Determination of zolpidem in human plasma by liquid chromatography-tandem mass spectrometry for clinical application.
Zolpidem (ZPD) is widely described for the short-term treatment of insomnia. We have developed and validated a simple and rapid liquid chromatography analytical method using tandem mass spectrometry (LC-MS/MS) for the quantification of ZPD in human plasma. Using dibucaine as an internal standard (IS), the analyte was extracted with methyl t-butyl ether (MTBE). Chromatographic separation of ZPD was performed on a reversed-phase Luna C18 column (50 mm × 2.0 mm i.d., 5 μm particles) with a mobile phase of 10mM ammonium formate buffer (pH 3.0)-methanol (15:85, v/v) at a flow rate of 250 μm/min. The total run-time was 2.5 min and the retention times of ZPD and IS were 0.66 and 0.74 min, respectively. The mass-to-charge transition monitored for quantification of ZPD and IS was 308.2→235.2 and 344.0→271.0, respectively. The lower limit of quantification (LLOQ) using 100 μL of human plasma was 0.05 ng/mL and the calibration curves were linear over a range of 0.05-200 ng/mL (r(2)>0.9964). The mean accuracy and precision for intra- and inter-run validation of ZPD were within acceptable limits. In the present LC-MS/MS method, we showed improved sensitivity for quantification of the ZPD in human plasma using lower volume of plasma compared with previously described analytical methods for ZPD. This validated method was successfully applied to a pharmacokinetic study in humans.
School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Ji-Yeong
JY
Byeon
School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Hye-In
HI
Lee
School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea; College of Pharmacy, Dankook University, Cheonan 330-714, Republic of Korea.
Yun-Jeong
YJ
Lee
School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Jung-Eun
JE
Lee
School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Se-Hyung
SH
Kim
School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Young-Hoon
YH
Kim
National Institute of Food and Drug Safety Evaluation, MFDS, Cheongwon-gun, 363-951, Republic of Korea.
Han-Sung
HS
Na
School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Choon-Gon
CG
Jang
School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea. Electronic address: sylee@skku.ac.kr.
Seok-Yong
SY
Lee
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D016014:Linear Models; D008297:Male; D011725:Pyridines / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D000077334:Zolpidem
D011725:Pyridines; D000077334:Zolpidem
Human plasma; LC–MS/MS; Pharmacokinetics; Zolpidem