Salting-out Assisted Liquid-liquid Extraction Research Articles

Salting-out assisted liquid-liquid extraction for the simple and rapid determination of veterinary antibiotic residues in aquatic products by HPLC-MS/MS

Based on salting-out assisted liquid-liquid extraction (SALLE) and high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS), a simple, rapid pretreatment without complex clean-up for the determination of 22 veterinary drug residues in aquatic products was developed and validated. In order to improve the efficiency of the method, the key procedural parameters of SALLE were fabricated. Na2EDTA-Mcllvaine buffer/ACN was used as the extraction solvent, anhydrous MgSO4 and NaCl as the extraction salts. The relationship between extraction efficiency and logD was initially evaluated during the optimization process. This study was well validated in various aquatic samples such as bass, large yellow croaker, carp, and shrimp, the limits of detection (LOD) and accuracy for all compounds ranged from 0.5 to 1.0 μg/kg, 71.4% to 120%. This method has the advantages of rapidity, simplicity, low cost, and high efficiency, and has broad potential for risk monitoring and evaluation of veterinary antibiotics in aquatic products.

An analytical method using salting-out assisted liquid-liquid extraction to quantify ceftriaxone from micro volumes of human serum.

Ceftriaxone (CTRX) is a commonly used cephalosporin antibiotic. It is suggested that monitoring plasma/serum concentrations is helpful for its safe use. This study aimed to develop and validate an analytical method for measuring CTRX concentrations in human serum according to International Conference on Harmonization guideline M10. Ten microliters of serum sample was purified using a salting-out assisted liquid-liquid extraction procedure with magnesium sulfate. The upper layer was then diluted threefold and analyzed using a liquid chromatography-tandem mass spectrometry-based method with a total run time of 12 min. The linear calibration curve was obtained over the concentration range 5-500 μg/ml. The within-run accuracy varied from 0.2 to 6.5%, and the precision was ≤8.0%. The between-run accuracy and precision ranged from 0.7% to 5.6% and ≤6.4%, respectively. Significant carryover was resolved by injecting four blanks after high-concentration CTRX samples. The recovery rates from spiked serum at low and high concentrations were 44.4 and 43.4%, respectively. Other factors, including selectivity, matrix effects, stability, dilution integrity and reinjection reproducibility also met the acceptance criteria. Serum concentrations in 14 samples obtained from two participants receiving 2 g/day of CTRX were successfully determined using this method.

Salt assisted liquid-liquid extraction combined with dispersive liquid-liquid microextraction for the determination of 24 regulated polycyclic aromatic hydrocarbons in human serum

Polycyclic aromatic hydrocarbons (PAHs) are persistent organic pollutants of great concern due to their carcinogenicity and mutagenicity. Their determination in human serum, particularly in at-risk populations, is necessary but difficult because they are distributed over a wide range of polarity and are present at trace level. A new method combining salting-out assisted liquid-liquid extraction (SALLE) and dispersive liquid-liquid microextraction with solidification of floating organic drop (DLLME-SFO) adapted to a reduced volume of sample (100 µl) was developed to determine 24 PAHs in human serum. Some key parameters of DLLME-SFO (volume of extraction solvent, ratio of extraction/dispersive solvent volumes, and salt addition) were first studied by applying it to spiked pure water. For its application to serum, a sample treatment step involving SALLE was optimized in terms of nature and content of salts and applied upstream of DLLME-SFO. It was applied to the extraction of 24 regulated PAHs from spiked serum followed by an analysis by liquid chromatography coupled with UV and fluorescence detection. The extraction recoveries ranged from 48.2 and 116.0 % (relative standard deviations: 2.0–14.6 %, n=5–9), leading to limits of quantification of PAHs in human serum from 0.04 to 1.03 µg/L using fluorescence detection and from 10 to 40 µg/L using UV detection. This final method combining SALLE and DLLME-SFO showed numerous advantages such as no evaporation step, high efficiency and low solvent-consumption and will be useful for monitoring PAHs in low volumes of serum.

Photodecomposition of antibiotics and their transformation products in wastewaters using ZnO and TiO2 with LED lamps

This work presents a study of the behavior of three antibiotics (erythromycin (ERY), clarithromycin (CLR) and sulfadiazine (SFZ)) and their transformation products (TPs), generated after photolytic and photocatalytic (TiO2 and ZnO) treatments, in wastewater irradiated with ultraviolet (UV) light emitting diode (LED) lamps as light source. For this, an analytical methodology for the antibiotics and their TPs was developed. Thus, samples were treated by salting-out assisted liquid–liquid extraction (SALLE) and identification and quantification was carried out by liquid chromatography with mass spectrometry (LC-MS) using quadrupole time-of-flight (QTOF) and triple quadrupole (QqQ) analyzers, respectively. The results revealed the presence of 14, 13 and 5 TPs of ERY, CLR and SFZ, respectively, being found in wastewaters spiked with the antibiotics during photo-oxidation treatment. Subsequently, the established analytical method was also applied, in a real case, to study of behavior of these pharmaceuticals (0.2 mg L-1) and their TPs generated in wastewater during a photocatalytic treatment. In this case, 13, 11 and 2 TPs were tentatively identified ERY, CLR and SFZ, respectively. In addition, the photocatalytic treatment using TiO2 exhibited higher degradation rate than photocatalytic treatment using ZnO. The ECOSAR programme was used to estimate the potential ecotoxicity of the identified TPs. The results showed a potential hazard of SFZ, CLR, ERY and some of their TPs to aquatic organisms. Furthermore, some of the identified TPs were found to be more toxic than their parent compounds.

Simultaneous determination of six catechins and caffeine in tea and wine using salting-out assisted liquid-liquid extraction and high-performance liquid chromatography with ultraviolet detection.

Catechins, renowned for their antioxidant properties and health benefits, are commonly present in beverages, particularly tea and wine. An efficient and cost-effective salting-out assisted liquid-liquid extraction (SALLE) method has been developed and validated for the simultaneous determination of six catechins and caffeine in tea and wine samples using high-performance liquid chromatography-ultraviolet (HPLC-UV). This method demonstrates outstanding performance: linearity (1-120µg/mL, r2>0.999), accuracy (96.5%-103.4% recovery), and precision (≤14.7% relative standard deviation), meeting validation requirements set by the US Food and Drug Administration. The reduced sample size (0.1g) minimizes matrix interferences and costs without compromising sensitivity. All analytes were detected in Camellia sinensis teas, with green tea displaying the highest total catechin content (47.5-100.1mg/mL), followed by white and black teas. Analysis of wine samples reveals the presence of catechin in all red and white wines, and epigallocatechin gallate in all red wine samples, highlighting the impact of winemaking processes on catechin content. The SALLE-HPLC-UV approach represents a green alternative by eliminating organic waste, surpassing conventional dilution methods in specificity and sensitivity for catechin determination. AGREEprep assessment emphasizes the strengths of the SALLE procedure, including material reusability, throughput efficiency, minimal sample requirements, low energy consumption, and the absence of organic waste generation.

Evaluation of Iron Chlorin e6 disappearance and hydrolysis in soil and garlic using salting-out assisted liquid-liquid extraction coupled with high-performance liquid chromatography and ultraviolet-visible detection

Iron Chlorin e6 (ICE6), a star plant growth regulator (PGR) with independent intellectual property rights in China, has demonstrated its efficacy through numerous field experiments. We innovatively employed salting-out assisted liquid-liquid extraction (SALLE) with HPLC-UV/Vis to detect ICE6 residues in water, soil, garlic seeds, and sprouts. Using methanol and a C18 column with acetonitrile: 0.1% phosphoric acid mobile phase (55:45, v:v), we achieved a low LOQ of 0.43 to 0.77 μg kg−1. Calibration curves showed strong linearity (R2 > 0.992) within 0.01 to 5.00 mg kg−1. Inter-day and intra-day recoveries (0.05 to 0.50 mg kg−1) demonstrated high sensitivity and accuracy (recoveries: 75.36% to 107.86%; RSD: 1.03% to 8.78%). Additionally, density functional theory (DFT) analysis aligned UV/Vis spectra and indicated ICE6's first-order degradation (2.03 to 4.94 days) under various environmental conditions, mainly driven by abiotic degradation. This study enhances understanding of ICE6's environmental behavior, aids in risk assessment, and guides responsible use in agroecosystems.

Application of salting-out assisted liquid-liquid extraction for the determination of oxytetracycline, tetracycline, tilmicosin, and tylosin in cow milk by liquid chromatography with photodiode array detection

This paper introduces a novel, sensitive, and rapid method for the quantification of oxytetracycline, tetracycline, tilmicosin, and tylosin residues in cow's milk. The method involves a two-step process of extraction and detection. The extraction process uses acetonitrile and salting-out assisted liquid-liquid extraction to isolate the antibiotics from the milk. The detection process employs Liquid Chromatography coupled with photo-diode array detector (PDA) to quantify the antibiotics. The method has been successfully applied to milk samples, demonstrating its effectiveness and potential for widespread use in residue analysis.•The calibration curves for the antibiotics were found to be linear within the range of 0.06–3.0 µg/mL to 0.1–3.0 µg/mL.•The limits of detection for oxytetracycline, tetracycline, tilmicosin, and tylosin were 0.03 µg/mL, 0.02 µg/mL, 0.04 µg/mL, and 0.02 µg/mL respectively.•The method demonstrated an average recovery rate of over 90% from milk samples with peak values reaching up to 0.100–0.200 µg/mL.

Method Development for Aminopyralid Residues Determination by Ultra-High-Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry in Straw

Straw is an important by-product of crop production. It has been widely used as a feed as well as in fruit–vegetable production due to exerting a positive impact on soil and crop health and quality in the operated areas. On the other hand, applied crop-specific herbicides throughout the production stage, like aminopyralid (AP), may lead to significant plant injuries in sensitive vegetable plantations, even in very low concentrations. This study aims to optimize a short time, minimal consumables needed, and sensitive extraction and analytical method for AP residues in straw material using LC-MS/MS. For this purpose, standard AP signal response in acetonitrile (MeCN) and 1% of formic acid (FA) in methanol (MeOH) were tested; several chromatographic separation and mass spectroscopy (MS) parameters were improved and optimized. Subsequently, two different extraction methods were performed in AP spiked straw samples, and extraction efficacies were compared. Salting-out assisted liquid–liquid extraction (SALLE) by acidified MeCN combined with final dissolution in MeOH with 1% of FA prior to the analysis was chosen as the most appropriate method for the straw matrix. This method was then used for validation and real sample test analysis. The limit of quantification (LOQ) of AP in this optimized extraction method achieved 10 ng/g straw with a recovery rate of 71% and 13% RSD. The method was also tested on field straw mulch samples and 22.54 ± 0.8 ng/g of AP was detected.

A highly efficient strategy for simultaneous analysis of eleven monomeric hydroxylated-stilbenes in grape wines and juices by a salting-out assisted liquid-liquid (SALLE) extraction combined with a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS).

Wine and grape juice are known to have hydroxylated stilbenes, a group of phytoalexins, health-promoting compounds. The determination of stilbene species in wine and grape juices remains a challenging task. Here, we propose an efficient strategy for the simultaneous determination of eleven hydroxylated stilbenes in grape wines and grape juice by a SALLE in conjunction with an HPLC-ESI-MS/MS. SALLE and HPLC-MS/MS conditions were optimized. The proposed method was validated and found to be a simple, sensitive, and reliable measure for the determination of the stilbene species in wine and grape juice. The SALLE was fast and efficient, taking only 10min. The HPLC-MS/MS was able to quickly separate and quantitate the eleven stilbenes (9min running). This method was successfully applied to determine the contents of stilbene species in commercial wine and grape juice in Korea. This method is advantageous in reduced sample preparation time, low labor, and cost efficiency.

Salting-Out Assisted Liquid-Liquid Extraction of Cr (VI) from Water Samples before Its Determination by UV-Vis Spectrophotometry

This paper proposes salting-out assisted liquid-liquid extraction for selective extraction of a Cr (VI)-1,5-diphenylcarbazide complex before its determination by UV-Vis spectrophotometry. Parameters affecting the complexation process and the extraction efficiency of the method were studied. The highest extraction efficiency was obtained when acetonitrile was used as an extraction solvent and (NH4)2SO4 was used as a salting-out agent. Under optimum conditions, the calibration curve constructed at seven concentration points exhibited good linearity from 0.015 to 2.00 mg/L with a 0.9973 coefficient of determination. The method’s detection and quantification limits were 0.004 and 0.015 mg/L, respectively. The method showed a 5.8 enrichment factor. The intra- and inter-day precisions studied at two concentration levels (0.75 and 1.25 mg/L) showed relative standard deviation values of <9.0. The recoveries of the spiked water samples at the two concentration levels ranged from 88.0 ± 2.5 to 97.3 ± 0.1%. The method demonstrated negligible interferences (<5%) for Fe, Cu, V, and Cd up to 500 mg/L. The findings showed that the method is simple, rapid, efficient, and selective to be used as an alternative procedure for analyzing Cr (VI) from water and other related matrices.

Simultaneous determination of bicyclol and its two active metabolites concentration in rat plasma and application for pharmacokinetics study of bicyclol and optimized bicyclol-nanoparticles

A simple, sensitive, and rapid LC-MS/MS method for the simultaneous determination of BIC, the M7 and M8 active metabolites in SD rat plasma was developed. After salting-out assisted liquid–liquid extraction (SALLE) with 5 M ammonium acetate solution, the sample was analyzed on a Sciences column (C18 3.0 × 100 mm, 3 μm) using a gradient elution at 40 ℃ within 7 min. The assay displayed excellent linearity in the range of 4–2000 ng/mL for M7 and BIC, and 1–500 ng/mL for M8. The results of this method exhibited that the precision, accuracy, matrix effect, recovery, and stability of BIC, M7 and M8 met all requirements for the quantitation in plasma samples. The pharmacokinetic result showed that the AUC(0→t) was calculated as 383.1 ± 164.4 (ng/mL·h) for BIC, and 5627 ± 1261 (ng/mL·h) for M7 after oral administration with BIC. Compared with BIC group, the pharmacokinetic parameters in BIC-NPs group were improved. Augmentation in AUC(0→t) (3.02-fold) and t1/2 (1.43-fold) for BIC. Meanwhile, double peak phenomenon was observed on the mean plasma concentration–time curves of M7 in BIC and BIC-NPs group. In conclusion, both BIC and BIC-NPs were metabolized to abundant M7 in SD rat, which would provide a basis for researching the treatment mechanism of liver injury.

A Highly Selective Analytical Method Based on Salt-Assisted Liquid-Liquid Extraction for Trace-Level Enrichment of Multiclass Pesticide Residues in Cow Milk for Quantitative Liquid Chromatographic Analysis.

In this study, a simple, inexpensive, selective, and fast salting-out assisted liquid-liquid extraction (SALLE) technique coupled with high-pressure liquid chromatography-diode array detection (HPLC-DAD) was developed for the extraction, preconcentration, and analysis of trace level seven multiclass pesticide residues in pasteurized and raw cow milk samples. The significant factors that affect the extent to which the target analytes are extracted, such as the type of extraction solvent and its volume, the type and concentration of salting-out salts, the pH of the solution, and the extraction time, have been investigated. Under optimum conditions, the correlation coefficient (r2) was obtained within a range of 0.9982-0.9997 for a broad linear range concentration of 2-1500 ng·mL-1. Reliable sensitivity was achieved with limits of detection (LODs) and limits of quantification (LOQs) ranging from 0.58-2.56 ng·mL-1 and 1.95-8.51 ng·mL-1, respectively. While precision with interday and intraday in terms of relative standard deviations (RSDs) was observed in the range of 1.97 - 7.88% and 4.52 - 8.04%, respectively. The results of the precision studies reveal that good repeatability and reproducibility (RSDs <9) were achieved, thus showing a low variability extraction of the developed method. Finally, the proposed and validated approach was effectively used to extract and determine pesticide residues in real milk matrices; however, the target analytes were not detected in all samples.

Salting-out assisted liquid–liquid extraction combined with LC–MS/MS for the simultaneous determination of seven organic UV filters in environmental water samples: method development and application

Organic UV filters (OUVFs), the active ingredient in sunscreens, are of environmental concern due to reported ecotoxicological effects in aquatic biota. Determining the environmental concentrations of these chemicals is essential for understanding their fate and potential environmental risk. Salting‐out assisted liquid–liquid extraction (SALLE) coupled with liquid-chromatography tandem mass spectrometry (LC–MS/MS) was developed for simultaneous extraction, separation, and quantification of seven OUVFs (2,4-dihydroxybenzophenone, 2,2′,4,4′-tetrahydroxybenzophenone, 4-methylbenzylidene camphor, butyl-methoxy-dibenzoyl methane, octocrylene, octyl methoxycinnamate, and oxybenzone). Method detection limits (MDLs) ranged from 11 to 45 ng/L and practical quantification limits (PQLs) from 33 to 135 ng/L. Method trueness, evaluated in terms of recovery, was 69–127%. Inter-day and intra-day variability was < 6% RSD. The coefficients of determination were > 0.97. The method was applied to river and seawater samples collected at 19 sites in and near Port Phillip Bay, Australia, and temporal variation in OUVF concentrations was studied at two sites. Concentrations of OUVF were detected at 10 sites; concentrations of individual OUVFs were 51–7968 ng/L, and the maximum total OUVF concentration detected at a site was 8431 ng/L. Recreational activity and water residence time at the site contributed to OUVF’s environmental presence and persistence. The benefits of the SALLE-LC–MS/MS method include its simple operation, good selectivity, precision over a wide linear range, and that obtained extracts can be directly injected into the LC–MS/MS, overall making it an attractive method for the determination of these OUVFs in environmental water matrices. To our knowledge, this is the first report of the occurrence of OUVFs in Port Phillip Bay, Australia.

A salting-out assisted liquid-liquid extraction method for 25 emerging pesticides in follicular fluid

Emerging pesticides of neonicotinoids (NEOs) and “Universal Pesticides” (UPs) are a growing global concern due to their growing commercial importance and potential risks to human health. The currently available analytical methods for these pesticides in biomonitoring were usually tailored for limited number of analytes, or were time consuming and costly. In this study, an efficient and sensitive method for the analysis of 16 NEOs and nine UPs in human follicular fluid (FF) was developed by using a salting-out assisted liquid-liquid extraction (SALLE) method and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Method performance was evaluated by calibration linearity (r > 0.99), sensitivity at limits of quantification (0.01–0.50 ng/mL), accuracy at relative recoveries (81–117%) and precision at relative standard deviations (≤16%). The developed method was further validated by analyzing 21 human FF samples that were collected from a hospital in Guangzhou, China. Among the 25 study analytes, two NEOs and six UPs had their detection rates over 85% and medians at 0.048–0.808 ng/mL in the FF samples. Considering the well-known toxicity of these pesticides and their metabolites, it is urgent to figure out exposure profiles of study pesticides and potential reproductive risk for women. To the best of our knowledge, this study is the first to develop and apply the SALLE method in the extraction of 16 NEOs and nine UPs simultaneously in human FF.

Salting-Out-Assisted Liquid-Liquid Extraction Method for the Determination of Nicotine from Oral Traditional and Innovative Tobacco Products Using UPLC-MS/MS.

In this study, we developed and validated a novel method that allows for the extraction and quantitation of nicotine from a variety of commercially available oral tobacco products including loose and pouched traditional moist smokeless tobacco products, and oral tobacco-derived nicotine (OTDN) lozenges, gums, and pouches. The method employed an extraction technique consisting of salting-out assisted liquid-liquid extraction using sodium hydroxide and acetonitrile in conjunction with ultra-high pressure liquid chromatography coupled to mass spectrometry. Accurate quantitation was obtained using nicotine methyl-d3 isotopically labeled internal standard. Chromatographic separation of nicotine and nicotine methyl-d3 internal standard was achieved using a Waters Acquity C18 column (50 mm × 2.1 mm i.d., 2.5 μm) with 10 mM ammonium acetate buffer (pH = 10) and acetonitrile as mobile phase A and B, respectively. Using a gradient elution and a flow rate of 0.4 mL/min for 5 min runtime, nicotine eluted at 1.74 min. The method was validated according to ICH guidelines for all the sample types with an accuracy for nicotine within 89-109%. Repeatability and intermediate precision were both estimated to be ≤7% relative standard deviation (% RSD). This method is applicable for a wide range of traditional moist smokeless and OTDN tobacco products.

An automated accelerated salting-out assisted solvent extraction (A-ASASE) of stilbenoids from Vitis vinifera L. branches: False proof or a proof of concept?

An automated accelerated salting-out assisted solvent extraction (A-ASASE) was developed. This approach made full use of the advantages of both the accelerated solvent extraction (ASE) technique and salting-out assisted liquid-liquid extraction (SALLE), without any modifications at the level of the ASE machine. The A-ASASE combined an automated extraction at high temperature/pressure and an enrichment step using eco-friendly solvents (e.g., water and ethanol) in a fully integrated and simple way that required only low volumes of solvents. The A-ASASE was successfully applied to extract three stilbenoids, namely E-resveratrol, E-ε-viniferin and E-vitisin B, present in grapevine (Vitis vinifera L.) by-products as a proof-of-concept compound. First, factors affecting the yield of stelbenoids by SALLE were investigated by means of a design of experiment model (DOE). The SALLE was then incorporated into ASE. Various ASE operational parameters such as temperature and number of cycles were also investigated. The highest yield was obtained using a mixture of ethanol-water, 70:30 (v/v) at 60 °C, using three extraction cycles and with 5 min of contact each. The highest yields of stilbenoids were 5.87 ± 0.28 g kg−1 dry weight, 3.27 ± 0.72 g kg−1 dry weight and 1.81 ± 0.2 g kg−1 dry weight for resveratrol, E-ϵ-viniferin and vitisin B, respectively. The A-ASASE was then successfully applied to extract the same stilbenoids from the roots of Vitis vinifera L. These results highlight the applicability of A-ASASE methodology to the extraction and enrichment of the extract in compounds of medium-high polarity in an easy and fully automated way.

Salting-out assisted acidic deep eutectic solvent Liquid-Liquid extraction of 2,3-Butanediol from aqueous Solutions: Green and sustainable approach

Biological fermentation is considered as a green and economically viable method to produce 2,3-Butanediol (BDO, potential alternative fuel). However, the downstream extraction of BDO is the challenging and cost determining factor in biological fermentation. Deep eutectic solvent (DSE) has been widely used in separation field for its simple preparation, cut-price, environment friendliness and recyclability. Nine acidic deep eutectic solvents (ADES) were prepared and used to construct a salting-out liquid–liquid extraction system to separate BDO from aqueous solution. The major experimental parameters influencing salting-out assisted liquid–liquid extraction (SALLE), including type and volume of ADES, salt type and concentration, BDO concentration, equilibrium time, equilibrium temperature, and solution pH, were investigated. Under optimal conditions when using TEAC/n-CA as extractant, the extraction efficiency, selectivity and distribution coefficient for 10% BDO were 92.3%, 281.0 and 121.1, respectively. Moreover, evaporation process was used to recover K3PO4, the recovery was more than 90%, and the extraction efficiency of BDO was still more than 92% after recycling 5 times. Meaningfully, the recovery and recycling of TEAC/n-CA were realized by adjusting hydrophobic capacity with pH-response characteristic of ADES, the recovery was more than 66%, and the extraction efficiency of BDO was still more than 91% after recycling 5 times. It demonstrated that salting-out agent and pH-responsive deep eutectic solvent could be repeatedly used. The novel processing offers a green and sustainable route for BDO extraction from aqueous solutions.

Determination of vitamin D3 in pharmaceutical products using salting-out assisted liquid–liquid extraction coupled with reversed phase liquid chromatography

A new method, based on salting-out assisted liquid–liquid extraction (SALLE) coupled with HPLC-UV, has been introduced for the analysis of vitamin D3, cholecalciferol, in different pharmaceutical dosage forms. The developed method has been optimized in terms of chromatographic and extraction parameters. Higher yields were obtained when the SALLE was carried out using 10 mL of acetonitrile and 7.5 g of sodium chloride. The proposed method was validated in terms of selectivity, linearity, accuracy, limit of detection (LOD), limit of quantification (LOQ), robustness, and stability of vitamin D3 in solution. The method was linear with r2 > 0.999 over the range of 100–300 IU.mL−1. Recovery was within 98.80–100.43 % of the nominal vitamin D3 and the relative standard deviation values of the precision study were less than 2.0%. The LOD and LOQ were 3.17 IU.mL−1 and 9.61 IU.mL−1, respectively. Vitamin D3 in different commercial pharmaceutical formulations was analyzed using the developed method and the results were compared to those obtained using the pharmacopeial USP method. Low deviations (0.2–5.9%) between the two methods were obtained, suggesting that the developed method could replace the well-known USP method. The SALLE method proposed in this study offers a simpler and more efficient approach, requires smaller volumes of less toxic organic solvents and significantly reduces the analysis time needed to determine vitamin D3. Consequently, the SALLE method can be a promising alternative to the conventional pharmacopeial method used for vitamin D3 determination in dosage forms as it resolves most of the drawbacks associated with the pharmacopeial method.

Monitoring of hydroxylated polycyclic aromatic hydrocarbons in human tissues: Targeted and untargeted approaches using liquid chromatography-high resolution mass spectrometry.

Hydroxylated polycyclic aromatic hydrocarbons are metabolites of persistent organic pollutants which are formed during the bioactivation process of biological matrices and whose toxicity is being studied. The aim of this work was the development of a novel analytical method for the determination of these metabolites in human tissues, known to have bioaccumulated their parent compounds. Samples were treated by salting-out assisted liquid-liquid extraction and the extracts were analyzed by ultra-high performance liquid chromatography coupled to mass spectrometry with a hybrid quadrupole-time-of-flight analyzer. The proposed method achieved limits of detection in the 0.15-9.0ng/g range for the five target analytes (1-hydroxynaphthalene, 1-hydroxypyrene, 2-hydroxynaphthalene, 7-hydroxybenzo[a]pyrene, and 9-hydroxyphenanthrene). The quantification was achieved by matrix-matched calibration using 2,2´-biphenol as internal standard. For all compounds, relative standard deviation, calculated for six successive analyses, was below 12.1%, demonstrating good precision for the developed method. None of the target compounds was detected in the 34 studied samples. Moreover, an untargeted approach was applied to study the presence of other metabolites in the samples, as well as their conjugated forms and related compounds. For this objective, a homemade mass spectrometry database covering 81 compounds was created and none of them was detected in the samples.

Evaluation of Environmentally Relevant Nitrated and Oxygenated Polycyclic Aromatic Hydrocarbons in Honey.

In this work, a novel analytical methodology for the extraction and determination of polycyclic aromatic hydrocarbon derivatives, nitrated (NPAH) and oxygenated (OPAH), in bee honey samples was developed. The extraction approach resulted in being straightforward, sustainable, and low-cost. It was based on a salting-out assisted liquid-liquid extraction followed by liquid chromatography-tandem mass spectrometry determination (SALLE-UHPLC-(+)APCI-MS/MS). The following figures of merit were obtained, linearity between 0.8 and 500 ng g-1 for NPAH and between 0.1 and 750 ng g-1 for OPAH compounds, coefficients of determination (r2) from 0.97 to 0.99. Limits of detection (LOD) were from 0.26 to 7.42 ng g-1 for NPAH compounds and from 0.04 to 9.77 ng g-1 for OPAH compounds. Recoveries ranged from 90.6% to 100.1%, and relative standard deviations (RSD) were lower than 8.9%. The green assessment of the method was calculated. Thus, the Green Certificate allowed a classification of 87 points. This methodology was reliable and suitable for application in honey samples. The results demonstrated that the levels of nitro- and oxy-PAHs were higher than those reported for unsubstituted PAHs. In this sense, the production chain sometimes transforms foods as direct carriers of contaminants to consumers, representing a concern and demonstrating the need for routine control.