A rapid and highly sensitive method for the determination of glimepiride in human plasma by liquid chromatography-electrospray ionization tandem mass spectrometry: application to a pre-clinical pharmacokinetic study.
A sensitive and specific liquid chromatography-positive electrospray ionization-tandem mass spectrometry method has been developed and validated for the determination of glimepiride (GPD) in human plasma. GPD and the internal standard (IS, glibenclamide) were extracted from a small aliquot of human plasma (200 microL) by a simple liquid-liquid extraction technique using ethyl acetate as extraction solvent. The compounds were separated on a YMC Propack, C18, 4.6x50 mm column using a mixture of ammonium acetate buffer, acetonitrile and methanol (30:60:10, v/v) as mobile phase at 0.5 mL/min on an API 4000 Sciex mass spectrometer connected to an Agilent HPLC system. Method validation and pre-clinical sample analysis was performed as per FDA guidelines and the results met the acceptance criteria. GPD and IS were detected without any interference from human plasma matrix. The method was proved to be accurate and precise at linearity range of 0.02-100.00 ng/mL with a correlation coefficient of 0.999. The method was robust with a lower limit of quantitation of 0.02 ng/mL. Intra- and inter-day accuracies for GPD were 88.60-113.50 and 96.82-103.93%, respectively. The inter-day precision was better than 12.21%. This method enabled faster and reliable determination of GPD in a pre-clinical study.
Chakradhar
Lagishetty
L
Department of Drug Metabolism and Pharmacokinetics, Discovery Research, Dr Reddy's Laboratories Ltd, Miyapur, Hyderabad-500 049, India.
Kallem
Rajareddy
R
Karthik
Arumugam
A
Sundari
Bala Tripura
BT
Ramesh
Srirangam
S
Mullangi
Ramesh
R
Srinivas
Nuggehally R
NR
D000085:Acetates / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D000284:Administration, Oral; D000818:Animals; D005591:Chemical Fractionation / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D005905:Glyburide / Q000592:standards; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007004:Hypoglycemic Agents / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D008297:Male; D000432:Methanol / Q000737:chemistry; D015394:Molecular Structure; D051381:Rats; D017208:Rats, Wistar; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*; D013453:Sulfonylurea Compounds / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods*
D000085:Acetates; D000097:Acetonitriles; D007004:Hypoglycemic Agents; D013453:Sulfonylurea Compounds; C057619:glimepiride; C007650:ethyl acetate; D005905:Glyburide; D000432:Methanol; C032159:acetonitrile