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INTRODUCTION: Aflatoxins, potent carcinogens produced by Aspergillus species, present significant health risks and commonly contaminate herbal products such as Chrysanthemum morifolium. Detecting these toxins in C. morifolium proves challenging due to the complex nature of the herbal matrix and the fluctuating levels of toxins found in different samples. OBJECTIVES: This study aimed to develop and optimize a novel method for the detection of aflatoxins in C. morifolium using dispersive liquid-liquid microextraction combined with high-performance liquid chromatography-fluorescence detection based on quality by design principles. METHODOLOGY: The method involved determining critical method attributes and parameters through the Plackett-Burman design, followed by optimization using the Box-Behnken design. Monte Carlo simulation was employed to establish a design space, which was experimentally verified. Method validation was performed to confirm accuracy, precision, and stability. RESULTS: The developed method exhibited excellent linearity (R2 > 0.9991) for aflatoxins B1, B2, G1, and G2 across a range of concentrations, with recovery rates between 85.52% and 102.01%. The validated method effectively quantified aflatoxins in C. morifolium under different storage conditions, highlighting the impact of temperature and storage time on aflatoxin production. CONCLUSION: This study successfully established a reliable and effective method for the detection of aflatoxins in C. morifolium, highlighting the importance of strict storage conditions to reduce aflatoxin contamination. Using a quality by design framework, the method demonstrated robustness and high analytical performance, making it suitable for routine quality control of herbal products.

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The determination of biogenic amines (BAs) in alcoholic beverages is crucial for assessing their health impact, ensuring beverage quality, and guaranteeing safety. Herein, a rapid one-pot derivatization/magnetic solid-phase extraction (OPD/MSPE) method was proposed using 6-aminoquinolinyl-N-hydroxysuccinimide carbamate as the derivatization reagent and magnetic hydroxyl-functionalized multi-walled carbon nanotubes as the extraction material. Integration of derivatization and extraction steps simplifies the sample preparation process, taking only three minutes and eliminating the need for centrifugation by utilizing magnetic sorbent. The resulting desorption solution was directly analyzed by high-performance liquid chromatography-fluorescence detection (HPLC-FLD) without any evaporation or reconstitution steps. The integrated OPD/MSPE-HPLC-FLD method demonstrates excellent linearity (R2 > 0.992), accuracy (relative recoveries: 85.1-109.2%), precision (RSDs≤9.7%) and detection limits (limits of detection: 0.3-2 ng/mL). It has been successfully applied to determine free BAs in various alcoholic beverages, including red wine, Baijiu, Huangjiu, and beer. This method enables rapid, sensitive and precise analysis of BAs in alcoholic beverages.

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This research introduces a novel approach for detecting sartan antihypertensive drug adulteration in herbal oral liquids using cotton fiber-supported liquid extraction (CF-SLE) combined with high-performance liquid chromatography-fluorescence detection (HPLC-FLD). Optimal extraction parameters were determined through systematic method development, establishing a sample solution with a pH of 3.0, using 200 mg of cotton fiber, ethyl acetate as the extraction solvent, and a solvent volume of 4 mL. These conditions demonstrated robust extraction efficiency and were further validated for precision and accuracy, with intra- and inter-day relative standard deviations consistently below 7.5 % and relative recoveries ranging from 88.5 % to 106.1 %. The method exhibited excellent linearity for sartans, with R² values greater than 0.993 across a concentration range of 10-2000 ng/mL. Detection limits were effectively established in the range of 2.6-3.1 ng/mL, indicating that the method's sensitivity is adequate for the intended screening purposes. This validated method was then applied to real sample analysis, confirming its potential for routine use in detecting illegal additives within complex herbal matrices, thereby ensuring consumer safety and supporting regulatory compliance.

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Structural analysis of O-glycans from mucins and characterization of the interaction of these glycans with other biomolecules are essential for a full understanding of mucins. Various techniques have been developed for the structural and functional analysis of glycans. While 9-fluorenylmethyl chloroformate (Fmoc-Cl) is generally used to protect amino groups in peptide synthesis, it can also be used as a glycan-labeling reagent for structural analysis. Fmoc-labeled glycans are strongly fluorescent and can be analyzed with high sensitivity using liquid chromatography-fluorescence detection (LC-FD) analysis as well as being analyzed with high sensitivity by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Fmoc-labeled glycans can be easily delabeled and converted to glycosylamine-form or free (hemiacetal or aldehyde)-form glycans that can be used to fabricate glycan arrays or synthesize glycosyl dendrimers. This derivatization allows for the isolation from biological samples of glycans that are difficult to synthesize chemically, as well as the fabrication of immobilized-glycan devices. The Fmoc labeling method promises to be a tool for accelerating O-glycan structural analysis and an understanding of molecular interactions. In this chapter, we introduce the Fmoc labeling method for analysis of O-glycans and fabrication of O-glycan arrays.

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To reveal O-glycan structures in mucins, it is necessary to release covalently bound O-glycans from the polypeptide backbone and derivatize to a form suitable for structural analysis. Various derivatization methods can now be applied in the analysis of O-glycans following the development of O-glycan release methods. Among the many derivatization methods available, we prefer to use fluorescent labeling with 2-aminobenzoic acid (anthranilic acid, 2AA). 2AA-labeled O-glycans can be detected with high sensitivity using liquid chromatography fluorescence detection (LC-FD) analysis because of the strong fluorescence. In addition, as 2AA has a carboxyl group that carries a negative charge, 2AA-labeled O-glycans can be analyzed with high sensitivity in negative ion mode mass spectrometry. Furthermore, because the negative charge of 2AA provides a driving force for electrophoresis, 2AA-labeled O-glycans can be analyzed using capillary electrophoresis (CE) and capillary affinity electrophoresis. High detection sensitivity and versatility are key advantages of the 2AA-labeling method. Here we present our preferred LC-FD and CE analytical methods for 2AA-labeled O-glycans.

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Consuming foods with excess sulfonamide residues threatens human health, underscoring the importance of their detection in food. This study presents an innovative one-pot derivatization/magnetic solid-phase extraction (OPD/MSPE) method for sulfonamides analysis. This approach integrates the derivatization and extraction steps into a single process. The sample solution, along with the derivatization reagent fluorescamine and the sorbent magnetic hydroxyl multi-walled carbon nanotubes, is mixed and vortexed for 3 min. This procedure simultaneously conducts derivatization and extraction, with easy phase separation using an external magnet. This streamlined sample preparation method is completed in only 5 min and, when combined with liquid chromatography-fluorescence detection (LC-FLD), demonstrates excellent linearity (R2 > 0.99) and satisfactory detection limits (0.004-0.04 ng/g) for the quantification of nine sulfonamides in honey samples. The proposed OPD/MSPE-LC-FLD method is distinguished by its simplicity, rapidity, high sensitivity, and specificity, making it an outstanding advancement in the field of food safety analysis.

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Synthetic cannabinoids are one of the most consumed new psychoactive substances, being absolutely necessary the development of analytical methodologies for the determination of these substances in biological fluids. In this study, a liquid chromatography with fluorescence detection (LC-FD) method has been developed for the analysis of 8 synthetic cannabinoids in oral fluids. The method has been validated in terms of linearity, precision and extraction recoveries, giving limits of detection as low as 0.7 µg L-1, and limits of quantification of 2.6 µg L-1. Different silica and polymeric commercial solid sorbents such as C18, Supel-Select HLB, EB2 ExtrabondⓇ and SampliQ-OPT were tested, concluding that Supel-Select HLB provided quantitative recoveries for the extraction of synthetic cannabinoids in oral fluids.•Analysis of synthetic cannabinoids in oral fluids.•Analytical procedure based on liquid chromatography with fluorescence detection.•Sample treatment based on solid phase extraction with HLB cartridges.

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Liquid-liquid microextraction (LLME) techniques have experienced a tremendous growth over the last years but still face major challenges related to the use of more efficient and environmentally friendly solvents. Supramolecular solvents (SUPRASs) have proved outstanding efficiency in LLME, but many of the experimental conditions required for SUPRAS formation and/or application cannot be considered green or experimentally convenient. This paper was intended to make greener both SUPRAS formation and their application to the LLME of low-concentration organic pollutants in environmental waters. For this purpose, a variety of SUPRASs were produced at room temperature by simply mixing alkyl phosphonates (A6-12PO3H- and A6-12PO3-2) and tetrahexylammonium (He4N+) ions in aqueous media. Among them, the SUPRASs produced from decyl hydrogen phosphonate (DePO3H-) and He4N+ allowed, for the first time, the development of SUPRAS-based LLMEs where the SUPRAS previously synthesized was added to the liquid sample, instead of being formed in situ as usual, which was proved particularly advantageous for analyses involving large sample/SUPRAS volume ratios. At near equimolar amounts of DePO3H- and He4N+, the amphiphile arranged in the SUPRAS as planar ribbons consisting of water (21 ± 3%, w/v) and DePO3H- and He4N+ in the concentration range 1.0-1.4 M. The application of these SUPRASs to LLMEs was proved by extracting carcinogenic polycyclic aromatic hydrocarbons (CPAHs) from drinking (tap and bottled) and natural (river, reservoir and underground) water (recoveries between 84 and 117% with standard deviations varying between 1 and 14%). The developed method was simple (it only required the addition of 500 µL of SUPRAS to 75 mL of sample, stirring and centrifugation), sensitive (method quantitation limits were below the maximum allowed limits set by the EU; were 0.6-7.1 ng L-1) and selective (SUPRAS extracts were directly analyzed by liquid chromatography-fluorimetry). This research proves that SUPRASs can be operationally used in LLMEs similarly to conventional solvents, which should favor their routine application in high-sample throughput laboratories.

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Herein, we successfully prepared magnetic Co/Ni-based N-doped 3D carbon nanotubes and graphene nanocomposites (CoNi@NGC) using a simple high-temperature calcination method. The CoNi@NGC nanocomposites were used as adsorbents to study their adsorption performances and underlying kinetic mechanisms for six types of bisphenol compounds (BPs) in water. They were also used as extractants, and acid-base effervescent tablets were used to enhance extractant dispersion with the aid of vigorous CO2 bubbling. Thus, a novel pretreatment method was developed, denoted effervescent reaction-assisted dispersive solid-phase microextraction (ER-DSM), which was combined with high performance liquid chromatography-fluorescence detection (HPLC-FLD) to rapidly quantify trace-level BPs in several drinks. The morphology and structure of the CoNi@NGC adsorbent were characterized in detail using scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), N2 adsorption and desorption (BET-BJH), X-ray photoelectron spectroscopy (XPS), and vibrating sample magnetometry (VSM). The CoNi@NGC nanocomposites were successfully doped with N and exhibited large specific surface areas (109.42 m2/g), abundant pores, and strong magnetic properties (17.98 emu/g).Key parameters were rigorously optimized to maximize the adsorption performance of CoNi@NGC, including adsorbent dosage, solution pH, temperature, and time. Under the constant conditions of pH=7, 5 mg of CoNi@NGC, initial BP concentrations of 5 mg/L, and 5 min of shaking at 298 K, the adsorption percentages of bisphenol M (BPM) and bisphenol A (BPA) reached respective maxima of 99.01% and 98.21%. Remarkably, those of bisphenol Z (BPZ), BPA, and BPM reached almost 100% after 90 min. The adsorption between the BPs and CoNi@NGC was mainly governed by hydrogen bonds, electrostatic interactions, and π-π conjugation. The entire adsorption process was consistent with Freundlich adsorption and a quasi-second-order kinetic equation, representing spontaneous adsorption. Via integration with HPLC-FLD, ER-DSM was used to rapidly extract and analyze trace-level BPs in six types of boxed drinks. Critical factors were optimized individually, including the type of eluent and elution time and volume, which influenced the enrichment effect. Under the optimized extraction conditions (pH=7, 5 mg CoNi@NGC, elution with 2 mL acetone for 6 min), the limits of detection and quantification of the novel extraction method were 0.06-0.20 and 0.20-0.66 µg/L, respectively. The intra- and inter-day precisions spanned the ranges 1.44%-4.76% and 1.69%-5.36%, respectively, and the recoveries in the actual samples were in the range 82.4%-103.7%. Moreover, the respective residual levels of BPA and BPB in peach juice samples were 2.09 and 1.37 µg/L. Regeneration studies revealed that the CoNi@NGC adsorbent could be reused at least five times, which significantly reduced the cost of evaluation. In summary, compared to other methods, this method displays the advantages of a high sensitivity, rapid extraction, and environmental friendliness, thereby exhibiting considerable potential for use in conventional monitoring of trace-level BPs in food matrices.

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A simple, sensitive and reproducible solid-phase extraction method using plastic cartridges containing a monolithic sorbent (m-SPE), coupled to reverse phase liquid chromatography analysis, aiming to determine fifteen polycyclic aromatic hydrocarbons in surface water samples, was developed. The sorbent was easily prepared through a thermal polymerization reaction by using a mixture of n-butyl methacrylate as non-polar monomer and ethylene glycol dimethacrylate as crosslinker contained in a typical Polypropylene syringe cartridge. The effect of different parameters (type of hydrophobic monomer, elution solvent, sample volume, sorbent amount and sorbent load capacity) on the extraction efficiency was optimized. The optimal conditions were achieved by using n-butyl methacrylate as monomer, tetrahydrofurane (THF) as solvent for sorbent cleaning, THF:acetone (1:1) as elution solvent, 25.00 mL of sample volume, 600 µL of the polymerization mixture and 60 µg L-1 as sample loading capacity. Finally, the sorbent charge capacity, the reusability of the cartridges and the extraction efficiency of the m-SPE monolith, as compared with a typical C8 cartridge, were evaluated. Under the optimized experimental conditions, enrichment factors were between 76 and 103, relative recovery factors from 78 to 103%, accuracy values in the range of 58 to 98%, and inter-batch reproducibility values from between 2 and 10%, were obtained. The limits of detection and quantification were obtained by two different procedures: the signal to noise (S/N) ratios (3 and 10, respectively) and the IUPAC convention. The lowest LOD and LOQ values, obtained with the S/N ratios, were between 0.02 and 1.00 µg L-1, respectively whereas with the IUPAC convention the values were between 0.07 and 5 µg L-1. Using this procedure, several PAHs could be detected in the surface water sample taken from a river stream located in La Plata city (Buenos Aires Province, Argentina).

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In this study, a fluorescent reagent, 4-((aminooxy)methyl)-7-hydroxycoumarin (AOHC), was for the first time applied to label the low-molecular-mass aldehydes (LMMAs) through reductive oxyamination reaction to afford single N,O-substituted oxyamine derivatives at room temperatures with derivatization efficiencies as high as 96.8%. In the following high-performance liquid chromatography with fluorescence detection analysis, 12 LMMAs, including furfurals, aromatic aldehydes, and aliphatic aldehydes, were baseline-separated on an ODS column and detected with low limits of detection (LODs) (0.2-50 nM), and good precisions (intraday relative standard deviations [RSDs] were 2.40-4.68%, and interday RSDs were 4.65-8.91%). This approach was then adopted to analyze six alcoholic beverages and five dairy products, and nine LMMAs with concentrations in the 0.28-798.16 µM range were successfully detected with excellent accuracies (recoveries were 92.2-106.2%). Finally, the results were statistically analyzed and discussed. The proposed method has several advantages, including high sensitivity, room-temperature labeling, and the avoidance of further extraction and/or enrichment procedures, demonstrating its great utility for monitoring LMMAs in various complex matrices.

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A fast, simple and efficient ultrahigh-performance liquid chromatography-fluorescence detection (UPLC-FLD) method for the determination of residues of albendazole (ABZ) and its three metabolites, albendazole sulfone (ABZ-SO2), albendazole sulfoxide (ABZ-SO), and albendazole-2-aminosulfone (ABZ-2NH2-SO2), in pig and poultry muscle (chicken, duck and goose) was established. The samples were extracted with ethyl acetate, and the extracts were further subjected to cleanup by utilizing a series of liquid-liquid extraction (LLE) steps. Then, extracts were purified by OASIS® PRiME hydrophilic-lipophilic balance (HLB) solid-phase extraction (SPE) cartridges (60 mg/3 mL). The target compounds were separated on an ACQUITY UPLC® BEH C18 (2.1 mm × 100 mm, 1.7 µm) chromatographic column, using a mobile phase composed of 31% acetonitrile and 69% aqueous solution (containing 0.2% formic acid and 0.05% triethylamine). The limits of detection (LODs) and limits of quantification (LOQs) of the four target compounds in pig and poultry muscle were 0.2-3.8 µg/kg and 1.0-10.9 µg/kg, respectively. The recoveries were all above 80.37% when the muscle samples were spiked with the four target compounds at the LOQ, 0.5 maximum residue limit (MRL), 1.0 MRL, and 2.0 MRL levels. The intraday relative standard deviations (RSDs) were less than 5.11%, and the interday RSDs were less than 6.29%.

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This paper reports a selective, sensitive, and miniaturized analytical method based on a molecularly imprinted graphene oxide (MIP-GO) composite as adsorbent for miniaturized tip solid-phase extraction (MTSPE) to determine naphthalene-derived plant growth regulators (PGRs) in apples. The proposed method combines the advantages of MIP-GOs (high selectivity), MTSPE (low consumption), and high-performance liquid chromatography-fluorescence detection (high sensitivity). Under optimized conditions, the method exhibited appreciable linearity (2.00-200 ng/g), low detection limits (0.21-0.53 ng/g), high accuracy (absolute recoveries: 87.6-99.5%), and high precision (relative standard deviations ≤ 3.0%), along with low consumption (0.5 mL sample solution and 2.0 mg adsorbent). In addition, the adsorption performance of the MIP-GO adsorbent did not decrease over ten months, highlighting the long storage and operational lifetime of the adsorbent. The proposed method was employed for the analysis of naphthalene-derived PGR residues in apples and exhibited promising potential for application in food safety analysis.

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In this study, a novel fluorescent labeling reagent 2-(9-acridone)-ethyl chloroformate (AEC-Cl) was designed, synthesized and applied for the determination of free amino acids by high-performance liquid chromatography with a fluorescence detector (HPLC-FLD). The free amino acids were rapidly and efficiently labeled by AEC-Cl in the presence of basic catalyst (pH 9.0) within 5 min at room temperature (25 °C). The derivatives exhibited excellent stability and fluorescence properties, with maximum excitation and emission wavelengths at 268 nm and 438 nm, respectively. Derivatives of 22 kinds of natural amino acids were completely separated by gradient elution on a Hypersil ODS C18 column. Under the optimal conditions, the calibration curves exhibited excellent linear responses, with correlation coefficients of R2 > 0.9994. The detection and quantification limits were in the range of 0.61-2.67 µg kg-1 and 2.07-8.35 µg kg-1, respectively. Therefore, AEC-Cl was successfully applied for the detection of trace levels of free amino acids in honey samples. Graphical abstract A novel fluorescent labeling reagent was applied for the determination of free amino acids in honey by high-performance liquid chromatography with a fluorescence detector.

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In this study, high-performance liquid chromatography with fluorescence detection (HPLC-FLD) has been used for the first time, for direct determination of warfarin and its major metabolite, 7-hydroxywarfarin, in rat plasma. The simple and sensitive method was developed using Fortis® reversed-phase diphenyl column (150 × 4.6 mm, 3 µm) and a mobile phase composed of phosphate buffer (25 mmol L-1)/methanol/acetonitrile (70:20:10, V/V/V), adjusted to pH 7.4, at a flow rate of 0.8 mL min-1. The diphenyl chemistry of the stationary phase provided a unique selectivity for separating the structurally related aromatic analytes, warfarin and 7-hydroxywarfarin, allowing their successful quantification in the complex plasma matrix. The method was linear over the range 0.01-25 µg mL-1, for warfarin and 7-hydroxywarfarin, and was found to be accurate, precise and selective in accordance with US FDA guidance for bioanalytical method validation. The method was sensitive enough to quantify 0.01 µg mL-1 of warfarin and 7-hydroxywarfarin (LLOQ) using only 100 µL of plasma. The applicability of this method was demonstrated by analyzing samples obtained from rats after oral administration of a single warfarin dose, and studying warfarin and 7-hydroxywarfarin pharmacokinetics.

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A method was developed for the rapid determination of bisphenol A (BPA) and eight structural analogs in children's plastic water bottles by online enrichment coupled with high performance liquid chromatography-fluorescence detection (HPLC-FLD). The correlation coefficients of the nine bisphenols were greater than 0.998. The limits of detection (LOQs) ranged from 0.13 ng/L to 66.7 ng/L. The recoveries ranged from 90.7% to 112.4% (RSD<11.3%, n=6). This method was applied to monitor nine bisphenols in children's water bottles. The results showed that except 4,4'-(9H-fluoren-9-ylidene)bisphenol (BPFL), all the remaining eight bisphenols were detected in different water bottles. The amounts of bisphenols leached increased with the increase of soaking time, and decreased after washing several times at 100℃. The proposed strategy is rapid, sensitive, reliable and eco-friendly, and is suitable for the simultaneous analysis of new bisphenols in water samples.

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A simple, rapid and low-cost determination method of benzo(a)pyrene in fried and baked foods was proposed by high performance liquid chromatography combined with vesicular coacervative supramolecular solvent (SUPRAS) extraction. The vesicular coacervate was composed of 1-octanol and tetrabutylammonium bromide. 200 mg of dried samples with 600 µL SUPRAS could be mixed to extract benzo(a)pyrene. Neither evaporation nor further clean-up steps for the extracts were needed. The overall sample treatment took approximately 30 min, and several samples could be simultaneously treated using conventional lab equipment. Then, benzo(a)pyrene was analyzed via liquid chromatography-fluorescence detection. Parameters affecting the extraction efficiency were investigated and optimized. The results showed good linearity of benzo(a)pyrene with the coefficients of determination (R 2) of more than 0.9999 in the range of 0.1-50.0 µg/kg. The limit of detection of the method was 0.11 µg/kg. Recoveries for spiked samples in the range of 1-10 µg/kg were between 89.86 and 100.01%, with relative standard deviations from 1.20 to 3.20%. Benzo(a)pyrene was present in food samples (including instant noodles, biscuits, rice crust and fried bread stick) at concentrations in the range of 0.08-0.39 µg/kg according to the proposed method. The proposed pretreatment method significantly reduces the analysis time. Furthermore, the solventless approach is in accordance with the green chemistry development trend and has significant application prospects.

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In this work, a novel Fe3O4@Cu3(btc)2-embedded polymerized high internal phase emulsion (Fe3O4@HKUST-1-polyHIPE) monolithic cake was synthesized, characterized and used as an adsorbent in the magnetic stir cake sorptive extraction (MSCSE) and determination of tetracycline antibiotics (TCs) in food samples by a combination of with high-performance liquid chromatography-fluorescence detection (HPLC-FLD). The prepared Fe3O4@HKUST-1-polyHIPE monolithic composites displayed a strong extraction ability and high column capacity due to enhanced interactions such as π-π interactions, hydrogen bonding, and electrostatic interactions. The extraction and desorption conditions were evaluated, and the calibration curves of four spiked TCs were linear (R2 ≥ 0.9991) in the range from 20 to 800 ng mL-1 for milk and egg samples, and 20 to 800 ng g-1 for chicken muscle and kidney samples. The limits of detection and the limits of quantification of the four TCs by using the proposed MSCSE-HPLC-FLD method were in the range of 1.9-4.6 and 5.5-13.9 ng mL-1 for milk and egg samples, and 1.8-3.7 and 5.3-13.0 ng g-1 for chicken muscle and kidney samples, respectively. The recoveries of the target TCs from spiked food samples were in the range from 86.6 to 110.7% with relative standard deviations lower than 7.0%. The proposed method was successfully applied for the determination of these four TCs in milk, egg, chicken muscle, and kidney samples.

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A new method was established for the determination of indoles (indole (IND), 3-methylindole (3-MI), indolyl-3-acetic acid (IAA) and indolyl-3-propionic acid (IPA)) in plasma by high performance liquid chromatography-fluorescence detection (HPLC-FLD). The analytes were separated simultaneously on a Shim-pack VP-ODS column (150 mm×4.6 mm, 4.6 µm) using 15 mmol/L sodium dihydrogen phosphate solution and methanol (48:52, v/v) as the mobile phases. The column temperature was 30 ℃, and the flow rate was 0.8 mL/min. The calibration curves of IND, 3-MI, IPA and IAA showed good linear relationships. The intra-day and inter-day relative standard deviations (RSDs) for the analytes were both less than 6.31%. The average recoveries of the analytes in plasma were 97.5%-107.0%. The method was successfully applied to the analysis of indoles in the plasma of healthy women of reproductive age (n=25) as controls and patients with polycystic ovarian syndrome (n=61). The results showed that the concentrations of indoles in the plasma were significantly different between the two groups, and IND was found to be a risk factor and a potential diagnostic biomarker for polycystic ovarian syndrome (PCOS). The method is simple, sensitive and suitable for use in clinical testing and laboratory research.

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A novel high-performance liquid chromatography-fluorescence analysis in combination with in situ degradation-derivatization (ISD-D) technique was developed for simultaneous determination of seven organophosphorus thioester pesticides (OPTPs) in tea. The ISD-D technique was based on degradation of OPTPs by a nucleophilic substitution reaction between phenylbutane-1,2,3-trione-2-oxime and OPTPs, which can give thiol degradation products (DPs). The thiol DPs obtained were derivatized with the novel derivatization reagent N-(4-(carbazole-9-yl)-phenyl)-N-maleimide (NCPM) in a syringe. Attractively, NCPM itself did not fluoresce, whereas the derivatives of the thiol DPs fluoresced intensely, with excitation and emission maxima at 290 nm and 368 nm, respectively, which extraordinary reduced the background interference and increased the detection sensitivity for thiol DPs. Excellent linearity (R2 > 0.995) for all OPTPs was achieved, with limits of detection and limits of quantitation ranging from 0.23 to 0.45 µg/kg and from 0.75 to 1.43 µg/kg, respectively. Satisfactory recoveries ranging from 90.5% to 96.0% were obtained for all OPTPs. The ISD-D technique provided a novel and sensitive strategy for quantitation of trace amounts of OPTPs in real samples. Graphical abstract ᅟ.

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