Rapid quantification of lamotrigine in human plasma by two LC systems connected with tandem MS.
A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS-MS) method has been developed and validated for the determination of lamotrigine in human plasma using multiplexing technique (two HPLC units connected to one MS-MS). Lamotrigine was extracted from human plasma by solid-phase extraction technique using Oasis Hydrophilic Lipophilic Balance (HLB) or N-vinylpyrrolidone and divinylbenzene cartridge. A structural analog, 3,5-diamino-6-phenyl-1,2,4-triazine, was used as an internal standard (IS). A BetaBasic C(8) column was used for the chromatographic separation of analytes. The mass transition [M+H](+) ions used for detection were m/z 256.0 --> 211.0 for lamotrigine and m/z 188.0 --> 143.0 for IS. The method involved a simple multiplexing, rapid solid-phase extraction without evaporation and reconstitution. The proposed method has been validated for a linear range of 0.025 to 10.000 microg/mL with a correlation coefficient > or = 0.9991. The limit of quantification for lamotrigine was 0.025 microg/mL, and limit of detection was 50.000 pg/mL. The intra-run and inter-run precision and accuracy were within 10.0% for intra-HPLC runs and inter-HPLC runs. The overall recoveries for lamotrigine and IS were 97.9% and 92.5%, respectively. Total MS run time was 1.4 min per sample. The validated method has been successfully used to analyze human plasma samples for applications in pharmacokinetic, bioavailability, bioequivalence, or in vitro in vivo correlation studies.
Shah
Hiten J
HJ
Bioanalytical Laboratory, Torrent Pharmaceutical Ltd., Bhat, Gandhinagar-382428, Gujarat, India. hitenshah@torrentpharma.com
Subbaiah
Gunta
G
Patel
Dasharath M
DM
Suhagia
Bhanubhai N
BN
Patel
Chhagan N
CN
D000927:Anticonvulsants / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D000077213:Lamotrigine; D057230:Limit of Detection; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014227:Triazines / Q000097:blood*
D000927:Anticonvulsants; D014227:Triazines; D000077213:Lamotrigine