Development and validation of a UPLC-MS/MS method for quantitation of droxidopa in human plasma: Application to a pharmacokinetic study.
In this study, a simple and sensitive ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for quantitation of droxidopa in human plasma for the first time. A simple plasma protein precipitation method using methanol containing 3% formic acid was selected, and the separation was achieved by an Acquity UPLC™ BEH Amide column (2.1mm×50mm, 1.7μm) with a gradient elution using acetonitrile, ammonium formate buffer and formic acid as mobile phase. The detection of droxidopa and benserazide (internal standard, IS) was performed using positive-ion electrospray tandem mass spectrometry via multiple reaction monitoring (MRM). The precursor-to-product ion transitions m/z 214.2→m/z 152.0 for droxidopa, and m/z 258.1→m/z 139.1 for IS were used for quantification. A lower limit of quantification of 5.00ng/mL was achieved and the linear curve range was 5.00-4000ng/mL using a weighted (1/x(2)) linear regression model. Intra-assay and inter-assay precision was less than 10.2%, and the accuracy ranged from 0.1% to 2.1%. Stability, recovery and matrix effects were within the acceptance criteria recommended by the regulatory bioanalytical guidelines. The method was successfully applied to a pharmacokinetic study of droxidopa in healthy Chinese volunteers.
Department of Pharmacy, The First People's Hospital of Lianyungang, Jiangsu, Lianyungang 222002, PR China.
Haidong
H
Wang
Department of Pharmacy, The First People's Hospital of Lianyungang, Jiangsu, Lianyungang 222002, PR China.
Guangsheng
G
Yang
Department of Pharmacy, The First People's Hospital of Lianyungang, Jiangsu, Lianyungang 222002, PR China.
Jinyu
J
Zhou
Institute of Clinical Pharmacy, Jining First People's Hospital,Jining Medical University, Jining, 272000, PR China.
Jiang
J
Pei
Department of Pharmacy, The First People's Hospital of Lianyungang, Jiangsu, Lianyungang 222002, PR China.
Qiangfeng
Q
Zhang
Department of Pharmacy, The First People's Hospital of Lianyungang, Jiangsu, Lianyungang 222002, PR China.
Xingfa
X
Song
Department of Pharmacy, The First People's Hospital of Lianyungang, Jiangsu, Lianyungang 222002, PR China. Electronic address: sunzx_715@163.com.
Zengxian
Z
Sun
D000328:Adult; D000818:Animals; D000978:Antiparkinson Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015103:Droxidopa / Q000097:blood*; D006801:Humans; D057230:Limit of Detection; D008297:Male; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
D000978:Antiparkinson Agents; D015103:Droxidopa
Droxidopa; Plasma concentrations; UPLC–MS/MS