ABSTRACT
A study of the simultaneous separation and determination of selected polyphenols (rutin, narirutin, naringin, hesperidin, neohesperidin, quercetin, naringenin, kaempferol and hesperetin) with reported effects in the treatment of depression and cardiac and neurodegenerative diseases was performed. An RP-ultrahigh-performance liquid chromatography (UHPLC)-ultraviolet method for analyte separation and determination was successfully developed and validated for a ZORBAX Eclipse Plus C18 RRHD analytical column. Separation was carried out in gradient elution mode with acetonitrile and water modified with 0.05% trifluoroacetic acid. In the selected working range, the method linearity was satisfactory, with coefficients of determination >0.99. The precision and accuracy did not exceed the acceptable limits of 15%. The method was used to compare 16 different SPE sorbents for medicinal plant sample preparation in terms of analyte recoveries and matrix purification. The analysis of real samples was carried out for Menthae piperitae (predominant analyte was rutin), Hypericum perforatum (predominant analyte was rutin), Salvia officinalis (predominant analyte was kaempferol) and their derived products, enabling a comparison of different plant materials. Additional confirmation by UPHLC coupled with tandem mass spectrometry was performed. For the chiral aglycones naringenin and hesperetin, the determination of individual enantiomers was also performed with a Chiralpak AD-3R analytical column.
Subject(s)
Plants, Medicinal/chemistry , Polyphenols/analysis , Polyphenols/isolation & purification , Chromatography, High Pressure Liquid/methods , Limit of Detection , Linear Models , Plant Extracts/chemistry , Polyphenols/chemistry , Reproducibility of Results , Tandem Mass Spectrometry/methodsABSTRACT
A quick and sensitive RP-UHPLC-ESI-MS/MS method for the separation of flavanone, naringenin and hesperetin enantiomers was developed. The separation of analytes was performed using a Chiralpak AD-3R column, and methanol was used as the mobile phase. Detection was carried out using a triple quadrupole tandem mass spectrometer with an electrospray ionisation source. Positive ionisation and multiple reaction monitoring (MRM) were used. The developed method showed satisfactory linearity with determination coefficients greater than 0.996 in the concentration ranges of 2.5-100.0ngmL(-1) for naringenin and flavanone enantiomers and 0.5-100.0ngmL(-1) for hesperetin enantiomers. The limits of quantification varied from 0.1 to 2.0ngmL(-1). The intra-day and inter-day precisions were below 15%, and the accuracy varied from -13.6% to 13.5%. The described method was successfully applied for the chiral separation and determination of flavonoid enantiomers in real samples of spices and herbal root. Due to the occurrence of the natural compounds in the forms of free aglycones and their glycosides, these samples were subjected to hydrolysis in order to obtain free aglycones from the glycosylated forms. Acid and enzymatic hydrolysis techniques were also compared. In the course of this study, the enzymatic hydrolysis technique was selected.
Subject(s)
Chemistry Techniques, Analytical/methods , Chromatography, High Pressure Liquid , Flavanones/isolation & purification , Hesperidin/isolation & purification , Tandem Mass Spectrometry , Flavanones/chemistry , Hesperidin/chemistry , Reproducibility of ResultsABSTRACT
A rapid and effective RP-UHPLC-DAD method for enantioseparation of three flavanones, i.e., flavanone, naringenin, and hesperetin, was developed and validated. Chromatographic separation of the analytes was performed using a Chiralpak AD-3R analytical column under reverse phase conditions with methanol as the mobile phase. The method was validated in the concentration range of 0.2 to 50 µg/mL for enantiomers of flavanone and 0.5 to 50 µg/mL for enantiomers of naringenin and hesperetin. The limits of quantification were between 0.03 to 0.5 µg/mL. Intraday and interday precision were below 14% and accuracy varied from 0.04 to 8.17%.
Subject(s)
Amylose/analogs & derivatives , Chromatography, High Pressure Liquid/methods , Flavanones/chemistry , Hesperidin/chemistry , Phenylcarbamates/chemistry , Amylose/chemistry , Molecular Structure , StereoisomerismABSTRACT
A procedure based on solid-phase extraction (SPE) followed by ultra-high-performance liquid chromatography (UHPLC) with UV detection has been developed for the analysis of multiple drugs in human urine. The compounds evaluated were aliskiren, prasugrel, rivaroxaban, prednisolone, propranolol, ketoprofen, nifedipine, naproxen, terbinafine, ibuprofen, diclofenac, sildenafil and acenocoumarol. Seventeen different solid phase extraction (SPE) cartridges were tested to evaluate their applicability for the isolation of drugs from human urine. Comparison were recovery of different drugs and reproducibility. The samples were analyzed by UHPLC using a Poroshell 120 EC-C18 column and acetonitrile -0.05% TFA in water as the mobile phase under gradient elution conditions. SPE combined with UHPLC-UV allowed the determination of drugs over a linear range of 0.01-30.0µg/mL, with limits of detection at 0.003-0.217µg/mL and precision of 0.8-7.1%. Phenyl (C6H5) sorbent was found to provide the most effective clean-up, removing the greatest amount of interfering substance and simultaneously ensuring analyte recoveries higher than 85.5% with relative standard deviations (RSD) <10%. The method was applied with good accuracy and precision in the determination of drugs in human urine obtained from patients treated with selected drugs.
