High-performance Liquid Chromatography With Fluorescence Detection Research Articles

Evaluation of the safety and quality of Brazil nuts (Bertholletia excelsa) using the tools of dna sequencing technology and aflatoxin profile.

Brazil nuts (BNs) result from sustainable extraction and are widely exploited in the Amazon region. Due to the production characteristics in the forest and the nutritional characteristics of these nuts, the occurrence of fungal contamination and the presence of aflatoxins are extensively discussed in the literature as a great aspect of interest and concern. This study aims to evaluate the microbial profile through DNA sequencing and amplification of 16S and ITS genes for bacterial and fungal analysis, respectively, and the presence of mycotoxins using high-performance liquid chromatography with fluorescence detection (HPLC-FD) from different fractions of the nuts processed. The BN samples, harvest A (HA) and harvest B (HB), from two different harvests were collected in an extractive cooperative in the Amazon region for microbiological analysis (from DNA extraction and amplification of 16S genes, bacteria analysis, and ITS for fungi) and mycotoxins (aflatoxins AFB1, AFB2, AFG1, and AFG2) using HPLC-FD/KobraCell®. The samples showed a very different microbiome and aflatoxin profile. Genera such as Rothia (HA) and Cronobacter (HB) were abundant during the analysis of bacteria; as for fungi, the genera Aspergillus, Fusarium, Penicillium, and Alternaria were also considered prevalent in these samples. Soil microorganisms, including those pathogenic and related to inadequate hygienic-sanitary production practices, as well as aflatoxins, were found in the samples. However, they were within the established limits permitted by Brazilian legislation. Nuts have a diverse microbiota and are not restricted to fungi of the genus Aspergillus. The microbiological and toxicological profile can vary significantly within the same nut in the same extraction region and can be exacerbated by global climate changes. Therefore, it is necessary to advance sanitary educational actions by applying good production practices and inspection programs to ensure the sustainability and quality of the BN production chain.

Anticancer Drug Extraction from Plasma Samples Using Three-Dimensional Polyoxometalate-Based Supramolecular Frameworks as Sorbents.

Four self-assembled inorganic-organic hybrid materials, namely, H{Na(H2O)3[Gd(PDA)(H2O)2]3[BW12O40]}·4H2O (1), H{Na(H2O)3[Tb(PDA)(H2O)2]3[BW12O40]}·3H2O (2), H{Na(H2O)3[Er(PDA)(H2O)3]3[BW12O40]}·H2O (3) (PDA = 1,10-phenanthroline-2,9-dicarboxylate), and [Pr3(H2O)13(pydc-OH)2][BW12O40]·12H2O (4) (pydc-OH = 4-hydroxy-2,6-pyridinedicarboxylate), were hydrothermally synthesized and structurally characterized. Hybrids 1-3 are isostructural and contain a Keggin unit, which is linked to lanthanoids to produce distinct trinuclear lanthanoid building blocks. The fragments are connected by anion-π and hydrogen bonding interactions to create 3D networks. In hybrid 4, a trimeric Pr-organic species bearing a Keggin unit forms a 2D coordination polymer, and then hydrogen bonding interactions between 2D layers lead to the formation of a 3D structure. These polyoxometalate-based frameworks were used as sorbents for the dispersive microsolid-phase extraction (D-μSPE) of two anticancer drugs (doxorubicin and epirubicin) in human plasma samples. Analytes were quantified and separated using high-performance liquid chromatography with fluorescence detection (HPLC-FLD). The method's linearity was between 0.8-500 ng mL-1 and 1.0-500 ng mL-1 for the antineoplastic drugs doxorubicin and epirubicin, respectively. The limits of detection (S/N = 3) were in the range of 0.2-0.3 ng mL-1, while the precision was in the range of 3.5-4.3%. Finally, human plasma samples from patients treated with doxorubicin or epirubicin were analyzed by using the D-μSPE-HPLC-FLD method.

Synthesis of Layer‐By‐Layer Decorated Graphene Oxide with Bismuth Telluride Nanoparticles‐modified Dendrimer: Application for Nonylphenol Removal from Aqueous Solutions

AbstractHerein, for the first time, the bismuth telluride (Bi2Te3) nanoparticles functionalized graphene oxide (GO) were synthesized for the removal of nonylphenol (NP). The surface of the GO‐Bi2Te3 nanocomposite was modified by hyperbranched dendrimer and characterized by different spectroscopic and morphological techniques such as Fourier transform infrared (FT‐IR) spectroscopy, X‐ray diffraction (XRD), field‐emission scanning electron microscope (FE‐SEM), energy dispersive X‐ray spectroscopy (EDX), and thermogravimetric analysis (TGA). In this paper, to enhance the capacity, the surface of GO‐Bi2Te3@GlyTsCl dendrimer was modified by 5‐amino‐2‐hydroxybenzoic acid (5‐ASA). The effects of parameters on the removal of NP were also studied. This study reports the utilization of nanoadsorbent (GO‐Bi2Te3@GlyTsCl/5‐ASA) for the removal of NP by high‐performance liquid chromatography with fluorescence detector (HPLC‐FLD). Under optimal conditions, the removal values for the method were in the range of 90–96 %. Additionally, the Langmuir model was found to be fit with the isotherms data. The best kinetic model fit for the adsorption of NP from nanoadsorbent was the pseudo‐second‐order model. Moreover, GO‐Bi2Te3@GlyTsCl/5‐ASA nanoadsorbent exposed good reusability even after six consecutive cycles (from 100 % to 93 %) with high stability. Consequently, synthesized GO‐Bi2Te3@GlyTsCl/5‐ASA composites could be potentially used as an efficient nanoadsorbent for NP removal in water samples.

Influence of UV light, ultrasound, and heat treatment on the migration of bisphenol A from polyethylene terephthalate bottle into the food simulant

This research examined the impacts of ultrasound, UV light, storage time, and temperature on the leaching of bisphenol A (BPA) from polyethylene terephthalate (PET) drinking water bottles in Turkey. The initial phase of the investigation encompassed the quantification of BPA in two distinct brands of bottled water. Samples were extracted by solid- phase extraction (SPE) and analyzed by high performance liquid chromatography with fluorescence detection (HPLC‐FLD). According to the results in the first part, the highest BPA levels were found in bottled water. In the second part of the study, 10 to 30 min of ultrasound treatment increased the BPA migration with increased time in simulants. In the first and second weeks of storage at 25 °C, the effect of storage on BPA migration was below the detection limit (<LOD) in simulants. However, at 35 °C and 45 °C, BPA concentration increased with increased temperature. According to these results, increasing the storage temperature to 35 °C and 45 °C increased the BPA pass faster than the increase in storage time. When simulants in PET bottles were exposed to UV light for 1 and 6 h, BPA transitions higher from 6 to 1 h was found in simulants. Except for first-brand water samples, the passing of bisphenol A was found to comply with the European Food Safety Authority (EFSA) criteria in this study.

Development, in-house validation and application of a method using high-performance liquid chromatography with fluorescence detection (HPLC-FLD) for the quantification of 12 ergot alkaloids in compound feeds.

Ergot alkaloids (EAs) are toxic substances naturally produced by Claviceps fungi. These fungi infest a wide range of cereals and grasses. When domestic animals are exposed to EAs through contaminated feeds, it is detrimental to them and leads to significant economic losses. For that reason, it is important to monitor feed for the presence of EAs, especially with methods enabling their determination in processed materials. Ergot alkaloids were extracted with acetonitrile, and dispersive solid phase extraction (d-SPE) was used for clean-up of the extracts. After evaporation, the extracts were reconstituted in ammonium carbonate and acetonitrile and subjected to instrumental analysis using high-performance liquid chromatography with fluorescence detection. The developed method was validated in terms of linearity, selectivity, repeatability, reproducibility, robustness, matrix effect, limits of quantification and detection and uncertainty. The EA content of 40 compound feeds was determined. All the assessed validation parameters fulfilled the requirements of Regulation (EU) 2021/808. At least one of the monitored alkaloids was determined in 40% of the samples. The EAs with the highest incidence rate were ergocryptine, ergometrinine and ergocornine. The total concentrations of EAs ranged from under the limit of quantification to 62.3 μg kg-1. The results demonstrated that the developed method was suitable for simultaneously determining twelve EAs in compound feed and could be used for routine analysis.

Synthesis and application of BODIPY-based fluorescent labeling tag for oligosaccharide and N-linked glycan analysis by high-performance liquid chromatography with fluorescence detection

BackgroundGlycosylation analysis is still challenging, not only because of the extreme structure complexity and conjugation diversity of glycans but also because of instrumental aspects such as the sensitivity limits of analyses. Therefore, glycan analysis by chromatographic methods is very often combined with fluorescence detection in addition to MS. The majority of fluorescent labeling employed before LC separation is based on 2-aminobenzamide, which has several disadvantages such as low labeling yield, poor fluorescence properties, and MS ionization efficiency. Therefore, even after several decades of development of new labels, there is still a need for new labeling tags with improved characteristics. ResultsWe present the application of a newly synthesized fluorescent label designed for oligosaccharide and glycan analysis by high-performance liquid chromatography with fluorescence detection (HPLC/FLD). The novel hydrazide derivative of dipyrrometheneboron difluoride (BODIPY) was synthesized from 2,4-dimethylpyrrole, methyl succinyl chloride, and boron trifluoride etherate followed by a reaction with hydrazine. The synthesized label was characterized by several analytical methods including NMR, UV/Vis and fluorescence spectroscopy, and mass spectrometry. The labeling reaction via hydrazone formation chemistry was optimized by labeling of maltooligosaccharide standards. The analysis of maltohexaose labeled by BODIPY-hydrazide followed by HPLC/FLD analysis provided the limit of detection in the low tens of femtomole. The presented method based on fluorescence detection is at least 30 times more sensitive than the standard approach employing labeling by 2-aminobenzamide. In addition, the labeling method by BODIPY-hydrazide was used for N-linked glycan profiling of several glycoproteins (ribonuclease B, immunoglobulin G) by RP-HPLC/FLD as well as HILIC/FLD analysis. SignificanceThis work represents the design, synthesis, and application of a new fluorescent label based on the BODIPY core and hydrazone formation chemistry for oligosaccharide and glycan analysis by HPLC/FLD. The proposed approach significantly improved the oligosaccharide and glycan analysis in comparison to the commonly used procedure employing 2-aminobenzamide.

In Situ Formation of a Relatively Transparent Ion-Associate Liquid Phase from an Aqueous Phase and Its Application to Microextraction/High-Performance Liquid Chromatography-Fluorescence Detection of Bisphenol A in Water.

The design of a simple approach enabling the detection of bisphenol A (BPA) in water samples without the need for large amounts of solvents is of utmost importance. This paper reports a simple method for the separation, concentration, and quantification of BPA in water samples using high-performance liquid chromatography with fluorescence detection (HPLC-FLD) after its microextraction into an in situ formed organic ion-associate (IA) liquid phase (LP). Novel IA phase components without conjugated double bonds, such as benzene rings, were investigated. Ethylhexyloxypropylamine hydrochloride and sodium dodecyl sulfate solutions were added to the water samples to form IAs. The aqueous phase and ion-associate liquid phase (IALP) were separated by centrifugation. The aqueous phase was removed, and the liquid phase was recovered and measured using HPLC-FLD or HPLC-electrochemical detection (ECD). The concentrated phase (IALP) had a relatively low viscosity and could be injected directly into the chromatograph without dissolving it in organic solvents. The detection limits for BPA by HPLC-FLD and HPLC-ECD were 0.009 and 0.3 µg L-1, respectively.

In Vitro Permeability Study of Homotaurine Using a High-Performance Liquid Chromatography with Fluorescence Detection Pre-Column Derivatization Method.

Homotaurine (HOM) is considered a promising drug for the treatment of Alzheimer's and other neurodegenerative diseases. In the present work, a new high-performance liquid chromatography with fluorescence detection (HPLC-FLD) (λex. = 340 nm and λem. = 455 nm) method was developed and validated for the study of substance permeability in the central nervous system (CNS). Analysis was performed on a RP-C18 column with a binary gradient elution system consisting of methanol-potassium phosphate buffer solution (pH = 7.0, 0.02 M) as mobile phase. Samples of homotaurine and histidine (internal standard) were initially derivatized with ortho-phthalaldehyde (OPA) (0.01 M), N-acetylcysteine (0.01 M) and borate buffer (pH = 10.5; 0.05 M). To ensure the stability and efficiency of the reaction, the presence of different nucleophilic reagents, namely (a) 2-mercaptoethanol (2-ME), (b) N-acetylcysteine (NAC), (c) tiopronin (Thiola), (d) 3-mercaptopropionic acid (3-MPA) and (e) captopril, was investigated. The method was validated (R2 = 0.9999, intra-day repeatability %RSD < 3.22%, inter-day precision %RSD = 1.83%, limits of detection 5.75 ng/mL and limits of quantification 17.43 ng/mL, recovery of five different concentrations 99.75-101.58%) and successfully applied to investigate the in vitro permeability of homotaurine using Franz diffusion cells. The apparent permeability (Papp) of HOM was compared with that of memantine, which is considered a potential therapeutic drug for various CNSs. Our study demonstrates that homotaurine exhibits superior permeability through the simulated blood-brain barrier compared to memantine, offering promising insights for enhanced drug delivery strategies targeting neurological conditions.

Highly selective and sensitive high performance liquid chromatography method for determination of fumonisins from feed samples with MIL-101 based dispersed micro-solid-phase extraction and precolumn fluorescence derivatization

A selective and rapid high-performance liquid chromatography with fluorescence detection (HPLC-FLD) method for determining fumonisins was established and validated, which based on dispersed microsolid-phase extraction (D-μ-SPE) and precolumn derivatization. The MIL-101 was prepared as sorbents to selectively enrich two fumonisins from feed samples. The obtained extracts were labeled by a novel derivatization reagent 2,5-dioxopyrrolidin-1-yl 4-(1-methyl-1H-phenanthro[9,10-d]imidazol-2-yl)benzoate (MPIBA-Su). There was no fluorescence from fumonisins itself, while their derivatives exhibited considerable fluorescence with a maximum excitation at λex 255 nm and a maximum emission at λem 457 nm. Key factors affecting extraction efficiency and derivatization efficiency were fully optimized. Excellent linearity was achieved with correlation coefficients > 0.9935 under optimal conditions. The detection limits (LODs) were between 2.4 and 3.5 ng·mL−1, and the quantitation limits (LOQs) were between 8.1 and 11.7 ng·mL−1. With recoveries in the range of 83.8% − 94.3%, the established method has successfully been applied to the quantitative analysis of fumonisins in feed samples.

Development and Comparative Evaluation of Two Different Label-Free and Sensitive Fluorescence Platforms for Analysis of Olaparib: A Recently FDA-Approved Drug for the Treatment of Ovarian and Breast Cancer.

Olaparib (OLA) is a PARP inhibitor drug which has been recently approved by the Food and Drug Administration (FDA) for the treatment of ovarian and breast cancer. A convenient analytical tool for the quantitation of OLA in its dosage form and plasma samples was urgently needed. This study describes, for the first time, the development of two different label-free and sensitive fluorescence-based platforms for the pharmaceutical and bioanalysis of OLA. These platforms were microwell-assisted with a fluorescence microplate reader (MW-FLR) and high-performance liquid chromatography with fluorescence detection (HPLC-FD). Both MW-FLR and HPLC-FD employed the native fluorescence of OLA as an analytical signal. The MW-FLR involved measuring the fluorescence signals in 96-well white-opaque plates. The HPLC-FD involved chromatographic separation of OLA and duvelisib (DUV), as an internal standard on a Nucleosil-CN HPLC column (250 mm length × 4.6 mm i.d., 5 µm particle diameter) with a mobile phase composed of acetonitrile: water (25:75, v/v) pumped at a flow rate of 1.7 mL/min. Elution of OLA and DUV was detected using a fluorescence detector. The optimal conditions of both MW-FLR and HPLC-FD were established, and they were validated according to the guidelines of the International Council for Harmonization for the validation of analytical procedures. The linear ranges of MW-FLR and HPLC-FD were 25-1000 and 5-200 ng/mL, respectively, with limits of detection of 15 and 1.7 ng/mL, respectively. The accuracy and precision of both platforms were confirmed as the recovery values were ≥98.2% and the relative standard deviations (RSD) were ≤2.89%. Both methodologies were satisfactorily applied to the quantitation of OLA in its commercial dosage form (Lynparza® tablets) and plasma samples with high accuracy and precision. The greenness of both MW-FLR and HPLC-FD was assessed using two different multiple parameter-based metric tools, and the results proved their greenness and adherence to the requirements of green analytical approaches. Both platforms have simple procedures and acceptable levels of analytical throughput. In conclusion, the proposed MW-FLR and HPLC-FD are valuable tools for routine use in quality control and clinical laboratories for the quantitation of OLA for the purposes of pharmaceutical quality control, pharmacokinetic studies, and bioequivalence testing.

Occurrence and risk characterization of aflatoxin M1 in milk samples from southeastern Iran using the margin of exposure approach.

This study aimed to investigate Aflatoxin-M1 (AFM1) contamination in pasteurized and raw milk samples consumed in Kerman and Rafsanjan in southeastern Iran. In this cross-sectional study, a total of 100 samples of raw (n = 67) and pasteurized (n = 33) milk were randomly collected from retail stores, supermarkets, and milk transport tankers in the winter of 2020 and the summer of 2021. The level of AFM1 contamination in the collected samples was evaluated by high-performance liquid chromatography with fluorescence detection (HPLC-FD). AFM1 was detected in 95% of samples and its median concentration was 17.38 ng/L. The median concentration of AFM1 in the pasteurized milk samples (24.89 ng/L) was significantly higher than in the raw milk samples (13.54 ng/L). The AFM1 contamination level in 20% (raw = 13% and pasteurized = 7%) of the samples was higher than the maximum permitted level (MPL) recommended by the European Union (i.e., 50 ng/L), whilst 4% (raw = 3% and pasteurized = 1%) of the samples was higher than the Iranian maximum standard limit (i.e., 100 ng/L). The hazard index (HI) was higher than 1 in 16%, 18%, and 35% of total milk samples for men, women, and children, respectively. The AFM1 contamination level in the milk samples collected in southeastern Iran was worrying. The margin of exposure (MoE) values were lower than 10,000 for children. Because aflatoxins are among the most potent carcinogens known, prevention of milk contamination in all stages from the farm to the table can considerably reduce the community's exposure to AFM1 and its consequent health risks.

AKR1B1 inhibition using NARI-29-an Epalrestat analogue-alleviates Doxorubicin-induced cardiotoxicity via modulating Calcium/CaMKII/MuRF-1 axis

The clinical use of doxorubicin (Dox) is narrowed due to its carbonyl reduction to doxorubicinol (Doxol) implicating resistance and cardiotoxicity. Hence, in the present study we have evaluated the cardioprotective effect of AKR1B1 (or aldose reductase, AR) inhibitor NARI-29 (epalrestat (EPS) analogue) and its effect in the Dox-modulated calcium/CaMKII/MuRF1 axis. Initially, the breast cancer patient survival associated with AKR1B1 expression was calculated using Kaplan Meier-plotter (KM-plotter). Further, breast cancer, cardiomyoblast (H9c2), and macrophage (RAW 264.7) cell lines were used to establish the in vitro combination effect of NARI-29 and Dox. To develop the cardiotoxicity model, mice were given Dox 2.5 mg/kg (i.p.), biweekly. The effect of AKR1B1 inhibition using NARI-29 on molecular and cardiac functional changes was measured using echocardiography, fluorescence-imaging, ELISA, immunoblotting, flowcytometry, High-Performance Liquid Chromatography with Fluorescence Detection (HPLC-FD) and cytokine-bead array methods. The bioinformatics data suggested that a high expression of AKR1B1 is associated with significantly low survival of breast cancer patients undergoing chemotherapy; hence, it could be a target for chemo-sensitization and chemo-prevention. Further, in vitro studies showed that AKR1B1 inhibition with NARI-29 has increased the accumulation and sensitized Dox to breast cancer cell lines. However, treatment with NARI-29 has alleviated the Dox-induced toxicity to cardiomyocytes and decreased the secretion of inflammatory cytokines from RAW 264.7 cells. In vivo studies revealed that the NARI-29 (25 and 50 mg/kg) has prevented the functional, histological, biochemical, and molecular alterations induced by Dox treatment. Moreover, we have shown that NARI-29 has prevented the carbonyl reduction of Dox to Doxol in the mouse heart, which reduced the calcium overload, prevented phosphorylation of CaMKII, and reduced the expression of MuRF1 to protect from cardiac injury and apoptosis. Hence in conclusion, AKR1B1 inhibitor NARI-29 could be used as an adjuvant therapeutic agent with Dox to prevent cardiotoxicity and synergize anti-breast cancer activity.

Comparison of aflatoxins and ochratoxin A contaminated in homemade and commercial doenjang manufactured by traditional and modified methods

Doenjang, a soybean-based fermented food that is traditionally consumed in Korea, is vulnerable to contamination by mycotoxins, such as aflatoxins (AFs) and ochratoxin A (OTA), during the fermentation process. In this study, a novel accurate high-performance liquid chromatography with fluorescence detector (HPLC-FLD) method was validated for rapid and simultaneous detection of AFs and OTA levels in doenjang. The HPLC-FLD method performs well in terms of sensitivity, selectivity, accuracy, and precision, with detection range of 0.03–0.13 μg/kg and quantification range of 0.10–0.40 μg/kg for AFs and OTA. The validated method was applied to 134 homemade and 157 commercial doenjang samples manufactured in Korea using both traditional (spontaneous fermentation-based) and modified (starter culture-based) methods. AFs and OTA were detected in 13.1% (0.16–273.35 μg/kg) and 28.5% (0.56–46.29 μg/kg) of the doenjang samples, respectively. There were no significant differences in the contamination rate or mean concentration between the homemade and commercial doenjang. By contrast, doenjang manufactured using the modified method exhibited significantly (p < 0.05) lower contamination rates and levels of AFs and OTA compared with those produced using traditional methods. The risk of exposure to AFs and OTA through consumption of doenjang manufactured by different methods was estimated by the margin of exposure approach and revealed that the health risk for Korean was lower in doenjang manufactured by modified method, regardless of homemade or commercial products. Thus, the use of selected fermentation starters for the modified method can reduce contamination by AFs and OTA in doenjang, whether manufactured at domestic or industry level. These findings are expected to improve the safety management of mycotoxin contamination in doenjang and similar soybean-based fermented foods.

Optimization and Validation of Dispersive Liquid-Liquid Microextraction for Simultaneous Determination of Aflatoxins B1, B2, G1, and G2 in Senna Leaves and Pods Using HPLC-FLD with Pre-Column Derivatization.

Dispersive liquid-liquid microextraction (DLLME) was optimized for the simultaneous extraction of aflatoxins (AFB1, AFB2, AFG1, and AFG2) from powdered senna leaves and pods. Detection was performed using high-performance liquid chromatography with fluorescence detection (HPLC-FLD) and pre-column derivatization. The parameters affecting the DLLME extraction efficiency were evaluated. Chloroform (200 µL) was used as an extraction solvent, 500 µL of distilled water was used as a dispersive solvent, and the extraction was performed at pH 5.6 with no salt added. The optimized method was validated using leaves and pods according to the European Commission guidelines. The linear range for all aflatoxins was 2-50 µg/kg, with values for regression coefficients of determination exceeding 0.995. The recoveries of spiked senna leaves and pods were in the ranges of 91.77-108.71% and 83.50-102.73%, respectively. The RSD values for intra-day and inter-day precisions were in the ranges of 2.30-7.93% and 3.13-10.59%, respectively. The limits of detection and quantification varied in the ranges of 0.70-1.27 µg/kg and 2.13-3.84 µg/kg, respectively. The validated method was successfully applied for the quantification of aflatoxins in 60 real samples of dried senna leaves and pods.

Polycyclic Aromatic Hydrocarbons (PAHs) in Roasted Pork Meat and the Effect of Dried Fruits on PAH Content.

Diet is one of the main factors affecting human health. The frequent consumption of heat-treated meat has been classified as both directly carcinogenic to humans and as a risk factor, especially in the case of cancers of the gastrointestinal tract. Thermally processed meat may contain harmful muta- and carcinogenic compounds, including polycyclic aromatic hydrocarbons (PAHs). However, there are natural ways to reduce the risk of diet-related cancers by reducing the formation of PAHs in meat. The purpose of this study was to determine changes in PAH levels in pork loin dishes prepared by stuffing the meat with dried fruits (prunes, apricots and cranberries) and baking it in a roasting bag. High-performance liquid chromatography with fluorescence detection (HPLC-FLD) was used to conduct a quantitative analysis of seven PAHs. Recovery results ranged from 61 to 96%. The limit of detection (LOD) was 0.003 to 0.006 ng/g, and the limit of quantification (LOQ) was 0.01 to 0.02 ng/g. Gas chromatography-mass spectrometry (GC-MS/MS) was used to confirm the presence of PAHs in food. The total PAH content of the roasted pork loin was 7.4 ng/g. This concentration decreased by 35%, 48% and 58% when the meat was roasted with apricots, prunes and cranberries, respectively. The cranberries also inhibited the formation of benzo(a)pyrene to the greatest extent. Thermally treating meat stuffed with dry fruits may be a simple and effective way to prepare foods with reduced levels of mutagens and carcinogens belonging to PAHs, and thus reduce the risk of cancer.

Determination of histamine level in commonly consumed frozen fish in Mauritania

The objective of the research was to evaluate the quality of most consumed fish products in Nouakchott-Mauritania, by quantifying histamine levels in frozen marine fish samples to assess the storage conditions and the handling methods for a fish product intended for exportation and local consumption. Histamine levels were determined by HighPerformance Liquid Chromatography with Fluorescence detection (HPLC-FLD) method in 207 samples of frozen different fish species collected from different wholesalers at the Nouakchott fish market. According to the 200 ppm limit required by the FDA, our results showed that 17.77% of samples of non-scombroid fish (Sardinella aurita) analyzed had histamine levels above the limit of 200 ppm, while samples of scombroid fish (Scomber japonicus and Sarda sarda) were all below the limit of 200 ppm. Results showed that more attention should be carried out while handling fishery products from the landing until they are available to consumers to ensure food safety.

Development of Novel Microwell-Based Spectrofluorimetry and High-Performance Liquid Chromatography with Fluorescence Detection Methods and High Throughput for Quantitation of Alectinib in Bulk Powder and Urine Samples

Background and Objectives: This study presents the development and validation of the 96-microwell-based spectrofluorimetric (MW-SFL) and high performance liquid chromatography (HPLC) with fluorescence detection (HPLC-FD) methods for the quantitation of alectinib (ALC) in its bulk powder form and in urine samples. Materials and Methods: The MW-SFL was based on the enhancement of the native fluorescence of ALC by the formation of micelles with the surfactant cremophor RH 40 (Cr RH 40) in aqueous media. The MW-SFL was executed in a 96-microwell plate and the relative fluorescence intensity (RFI) was recorded by utilizing a fluorescence plate reader at 450 nm after excitation at 280 nm. The HPLC-FD involved the chromatographic separation of ALC and ponatinib (PTB), as an internal standard (IS), on a C18 column and a mobile phase composed of methanol:potassium dihydrogen phosphate pH 7 (80:20, v/v) at a flow rate of 2 mL min-1. The eluted ALC and PTB were detected by utilizing a fluorescence detector set at 365 nm for excitation and 450 nm for emission. Results: Validation of the MW-SFL and HPLC-FD analytical methods was carried out in accordance with the recommendations issued by the International Council for Harmonization (ICH) for the process of validating analytical procedures. Both methods were efficaciously applied for ALC quantitation in its bulk form as well as in spiked urine; the mean recovery values were ≥86.90 and 95.45% for the MW-SFL and HPLC-FD methods, respectively. Conclusions: Both methodologies are valuable for routine use in quality control (QC) laboratories for determination of ALC in pure powder form and in human urine samples.

Comprehensive evaluation of solvent in dispersive liquid-liquid microextraction for determination of itraconazole and hydroxy itraconazole by high performance liquid chromatography with fluorescence detection

This paper aims to propose a multicriteria decision analysis (MCDA) method to evaluate and select solvent in a dispersive liquid-liquid microextraction (DLLME) approach. The DLLME is applied to the determination of itraconazole and hydroxy itraconazole in plasma by high performance liquid chromatography with fluorescence detection (HPLC-FLD). To achieve this goal, extraction efficiency, chromatographic resolution, and greenness of solvent were identified as the three indicators in MCDA. Then, Technique for Order of Preference by Similarity to Ideal Solution (TOPSIS) was employed to evaluate and select the solvent. Weight assignment was set up by integrating the subjective scoring from experienced analysts and the objective approach using Shannon’s entropy weight method. Under chosen parameters (extraction solvent: 1-decanol 40 μL, dispersing solvent: methanol 400 μL), the method was validated satisfactorily and applied successfully to the analysis of itraconazole and hydroxy itraconazole in real human plasma samples. The results show that the comprehensive weighted TOPSIS analysis is a promising tool to choose experimental conditions toward an integrative goal. It can provide analysts with a decision making reference to maintain a balance between the analytical performance and eco-friendliness.

Human exposure to ochratoxin A and its natural occurrence in spices marketed in Chile (2016–2020): A case study of merkén

The potential ingestion of food products contaminated with ochratoxin A (OTA) is cause of concern for public health. Currently, several detection methods are available to detect OTA in singles spices but not in spice mixtures such as merkén. The present study modified, optimized, and validated a method for successful OTA quantification in merkén, paprika, and chili. A total of 321 merkén (n: 256), paprika (n: 39), and chili (n: 26) samples were collected from different retail locations in Chile over a 5-year period (2016–2020). OTA levels were determined by high performance liquid chromatography with fluorescence detection (HPLC-FLD). Validation parameters that included the limit of detection (LOD: 0.3 ng g−1), limit of quantification (LOQ: 1.0 ng g−1), linearity (0.9975), recovery in a fortified blank matrix (5 ng g−1: 81–105%), and relative standard deviation (RSD: 0.49–6.87%) were satisfactory. The presence of OTA in merkén, paprika, and chili samples was 66%, 59%, and 61%, respectively. Maximum OTA levels in merkén, paprika, and chili exceeded the limit established by national regulations of 5 ng g−1 by 77-fold (387.9 ng g−1), 59-fold (297 ng g−1), and 14-fold (71.6 ng g−1), respectively. The estimated daily intake (EDI) values were considerably higher in 2017 than 2020 (paprika ˃ chili ˃ merkén). The EDI values were also lower than the Benchmark dose level (BMDL10; 4730 ng kg−1 bw day) for non-neoplastic effects. Our study pinpointed the importance for continuously monitoring and regulating the OTA contamination in agricultural commodities such as spices. More investigations should be generated to modify the current regulation of OTA in spices.

Flowing on-line preparation of deglycosylation, labeling and purification for N-glycan analysis

The current in-solution analysis of N-glycans suffers from several disadvantages including tedious de-glycosylation time and multi-step pre-treatment procedures. Here, an ultra-simple flowing on-line analysis of labeled N-glycans for high-performance liquid chromatography with fluorescence detection (HPLC-FLD) was developed for eliminating the deficiencies. This on-line analysis consisted of an immobilized enzyme reactor (IMER) of PNGase F for efficient release of N-glycans, labeling of released N-glycans and following purification of derivatives on microfluidic chip. Notably, efficient preparations for all type of N-glycans were completed within ∼30 min. To our best knowledge, this is the first time to integrated the whole preparation of N-glycan deglycosylation, labeling and purification only by a simple fluidic flow with our developed device. Good reproducibility and stability were achieved with the relative standard deviation (RSD) less than 10%. Furthermore, the glycome studies with human serum revealed a good adaptability for biological samples. Our work provides an efficient N-glycomic strategy that can be applied to further multilayered clinical analysis.