A Rapid and Highly Sensitive UPLC-MS-MS Method for the Quantification of Zolpidem Tartrate in Human EDTA Plasma and its Application to Pharmacokinetic Study

Abstract

A rapid and high sensitive liquid chromatography-tandem mass spectrometry (LC-MS-MS) method was developed and validated for the quantification of zolpidem in human EDTA plasma using ondansetron (IS) as an internal standard. The analyte and IS were extracted from human plasma using ethyl acetate and separated on a C18 column (Inertsil-ODS, 5 µm, 4.6 × 50 mm) interfaced with a triple quadrupole tandem mass spectrometer. The mobile phase, which consisted of a mixture of methanol and 20 mM ammonium formate (pH 5.00 ± 0.05; 75:25 v/v), was injected at a flow rate of 0.40 mL/min. The retention times of zolpidem and IS were approximately 1.76 and 1.22. The LC run time was 3 min. The electrospray ionization source was operated in positive ion mode. Multiple reaction monitoring used the [M + H](+) ions m/z 308.13 → 235.21 for zolpidem and m/z 294.02 → 170.09 for the ondansetron, respectively. Five freeze-thaw cycles was established at -20 and -70°C.The linearity of the response/concentration curve was established in human EDTA plasma over the concentration range 0.10-149.83 ng/mL. The lower detection limit [(signal-to-noise (S/N) > 3] was 0.04 ng/mL and the lower limit of quantification (S/N > 10) was 0.10 ng/mL. This LC-MS-MS method was validated with intra-batch and inter-batch precision of 0.52-8.66.The intra-batch and inter-batch accuracy was 96.66-106.11. Recovery of zolpidem in human plasma was 87.00% and IS recovery was 81.60%. The primary pharmacokinetic parameters were T(max) (h) = (1.25 ± 0.725), C(max) (ng/mL) (127.80 ± 34.081), AUC(0→t), = (665.37 ± 320.982) and AUC(0→∞), 686.03 ± 342.952, respectively.

Comparative bioavailability of two zolpidem hemitartrate formulations in healthy human Brazilian volunteers using high‐performance liquid chromatography coupled to tandem mass spectrometry

“…After oral administration of the zolpidem hemitartrate 5 mg (orodispersible tablet 5 mg; sublingual tablet 5 mg) to the volunteers, the peak plasma concentrations ( C max ) of zolpidem for both formulations were different from those reported in the literature [dose 5 mg, C max 50 ng/ml (Byeon et al, ); dose 10 mg, C max 120 ng/ml (Reddy et al, ); dose 10 mg, C max 107 ng/ml (Nirogi et al, )]. At the same dose (5 mg), the C max of our study (97 ng/ml) was ~2× bigger.…”

“…Zolpidem has been determined in several biological matrices (oral fluid, hair, plasma, nails, serum, urine and blood) and by several methods: capillary electrophoresis (Hempel & Blaschke, 1996), radioimmunoassay (de Clerck & Daenens, 1997), reversed-phase chromatography with fluorescence (Ptacek, Macek, & Klima, 1997), gas chromatography (Gaillard, Gay-Montchamp, & Ollagnier, 1993;Stanke, Jourdil, Lauby, & Bessard, 1996), gas chromatography-mass spectrometry (GC-MS), HPLC with fluorimetric or UV detection (Ascalone, Flaminio, Guinebault, Thenot, & Morselli, 1992;Debailleul, Khalil, & Lheureux, 1991;Durol & Greenblatt, 1997;Guinebault, Dubruc, Hermann, & Thenot, 1986;Nirogi, Kandikere, Shrivasthava, & Mudigonda, 2006;Tracqui, Kintz, & Mangin, 1993), GC/MS (Keller, Schneider, & Tutsch-Bauer, 1999;Lichtenwalner & Tully, 1997;Wang, Sun, & Lau, 1999), LC/MS (Giroud, Augsburger, Menetrey, & Mangin, 2003), LC/MS/MS (Byeon et al, 2015;Jang et al, 2013;Jeong et al, 2015;Miyaguchi, 2013;Moretti et al, 2018), UHPLC/MS/MS (Eliassen & Kristoffersen, 2014;Marin, Roberts, Wood, & McMillin, 2012), UHPLC-Q-TOF-LC/MS (Kronstrand et al, 2018) and UPLC-MS-MS (Reddy, Bapuji, Rao, Himabindu, & Ravinder, 2012). The HPLC or UPLC coupled to tandem mass spectrometry in plasma has an LOQ of 0.1-1.8 ng/ml, RT 0.7-1.76 min, run time 2.5-4.5 min, liquid-liquid extraction, 100 μl of plasma (Byeon et al, 2015;Marin et al, 2012;Reddy et al, 2012). Considering the purpose of the study (bioequivalence), the LOQ (1 ng/ml) was adequate, since it was 1.03% of mean C max (97 ng/ml).…”

Bioequivalence of zolpidem hemitartrate: new formulations of 5 mg SL, 10 mg and 12.5 mg XR and comparison of their bioavailabilities

“…As in other published works, the analytical selected technique of the study to quantify zolpidem in human plasma was LC-MS/MS [29,37,38].…”

Development and Validation of a Sonication-Assisted Dispersive Liquid–Liquid Microextraction Procedure and an HPLC-PDA Method for Quantitative Determination of Zolpidem in Human Plasma and Its Application to Forensic Samples

“…Several analytical methods have been reported for the quantification of zolpidem in blood, including HPLC with fluorescence detection [ 2 , 10 ], HPLC-DAD [ 11 ], GC/MS [ 12 ], GC/MS/MS [ 13 ], LC/MS [ 8 , 14 ], UPLC/MS [ 15 ], LC/MS/MS [ 16 , 17 , 18 , 19 , 20 , 21 ], and UHPLC-MS/MS [ 5 , 22 , 23 ]. The most commonly used techniques for zolpidem extraction from biological matrices are liquid–liquid extraction (LLE) [ 2 , 5 , 14 , 16 , 19 , 20 , 21 ] and solid-phase extraction (SPE) [ 8 , 12 , 15 ].…”