Liquid Chromatography-fluorescence Detection Method Research Articles

High-Performance Liquid Chromatography-Fluorescence Detection Method for Ochratoxin A Quantification in Small Mice Sample Volumes: Versatile Application across Diverse Matrices Relevant for Neurodegeneration Research.

Ochratoxin A (OTA) is a mycotoxin commonly found in various food products, which poses potential health risks to humans and animals. Recently, more attention has been directed towards its potential neurodegenerative effects. However, there are currently no fully validated HPLC analytical methods established for its quantification in mice, the primary animal model in this field, that include pivotal tissues in this area of research, such as the intestine and brain. To address this gap, we developed and validated a highly sensitive, rapid, and simple method using HPLC-FLD for OTA determination in mice tissues (kidney, liver, brain, and intestine) as well as plasma samples. The method was rigorously validated for selectivity, linearity, accuracy, precision, recovery, dilution integrity, carry-over effect, stability, and robustness, meeting the validation criteria outlined by FDA and EMA guidelines. Furthermore, the described method enables the quantification of OTA in each individual sample using minimal tissue mass while maintaining excellent recovery values. The applicability of the method was demonstrated in a repeated low-dose OTA study in Balb/c mice, which, together with the inclusion of relevant and less common tissues in the validation process, underscore its suitability for neurodegeneration-related research.

Phenyl-imidazole based and nitrogen rich hyper-crosslinked polymer for sensitive determination of aflatoxins

Using heterocyclic phenyl-imidazole (2-phenylimidazole, 4,5-diphenylimidazole and 2,4,5-triphenylimidazole) as monomer, three novel hyper-crosslinked polymers (PI-HCP, DPI-HCP and TPI-HCP) with large specific surface area and excellent chemical stability were fabricated by Friedel-Crafts reaction. With PI-HCP as solid phase extraction adsorbent, a high performance liquid chromatography-fluorescence detection method was developed for sensitive determination of aflatoxins B1, B2, G1 and G2 in rice and fragrant rice samples. Under optimized conditions, the established method provided wide linear range, low detection limit and satisfactory recovery. Moreover, the adsorption mechanism was also explored. The polar interaction, hydrogen bonding and π-π stacking interactions between PI-HCP and AFs might be the main adsorption mechanisms. This work provides a powerful tool for simultaneous determination of four AFs in food grains with great application potential in AFs monitoring.

Core-shell magnetic porous organic polymer for magnetic solid-phase extraction of fluoroquinolone antibiotics in honey samples followed by high-performance liquid chromatography with fluorescence detection.

By monomer-mediated in-situ growth synthesis strategy, with hydroquinone and 1,3,5-tris(4-aminophenyl)benzene as monomers, a core-shell magnetic porous organic polymer was synthesized through a simple azo reaction. Based on this, a magnetic solid-phase extraction-high-performance liquid chromatography-fluorescence detection method was proposed for the analysis of fluoroquinolones in a honey sample. With ofloxacin, ciprofloxacin, enrofloxacin, lomefloxacin, and difloxacin as target analytes, factors affecting the extraction efficiency had been optimized. The LODs were 1.5-5.4ng/L (corresponding to 0.23-0.81μg/kg in honey). The linear range was 0.005-20 μg/L for difloxacin, 0.01-20 μg/L for ofloxacin, ciprofloxacin and lomefloxacin, and 0.02-20 μg/L for enrofloxacin. The enrichment factor was 84.4-91.7-fold with a high extraction efficiency of 84.4-91.7%. The method was assessed by the analysis of target fluoroquinolones in honey samples, and the recoveries for the spiked samples were 79.3-95.8%. The results indicated that the established magnetic solid-phase extraction-high-performance liquid chromatography-fluorescence detection method is efficient for the analysis of trace fluoroquinolones in honey.

Column-switching high-performance liquid chromatography-fluorescence detection method for malic acid enantiomers in commercial wines

Determination of malic acid (MA) in commercial drinks is imperative to assure the quality of taste and safety of the product to human health. In this study, a column-switching high-performance liquid chromatography (HPLC)-fluorescence detection method was employed to determine the content of d- and l-MA in wines. The developed method successfully detected both MA enantiomers in several wines with a pre-column fluorescence derivatization procedure. Additionally, the present HPLC system provided sufficient data with a small sample quantity. Moreover, the d-MA peak was detected in the chromatogram of wine samples without acidulants. Thus, we believe that the proposed method would benefit the wine-brewing process and facilitate quality certification in the wine industry.

A new magnetic Fe3O4@SiO2@TiO2-APTMS-CPA adsorbent for simple, fast and effective extraction of aflatoxins from some nuts

A novel modified magnetic adsorbent was synthesized by binding 3-aminopropyltrimethoxysilane and 1,8-bis (3-chloropropoxy) anthracene-9,10-dione to magnetic nanoparticles. The adsorbent has been characterized by transmission electron microscopy, scanning electron microscopy, energy-dispersive X-ray spectroscopy, X-ray diffraction, fourier transform infrared spectroscopy. The extraction parameters of nanoparticle amount, pH, vortex time, and desorption conditions (solvent, volume, time) were optimized by combining aflatoxin (AFB1, AFB2, AFG1, AFG2) analyzes in samples, high-performance liquid chromatography-fluorescence detection method, and post-column derivatization. The proposed method was compared with the conventional immunoaffinity column method. Under optimum conditions, the detection limits (LODs) of the aflatoxins range from 0.05 μg kg−1 to 0.15 μg kg−1. The recovery of the four aflatoxins for the added nut samples ranged from 87.7 % to 97.5 %. In addition, the intra-day and inter-day relative standard deviations were <7.1 %. The satisfactory results show that the synthesized adsorbent has advantages such as good sensitivity and accuracy, environmental friendliness, simplicity, easy of application, reliability, and speed in the extraction of aflatoxins in some nuts.

Eco-Friendly Fluorescent ELISA Based on Bifunctional Phage for Ultrasensitive Detection of Ochratoxin A in Corn.

Conventional enzyme-linked immunosorbent assay (ELISA) is commonly used for Ochratoxin A (OTA) screening, but it is limited by low sensitivity and harmful competing antigens of enzyme-OTA conjugates. Herein, a bifunctional M13 bacteriophage with OTA mimotopes fused on the p3 protein and biotin modified on major p8 proteins was introduced as an eco-friendly competing antigen and enzyme container for enhanced sensitivity. Mercaptopropionic acid-modified quantum dots (MPA-QDs), which are extremely sensitive to hydrogen peroxide, were chosen as fluorescent signal transducers that could manifest glucose oxidase-induced fluorescence quenching in the presence of glucose. On these bases, a highly sensitive and eco-friendly fluorescent immunoassay for OTA sensing was developed. Under optimized conditions, the proposed method demonstrates a good linear detection of OTA from 4.8 to 625 pg/mL and a limit of detection (LOD) of 5.39 pg/mL. The LOD is approximately 26-fold lower than that of a conventional horse radish peroxidase (HRP) based ELISA and six-fold lower than that of a GOx-OTA conjugate-based fluorescent ELISA. The proposed method also shows great specificity and accepted accuracy for analyzing OTA in real corn samples. The detection results are highly consistent with those obtained using the ultra-performance liquid chromatography-fluorescence detection method, indicating the high reliability of the proposed method for OTA detection. In conclusion, the proposed method is an excellent OTA screening platform over a conventional ELISA and can be easily extended for sensing other analytes by altering specific mimic peptide sequences in phages.

Adsorption of aflatoxins and ochratoxins in edible vegetable oils with dopamine-coated magnetic multi-walled carbon nanotubes

A new, green, and cost-effective magnetic solid-phase extraction of aflatoxins and ochratoxins from edible vegetable oils samples was developed using polydopamine-coated magnetic multi-walled carbon nanotubes (PDA@Fe3O4-MWCNTs) as the absorbent. PDA@Fe3O4-MWCNTs nanomaterials were prepared by chemical co-precipitation and in situ oxidation and self-polymerization of dopamine and was characterized. Factors affecting MSPE and the adsorption behavior of the adsorbent to mycotoxins were studied, and the optimal extraction conditions of MSPE and the complexity of the adsorption process were determined. Based on this, the magnetic solid-phase extraction-high-performance liquid chromatography-fluorescence detection method (MSPE-HPLC-FLD) was established for determining six mycotoxins [aflatoxin B1 (AFB1), AFB2, AFG1, and AFG2, and ochratoxin A (OTA) and OTB)] in vegetable oils. The recovery was 70.15%~89.25%, and RSD was ≤6.4%. PDA@Fe3O4-MWCNTs showed a high affinity toward aflatoxins and ochratoxins, allowing selective extraction and quantification of aflatoxins and ochratoxins from complex sample matrices.

Determination of Ochratoxin-A in the brain microdialysates and plasma of awake, freely moving rats using ultra high performance liquid chromatography fluorescence detection method

In this study, a simple, efficient and rapid Ultra High Performance Liquid Chromatography method with fluorescence detection (UHPLC-FLD) has been developed and validated for the determination of Ochratoxin-A (OTA) in rat brain microdialysates and plasma samples. Six adult male wistar rats were used in the study and a single dose (5 mg/kg b.w.) of OTA was given by intraperitoneal (i.p.) injection. Rat blood and microdialysate samples were collected simultaneously after i.p. injection in awake freely moving rats, over a twelve-hour period. An UHPLC analysis was performed on a Zorbax Eclipse Plus C8 (150 mm × 3.0 mm ID × 1.8 μm particles) column with a mobile phase of acetonitrile:water:phosphoric acid (50:50:0.1, v/v) using a flow rate of 0.6 mL/min. The fluorescence detector was set at 330 nm excitation and 460 nm emission wavelengths. Diflunisal (DIF) was used as an internal standard (IS). OTA and IS were separated within 5 min under these conditions. The method was validated in terms of linearity, precision, accuracy, limit of detection, limit of quantification, and stability. Calibration curves obtained with spiked biological matrices show good linearity with high correlation coefficients. The intra- and inter-day assay variability was <5% for the OTA. The limit of detection and the limit of quantification values were found to be 0.490 ng/mL and 1.48 ng/mL for plasma; 0.0900 ng/mL and 0.270 ng/mL for microdialysate samples, respectively. This method was successfully applied for the monitoring of OTA levels in the rat brain and plasma samples.

Development of Graphene Oxide Functionalized Cotton Fiber Based Solid Phase Extraction Combined with Liquid Chromatography-Fluorescence Detection for Determination of Trace Auxins in Plant Samples

In this work, a convenient and sensitive solid phase extraction (SPE) method was developed based on graphene oxide functionalized cotton fiber (GO-CF), a SPE sorbent followed by liquid chromatography-fluorescence detection (LC-FLD) for determination of trace indole acetic acid, indole propionic acid, indole butyric acid and naphthalene acetic acid. GO-CF was synthesized via covalent bonding method on the surface of cotton fiber, and exhibited higher extraction capacity in comparison with that of cotton fiber. The conditions affecting the extraction efficiency, including sample pH, elution solvent, elution volume, extraction time, and sample volume, were optimized to obtain high extraction recoveries. Under optimal conditions, the calibration curves were linear over the concentration ranging from 2 to 200 ng mL−1 with correlation coefficients between 0.9987 and 0.9998 for all the analytes. The limits of detection were lower than 0.371 ng mL−1 for the four auxins. The intra- and inter-day relative standard deviations (RSDs) were less than 3.5 and 7.1%, respectively. The mean recoveries ranged from 73.5 to 105.4% with RSDs lower than 12.6%. The developed GO-CF–SPE–LC-FLD method has been successfully applied to the analysis of four auxins in real plant samples, including mung bean stalk, mung bean sprout, and tobacco leaf samples.

An easy-to-use liquid chromatography method with fluorescence detection for the simultaneous determination of five neuroactive amino acids in different regions of rat brain

Introduction: To comprehend the normal function and pathological characteristics of certain neurological disorders it is important to evaluate the neuroactive amino acids levels in animal models.Methods: This work describes a simple liquid chromatography-fluorescence detection (LC-FLD) method for the simultaneous determination of aspartic acid (Asp), glutamic acid (Glu), glutamine (Gln), taurine (Tau) and γ-aminobutyric acid (GABA), using methyl-L-arginine as internal standard, in samples of rat brain tissue. The five target analytes (Asp, Glu, Gln, Tau and GABA) were determined in a single chromatographic run of less 11 min after a derivatization step with o-phthalaldehyde. The derivatives were separated on a reversed-phase C18 column and detected by fluorescence at excitation and emission wavelengths of 340 and 448 nm, respectively.Results: The method was validated in accordance with international guidelines on bioanalytical methods validation and it presented limits of quantification in the range of 25–50 ng mL−1 and calibration curves with determination coefficients (r2) equal to or higher than 0.9920. In addition, the precision (coefficient variation, %) and accuracy (bias, %) of the method meet the established criteria, and the stability of the analytes at the sample handling and storage conditions was demonstrated.Discussion: Unlike other similar bioanalytical assays, the current method was validated using diluted biological matrix, which is advantageous in order to ensure the derivatization process integrity. Moreover, this LC-FLD method was successfully applied for the determination of the compounds of interest in different rat brain tissue regions (frontal cortex, amygdala, hippocampus, cerebellum and striatum). Thus, this bioanalytical assay represents a useful tool to support multiple nonclinical studies in the field of neurosciences, requiring the quantitative profiling and pattern analysis of neuroactive amino acids.

Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography.

The profiling analysis of catecholamines and their metabolites in brain tissue offers a crucial key to understand their functions in the body and the opportunity to follow up neural diseases. A rapid and simple liquid chromatography-fluorescence detection (LC-FLD) method was developed and validated for simultaneously measuring several catecholamines and endogenous related compounds in the rat brain tissue samples. The target analytes measured in this bioanalytical assay were levodopa (L-DOPA), dopamine (DA), norepinephrine (NE), epinephrine (E), 3-O-methyldopa (3-O-MD), and homovanillic acid (HVA), being the 3,4-dihydroxybenzylamine (DHBA) used as internal standard (IS). The six analytes (L-DOPA, DA, NE, E, 3-O-MD and HVA) can be determined in a single chromatographic run of less than 12min, and all the compounds (analytes and IS) were detected using their native fluorescence and monitored at excitation/emission wavelengths of 279nm/320nm, respectively. The chromatographic and detection conditions were experimentally optimized and then several validation parameters (linearity, limits of quantification and detection, precision and accuracy, recovery, stability and selectivity) were examined. In accordance with the international guidelines of the Food and Drug Administration and European Medicines Agency the method described herein exhibited limits of quantification in the range of 2-25ngmL-1, linearity in wide concentration ranges (r2≥0.994), and acceptable precision (coefficient variation ≤8.76%) and accuracy (bias ±14.65%) levels. Since the bioanalytical procedure does not involve pre-purification or derivatization of the sample, the absolute recovery was found to be around 100%. Moreover, the developed LC-FLD method was successfully applied for the determination of the compounds of interest in tissue samples of different rat brain regions (cerebellum, amygdala, cortex, hippocampus, striatum, mesencephalon, medulla oblongata, substantia nigra and ventral tegmental area). Hence, this assay represents a valuable bioanalytical tool to support several pre(non)clinical studies in the broad field of neurosciences, requiring the quantitative analysis of these bioamines and their metabolites.

Simultaneous determination and identity confirmation of thiodicarb and its degradation product methomyl in animal-derived foodstuffs using high-performance liquid chromatography with fluorescence detection and tandem mass spectrometry.

A high-performance liquid chromatography-fluorescence detection method was developed for the simultaneous determination of thiodicarb and its degradation product methomyl in animal-derived food products, including chicken muscle, beef, pork, table eggs, and milk. Thiodicarb is known to degrade during analysis; therefore, a thorough investigation was carried out, revealing that thiodicarb degrades to methomyl immediately after spiking into a matrix of animal-derived food products. Consequently, thiodicarb was determined as the sum of the parent compound and methomyl. Samples were extracted with acetonitrile and sodium salts, and purified using solid-phase extraction (SPE). The limits of detection (LODs) and quantification (LOQs) were 0.0013 and 0.004mg/kg, respectively, for both analytes in various matrices. Seven-point external calibration curves were obtained, and they showed excellent linearity with determination coefficients (R2)≥0.999 for all tested matrices. The method was validated at three fortification levels (LOQ, LOQ×2, and LOQ×10) in triplicate with average recoveries ranging from 84.24 to 112.8% (for methomyl) and relative standard deviations (RSDs)≤6.5% in all matrices. The converted recoveries of thiodicarb in various matrices ranged from 74.80 to 107.80% with RSDs≤4.5%. The identities of both compounds in standard solutions and for recovery were confirmed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The developed method was further validated by accurate reproduction at another laboratory. Finally, the method was applied to market samples collected from different areas (and, in the case of milk, different brands), and none of the samples tested positive for thiodicarb or methomyl. In conclusion, the developed method can be successfully applied for a single-run analysis of thiodicarb and methomyl in livestock products.

Analysis and dissipation of the antiparasitic agent ivermectin in cattle dung under different field conditions.

Cattle treated with the veterinary parasiticide ivermectin fecally excrete residues. The authors report the exposition and dissipation characteristics of these residues in dung of ivermectin-treated cattle and in soil beneath this dung on pastures in Canada, France, Switzerland, and The Netherlands. Residues were quantified for dung collected from cattle after 3 d, 7 d, 14 d, and 28 d posttreatment and subsequently exposed in the field for up to 13 mo. The authors optimized a high-performance liquid chromatography-fluorescence detection method to detect ivermectin residues in dung and soil matrices. They showed that a solid phase extraction and purification step generally can be eliminated to reduce the time and cost of these analyses. They also found that the addition of water to relatively dry samples improves the extraction efficiency of residues. They then analyzed the field samples to document differences in ivermectin dissipation in cattle dung among sites, with 50% dissipation times of up to 32 d and 90% dissipation times >396 d. They further showed that the dissipation characteristics of residues are comparable between dung of ivermectin-treated cattle and dung to which ivermectin has been added directly. Lastly, they report the first use of a desorption electrospray ionization-high-resolution-mass spectrometric method to detect residues of metabolites in a dung matrix. Environ Toxicol Chem 2016;35:1924-1933. © 2016 SETAC.

Development and Validation of a LC-FD Method for the Simultaneous Determination of Eight Bisphenols in Soft Drinks

Bisphenol A (BPA) is a chemical widely used as a monomer in the production of polymers of plastics. It acts as an endocrine-disrupting agent and thus its contaminations of food and beverage should be carefully monitored in order to assess consumers’ risk. In this study, we propose a liquid chromatography-fluorescence detection (LC-FD)-validated method for the simultaneous determination of BPA and seven analogues, i.e., bisphenol AF, bisphenol B, bisphenol E, bisphenol F, BPA diglycidyl ether, bisphenol F diglycidyl ether, and Bisphenol M in soft drinks. A one-step solid-phase extraction (SPE) was effective at reducing the interferences, obtaining good purification of the samples, and consequently good recoveries of all analytes. The separation was obtained on a C18 column by using acetonitrile/water 55:45 (v/v) under isocratic conditions. Method validation was performed according to the European Commission Decision 2002/657/EC criteria, providing good results regarding the analytical parameters of linearity, selectivity, sensitivity, precision, recovery, decision limit (CCα), detection capability (CCβ), limit of detection (LOD), limit of quantification (LOQ), stability, and robustness. The method allows the detection of BPA and BADGE at levels much lower than their legal limits in the food, which are 0.06 and 9.00 mg kg−1, respectively.

Sensitive Estrogens Determination in Wastewater Samples by HPLC and Fluorescence Detection

A sensitive high-performance liquid chromatography-fluorescence detection method (HPLC-FLD) was developed and validated for 17β-estradiol (E2) and 17α-ethinylestradiol (EE2) determination in wastewater samples. Sample preparation was performed by solid-phase extraction (SPE) on Hypersep C18 cartridges. Conditioning and elution solvents and sample volume were investigated by a 23 factorial design for both analytes. The pre-concentration factor was 2500 times, allowing to quantify 5.0 ng L-1 for both hormones. The SPE-HPLC-FLD method was successfully applied to detect and quantify trace amount of target compounds in raw and treated wastewater samples. E2 was found below limit of quantification in influent and effluent samples. Nevertheless, the recoveries of E2 in standard addition experiments were between 41 and 72%, with relative standard deviations between 4 and 16%. An important matrix effect was observed in the determination of EE2, with fluorescence signal suppression.

Highly sensitive and selective detection of human carboxylesterase 1 activity by liquid chromatography with fluorescence detection.

Human carboxylesterases 1 (hCE1), one of the most important human drug metabolizing enzymes, catalyzes the hydrolysis of a large number of structurally diverse of endogenous and exogenous substrates. However, a practical, reliable and sensitive method for the precise measurement of hCE1 activities in complex biological samples has been rarely reported. In this study, a liquid chromatography-fluorescence detection (LC-FD) based method was developed for highly selective and sensitive measurement of hCE1 activities in human tissue and cell preparations. This method was based on the fluorimetric detection of HMBT, the hydrolyzed product of BMBT which was a newly developed specific probe substrate for hCE1. The developed LC-FD method was fully validated in terms of specificity, sensitivity, linearity, precision, recovery and stability. With the help of LC separation, most polar endogenous compounds in biological samples could be eluted in the column dead time, which is very beneficial for accurate determination of hCE1 activities in complex biological samples. The lower limit of quantification for HMBT (product of hCE1) of this LC-FD based method was as low as 20nM, which was quite lower than other reported methods. The method also exhibited good precision, both intra- and inter- assay variances were both lower than 2.5%. Furthermore, the newly developed method was successfully applied to measure hCE1 activity in human liver preparations from individual donors (n=12), as well as in homogenates from eleven different human cell lines. All these findings combined with this practical method are very helpful for the deep understanding of the expression and function of hCE1 in human biological samples.

Occurrence and Source Appraisal of Polycyclic Aromatic Hydrocarbons (PAHs) in Surface Waters of the Almendares River, Cuba.

In this work, 14 polycyclic aromatic hydrocarbons (PAHs) included in the United States Environmental Protection Agency pollutant priority list were analyzed in the surface water of the upper urbanized part of Almendares River, the most important water course in Havana, Cuba. Surface water from five sampling sites was collected at the end of dry season and analyzed by high-performance liquid chromatography-fluorescence detection method after solid phase extraction procedure. Total PAHs concentrations varied from 836 to 15 811ngL(-1) with a geometric mean value of 2512ngL(-1). PAH typology was dominated by low molecular-weight PAHs (2- to 3-ring components). Pollutant source appraisal was determined by diagnostic ratios method in five sampling sites. Factor analysis of normalized samples was used to concentration identified two factors as the main significant pollutant sources and to cluster similar sampling sites corresponding to petrogenic and combustion inputs, respectively. Ecological risks were considered. For animal aquatic life, acute toxicity values exceed the permissible values in the more-polluted sampling sites.

Development and validation of a high-performance liquid chromatography-fluorescence detection method for the accurate quantification of colistin in human plasma.

Recently, colistin has become one of the most important drugs for treating infections caused by multidrug-resistant Gram-negative bacteria. Therapeutic drug monitoring is recommended to ensure the safety and efficacy of colistin and to improve clinical outcomes. This study developed an accurate and sensitive high-performance liquid chromatography-fluorescence detection (HPLC-FLD) method for the quantification of colistin in human plasma. The sample preparation included protein precipitation using trichloroacetic acid (TCA) and methanol, followed by in-solid phase extraction (In-SPE) derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl). A Poroshell 120 EC-C18 2.1×100mm (2.7μm) column was used in the HPLC method with a mobile phase composed of acetonitrile (ACN), tetrahydrofuran (THF), and deionized (DI) water (82%, 2%, 16% (v/v), respectively). Polymyxin B1 was used as the internal standard. The total analysis time was 22min under optimal separation conditions. The HPLC-FLD method was validated over a therapeutic range of 0.3-6.0μgmL(-1). The intra-day and inter-day precisions for colistin A and colistin B were below 9.9% and 4.5% relative standard deviations, respectively. The accuracy test results were between 100.2 and 118.4%. The extraction recoveries were between 81.6 and 94.1%. The method was linear over the test range, with a 0.9991 coefficient of determination. The limit of detection was 0.1μgmL(-1). The validated HPLC-FLD method was successfully applied to quantify the colistin concentrations in 2 patient samples for therapeutic drug monitoring.

Determination of Aminothiol Adsorption Properties of Titanium(IV) Oxide Nanoparticles Using High-Performance Liquid Chromatography Fluorescence Detection

The adsorptive properties of eight forms of nano-TiO2 were evaluated when they were exposed to the toxic thiol-containing amino acid homocysteine. Homocysteine is an amino acid known to cause cardiovascular toxicity and neurodegenerative disorders. A homocysteine standard solution was combined with dispersions of each TiO2 nanoparticle under physiological conditions, and then a high-performance liquid chromatography fluorescence detection method optimized for quantification of total, unbound homocysteine was utilized. The nano-sized anatase nanoparticle removed 2.92 μg of homocysteine per microgram of nanoparticle, whereas amorphous TiO2 only removed 3.65 x 10-3 μg of homocysteine per microgram of nanoparticle. Other nanoparticles produced values between the anatase and amorphous forms. Surface chemistries are distinguishing characteristics of these nanoparticles and are responsible for their individual physicochemical properties. Variations in results were associated with the unique combinations of size ...

Assessment of a semiquantitative liquid chromatography-fluorescence detection method for the determination of paralytic shellfish poisoning toxin levels in bivalve molluscs from Great Britain.

AOAC Official Method 2005.06 precolumn oxidation LC-fluorescence detection method has been used for many years for the detection and quantitation of paralytic shellfish poisoning (PSP) toxins in bivalve molluscs. After extensive single- and multiple-laboratory validation, the method has been slowly gaining acceptance worldwide as a useful and practical tool for official control testing. In Great Britain, the method has become routine since 2008, with no requirement since then for reverting back to the bioassay reference method. Although the method has been refined to be semiautomated, faster, and more reproducible, the quantitation step can be complex and time-consuming. An alternative approach was developed to utilize the qualitative screening results for generating a semiquantitative results assessment. Data obtained over 5 years enabled the comparison of semiquantitative and fully quantitative PSP results in over 15 000 shellfish samples comprising eight different species showed that the semiquantitative approach resulted in over-estimated paralytic shellfish toxin levels by an average factor close to two in comparison with the fully quantified levels. No temporal trends were observed in the data or relating to species type, with the exception of surf clams. The comparison suggested a semiquantitative threshold of 800 microg saxitoxin (STX) eq/kg should provide a safe limit for the determination of samples to be forwarded to full quantitation. However, the decision was taken to halve this limit to include an additional safety factor of 2, resulting in the use of a semiquantitative threshold of 400 microg STX eq/kg. Implementation of the semiquantitative method into routine testing would result in a significant reduction in the numbers of samples requiring quantitation and have a positive impact on the overall turnaround of reported PSP results. The refined method would be appropriate for any monitoring laboratory faced with high throughput requirements.