product tests.

660. Evaluation of Qvella’s FAST-Prep™ Liquid Colony™ for Early Antimicrobial Sensitivity Testing of Positive Blood Culture by Disk Diffusion Method

BackgroundConventional antimicrobial susceptibility testing (AST) of microorganisms from positive blood cultures (PBC) can take ≥ 2 days. In order to improve the turnaround time for AST on a PBC, CLSI and EUCAST have made efforts to standardize procedures for disk diffusion (DD) direct from a PBC. Qvella Corporation (Richmond Hill, ON, Canada) has recently developed FAST-Prep, an automated centrifugal sample preparation system that rapidly delivers a Liquid Colony consisting of a purified, concentrated, viable cell suspension directly from a PBC. This study was performed to investigate the feasibility of DD AST off of a PBC using a FAST-Prep Liquid Colony.MethodsContrived PBC samples were prepared by spiking 6 species of Gram-positive and 4 species of Gram-negative bacteria (3-5 strains per species) into FA® Plus bottles and incubating in the BACT/ALERT® VIRTUO® System (bioMerieux, Durham, NC). After positivity, 3 mL of PBC was added to the FAST-Prep cartridge. After 20 minutes of processing in the FAST-Prep instrument, the Liquid Colony was removed from the cartridge and a 0.5 McFarland sample was prepared for DD AST. In parallel, the DD AST from a PBC was performed using 4 drops of PBC (CLSI direct method). Both methods were compared to conventional colony-based DD AST. After 16-18 hours of incubation zone diameters and S/I/R interpretations were determined. Categorical agreement (CA) and errors for both DD AST methods were calculated. In addition, colony plate counting was performed on 0.5 McFarland suspensions of Liquid Colony and the plate colony to determine biomass recovery and sample purity.ResultsCA for a FAST-Prep DD AST for Gram-positive and Gram-negative bacteria was 95.6% and 98.6%, respectively, compared to CA for CLSI DD AST of 77.2% and 81.9%, respectively. Biomass in the Liquid Colony was 7.2x108 and 1.2x109 CFU for Gram-positive and Gram-negative bacteria, respectively. Cell concentration in the 0.5 McFarland suspension of the Liquid Colony was 3.7x107 and 5.9x107 CFU/mL for Gram-positive and Gram-negative bacteria, respectively, which was similar to the concentration for the reference colony suspension.ConclusionThe results support the potential role of FAST-Prep in providing a Liquid Colony for use in rapid AST.DisclosuresSusan M. Novak-Weekley, PhD, D(ABMM), Qvella (Employee, Shareholder) Aye Aye Khine, PhD, Qvella (Employee, Shareholder) Tino Alavie, PhD, Qvella (Employee) Namidha Fernandez, MS, Qvella (Employee) Laxman Pandey, MS, Qvella (Employee) Abdossamad Talebpour, PhD, Qvella (Employee, Shareholder)

The problem of determining the antioxidant additive in transformer oil by chromatographic methods

THE PURPOSE. Transformer oil is widely used in high-voltage electrical equipment and acts as a dielectric and also as a cooling medium. It consists of a complex mixture of hydrocarbon compositions with various impurity compounds. During the operation of oil -filled electrical equipment under the influence of the technological operating conditions of high -voltage devices, aging of both transformer oil and solid insulation occurs. In this case, the dielectric properties deteriorate, which can damage the transformer equipme nt. METHODS. To increase the service life of oil-filled electrical equipment, an antioxidant additive is introduced into the transformer oil at the stage of its manufacture, which is most often used as ionol, the concentration of which must be constantly controlled by various instrumental methods. To determine the antioxidant additive in transformer oil, a sample preparation system is used based on liquid-phase extraction with selective solvents, which are used as aliphatic alcohols of various physical and chemical nature. RESULTS. It has been established that ethyl alcohol, which is widely used in the sample preparation system for liquid extraction of ionol, contains a significant amount of water, which has a negative effect on the process of ionol extracti on from transformer oil. In addition, the composition of ethyl alcohol contains impurity substances that can have a negative effect on the process of liquid-phase extraction of ionol from transformer oil. The generalized data on the physicochemical properties of extractants used to extract ionol from transformer oil are presented. CONLUSION. It has been shown that butanol is the most promising organic solvent for the extraction of ionol, which has a higher boiling point compared to ethyl alcohol and contains a small amount of water. It was found that the selectivity of the chromatographic separation of butanol components with an elevated column temperature increases. At the same time, the separation efficiency increases as the number of theoretical plates per meter of chromatographic column increases. As the temperature of the capillary column increases, the symmetry of the chromatographic peaks of the separated components also increases.

Nucleic acid sample preparation from whole blood in a paper microfluidic device using isotachophoresis

Nucleic acid amplification tests (NAATs) are a crucial diagnostic and monitoring tool for infectious diseases. A key procedural step for NAATs is sample preparation: separating and purifying target nucleic acids from crude biological samples prior to nucleic acid amplification and detection. Traditionally, sample preparation has been performed with liquid- or solid-phase extraction, both of which require multiple trained user steps and significant laboratory equipment. The challenges associated with sample preparation have limited the dissemination of NAAT point-of-care diagnostics in low resource environments, including low- and middle-income countries. We report on a paper-based device for purification of nucleic acids from whole blood using isotachophoresis (ITP) for point-of-care NAATs. We show successful extraction and purification of target nucleic acids from large volumes (33 µL) of whole human blood samples with no moving parts and few user steps. Our device utilizes paper-based buffer reservoirs to fully contain the liquid ITP buffers and does not require complex filling procedures, instead relying on the natural wicking of integrated paper membranes. We perform on-device blood fractionation via filtration to remove leukocytes and erythrocytes from our sample, followed by integrated on-paper proteolytic digestion of endogenous plasma proteins to allow for successful isotachophoretic extraction. Paper-based isotachophoresis purifies and concentrates target nucleic acids that are added directly to recombinase polymerase amplification (RPA) reactions. We show consistent amplification of input copy concentrations of as low as 3 × 103 copies nucleic acid per mL input blood with extraction and purification taking only 30 min. By employing a paper architecture, we are able to incorporate these processes in a single, robust, low-cost design, enabling the direct processing of large volumes of blood, with the only intermediate user steps being the removal and addition of tape. Our device represents a step towards a simple, fully integrated sample preparation system for nucleic acid amplification tests at the point-of-care.

A New Type of MgFe2O4@CuS-APTES Nanocarrier for Magnetic Targeting and Light-Microwave Dual Controlled Drug Release.

IntroductionCancer is a major health problem worldwide, and the most extensive treatment can be obtained by using chemotherapy in the clinic. However, due to the low selectivity of cancer cells, chemotherapy drugs produce a series of grievous side effects on normal cells.MethodsIn this research, we developed novel nanocarriers for magnetically targeted near-infrared (NIR) light-electromagnetic wave dual controlled drug delivery based on MgFe2O4@CuS nanoparticles (NPs) modified with aminopropyltriethoxysilane (APTES) in response to magnetic, NIR light, and electromagnetic wave irradiation. Synthesis and characterization of MgFe2O4@CuS-APTES NPs was carried out using X-ray diffraction measurements, scanning electron microscopy, transmission electron microscopy, photoluminescence emission spectra, UV-1800 spectrophotometer, N5230A vector network analyzer, MDS-6 microwave sample preparation system, and superconducting quantum interference device. In addition to that mentioned above, we also explored many other sides, such as the drug-loading, drug-controlled release efficiency, elect99omagnetic wave thermal effect and photo-thermal effect.ResultsThe results showed that APTES-modified MgFe2O4@CuS NPs had 37% high drug-loading capacity and high electromagnetic wave thermal conversion ability and NIR-light thermal conversion ability. In addition, ibuprofen (IBU) release from the MgFe2O4@CuS-APTES-IBU depends on the electromagnetic wave (2.45 GHz) and 1060 nm NIR light irradiation. After five cycles, the drug-release percentage was 90% and 66% separately, and could be adjusted by the time and cycles times of electromagnetic wave and NIR light irritation. Electromagnetic wave irradiation compared with NIR light irradiation, has a higher drug release rate and better penetration. Therefore, choosing different stimulation methods according to the treatment needs of the disease, we can achieve accurate personalized treatment of the disease.DiscussionOur findings indicate that multifunctional APTES modified MgFe2O4@CuS NPs could be used for the first time as a new drug carrier for “location-timing-quantification” drug release with magnetic targeting and dual control of NIR light-electromagnetic waves.

Research progress of electrically-driven force based online rapid separation and enrichment techniques

Sample preparation is a critical step in complex sample analysis, which enables analyte isolation and preconcentration from a complex matrix. Therefore, sample preparation is an effective approach to enhance the sensitivity, selectivity, and accuracy of the analytical method. However, the transfer of target analytes from a random state in the original sample matrix to a highly ordered pre-analysis state involves an entropy reduction process that cannot occur spontaneously. Therefore, sample preparation is always a time-consuming, labor-intensive, and error-prone process. Introducing additional energy or reducing the entropy of the system can enhance the separation and enrichment effects as well as accelerate sample preparation. The introduction of an electric field into an online sample preparation system can not only introduce additional energy into the system, but also drive the directional migration of the sample among the separation, enrichment, and detection processes, ensuring that the entropy reduction progresses smoothly. These advantages of electrically-driven force based online separation and enrichment techniques make them effective for accelerating sample preparation. Typically, there are four acceleration strategies in electrically-driven force based online separation and enrichment techniques: (1) the additional energy of the electric field is added into the system to accelerate mass transfer and energy exchange; (2) electrically-driven flows, including electroosmotic flow and electrophoretic flow, are applied to drive the directional migration of the sample among the separation, enrichment, and detection processes, ensuring that sample preparation and analysis are executed smoothly; (3) the online integration technique is applied to enhance the automaticity of the entire sample preparation and analysis processes, and reduce errors from manual operation; (4) device miniaturization or size reduction methods such as microextraction are applied to enhance the sample preparation efficiency and reduce the time consumed. This review summarizes the progress in electrically-driven force based online rapid separation and enrichment techniques in the last ten years. In this specific research area, more than one hundred research papers are published each year, and can be classified into three types based on the electrically-driven force based online rapid separation and enrichment techniques considered: capillary, microchip, and membrane extraction. Among these, over 50% of the studies focused on electrically-driven capillary force based online rapid separation and enrichment techniques. By applying a high-voltage electric field at the two ends of a capillary, charged species in the capillary can migrate along the direction of the medium. This makes the electrically-driven capillary technique not only fast, highly efficient, and low-cost, but also effective for rapid sample preparation. Typically, two modes of electrically-driven capillary force based online rapid separation and enrichment techniques are employed: online capillary electrophoretic separation and enrichment, and online microextraction-capillary electrophoresis. Device miniaturization from the capillary to microchip through microelectronic mechanical systems and microfluidics enables small-amount sample preparation and analysis, and is also advantageous due to being rapid and efficient, as well as low energy- and sample-consuming. The specific easy-integration trait of microchip devices enables the online integration of multi-step sample preparation and analysis. On the other hand, the controllable electrically-driven force can be used for both, directional flow transfer between different functional units in the microchip as well as for dynamic control of the electrically fluid pump and valve. Online microchip electrophoretic separation and enrichment, and online microextraction-microchip electrophoresis, are two common modes of microchip electrically-driven force based online rapid separation and enrichment techniques. In electrically-driven membrane extraction online separation and enrichment techniques, a supporting liquid membrane is used to eliminate the matrix interference, enabling large-scale real sample application. These techniques have received increasing attention in the research area of electrically-driven force based online separation and enrichment. Overall, rapid separation and enrichment techniques are highly desired in complex sample analysis, and electrically-driven force based online approaches offer significant application potential, especially in food, the environment, and medicine.