Molecularly imprinted dispersive micro solid-phase extraction and tandem derivatization for the determination of histamine in fermented wines.

Histamine causes allergic reactions and can serve as an indicator for assessing food quality. This study designed and developed a dispersive micro solid-phase extraction (D-µSPE) method that combined the advantages of dispersive liquid-liquid extraction and solid-phase extraction (SPE). Molecularly imprinted polymers (MIPs) were employed as the solid phase in the D-µSPE method to extract histamine in wine samples. We used microwave energy to significantly reduce the synthesis time, achieving an 11.1-fold shorter synthesis time compared to the conventional MIP synthetic method. Under optimized D-µSPE conditions, our results showed that the dispersive solvent could effectively increase the adsorption performance of MIPs in wine samples by 97.7%. To improve the sensitivity of histamine detection in gas chromatography-mass spectrometry, we employed the microwave-assisted tandem derivatization method to reuse excess derivatization reagents and reduce energy consumption and reaction time. Calibration curves were constructed for wine samples spiked with 0-400 nmol histamine using the standard addition method, resulting in good linearity with a coefficient of determination of 0.999. The intra- and inter-batch relative standard deviations of the slope and intercept were < 0.7% and < 5.3%, respectively. The limits of quantitation and detection were 0.4 nmol and 0.1 nmol, respectively. The developed method was successfully applied to analyze the histamine concentration in 10 commercial wine samples. In addition, the AGREEprep tool was used to evaluate the greenness performance of the developed method, which obtained a higher score than the other reported methods.

Simultaneous derivatization and liquid-solid phase transition microextraction of six biogenic amines in foods followed by narrowbore liquid chromatography-ultraviolet detection.

Biogenic amines are quality control criteria for foods that are potentially toxic to humans. In this study, amidation derivatization for biogenic amines and liquid-solid phase transition microextraction were carried out simultaneously for food sample pretreatment. The derivatization reaction was executed in one pot with coumarin-3-carboxylic acid as the derivatizing reagent and (1-cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate as the coupling agent. Liquid-solid phase transition microextraction was achieved by the salting-out effect, using a phase change salt (1 M disodium hydrogen phosphate) solution. The combined derivatization and microextraction process was completed within 3 min at 30 °C, and the liquid top phase was easily obtained by placing the tube in an ice bath. Finally, a narrowbore liquid chromatograph coupled with a UV detector was used to determine the levels of six biogenic amines. The coupling agent-assisted derivatization and liquid-solid phase transition microextraction parameters were also investigated. The quantitative linear ranges were 3-400 µM for histamine, putrescine, spermidine, cadaverine, and tyramine and 5-400 µM for spermine, and the detection limit was 1 µM. The relative standard deviations of the intra- and inter-batches were <5.3% and 8.4%, respectively, while the relative error was <4.5% for both. We successfully applied this simultaneous derivatization-microextraction method to determine the biogenic amines in fermented foods.

Ultrasound-Vortex-Assisted Dispersive Liquid-Liquid Microextraction Combined with High Performance Liquid Chromatography-Diode Array Detection for Determining UV Filters in Cosmetics and the Human Stratum Corneum.

This study explores the amounts of common chemical ultraviolet (UV) filters (i.e., avobenzone, bemotrizinol, ethylhexyl triazone, octocrylene, and octyl methoxycinnamate) in cosmetics and the human stratum corneum. An ultrasound-vortex-assisted dispersive liquid-liquid microextraction (US-VA-DLLME) method with a high-performance liquid chromatography-diode array detector was used to analyze UV filters. A bio-derived solvent (i.e., anisole) was used as the extractant in the US-VA-DLLME procedure, along with methanol as the dispersant, a vortexing time of 4 min, and ultrasonication for 3 min. The mass-transfer rate of the extraction process was enhanced due to vortex-ultrasound combination. Various C18 end-capped columns were used to investigate the separation characteristics of the UV filters, with XBridge BEH or CORTECS selected as the separation column. Calibration curves were constructed in the 0.05-5 µg/mL (all filters except octocrylene) and 0.1-10 µg/mL (octocrylene) ranges, and excellent analytical linearities with coefficients of determination (r2) above 0.998. The developed method was successfully used to analyze sunscreen. Moreover, experiments were designed to simulate the sunscreen-usage habits of consumers, and the cup method was used to extract UV filters from the human stratum corneum. The results suggest that a makeup remover should be employed to remove water-in-oil sunscreens from skin.

Microwave-assisted derivatization combined with coacervative extraction for determining glutathione in biomatrix samples, followed by capillary liquid chromatography.

In this study, an ecofriendly analytical method was developed for determining glutathione (GSH) levels in biomatrix samples. 9-(bromomethyl)acridine was used for the first time as a derivatization reagent in GSH analysis. Microwave-assisted derivatization reduced the reaction time to 1 min. After derivatization, coacervative extraction was employed to extract GSH derivative from the complex biomatrix and to increase sensitivity. Because the negatively charged group of the GSH derivative was neutralized by the extracting agent Aliquat 336, aggregates formed without any coacervating agents. Furthermore, capillary liquid chromatography coupled with ultraviolet detection was applied to decrease waste generation and increase selectivity. This method successfully quantified GSH levels in various biomatrices, including erythrocytes, HaCaT cells, BALB/3T3 cells, and 3T3-L1 fibroblasts. This method only required a low sample volume (≤10 µL). A standard addition method was utilized to spike the biomatrix samples with 0-4.8 nmol GSH to construct calibration curves. The proposed method performed well, with a determination coefficient of 0.999 and relative standard deviations of less than 6.59% for the slope and the intercept, as determined by linear regression analysis. The limit of detection of GSH in the standard solution was 800 nM or 0.4 pmol. Compared to non-derivatized GSH, the proposed method for detecting derivatized GSH provides 750-fold greater sensitivity.

Enantioselective determination of aspartate and glutamate in biological samples by ultrasonic-assisted derivatization coupled with capillary electrophoresis and linked to Alzheimer's disease progression.

A simple cyclodextrin-mediated capillary zone electrophoresis method equipped with a laser-induced fluorescence detector was developed for chiral analysis of the excitatory amino acids aspartate (Asp) and glutamate (Glu). Plasma and cerebrospinal fluid (CSF) samples were pretreated with centrifugal filter devices before analysis to remove high-molecular-weight proteins (molecular weight cut-off: 3000) and then derivatized using 10 mM 6-carboxyfluorescein N-hydroxysuccinimide ester in DMSO under sonication at 25 °C for 2 h. After the derivatization reaction, reacted samples were diluted 100-fold with 5-([4,6-dichlorotriazin-2-yl]amino)fluorescein hydrochloride (DTAF) solution and then hydrodynamically subjected to capillary electrophoresis (0.5 psi for 5 s, injection volume 8.27 nL). The separation buffer consisted of 50 mM borate buffer (pH 9.0) with 6 mM γ-CD and 0.1% polyvinylpyrrolidone, and the separation voltage was set at 20 kV. In the linearity calculations for the determination of d/l-Asp and d/l-Glu in plasma and CSF, a standard addition method was utilized to spike solutions with 0-20.0 µg mL-1l-Glu and 0-2.0 µg mL-1d-Glu and d/l-Asp to construct calibration curves. Correlation coefficients were above 0.998 for every analyte. The limits of detection (S/N = 3) for d/l-Asp and d/l-Glu standard solutions were 0.85-0.96 µg mL-1. The proposed method was applied successfully to determine d/l-Asp and d/l-Glu concentrations in the plasma and CSF samples of 26 patients with Alzheimer's disease (AD), and the association between these concentrations and disease severity was investigated. Statistical analysis showed a moderately negative correlation (r = -0.158) between plasma l-Asp concentration and AD severity.

Tandem derivatization combined with salting-out assisted liquid-liquid microextraction for determination of biothiols in urine by gas chromatography-mass spectrometry.

Detection of polar organic compounds (POCs) using gas chromatography (GC) is not straightforward due to high polarity, hydrophilicity, and low volatility of POCs. In this study, we report a tandem microwave-assisted derivatization method combined with salting-out assisted liquid-liquid microextraction (SALLME) to modify successively the polar groups of POCs in protic and aprotic solvents. Biothiols (cysteine and homocysteine) served as a proof of concept for this method because they possess three polar groups (thiol, amine, and carboxyl); the derivatizing reagent was 3,4,5-trifluorobenzyl bromide (Br-TFB) for alkylation. The solubility of the POCs in the protic or aprotic reaction medium affected the number of TFB molecules attached. Using the tandem derivatization with Br-TFB, the thiol and amine groups of biothiols were alkylated in the protic system, and the carboxylic groups of biothiols were alkylated in the aprotic system. The developed method was then successfully applied to measure biothiols in human urine. Because of the complex urine matrix and the lack of urine samples without endogenous biothiols, the standard addition method was utilized to avoid the matrix effect, check the recovery, and calculate the initial biothiol content in the urine. Regarding the linearity of the standard addition curves, the coefficient of determination was >0.996, and the linear regression showed satisfactory reproducibility with a relative standard deviation <3.9% for the slope and <8.8% for the intercept. The levels of cysteine and homocysteine in healthy human urine ranged from 28.8 to 111µmolL-1 and from 1.28 to 3.73µmolL-1, respectively. The proposed method effectively increased the sensitivity of GC-MS assays of water-soluble compounds in human urine.

Determination of tranexamic acid in various matrices using microwave-assisted derivatization followed by dispersive liquid-liquid microextraction.

Tranexamic acid (TA) is widely used to treat medical hemorrhagic conditions and as a whitening agent in cosmetic products. The aim of this study was to develop a micro-analytical method of determining TA in widely varying sample matrices, including pharmaceuticals, cosmetics, extracts of the stratum corneum, human keratinocyte cells, and human plasma. Using dansyl chloride as the derivatizing reagent in the pretreatment reduced the reaction time to within 4min in combined microwave-assisted reaction. To prevent excess dansyl chloride and sample matrices from damaging the column, 4µL chloroform was used to remove excess derivatizing agent and interference through dispersive liquid-liquid microextraction (DLLME) method. After pretreatment, 1µL of sample solution was injected in capillary liquid chromatography coupled with ultraviolet detector (CapLC-UV). By coupling the proposed method with cylinder-sampling method, TA was successfully extracted from the stratum corneum. The calibration curves for the standard solution and human plasma were in the ranges of 0.1-50µM and 5-500µM, respectively. Both calibration curves had correlation coefficients (r(2)) of 0.999. The limits of detection were 0.03pmol in standard solution and 3pmol in plasma. Compared to non-derivatized TA, the use of CapLC-UV for detecting derivatized TA provides a 1000-fold sensitivity improvement.