Salting‐out assisted liquid–liquid extraction method optimized by design of experiments for the simultaneous high‐performance liquid chromatography analysis of perampanel and stiripentol in mouse matrices

Hemp‐based materials have gained interest as alternative feed ingredients for livestock. However, safety concerns arise regarding the transfer of cannabinoids from the plant to the animals. Addressing these concerns requires the use of methods capable of detecting and quantifying cannabinoids in livestock. In this study, a fast and sensitive method was developed for quantification of cannabinoids and cannabinoid metabolites in cattle plasma using liquid chromatography‐tandem mass spectrometry (LC‐MS/MS). The extraction of cannabinoids from the plasma matrix was achieved by combining the Captiva Enhanced Matrix Removal‐Lipid clean‐up and salting‐out assisted liquid–liquid extraction procedure. The developed method underwent validation using various analytical parameters, and the results demonstrated good accuracy, precision, specificity, and high sensitivity. The method was applied to real plasma samples obtained from cattle fed hemp for 2 weeks, and successfully detected various cannabinoids, including delta‐9‐tetrahydrocannabinol. Furthermore, the study revealed that 7‐carboxy cannabidiol, a metabolite of cannabidiol, was the predominant cannabinoid present in the cattle plasma throughout the feeding period, which could remain detectable for weeks after the hemp feeding had ended.

Section: Resultsmentioning

confidence: 99%

Fast and sensitive LC‐MS/MS method for quantification of cannabinoids and their metabolites in plasma of cattle fed hemp

Xu,

Murphy

2023

“…Contrary to what was expected, only a high concentration of the stock solution of DF was degraded at 30 days, and it may be due to the high amount of DF in the solution. Stability results of QCs of DF have shown stability for 24 h according to European Medicine Agency and FDA [1–31] guidelines, which only recommends storing DF for 24 h without freezing. Freeze‐thaw stability (three cycles) in human plasma [16] has shown stability at –80°C.…”

Section: Resultsmentioning

confidence: 99%

“…Based on our experience in fluoroquinolone quantification, different preliminary experiments were carried out to obtain the best recovery. As DF is a neutral molecule at pH = 5.2, if pH is not correct, it would be impossible to extract DF using liquidliquid extraction [5,29,30]. DF was treated as other fluoroquinolone compounds adding ACN to precipitate proteins, but results were not satisfactory.…”

Section: Hplc Methods Development and Optimizationmentioning

confidence: 99%

A novel liquid chromatography‐fluorescence method for the determination of delafloxacin in human plasma

Hernandis

Escudero

Marı́n

2021

Delafloxacin is a novel fluoroquinolone antibiotic that was approved by the European Medicine Agency to treat bacterial infections of the skin and underlying tissues, and community‐acquired pneumonia. Despite being in the market since 2019 in the European Union, there is no published liquid chromatography‐fluorescence method for delafloxacin quantification in biological samples. A novel, rapid, and sensitive high‐performance liquid chromatographic method was developed to determine delafloxacin in human plasma using its native fluorescence. Plasma delafloxacin concentrations were determined by reverse‐phase chromatography with fluorescence detection at 405/450 nm of excitation/emission wavelengths. Delafloxacin was separated on a Kromasil C18 column 250 × 4.6 mm id, 5 µm using isocratic elution. The mobile phase was a mixture of 0.05% trifluoroacetic acid/acetonitrile (52/48). Retention times were 5.4 and 11.6 min for delafloxacin and valsartan (internal standard), respectively. Regression calibration curves were linear over the range of 0.1–2.5 µg/mL. The lower limit of detection was 0.05 µg/mL, and the lower limit of quantification was 0.1 µg/mL. Accuracy and precision were always <11%, and the limit of quantification was <16%. Mean recovery was 98.3%. This method can be applied to determine delafloxacin in human plasma and could be useful to perform pharmacokinetic studies.

“…LLME F I G U R E 1 Chemical structures of the studied antivirals, antidiabetics, and β-blockers can be classified into homogeneous and heterogeneous types depending, on the extractant miscibility in water. Homogeneous liquid-liquid microextraction [5,6] involves using a water miscible organic solvent, such as acetonitrile, ethanol, or acetone, that can be separated from the aqueous sample using salts (in salting-out induced LLME; SALLME) [7][8][9] or sugars (in sugaringout induced LLME; SULLME) [10][11][12][13]. Heterogeneous LLME is similar to conventional LLE in using a water immiscible organic solvent, but the amount employed is very small (30-200 µL instead of 3-10 mL).…”

Section: Introductionmentioning

confidence: 99%

Sugaring‐out induced homogeneous liquid‐liquid microextraction as an alternative mode for biological sample preparation: A comparative study

Abdallah

Hammad

Bedair

et al. 2021

Miniaturization of liquid‐liquid extraction is a growing field of sample preparation to reduce solvent consumption, protect the environment, and preserve operators’ health. In this work, four different modes of liquid‐liquid microextraction have been compared including dispersive liquid‐liquid microextraction, binary and ternary salting‐out, and sugaring‐out induced liquid‐liquid microextraction. The extraction efficiency was evaluated by the enrichment factors of 14 different drugs from three pharmacological classes. Compared with the other modes, sugaring‐out induced liquid‐liquid microextraction was found to be the most efficient and, thus, it was applied for sample preparation of the antivirals in human plasma. Method optimization was performed using response surface methodology for the sugar type and amount (in mg), the sample pH, the equilibration time (in min), and the extractant volume (in µL). The method was then validated and found linear in the concentration range of 0.10‐10 µg/mL for daclatasvir, 0.05‐10 µg/mL for velpatasvir, and 0.20‐10 µg/mL for ledipasvir, with correlation coefficients in the range 0.996‐0.999. These results shows that sugaring‐out induced liquid‐liquid microextraction could be a more efficient microextraction mode for preparation of biological samples. Compared with other types of microextraction, sugaring‐out induced liquid‐liquid microextraction is greener, simpler, and cost‐effective, with less tendency to affect the sample pH.