Analytical Recovery Research Articles

In-Syringe Vortex-Assisted Liquid-Liquid Microextraction Based on Natural Deep Eutectic Solvent for Simultaneous Determination of the Two Anticancer Polyphenols Chrysin and Resveratrol.

The simultaneous determination of multiple anticancer drugs in combination therapy poses a significant analytical challenge due to their complex nature and low concentrations. In this study, we propose an in-syringe vortex-assisted liquid-liquid microextraction (IS-VA-LLME), based on a green natural deep eutectic solvent (NaDES) for the simultaneous determination of two coadministered anticancer drugs (resveratrol and chrysin) prior to the HPLC-UV analysis, for the first time. The key parameters affecting the extraction efficiency, such as extraction solvent, vortex time, pH, and ionic strength were optimized. Under optimal conditions, the method demonstrates good linearity over the range of 0.05-15.0 μg/mL for RVT and 0.50-15.0 μg/mL for CHR with low limits of detection (LODs) of 16.78 and 161.60 ng/mL for RVT and CHR, respectively, confirming the high sensitivity of the method. The interday and intraday precision values, expressed as %RSDs, are below 2.0%, indicating good repeatability and reproducibility. Furthermore, the proposed method could be efficiently applied for the determination of the two drugs in human plasma and river water. The obtained results show satisfactory % recoveries (97.80%-102.04%), highlighting the accuracy and reliability of the developed method. The sustainability of the method was comprehensively evaluated using seven different tools. In conclusion, the developed IS-VA-LLME-NaDES allows for enhanced extraction efficiency, reduced extraction time, and improved recovery of the target analytes. This method holds great promise for applications in clinical and environmental research, enabling the precise quantification of these anticancer drugs in complex matrices.

A New LC-MS/MS-Based Method for the Simultaneous Detection of α-Tocopherol and Its Long-Chain Metabolites in Plasma Samples Using Stable Isotope Dilution Analysis

Background: Our study presented a novel LC-MS/MS method for the simultaneous quantification of α-tocopherol (α-TOH) and its phase II metabolites, α-13′-COOH and α-13′-OH, in human serum using deuterium-labeled internal standards (d6-α-TOH, d6-α-13′-COOH, d6-α-13′-OH). Methods: The method addresses the analytical challenge posed by the significantly different concentration ranges of α-TOH (µmol/L) and its metabolites (nmol/L). Previous methods quantified these analytes separately, which caused an increase in workflow complexity. Results: Key features include the synthesis of stable isotope-labeled standards and the use of a pentafluorophenyl-based core-shell chromatography column for baseline separation of both α-TOH and its metabolites. Additionally, solid phase extraction (SPE) with a HybridSPE® material provides a streamlined sample preparation, enhancing analyte recovery and improving sensitivity. By utilizing deuterium-labeled standards, the method compensates for matrix effects and ion suppression. This new approach achieves precise and accurate measurements with limits of detection (LOD) and quantification (LOQ), similar to previous studies. Calibration, accuracy, and precision parameters align well with the existing literature. Conclusions: Our method offers significant advantages in the simultaneous analysis of tocopherol and its metabolites despite concentration differences spanning up to three orders of magnitude. In contrast to earlier studies, which required separate sample preparations and analytical techniques for tocopherol and its metabolites, our approach streamlines this process. The use of a solid-phase extraction procedure allows for parallel sample preparation. This not only enhances efficiency but also significantly accelerates pre-analytical workflows, making the method highly suitable for large-scale studies.

Lipid and Corticosteroid Biomarkers Under the Influence of Bisphosphonates.

Detecting the use of bisphosphonates (BPs) in equine athletes is of interest to regulators and laboratories due to the threat to welfare issues for the potential to provide analgesic effects and manipulating bone structure. The detection of BPs in biological matrices is challenging due to erratic biological elimination and inconsistent analytical recoveries. Therefore, complementary approaches are needed to provide evidence of their misuse in racehorses. BPs have two sub-classes: nitrogenous and non-nitrogenous. This study investigated plasma elimination following administration of one example from each sub-class, together with changes in endogenous eicosanoid and corticosteroids. Zoledronic acid (ZA) and tiludronic acid (TA) were administered by IV infusion to 8 thoroughbred horses with an 11-month washout period between each administration. Sample preparation for quantification of BPs by liquid chromatography-tandem mass spectrometry (LC-MS/MS) utilised a two-step solid phase extraction (SPE) consisting of polymeric reversed-phase followed by weak anion exchange prior to derivatisation using trimethyl orthoacetate. Endogenous biomarkers were analysed after protein precipitation and SPE with polymeric reversed-phase prior to liquid chromatography-high resolution mass spectrometry (LC-HRMS) using data independent acquisition. The LC-MS/MS analysis showed ZA was undetectable after 8 h post-administration while TA was detected up to the final collection point of 28 days post-administration. The LC-HRMS analysis utilised targeted (i.e., prior inclusion list of compounds) approaches to monitor level changes of eicosanoid and corticosteroid biomarkers. Putative biomarkers were identified and now subject to validation for translation into routine sample analysis for improved retrospectivity to detecting BP misuse in equine plasma.

12260 Assessing The Impact Of Saliva Collection Devices On Analyte Recovery

Abstract Disclosure: D. Marshall: None. M. Dolan: None. M. Mctaggart: None. B.G. Keevil: None. Introduction: Analysis of saliva is gathering momentum, often conveying advantages over regular venepuncture, offering a convenient way of collecting multiple samples throughout the day with relative ease from the comfort of a patient’s home or workplace. The advent of more sensitive mass spectrometry equipment has opened the possibility of measuring analytes previously unsuitable, offering a multitude of steroid hormones for diagnosis and monitoring of a range of endocrine disorders. Traditionally, samples are collected onto a Sarstedt Salivette device comprising of a synthetic swab enclosed within a polypropylene tube. For certain analytes, the Salivette device is known to impede analyte recovery, resulting in the requirement for a passive drool sample which can be difficult and unpleasant to collect for some patients. Methods Previous work has been undertaken showing the SalivaBio Oral Swab (SOS) to have less of an impact on analyte recovery. Here, through addition of low, medium and high spikes and subsequent LC-MS/MS analysis of 19 different steroid hormones (11-dehydrocorticosterone, 11-deoxycorticosterone, 11-deoxycortisol, 11-hydroxyandrostenedione, 11-hydroxytestosterone, 11-ketoandrostenedione, 11-ketotestosterone, 17-hydroxyprogesterone, 18-hydroxycortisol, 18-oxocortisol, 21-deoxycortisol, aldosterone, androstenedione, corticosterone, cortisol, cortisone, dexamethasone, testosterone and tetrahydro11-deoxycortisol) we aimed to assess a broader repertoire of analytes and whether saliva collection devices impact on analyte recovery. Results With an acceptability criterion of 10% difference from baseline passive drool samples, Salivette devices were acceptable for 12 out of 18 analytes, SOS were acceptable for 16 analytes. Both devices were unacceptable for measurement of androstenedione, testosterone and 11-deoxycorticosterone. When 10% is deemed as an acceptable difference, the SOS performs better than the Salivette device for the steroid hormones investigated in this study. This may have implications for research studies, patient diagnosis and monitoring of endocrine disorders where saliva collection devices were previously deemed unacceptable due to issues with analyte recovery. Presentation: 6/2/2024

Liquid-Liquid Extraction Solvent Selection for Comparing Illegal Drugs in Whole Blood and Dried Blood Spot with LC-MS/MS.

This study focused on the simultaneous detection of amphetamine, 3,4-methyl enedioxy methamphetamine, morphine, benzoylecgonine, and 11-nor-9-carboxy- tetrahydrocannabinol (Δ9-THC-COOH) in whole blood and DBS. It is aimed to select a solvent mixture for liquid-liquid extraction (LLE) technique employing LC-MS/MS. The obtained DBS results were compared with the whole blood samples results. A simple, rapid, and reliable LC-MS/MS method was developed and validated for all analytes in whole blood and DBS. LC was performed on a Hypersil Gold C18 column an initial step with a gradient of 0.01 % formic acid, 5 mM ammonium format buffer in water, and acetonitrile at 0.3 ml/min with 7.5-min runtime. A methanol:acetonitrile (40:60 v/v) mixture was selected for both matrices. LOQ values were 10-25 ng/mL; linear ranges were LOQ-500 ng/ml for all analytes; correlation coefficients were greater than 0.99, and all calibrator concentrations were within 20%. Analytical recovery in blood and DBS ranged from 84.9-113.2% of the expected concentration for both intra and-inter day. Analytes were stable for 1, 10, and 30 days after three freeze/thaw cycles. It was determined that the variances of the results obtained with the two matrices in the comparison study were equal for each analyte, and the results were highly correlated (r=0.9625). A sensitive, accurate, and reliable chromatographic method was developed to determine amphetamine, MDMA, morphine, benzoylecgonine, and cannabis, by performing the same preliminary steps with whole blood and dried blood spots. It was observed that the results obtained in these two matrices were compatible and interchangeable when statistically compared.

Rapid bromate determination using short-column ion chromatography-mass spectrometry: Application to bromate quantification during ozonation

A streamlined, precise, and dependable method for the concurrent quantification of bromate has been established, employing ion chromatography in conjunction with electrospray ionization tandem mass spectrometry (IC-ESI-MS/MS). In an effort to expedite sample analysis duration, the AG18 short column was utilized for rapid separation of the specified analytes. Utilizing a blend of water and acetonitrile as the mobile phase within an optimized gradient elution setting, it was possible to achieve satisfactory resolution in under 3 minutes. Specifically, the mixing of acetonitrile in water samples improves the sensitivity and immunity of the method. This phenomenon may be attributed to the similar proportions of acetonitrile in the sample and the mobile phase, which effectively reduces the viscosity and surface tension of the droplets during the ESI process. Detection limits and quantification thresholds were 0.14 µg/L and 0.41 µg/L, respectively. Analyte recoveries, when spiked into tap water between 2.5 µg/L to 50 µg/L, ranged from 69.5 % to 116.7 %; precision metrics, measured as intra- and inter-day relative standard deviations, spanned 5.1–7.9 %. The devised approach encountered minimal interference from prevalent water matrix constituents, including chloride (50–200 mg/L), sulfate (25–200 mg/L), bicarbonate/carbonate (1–6 mM), and natural organic matter (1–8 mg/L). In environmental contexts, this methodology effectively tracked the production of bromate in controlled ozonation experiments conducted in the laboratory. Segmented ozone dosing reduced bromate production when the solution contained low concentrations of DOC, which provides an alternative strategy for controlling bromate production.

B-027 CLSI-Based Result Precision on the cobas ISE neo and cobas c 703 Analytical Units Across Two Sites in Europe

Abstract Background Novel cobas® pro integrated solutions analytical units, the cobas® ISE neo and cobas c 703 analytical units (all Roche Diagnostics International Ltd, Rotkreuz, Switzerland), were assessed at two sites in Europe (Aalst, Belgium and Heidelberg, Germany). During the system design validation, precision was evaluated under intended use conditions in a routine simulation setting. Methods Quality control (QC) materials at two analyte concentrations were measured per site for selected assays (ion selective electrodes [ISEs], general chemistries and specific proteins, and immunochemistry assays); analyte recovery per QC was closely monitored. Precision was evaluated at both study sites based on Clinical and Laboratory Standards Institute EP05-A3 guidelines over 21 days for 53 applications representing the entire assay menu: six ISE serum/plasma and urine applications; 40 clinical chemistry serum/plasma and urine applications (four were measured using cobas c 503 at one site only); and seven immunochemistry serum/plasma applications. Reproducibility under various stressed routine-like conditions, including provocations, was tested using routine simulation imprecision experiments. To test the interaction of new and existing analytical units under routine conditions, two test configurations were used: the first configuration comprised one cobas ISE neo, two cobas c 703, and one cobas e 801 analytical units; and the second configuration comprised one cobas ISE neo, one cobas c 703, one cobas c 503, and one cobas e 801 analytical units. The precision of batch type measurements was compared with the precision under routine simulated random access conditions. Random access coefficients of variance (CVs) that exceeded 1.5× reference batch CVs, or single measurements deviating by >10% from the batch mean, triggered in-depth analysis of the system components that contributed to the result. CVs for intermediate precision comprised measurements from ≤4 cobas c 703 reagent disks, dependent on the system set-up and application assignment. CVs for reproducibility were calculated across four ISE measuring units, ≤3 cobas c 703 reagent disks, or ≤4 cobas e 801 measuring cells. Results Using four analytical units, n=511 CVs for repeatability, intermediate precision, and reproducibility were calculated. For repeatability, the median CV for ISE (n=24) was 0.8%, for clinical chemistry (cobas c 703: n=146; cobas c 503: n=8) was 0.9%, and for immunochemistry (n=28) was 1.0%. For intermediate precision, the median CV for ISE (n=24) was 1.1%, for clinical chemistry (cobas c 703: n=146; cobas c 503: n=8) was 1.2%, and for immunochemistry (n=28) was 1.5%. For reproducibility, the median CV for ISE (n=12) was 0.9%, for clinical chemistry (cobas c 703: n=73) was 1.6%, and for immunochemistry (n=14) was 1.7%. Conclusions The novel cobas ISE neo and cobas c 703 analytical units demonstrated robust precision, comparable to the existing cobas pro integrated solutions analytical units, over a prolonged period.

B-295 Sensitive and Rapid Homogeneous Immunoassay for the Detection of Zolpidem and its Major Metabolite in Urine

Abstract Background Zolpidem, a schedule IV controlled substance under the Controlled Substances Act, is commonly known as Ambien and Ambien CR, a prescription only treatment for insomnia. Zolpidem is one of the most common prescribed sleep aids in the U.S. due to its rapid sedative effects. The recommended duration of treatment is between two days and four weeks, as long-term use of zolpidem increases the risk of habit formation. Addiction to zolpidem can cause severe side effects, including memory loss, delusion, and in extreme cases, fatal overdose. From 2005-2010, Zolpidem-related emergency room visits doubled, underscoring the need for accurate testing as zolpidem testing plays an important role in cases of misuse, diversion, and forensics. The sole commercially available homogeneous immunoassay for the detection of zolpidem in urine has a relatively high cutoff at 20 ng/mL and poor cross-reactivity (0.02%) to major metabolite zolpidem 4-carboxylic acid. ARK Diagnostics has developed the ARK™ Zolpidem Assay to detect zolpidem at a cutoff concentration of 10 ng/mL with high cross-reactivity to its metabolite, Zolpidem phenyl 4-carboxylic acid and no cross-reactivity to zaleplon and zopiclone at concentrations below 100,000 ng/mL. Methods The ARK™ Zolpidem Assay is a liquid stable homogeneous enzyme immunoassay, consisting of two reagents, with a cutoff concentration of 10 ng/mL and semi-quantitative range up to 40 ng/mL. Preliminary performance characterization for this assay was evaluated on the Beckman Coulter AU680 Automated Clinical Chemistry Analyzer. Precision, analytical recovery, specificity, Histogram Overlap Analysis of ±50% controls and the cutoff, and method comparison with LC-MS/MS were evaluated. Results In semi-quantitative mode, total precision ranged from 2.6 to 3.1% CV. Spiked recovery of zolpidem ranged from 88.6% (2.5 ng/mL) to 96.8% (10 ng/mL). The major metabolite, zolpidem phenyl-4-carboxylic acid, showed an approximate equivalence to the 10 ng/mL zolpidem cutoff at 30 ng/mL (33.3% cross-reactivity). Histogram overlap analysis showed no overlap between cutoff and control levels. Method correlation with LC-MS/MS using authentic urine samples showed an excellent agreement with a specificity of 100% and sensitivity of 100% (15 positives and 100 negatives). Conclusions The ARK™ Zolpidem Assay measures zolpidem and its major metabolite zolpidem phenyl-4-carboxylic acid in human urine with acceptable performance. The assay is sensitive, rapid, and applicable to a wide range of clinical chemistry analyzers.

B-182 Development and validation of an LC-MS/MS assay for simultaneous measurement of serum androstenedione and 17-hydroxyprogesterone in serum

Abstract Background Androstenedione and 17-hydroxyprogesterone are measured via immunoassays or high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) in clinical laboratories. Immunoassays have limited sensitivity and specificity for certain steroid hormones testing. We aimed to develop and validate a sensitive, accurate, and specific in-house LC-MS/MS assay to simultaneously measure both 17-hydroxyprogesterone and androstenedione in serum. Methods 13C-labeled androstenedione and 17-hydroxyprogesterone internal standards were added to calibrator, control, and patient samples. A hexane/ethyl acetate solution was used in a liquid-liquid extraction procedure. The organic layer was then removed and dried followed by reconstituting the extracts in 2.5 mM ammonium formate in 50% methanol. The HPLC mobile phase gradient began with 47%, 2.5 mM ammonium formate in water and 53%, 2.5 mM ammonium formate in methanol. This progressed to 56% of the methanol phase over three minutes. The mixtures were separated though an Agilent 1260 Infinity HPLC system with a Poroshell 120 EC-C18 analytical column (2.1 × 50 mm × 2.7 µm). The separated HPLC eluents then entered an Agilent 6460C QQQ mass spectrometer in positive ion mode through an electrospray ionization source where protonated androstenedione (m/z = 287.2) with the protonated androstenedione-[2, 3, 4-13C3] internal standard (m/z = 290.2) and protonated 17-hydroxyprogesterone (m/z = 331.2) with the protonated 17-hydroxyprogesterone-[2, 3, 4-13C3] (m/z = 334.2) were mass-selected for collision-induced dissociation. For androstenedione, respective quantifier and qualifier m/z values of 287.2 > 97.1 and 287.2 > 109.1 were used while respective quantifier and qualifier m/z values of 331.2 > 97.1 and 331.2 > 109.1 were used for 17-hydroxyprogesterone. Validation studies were performed including imprecision, lower limit of quantitation (LLOQ), analytical measurement range (AMR), interference, specificity, carryover, extraction recovery, matrix effect, and comparison with a reference laboratory LC-MS/MS method. The total allowable error limits used in this study were 23.5% for androstenedione and 30.2% for 17-hydroxyprogesterone. Results Total imprecision results across three concentrations yielded a maximum CV of 4.6 % for androstenedione and 6.5% for 17-hydroxyprogesteron. The AMRs of androstenedione and 17-hydroxyprogesterone were 0.2 to 13.4 and 0.1 to 22.0 ng/mL without significant carryovers. No significant interferences were observed from 11-deoxycorticosterone (700 ng/dL), testosterone (2500 ng/dL), DHEA (1000 ng/mL), bilirubin (30.1 mg/dL), hemoglobin (1032 mg/dL), or triglycerides (1858 mg/dL). The extraction recoveries of androstenedione and 17-hydroxyprogesterone were 98% and 81%, respectively. No significant matrix effects were identified. In comparison to the reference lab LC-MS/MS method, the in-house LC-MS/MS assay demonstrated a 0.523% bias for androstenedione and a -3.77% bias for 17-hydroxyprogesterone with both exhibiting excellent correlations with the reference lab measurements. More specifically, our results (as a function of values obtained from the reference lab) followed the relationship of y = 1.039x - 0.092 (R = 0.9951) for androstenedione and y = 0.957x + 0.01 (R = 0.9974) for 17-hydroxyprogesterone. Conclusions An LC-MS/MS assay is developed for simultaneous detection of serum androstenedione and 17-hydroxyprogesterone. The assay demonstrates acceptable imprecision, AMR, and accuracy. No significant carryover and interference from other structurally similar steroid hormones and other common interferents are identified.

Development of an ultrasensitive electrochemical aptasensor based on aptamer/rGO/SPGE for the detection of Botulinum neurotoxin A (BoTN/A) in food samples

Consumption of Botulinum neurotoxin A (BoTN/A) contaminated foods has increased the public health issues which necessitates the development of rapid and simple diagnostic tools for their sensitive detection. Electrochemical aptasensors can replace existing biosensors and traditional methods of BoTN/A detection in foods. These aptasensors are simple, quick response, economic, precise, sensitive, accurate and reproducible. In the present study, an electrochemical aptasensor was constructed by immobilizing BoTN/A specific aptamer and reduced graphene oxide (rGO) onto screen printed gold electrode (SPGE) for the detection of BoTN/A in spiked food samples. The specificity of this aptasensor was achieved by conjugating BoTN/A specific aptamer on rGO/SPGE. Test sample containing BoTN/A form aptamer-BoTN/A complex onto SPGE which was directly proportional to BoTN/A concentration. Formation of aptamer-BoTN/A complex reduced the surface of working SPGE for redox reactions. Thus decreased the current peaks that was taken as measuring signal of BoTN/A in test food sample. BoTN/A aptasensor showed optimum response at pH 6.5 and 30 °C temperature. The responsetime of BoTN/A/aptamer/rGO/SPGE was 6 s. In optimum conditions aptamer/rGO/SPGE BoTN/A aptasensor exhibited wide linearity ranges from 1-100 pM and 1 pM limit of detection (LOD). The analytical recoveries of aptamer/rGO/SPGE BoTN/A aptasensor at 5 pM and 10 pM BoTN/A were 98.5 % and 99.8 % respectively. The proposed aptasensor showed unique sensitivity, reproducibility and accuracy. Moreover, aptamer/rGO/SPGE aptasensor was used for the detection of BoTN/A in five different types of food samples.

Systematic Comparison of Extract Clean-Up with Currently Used Sorbents for Dispersive Solid-Phase Extraction

Dispersive solid-phase extraction (dSPE) is a crucial step for multiresidue analysis used to remove matrix components from extracts. This purification prevents contamination of instrumental equipment and improves method selectivity, sensitivity, and reproducibility. Therefore, a clean-up step is recommended, but an over-purified extract can lead to analyte loss due to adsorption to the sorbent. This study provides a systematic comparison of the advantages and disadvantages of the well-established dSPE sorbents PSA, GCB, and C18 and the novel dSPE sorbents chitin, chitosan, multi-walled carbon nanotube (MWCNT), and Z-Sep® (zirconium-based sorbent). They were tested regarding their clean-up capacity by visual inspection, UV, and GC-MS measurements. The recovery rates of 98 analytes, including pesticides, active pharmaceutical ingredients, and emerging environmental pollutants with a broad range of physicochemical properties, were determined by GC-MS/MS. Experiments were performed with five different matrices, commonly used in food analysis (spinach, orange, avocado, salmon, and bovine liver). Overall, Z-Sep® was the best sorbent regarding clean-up capacity, reducing matrix components to the greatest extent with a median of 50% in UV and GC-MS measurements, while MWCNTs had the largest impact on analyte recovery, with 14 analytes showing recoveries below 70%. PSA showed the best performance overall.

Reshaping Capillary Electrophoresis With State-of-the-Art Sample Preparation Materials: Exploring New Horizons.

Capillary electrophoresis (CE) is a powerful analysis technique with advantages such as high separation efficiency with resolution factors above 1.5, low sample consumption of less than 10µL, cost-effectiveness, and eco-friendliness such as reduced solvent use and lower operational costs. However, CE also faces limitations, including limited detection sensitivity for low-concentration samples and interference from complex biological matrices. Prior to performing CE, it is common to utilize sample preparation procedures such as solid-phase microextraction (SPME) and liquid-phase microextraction (LPME) in order to improve the sensitivity and selectivity of the analysis. Recently, there have been advancements in the development of novel materials that have the potential to greatly enhance the performance of SPME and LPME. This review examines various materials and their uses in microextraction when combined with CE. These materials include carbon nanotubes, covalent organic frameworks, metal-organic frameworks, graphene and its derivatives, molecularly imprinted polymers, layered double hydroxides, ionic liquids, and deep eutectic solvents. The utilization of these innovative materials in extraction methods is being examined. Analyte recoveries and detection limits attained for a range of sample matrices are used to assess their effects on extraction selectivity, sensitivity, and efficiency. Exploring new materials for use in sample preparation techniques is important as it enables researchers to address current limitations of CE. The development of novel materials has the potential to greatly enhance extraction selectivity, sensitivity, and efficiency, thereby improving CE performance for complex biological analysis.

A Simple Dispersive Liquid-Liquid Microextraction for the Determination of Copper II in the Water Samples of the Halfaya Oil Field in Maysan

Background: Copper, an important trace element for life on Earth, is crucial to many metabolic processes. This includes its role in the production and breakdown of nucleic acids and the metabolism of proteins, lipids, and carbohydrates. Objective: We aim to integrate DLLME with a spectrophotometric method to establish a novel approach for quantifying trace amounts of copper in water samples. Methods: In this study, a new ligand known as a 2-(4 aminophenol) benzimidazole azo derivative was successfully synthesized. This ligand was created by converting 2-(4-aminophenyl) benzimidazole into a diazonium salt and coupling it with 8-hydroxyquinoline in an alkaline environment. The resulting product was a brown azo dye, which served as the ligand for detecting copper (II) in aqueous samples. Results: To analyze copper (II) in both pure and aqueous samples, we developed and validated micro-extraction techniques combined with UV-Vis measurement. Specifically, we used the DLLME (dispersive liquid-liquid microextraction) method to separate, enrich, and assess copper (II) in both its pure form and in aqueous samples. UV-Vis spectroscopy at a wavelength of 503 nm was used for this purpose. We considered several variables during the experiment, including the type and volume of the dispersive solvent, the extraction solvents, the temperature, the reaction duration, and the centrifuging time. By optimizing these conditions, we achieved linear procedures within the concentration range of (1.0–25.0) and (2.0–25.0) μg/mL for spectroscopy and DLLME methods, respectively. For the spectroscopic and DLLME approaches, the coefficient of determination (R2) was found to be 0.9963 and 0.9979 for the spectrophotometric and DLLME methods, respectively. The limit of detection (LOD) for copper (II) was found to be 0.23 and 0.04 μg/mL, respectively. Moreover, the recovery of the target analyte in aqueous samples was found to be between 95.6% and 101% for the spectroscopy method and between 102.4% and 108.2% for the DLLME method. Conclusions: These results demonstrate the effectiveness and accuracy of both methods in analyzing copper (II) in aqueous samples.

Development and validation of multiresidue analysis method for biomonitoring of pesticides and metabolites in human blood and urine by LC-QToF-MS

The widespread use of pesticides and their consequential presence in the environment is a growing concern due to the harmful health effects associated with pesticide exposure. For clinical and toxicology laboratories, a method for simultaneously determining these compounds and their metabolic products in body fluids, such as blood and urine, is important. In the present study, a rapid, sensitive and simultaneous LC-QToF-MS method for detecting multiclass pesticides and metabolites in blood and urine samples has been developed and validated. Four sample preparation procedures, protein precipitation and three different variants of QuEChERS-based extraction were evaluated to find a suitable, simple, and effective sample pretreatment technique. The final optimized sample preparation method (acetonitrile; 400 μl, MgSO4; 40 mg and NaCl; 10 mg) was validated for accuracy, precision, matrix effect, recovery, stability, carryover, and dilution integrity. Analyte recoveries ranged from 75.40 to 113.54 % while accuracy was evaluated in the range of 71.41–108.26 % and precision (%RSD) in the range of 0.01 %–16.85 %. The limit of quantification (LOQ) for all compounds was established in the range of 0.82–7.05 ng mL-1. The developed reliable, robust, and sensitive method was successfully applied for the quantification of target pesticides and metabolites in human blood and urine samples. Evaluated samples resulted in detection of eleven analytes (seven pesticides and four metabolites).

Analysis of PFAS and further VOC from fluoropolymer-coated cookware by thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS)

Per- and polyfluoroalkyl substances (PFAS) are used in the production of PTFE based coatings for cookware. In this study, emission of PFAS and further volatile organic compounds (VOC) from kitchenware articles were investigated. First, method development for thermal extraction of baking trays, frying pans and baking mats at 250 °C was done by testing three different extraction devices. A thermal desorption oven showed the best blank and highest recoveries of PFAS analytes (70–101% for 12 perfluorocarboxylic acids (PFCAs), 2 fluorotelomer alcohls (FTOHs), 3 per- and polyfluoroether carboxylic acids (PFECAs), 1 polyfluoroether (PFE)). Second, 18 cookware samples, a PTFE micro powder and 2 lab made coating strips have been investigated. No PFAS were detected in 12 samples (limits of detection: 1–13 ng/dm2). PFCAs (C5–C23) were detected in a baking tray in amounts up to 34 ng/dm2. A baking mat contained 3 ng/dm2 PFOA. FTOHs were not detected in the samples. A PFECA (bC7O2) and its hydride (bC6O2H PFE) were detected in one coating intended for use in frying pans. The hydrides of the PFECA mixture, b(C3O1)nC3 PFECA (Krytox 157FSH), were detected in five baking trays. The PFAS target analytes were not detectable in the five investigated frying pans. Analysing further VOC in the emissions of coatings, 175 compounds could be identified, including alkanes, alkenes, aromatic substances, esters, aldehydes, ketones, ethers, alcohols, carboxylic acids, siloxanes and sulphur, nitrogen, as well as chlorine containing compounds (< 10 µg/dm2). The identified substances cannot be connected to the basic coating polymer of the kitchenware articles, which were PTFE and PES. All samples have undergone a threefold thermal extraction. No substances could be detected in the second and third consecutive extraction, which means that a removal and no new formation of the investigated PFAS as well as the further VOC at 250 °C has occurred.

Electropolymerized InZnSe@PtAg quantum dots@molecularly imprinted polymer on screen-printed carbon electrodes for the ultrasensitive detection of NS1 dengue virus protein with smartphone-based sensing in saliva

According to the World Health Organization (WHO), an estimated 100–400 million cases of dengue virus (DENV) infections are recorded annually with half of the global population being at risk of infection. Currently, there is no known treatment for DENV; however, early, rapid and accurate (sensitive and selective) detection can help to alleviate fatality rates. This work reports on the novel development of a point-of-care electrochemical biosensor for DENV using nanostructured InZnSe@PtAg quantum dots (QDs)-molecularly imprinted polymer (MIP) with smartphone-based detection functionality. Highly conductive InZnSe@PtAg QDs were newly synthesized in the presence of metal precursors, organic surfactants and ligands and surface capped with glutathione (GSH) using a ligand exchange reaction. PtAg was used as an electroactive shell layer on the InZnSe QDs core surface to increase the QDs conductivity. The GSH-InZnSe@PtAg QDs were drop-casted onto screen-printed carbon electrodes (SPCEs) and electropolymerized using cyclic voltammetry (CV) in the presence of o-phenylenediamine and the template DENV. The robust electropolymerization process allowed the overcoating of the MIP layer on the QDs/SPCE, where specific DENV size and shape cavities were created. Under optimal experimental conditions, DENV was rapidly, selectively and ultra-sensitively detected. Using differential pulse voltammetry (DPV), quantitative rebinding of DENV on the MIP@QDs/SPCE surface led to a steady decrease of the anodic peak current and a limit of detection of 1.36 pg/mL was obtained for DENV detection. Using a hand-held smartphone-based potentiostat, DENV was successfully detected in human saliva with satisfactory analytic recoveries.

SWIEET-a salt-free alternative to QuEChERS.

The efficient extraction of various analytes from a wide spectrum of matrices with organic solvents is still a great challenge in analytical chemistry. Especially polar and charged compounds are hard to extract in combination with neutral analytes of intermediate to low polarity. The QuEChERS method is often chosen and has been adapted not only to the analysis of food samples, but also to environmental matrices (soil, wastewater) or biota. In this study, we overcome major drawbacks of QuEChERS such as low recoveries of charged analytes and impairment of downstream analysis by high salt loads. The new extraction method, applicable to liquid and solid samples, is called SWIEET (sugar water isopropanol ethyl nitrile extraction technique). Phase separation of the otherwise miscible extraction solvents water and acetonitrile is achieved by sugaring-out instead of salting-out. Extraction efficiencies were greatly improved by adding isopropanol to the acetonitrile phase. The concentrations of the additives glucose and isopropanol, as well as temperature, were optimized by a design of experiment. Further improvement was achieved through electro- or double-extractions. For all sample types tested (surface water, wastewater treatment plant effluent, tomato, soil, and oats), recoveries and precision were higher with SWIEET than with the established QuEChERS method. From wastewater treatment plant effluent, 75% recovery on average were achieved with our SWIEET method compared to 37% with QuEChERS for a model analyte mixture with polarities of logDpH7 = - 5.7 - 3.5. Higher recoveries and lower standard deviations compared to QuEChERS were achieved especially for polar and charged analytes such as metformin. Handling proved to be easy, since there was no additional solid phase and no tedious weighing of salts.

Green and sensitive method combining ultrasonic solvent extraction, stir bar sorptive extraction and thermal desorption-gas chromatography–tandem mass spectrometry for determination of organotin compounds in sediment

A method integrating ultrasonic solvent extraction (USE) and stir bar sorptive extraction (SBSE) with thermal desorption-gas chromatography–tandem mass spectrometry (TD-GC–MS/MS) was developed and validated for determining critical organotin compounds (OTCs)—tributyltin (TBT) and triphenyltin (TPhT)—in sediment. In preparing a 0.5 g sediment sample, two 1-min cycles of US bath solvent extraction (using equal parts CH3COOH and MeOH), were followed by derivatization with NaBEt4 in the presence of a methanolic-aqueous solution of CH3COONa and SBSE for 1 h, achieving analyte recoveries of 64–94 %. Matrix-matched internal standard calibration and the use of an isotopically labelled recovery standard corrected for analyte losses and matrix effect (ME), ensuring accurate results for triorganotins. However, the method was not suitable for mono- and diorganotins due to low and anomalous recoveries caused by ME. Miniaturizing and optimizing the sample preparation reduced chemicals use and waste, contributing to the method being deemed ’an excellent green analysis’ by the Analytical Eco-Scale assessment. The method achieved a limit of quantification (LOQ) of 0.081 ng g−1 and an expanded uncertainty (U) value of 15 % for TBT, meeting the environmental quality standards (EQS) criteria of the European Union Water Framework Directive. Its applicability was demonstrated through the analysis of actual sediment samples and natural matrix certified reference materials.

Disposable glutamate biosensor based on platinum nanoparticles, carbon quantum dots and poly-L-aspartic acid

This study reports the design and development of a disposable amperometric biosensor for the determination of L-glutamate. Glutamate oxidase (GlOx) was immobilized onto a screen-printed carbon electrode (SPE) modified with poly-L-Aspartic acid (PAsp), carbon quantum dots (CQD), and platinum nanoparticles (PtNP) for the construction of the biosensor. The surface composition of the modified SPE was optimized using the one variable at a time method. The morphological properties of the biosensor were characterized by scanning electron microscopy and energy-dispersive X-ray spectroscopy. The electrochemical behavior of the modified electrodes was studied by cyclic voltammetry. Under the optimized experimental conditions the linear working range, detection limit and sensitivity of the GlOx/PtNP/CQD/PAsp/SPE were found to be 1.0 − 140 µM, 0.3 µM and 0.002 µA µM−1, respectively. The GlOx/PtNP/CQD/PAsp/SPE biosensor also exhibited good measurement repeatability. The as-developed biosensor was applied for the determination of L-glutamate in spiked serum samples and the average analytical recovery of added glutamate was 98.9 ± 3.9%.

Development and validation of a UPLC-MS/MS method for rapid and simultaneous quantification of BPI-460372 and its metabolites BPI-460444 and BPI-460456 in human plasma

In cancer development and progression, the Hippo signaling pathway functions. The transcriptional enhanced associate domain (TEAD) stands out as a pivotal transcription factor within this pathway, and the suppression of TEAD represents a promising approach for cancer treatment. The primary aim of the study was to establish an analytical method for the concurrent quantification of a novel TEAD target inhibitor, BPI-460372, and its principal metabolites, BPI-460444 and BPI-460456, in human plasma. The chromatographic separation utilized a XSelect™ HSS C18 column (2.1 × 100 mm, 2.5 µm), while quantification was conducted on a SCIEX API 4000 mass spectrometer. 22 plasma samples were tested via the developed method. The calibration curve for BPI-460372 exhibited linearity from 2 to 2000 ng/mL, while its metabolites BPI-460444 and BPI-460456 had linearity between 1 and 1000 ng/mL (r > 0.99). The precision (RSD) was ≤ 17.1 %, and the accuracy (RE) fell within the range of −17.7 % to 15.0 %, all meeting acceptance criteria. The matrix effect was from 101.0 % to 105.8 %. The extraction recovery of analytes fell within the range of 96.8 % to 104.1 % with an RSD of less than 7.4 %. The developed method was effectively utilized in an advanced solid tumor patient, and the concentration trends of the three analytes in plasma were found to be largely consistent. The established analytical method showed great sensitivity, simplicity, accuracy, and reliability for the rapid and simultaneous analysis of the TEAD target inhibitor BPI-460372, alongside its major metabolites BPI-460444 and BPI-460456 in human plasma. This analytical method provided essential support for future clinical investigations and pharmacokinetic analysis.