Solid-phase Extraction Liquid Chromatography Research Articles

Application of Solid-Phase Extraction and High-Performance Liquid Chromatography with Fluorescence Detection to Analyze Eleven Bisphenols in Amniotic Fluid Samples Collected during Amniocentesis.

Amniocentesis involves taking a sample of the amniotic fluid in order to perform a karyotype test and diagnose any genetic defects that may affect the fetus. Amniotic fluid has been collected from patients with an indication for amniocentesis in the 15–26th week of pregnancy. A simple and sensitive high-performance liquid chromatography with fluorescence detection (HPLC-FLD) method for identification and quantification of eleven selected bisphenols in amniotic fluid samples is proposed. The proposed method involved protein precipitation using acetonitrile, and next the extraction and concentration of analytes by solid-phase extraction (SPE). The solid-phase extraction (SPE) procedure with application of Oasis HLB SPE columns performed well for the majority of the analytes, with recoveries in the range of 67–121% and relative standard deviations (RSD%) less than 16%. The limits of detection (LODs) and quantification (LOQs) of all the investigated analytes were in the range of 0.8–2.5 ng mL−1 and 2.4–7.5 ng mL−1 (curves constructed in methanol) and 1.1–5.2 ng mL−1 and 3.2–15.6 ng mL−1 (curves constructed in the amniotic fluid), respectively. The method was validated at the following two concentration levels: 10 ng mL−1 (2 × LOQ) and 20 ng mL−1 (4 LOQ). The results confirm the validity of the SPE procedure and HPLC-FLD method for identification and quantification of bisphenols in amniotic fluid samples collected during an amniocentesis. The result obtained show that HPLC-FLD is a useful method for determination of bisphenol residues at nanogram per milliliter concentrations in amniotic fluid samples. Residues of five analytes (BADGE·2H2O, BPAF, BADGE, BADGE·H2O·HCl and BADGE·2HCl) were detected in amniotic fluid samples. Additionally, the harmfulness of bisphenols as potential pathogens that may cause karyotype disorders and contribute to preterm birth was estimated.

Determination of ten iodinated X-ray contrast media by solid-phase extraction and ultra-high performance liquid chromatography coupled with high-resolution orbitrap mass spectrometry

A highly sensitive method for the simultaneous determination of ten iodinated X-ray contrast media (ICM) in environmental water samples was developed, in which samples were pretreated using solid-phase extraction and analyzed by ultra-high performance liquid chromatography coupled with high-resolution orbitrap mass spectrometry. A combination of two ENVI-Chrom P cartridges was selected due to its high recoveries of ICM. The optimized sample pH, flow rate, eluent type, and volume of eluent were 2.0, 6 mL/min, MeOH/ACN (50:50, v/v), and 6 mL, respectively. The developed method was used for the tap, surface, and waste water sample analysis, and exhibited satisfactory accuracy (80.8-118.5%), high precision (0.1-11.6%), and low detection limits (0.3-1.0 ng/L). These results indicate that this method is highly effective for the simultaneous determination of trace-level ICM in the aquatic environment.

Preparation of Fe3O4@SW-MIL-101-NH2 for selective pre-concentration of chlorogenic acid metabolites in rat plasma, urine, and feces samples

An innovative sandwich-structural Fe-based metal-organic framework magnetic material (Fe3O4@SW-MIL-101-NH2) was fabricated using a facile solvothermal method. The characteristic properties of the material were investigated by field emission scanning electron microscopy, transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, X-ray powder diffraction, vibrating sample magnetometry, and Brunauer-Emmett-Teller measurements. Fe3O4@SW-MIL-101-NH2 is associated with advantages, such as robust magnetic properties, high specific surface area, and satisfactory storage stability, as well as good selective recognition ability for chlorogenic acid (CA) and its metabolites via chelation, hydrogen bonding, and π-interaction. The results of the static adsorption experiment indicated that Fe3O4@SW-MIL-101-NH2 possessed a high adsorption capacity toward CA and its isomers, cryptochlorogenic acid (CCA) and neochlorogenic acid (NCA), and the adsorption behaviors were fitted using the Langmuir adsorption isotherm model. Then, a strategy using magnetic solid-phase extraction (MSPE) and ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF MS/MS) was developed and successfully employed for the selective pre-concentration and rapid identification of CA metabolites in rat plasma, urine, and feces samples. This work presents a prospective strategy for the synthesis of magnetic adsorbents and the high-efficiency pretreatment of CA metabolites.

Characteristics and Risk Assessment of Antibiotic Contamination in Chishui River Basin, Guizhou Province, China

To preliminarily explore the antibiotic concentration distribution characteristics of Guizhou Chishui River basin surface water and potential ecological risks, we used solid phase extraction liquid chromatography tandem mass spectrometry (SPE-HPLC- MS) to analyze 21 types of antibiotics in surface water samples. Twelve types of antibiotics were detected in the Chishui River basin surface water, and the total concentrations of ofloxacin, sulfadiazine, and trimethoprim ranged from ND-1166.97 ng·L-1, with a detection rate of 100%. On average, the highest concentration of the three types of antibiotics detected were ofloxacin (221.59 ng·L-1), tetracycline (13.18 ng·L-1), and sulfadiazine (4.11 ng·L-1), and the antibiotic concentration distribution showed the following order of characteristics:downstream (359.41 ng·L-1) > midstream(224.59 ng·L-1) > upstream (179.72 ng·L-1). The ecological environment risk assessment results indicated the largest risk for downstream W21, tetracycline, doxycycline, enrofloxacin, norfloxacin, and erythromycin. The risk quotient revealed that lincomycin had a medium-risk level, and ofloxacin had a high-risk level. This shows that antibiotics in the waters of the Chishui River basin may cause certain ecological risks.

Simultaneous Determination of Selected Pesticides And/Or Their Metabolites in Urine by Off-Line Solid Phase Extraction and Ultra-High Performance Liquid Chromatography / Hybrid Quadrupole Time-of-Flight Mass Spectrometry

Simultaneous Determination of Selected Pesticides And/Or Their Metabolites in Urine by Off-Line Solid Phase Extraction and Ultra-High Performance Liquid Chromatography / Hybrid Quadrupole Time-of-Flight Mass Spectrometry

Evaluation of the diffusion of triamcinolone and demeclocycline through the dentinal tubules and apical foramen: A mass spectrometry study.

Aim:The aim of this study was to investigate the diffusion of triamcinolone and demeclocycline from an endodontic paste when used unmodified, versus when combined in equal parts with a calcium hydroxide paste, in terms of diffusion through the dentinal tubules versus through the apical foramen.Methodology:Medicaments were placed in endodontically prepared roots that were kept in vials of Milli-Q water. The five experimental groups in the study were (1) control – no medicament, (2) medicament containing triamcinolone and demeclocycline (T&D) and occluded apex, (3) T&D paste and patent apex, (4) T&D + calcium hydroxide (Ca(OH)2) occluded apex, and (5) T&D + Ca(OH)2 and patent apex. The triamcinolone and demeclocycline concentrations were measured with solid-phase extraction and ultra-high performance liquid chromatography–mass spectrometry, after 1, 3, 8, and 24 h, and after 1 week.Results:Most of the triamcinolone and demeclocycline diffused through the apical foramen, with sparse diffusion through the dentinal tubules. The T&D paste mixed with Ca(OH)2 in equal amounts showed greater than the expected 50% reduction in the diffusion of triamcinolone and demeclocycline from mass dilution alone (89% and 80%, respectively).Conclusions:These results stress the importance of maintaining apical patency, for allowing diffusion of active components of the drugs to target tissues in the periapical environment.

Non-Steroidal Anti-Inflammatory Drugs Current Challenges and Future Perceptions

An analytical methodology for the simultaneous determination of seven pharmaceuticals and two metabolites belonging to the non-steroidal anti-inflammatory drugs (NSAIDs) and analgesics therapeutic groups was developed based on off-line solid-phase extraction and ultra-high performance liquid chromatography coupled to tandem mass spectrometry 1 (SPE–UHPLC–MS/MS). Extraction conditions were optimized taking into account parameters like sorbent material, sample volume and sample 1,2 PH. Method detection limits (MDLs) ranging from 0.02 to 8.18 ng/L were obtained 2,3. This methodology was successfully applied to the determination of the selected pharmaceuticals in seawater samples of Atlantic Ocean in the Northern Portuguese coast. All the pharmaceuticals have been detected in the seawater samples, with pharmaceuticals like ibuprofen, acetaminophen, ketoprofen and the metabolite hydroxy ibuprofen being the most frequently detected at concentrations that can reach some hundreds of ng/L 4.

Impact of Artificial Infiltration on the Removal of Nonsteroidal Anti-Inflammatory Drugs during Treatment of Surface Water

The content of pharmaceuticals in natural waters is steadily increasing. Especially nonsteroidal anti-inflammatory drugs (NSAIDs) are often detected in natural waters due to their widespread use. This group of compounds includes commonly used representatives, such as paracetamol and ketoprofen. The quality of natural waters determines the processes applied for the treatment of drinking water. The methods used in order to remove pharmaceuticals from treated water include adsorption and biologically active filtration. Both processes also occur during artificial infiltration (forced flow of intake surface water through the ground to the collecting wells) at surface water intakes. The processes, which occur in the soil, change the water quality characteristics to a great extent. The goal of the study was to evaluate the removal efficiency of paracetamol and ketoprofen in the process of artificial infiltration used as a pre-treatment of surface water. The studies were conducted at a field experimental installation located at the technical artificial infiltration intake. The experimental installation consisted of three metering wells (piezometers) which were located on the way between the bank of the infiltration pond and the collecting well. The collected water samples allowed to evaluate the change of selected NSAIDs concentrations during the passage of water through the ground. The analysis procedure included solid phase extraction (SPE) and high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Removal of the studied NSAIDs in the infiltration process occurred with variable effectiveness throughout the year. Paracetamol was removed with annual efficiency equal to 42%, although no significant removal of ketoprofen was observed.

Detection and quantification of multiclass antibiotic residues in poultry products using solid-phase extraction and high-performance liquid chromatography with diode array detection

This article describes the initial study on the simultaneous determination of multiclass antibiotic residues in imported and local frozen poultry specimens, including turkey gizzard and muscle tissues, and chicken muscle tissues, commonly consumed in Ogun State, Nigeria. Minced tissues were treated with phosphate buffer adjusted to pH 7 that was cleaned using C18 SPE-column (Supelclean™) cartridge. For the determination of six antibiotic residues including fluoroquinolones, sulfonamides, and macrolides, a solid-phase extraction method was used, followed by extract analysis using high-performance liquid chromatography–diode array detection (HPLC–DAD). The coefficient of determination (R2) for the external standards for all the analytes ranged between 0.963 and 0.999. The limit of detection (LOD) and quantification (LOQ) ranged between 5.37 – 55.4 μg/kg, and 17.9–185 μg/kg, respectively. Enrofloxacin, sulfadimethoxine, sulfamerazine, and tylosin showed high concentration levels in the frozen poultry beyond acceptable maximum residue limits (MRLs). The six drugs considered in this study were present at higher concentrations in domestic chicken tissues than the permissible level. This suggests that farmers do not observe the cessation period before poultry birds previously treated with antibiotics are sold to consumers thus exposing them to potentially hazardous antibiotic residues.

Combination of pipette tip solid phase extraction and high performance liquid chromatography for determination of plant growth regulators in food samples based on the electrospun covalent organic framework/polyacrylonitrile nanofiber as highly efficient sorbent

Facile and sensitive determination of plant growth regulators (PGRs) in food samples is important but still remains great challenge. Herein, a pipette tip solid phase extraction (PT-SPE) method was developed for fast and sensitively detecting PGRs. The PT-SPE adsorbent was prepared by integrating a novel covalent organic framework (COF) of schiff base network 3 (SNW-3) and polyacrylonitrile (PAN) through electrospinning. The SNW-3 can easily adsorb PGRs with high special affinity through electrovalent bands between the ammonium ions of SNW-3 and the carboxy groups of PGRs. The polymer of PAN acts as scaffold material for SNW-3, which can lower seepage pressure hence accelerates adsorption/desorption kinetics. By combination with HPLC-DAD, a satisfactory method was successfully developed for simultaneous determination of ten PGRs in watermelon. Good analytical performances were achieved with this proposed method, including good linearity (5–500 ng/mL) with high correlation coefficients (R ≥ 0.9981), low limits of detection (S/N = 3, 0.24–3.19 ng/mL) and limits of quantification (S/N = 10, 1.65–5.72 ng/mL), satisfactory precision (intra-day RSDs ≤ 2.7%, inter-day RSDs ≤ 3.7%), and high accuracy (recovery: 82.8–113.0%). The method developed in this study shows high potential for design of high target-affinity adsorbents for food sample preparing.

Biochar for the removal of detected micropollutants in South African domestic wastewater: a case study from a demonstration-scale decentralised wastewater treatment system in eThekwini

The presence of micropollutants, such as pharmaceuticals and personal care products, in surface and ground water has escalated globally, leading to adverse effects on aquatic organisms in receiving waters. Untreated or inadequately treated wastewater is the main source of micropollutants entering the environment. In South Africa, the consumption of antibiotics and antiretroviral drugs is relatively higher than other nations; however, little data exists on the identification and remediation of micropollutants in domestic wastewater. In this study, a novel method to detect and measure 71 micropollutants using online solid phase extraction liquid chromatography coupled to tandem mass spectrometry was developed. To test the method in the South African context, grab samples of the influent and anaerobically treated effluent (AF effluent) from a demonstration-scale decentralised wastewater treatment system in eThekwini (Durban) were taken over 3 consecutive days at 2 time points. The presence of 24 micropollutants was detected in the raw wastewater, with analgesics/anti-inflammatory drugs, antiretrovirals, and antibiotics showing the highest concentrations and with the majority of compounds still present in the AF effluent. One antibiotic, ciprofloxacin, exceeded its predicted no-effect concentration in all influent and AF effluent samples. This suggests that the anaerobic treatment of the raw wastewater was not effective in removing micropollutants. Preliminary data from lab-scale adsorption experiments using biochar produced from a set of 4 feedstocks – olive residues, tomato residues, rice husks, and the African palm tree Raphia farinifera – showed average removal rates for 4 compounds of up to 62%. The application of biochar is thus recommended as a secondary treatment step in decentralised wastewater treatment for the removal of micropollutants in South Africa.

Taste-Active Dipeptides from Hydrolyzed Mushroom Protein Enhance Saltiness.

An activity-guided fractionation approach applied to thermally treated, enzymatically hydrolyzed mushroom, Agaricus bisporus L., protein led to the identification of several saltiness- and kokumi-enhancing peptides. The identification was accomplished by employing a combination of solid-phase extraction (SPE), gel-permeation chromatography (GPC), and semipreparative reverse-phase high-performance liquid chromatography (RP-HPLC), coupled with sensory analysis. As a result, this study led to the identification of a collection of common mushroom derived tastants, including 5'-mononucleotides and free amino acids, along with several taste-modulating pyroglutamyl dipeptides, including pyroglutamylcysteine (pGlu-Cys), pyroglutamylvaline (pGlu-Val), pyroglutamylaspartic acid (pGlu-Asp), pyroglutamylglutamic acid (pGlu-Glu), and pyroglutamylproline (pGlu-Pro). The taste-modulating thresholds for the pyroglutamyl dipeptides were calculated in a model mushroom broth containing natural concentrations of guanosine 5'-monophosphate and 14 amino acids, all with dose-over-threshold (DoT) factors ≥1. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed to quantitate the pyroglutamyl dipeptides, and their concentrations ranged from 2 to 58 μmol/L; however, they were determined to be present in the hydrolysate below their individual taste-modulating thresholds. Despite being present below their individual thresholds, when the dipeptides were collectively added to a model mushroom broth at their natural concentrations (143 μmol/L combined), both salty (p = 0.0061) and kokumi (p = 0.0025) taste attributes were significantly enhanced, demonstrating a synergistic subthreshold taste-modulating effect. This study lays the groundwork for future investigations on the saltiness-enhancing potential of mixtures of subthreshold levels of pyroglutamyl dipeptides found in mushrooms and other sources.

Application of Solid Phase Extraction and High-Performance Liquid Chromatography with Fluorescence Detection to Analyze Bisphenol A Bis (2,3-Dihydroxypropyl) Ether (BADGE 2H2O), Bisphenol F (BPF), and Bisphenol E (BPE) in Human Urine Samples.

In this study, we propose a simple, cost-effective, and sensitive high-performance liquid chromatography method with fluorescence detection (HPLC-FLD) for the simultaneous determination of the three bisphenols (BPs): bisphenol A bis (2,3-dihydroxypropyl) ether (BADGE 2H2O), bisphenol F (BPF), and bisphenol E (BPE) in human urine samples. The dispersive solid phase extraction (d-SPE) coupled with solid phase extraction (SPE) procedure performed well for the analytes with recoveries in the range of 74.3–86.5% and relative standard deviations (RSD%) less than 10%. The limits of quantification (LOQs) for all investigated analytes were in the range of 11.42–22.35 ng mL−1. The method was validated at three concentration levels (1 × LOQ, 1.5 × LOQ, and 3 LOQ). During the bisphenols HPLC-FLD analysis, from 6 min a reinforcement (10 or 12) was used, therefore analytes might be identified in the small volume human urine samples. The results demonstrated clearly that the approach developed provides reliable, simple, and rapid quantification and identification of three bisphenols in a urine matrix and could be used for monitoring these analytes.

Development, validation, and application of a new method for the quantitative determination of monohydrogen-substituted perfluoroalkyl carboxylic acids (H–PFCAs) in surface water

Per- and polyfluoroalkyl substances (PFASs) are a large and diverse class of chemicals. While some have been phased out internationally due to concerns over their human and environmental health risks, novel alternative PFASs continue to be manufactured and detected in environmental samples. The occurrence and fate of these alternatives remain poorly understood. The present study investigated the occurrence of an emerging class of PFAS alternative, the monohydrogen-substituted perfluoroalkyl carboxylic acids (H–PFCAs), in conjunction with the more well-known PFCAs. A weak anion exchange solid phase extraction-liquid chromatography tandem mass spectrometry method for quantitative determination of H–PFCAs in surface water was developed, validated, and applied on samples collected from the Netherlands. To improve chromatography, especially for short-chain (H-)PFCAs, an ion-pairing agent, tetrabutylammonium hydrogen sulphate, was used. The method was validated for linearity (R2 > 0.99), instrumental detection limits (0.01–0.09 ng/mL), method detection limits (0.03–0.75 ng/mL), matrix effects (<20%), percent absolute- and relative recovery (57–121%), trueness (130-80%), repeatability (<20%), and within-lab reproducibility (<20%). Eleven out of fourteen PFASs showed acceptable results. Application of the newly validated method to surface water throughout the Netherlands revealed trace levels of H–PFCAs (including two new H–PFCAs) and high concentrations of PFCAs.

Illicit drug use in Reykjavik by wastewater-based epidemiology

Estimation of illicit drug use on a community level by wastewater-based epidemiology (WBE) is both an objective and reliable way to establish near real-time results. Wastewater samples were collected at eleven timepoints in Reykjavik from 2017 to 2020. The use of commonly abused illicit drugs in Iceland (amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine (MDMA), cocaine, and cannabis) was estimated. Solid phase extraction and ultra-high performance liquid chromatography coupled to tandem mass spectrometry was used for analysis. Estimated amphetamine and methamphetamine use showed signs of an increase from 2017 to 2020 with amphetamine being the dominant stimulant on the market. MDMA use remained stable from 2017 to 2020. Results showed a large increase in cocaine use from 2017 to 2019 but interestingly, a marked decrease in 2020 during the COVID-19 pandemic. Cannabis use was stable from 2017 to 2019 but showed signs of an increase during the pandemic in 2020. Results by WBE corresponded with data based on two other indicators of drug use, seizure data and driving under the influence cases. Both temporal and spatial trends in illicit drug use were successfully estimated by using WBE, complimenting other indicators which provided a comprehensive picture of drug abuse in Reykjavik.

Characterization of Five Portuguese Wastewater Treatment Plants: Removal Efficiency of Pharmaceutical Active Compounds through Conventional Treatment Processes and Environmental Risk

Due to the high consumption and incorrect disposal of pharmaceutical active compounds (PhACs), they are recognized as contaminants of emerging concern. Wastewater treatment plants (WWTPs) may be inefficient in removing PhACs, therefore discharging them into surface waters. The removal efficiencies of five WWTPs located in the south of Portugal (Alentejo) were evaluated in 2020. Twenty-six PhACs were analyzed in wastewater influents, effluents, and surface waters, upstream and downstream of the WWTPs by solid-phase extraction (SPE) and ultra-performance liquid chromatography coupled with tandem mass detection (UPLC-MS/MS). The most representative PhACs in influents were acetaminophen, caffeine, naproxen, ibuprofen, and diclofenac with minimum-maximum concentrations of 49–225 µg/L, 26–46 µg/L, 5.9–13 µg/L, 5.2–22 µg/L, and 1.3–2.5 µg/L, respectively. For effluents, it was acetaminophen, caffeine, and diclofenac with minimum-maximum concentrations of 0.054–7.8 µg/L, 0.084–4.8 µg/L, and 0.28–3.3 µg/L, respectively. The highest removal efficiencies were observed for acetaminophen, sulfadiazine, cortisone, testosterone, metoprolol, and propranolol (100%). The lowest removal efficiencies were observed for carbamazepine (2.7%) and diclofenac (−13.2%). The risk quotient of sulfamethoxazole and diclofenac were higher than 1 for receiving waters, indicating they probably pose high risks to aquatic organisms.

N-Terminally Truncated and Pyroglutamate-Modified Aβ Forms Are Measurable in Human Cerebrospinal Fluid and Are Potential Markers of Disease Progression in Alzheimer's Disease.

Alzheimer’s disease (AD) is a pathology characterized by the accumulation in the brain of intracellular and extracellular amyloid-β (Aβ) aggregates, especially of Aβ1–40 and Aβ1–42 peptides. It is known that N-terminally truncated or modified Aβ forms also exist in AD brains and cerebrospinal fluid (CSF), and they play a key role in the pathogenesis of the disease. Herein, we developed an antibody-free method based on Solid-Phase Extraction and Electrospray Ionization Liquid Chromatography Mass Spectrometry for the identification and quantitation in human CSF of Aβ isoforms. In human CSF, we could detect and quantify a panel of 19 Aβ isoforms, including N-terminally truncated and pyroglutamate-modified forms, never quantified before in CSF. Among these, we identified novel N-terminally truncated Aβ species: four bound to copper and two phosphorylated forms, which were found to be the most common proteoforms in human CSF along with Aβ1–40, Aβ3–40, and AβpE11–42. We tested the newly developed and validated method in a pilot study on CSF from elderly individuals with subjective memory complaints (SMCs, n = 9), mild cognitive impairment (MCI, n = 18), and AD (n = 15); along with Aβ1–42, five N-terminally truncated forms (Aβ11–40, Aβ3–42, AβpE11–42, AβpE3–40, and Aβ4–40 Cu2+) are altered in AD/MCI. Thus, we demonstrated that N-terminally truncated and pyroglutamate-modified Aβ can be quantified in human CSF, and five of them, along with Aβ1–42, are potential markers of AD progression. The described method could represent a useful tool for patients’ stratification and monitoring. Moreover, the newly identified Aβ CSF species might represent new potential therapeutic targets.

Determination of macrolides in animal tissues and egg by multi-walled carbon nanotube-based dispersive solid-phase extraction and ultra-high performance liquid chromatography–tandem mass spectrometry

A simple and reliable analytical method was developed for the simultaneous determination of 11 macrolides in swine, chicken, bovine, and sheep tissues (muscle, liver, kidney, and fat), as well as eggs. Samples were extracted using a mixture of acetonitrile, ethyl acetate, and methanol; dispersive solid-phase extraction purification was then performed using multi-walled carbon nanotubes as the sorbent. The analytes were separated through ultra-high performance liquid chromatography and detected by electrospray ionization on a triple quadrupole mass spectrometer. The average recoveries ranged from 83.5% to 111.4%; the corresponding intra-day and inter-day relative standard deviations were less than 13.6% and 16.4%, respectively. The limit of detection and quantification of the eggs were 0.1–0.6 and 2.0 μg/kg, respectively. For other tissues, the limits of detection and quantification were 0.1–2.0 μg/kg and 5.0 μg/kg, respectively. The proposed method was successfully employed for the analysis of real samples to demonstrate its applicability.

Extraction and determination of the Pimelea toxin simplexin in complex plant-polymer biocomposites using ultrahigh-performance liquid chromatography coupled with quadrupole Orbitrap mass spectrometry.

In the present paper, we describe how a robust and fundamental methodology was developed for extraction and determination of a principal natural toxin compound, simplexin, from a series of bulk biocomposites. These complex matrices were fabricated by direct encapsulating either ground plant particles or an ethanolic crude extract of the Australian toxic pasture plant Pimelea trichostachya in the biodegradable polymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate). Proton nuclear magnetic resonance spectroscopy was initially employed to examine the chemical compositions of these complicated systems. Then, a more sensitive strategy was developed and validated by combining solid-phase extraction and ultrahigh-performance liquid chromatography hyphenated with a quadrupole Orbitrap mass spectrometer for the quantification of simplexin embedded in different biocomposites. Satisfactory linearity (R2 > 0.99) and recovery ranges (86.8-116%) with precision (relative standard deviations) of between 0.2 and 13% (n = 3) were achieved from seven biocomposites. The established protocol was further shown to be accurate and reliable in confirming the homogeneous distribution of the simplexin in different biocomposite formulations. A limited mass transfer of simplexin (< 3.5%) from one of the biocomposites into a simulated but sterilized in vitro rumen environment after a 10-day incubation was also revealed by utilizing the method. This quantitative analysis of targeted natural product within plant material-integrated polymeric platforms has potential application when controlled release is required in the bovine rumen and other biological systems. Graphical abstract.

Determination of Paralytic Shellfish Toxins in Bivalve Mollusks by Amino-Modified Multiwalled Carbon Nanotube (MWCNT) Solid-Phase Extraction (SPE) and High-Performance Liquid Chromatography–Tandem Mass Spectrometry (HPLC–MS/MS)

An amino-modified multiwalled carbon nanotube (MWCNT) solid-phase extraction-based hydrophilic interaction liquid chromatography–tandem mass spectrometry (HILIC–MS/MS) method was developed for simultaneous determination of 13 paralytic shellfish toxins (PSTs) in bivalve mollusks. Shellfish samples were extracted with 1% acetic acid followed by cleanup using optimized MWCNT-NH2 based solid-phase extraction. Absorbed PSTs were eluted with 2 mL 1% acetic acid–acetonitrile (70/30, v/v). The separation was performed on a Waters BEH Amide column within 7.5 min utilizing 0.1% formic acid–acetonitrile and 2 mM ammonium acetate containing 0.1% formic acid as the mobile phase. The analytes were quantified by matrix-matched external standard curves with correlation coefficients® exceeding 0.997 acquired in multiple reaction-monitoring mode. The limits of detection and quantitation in shellfish samples were from 2.76 to 9.8 μg/kg and 9.11 to 39.3 μg/kg, respectively. Recoveries at three spiked levels ranged from 71.4% to 97.8% with relative standard deviations less than 12.6%. The developed method was satisfactorily applied to the analysis of shellfish.