Sensitive and rapid determination of amantadine without derivatization in human plasma by LC–MS/MS for a bioequivalence study

Abstract

A highly sensitive, rapid and rugged liquid chromatography-tandem mass spectrometry (LC-ESI-MS/MS) method was developed for reliable estimation of amantadine (AMD), an antiviral drug in human plasma. The analyte and internal standard (IS), amantadine-d6 (AMD-d6), were extracted from 200 µL plasma by solid phase extraction on Phenomenex Strata-X-C 33 µ cartridges. Chromatography was performed on Synergi™ Hydro-RP C18 (150 mm × 4.6 mm, 4 µm) analytical column using a mixture of acetonitrile and 10 mM ammonium formate, pH 3.0 (80:20, v/v) as the mobile phase. Detection and quantitation was done by multiple reaction monitoring in the positive ionization mode for AMD (m/z 152.1 → 135.1) and IS (m/z 158.0 → 141.1) on a triple quadrupole mass spectrometer. The assay was linear in the concentration range of 0.50–500 ng/mL with correlation coefficient (r2) ≥ 0.9969. The limit of detection of the method was 0.18 ng/mL. The intra-batch and inter-batch precisions were ≤ 5.42% and the accuracy varied from 98.47% to 105.72%. The extraction recovery of amantadine was precise and quantitative in the range of 97.89%–100.28%. IS-normalized matrix factors for amantadine varied from 0.981 to 1.012. The stability of AMD in whole blood and plasma was evaluated under different conditions. The developed method was successfully applied for a bioequivalence study with 100 mg of AMD in 32 healthy volunteers. The reproducibility of the assay was determined by reanalysis of 134 subject samples.

Covalent cucurbit[7]uril–dye conjugates for sensing in aqueous saline media and biofluids

“…for point-of-care use) that enables the routine quantication of the amantadine concentration in noninvasively available biouids such as urine and saliva, may enable individual drug-dose adjustments and could contribute to improved life quality. To date, the most widely used detection methods for amantadine are chromatographic methods such as LC/GC/HPLC-MS, [83][84][85] near-infrared spectroscopy, 86 piezoelectric immunosensor, 87 and electrochemical techniques, e.g., potentiometry. 88,89 Each of these methods has its strengths, however, all of them are largely constrained by complicating and long-lasting operating procedures and expensive equipment.…”

Comparison of LC-MS3 and LC-MRM Methods for Quantifying Amantadine and Its Application in Therapeutic Amantadine Monitoring in Human Plasma

“…Currently, there are several TDM analytical methods for AMT, including immunoassays, which have been used for a long time [ 11 , 12 ], high-performance liquid chromatography with fluorescence detection (HPLC-Flu) [ 13 , 14 ], high-performance liquid chromatography combined with ultraviolet detection (HPLC-UV) [ 15 ], and liquid chromatography combined with mass spectrometry (LC-MS/MS) [ 16 , 17 ]. AMT has no particular UV absorption and fluorescence characteristics; thus, it requires derivatization by HPLC-UV and HPLC-Flu to enhance its sensitivity.…”

Development and validation of simple simultaneous analysis for amlodipine and glibenclamide by nonderivatization high-performance liquid chromatography-fluorescence

“…[12] The analytical method using HPLC without derivatization is also capable of producing a low detection limit value of 50 pg/ml, 0.18 ng/ml, and 0.4 ng/ml. [131415]…”