HPLC-MS-MS Method Development and Validation of Antileishmanial Agent, S010-0269, in Hamster Serum.
A rapid, sensitive and simple high-performance liquid chromatography-tandem mass spectrometry method was developed and validated for the quantification of the antileishmanial agent, S010-0269, in hamster serum. A Discovery HS C-18 column (5 μm, 50 × 4.6 mm) maintained at 40°C was utilized for chromatographic separation with mobile phase [acetonitrile: aqueous ammonium acetate (0.01 M) buffer (85:15, v/v)] at a flow rate of 0.6 mL/min. The method requires low serum volume (20 µL) with a run time of 3.5 min. Excellent linear relationships (r ≥ 0.99) were obtained between the measured and added concentration over a range of 1-200 ng/mL. Validation parameters (accuracy, specificity, precision, recovery, matrix effect and stability) were assessed as per FDA guidelines. The precision and accuracy were acceptable as indicated by relative standard deviation ranging from 2.3 to 13.6% and bias values ranging from 1.5 to 6.5%, respectively. Moreover, the compound was found stable in hamster serum even after 30 days of storage at -80°C and being subjected to two freeze-thaw cycles. The validated method was successfully applied to the pharmacokinetic study after 10 mg/kg oral dose of S010-0269 in hamsters.
Pharmacokinetics & Metabolism Division, CSIR-Central Drug Research Institute, Jankipuram Extension, Sitapur Road, Lucknow 226 031, India Academy of Scientific and Innovative Research, New Delhi, India.
Abhisheak
A
Sharma
Pharmacokinetics & Metabolism Division, CSIR-Central Drug Research Institute, Jankipuram Extension, Sitapur Road, Lucknow 226 031, India Academy of Scientific and Innovative Research, New Delhi, India.
Swati
S
Jaiswal
Pharmacokinetics & Metabolism Division, CSIR-Central Drug Research Institute, Jankipuram Extension, Sitapur Road, Lucknow 226 031, India Academy of Scientific and Innovative Research, New Delhi, India.
Mahendra
M
Shukla
Medicinal & Process Chemistry Division, CSIR-Central Drug Research Institute, Lucknow 226 031, India.
Moni
M
Sharma
Academy of Scientific and Innovative Research, New Delhi, India Medicinal & Process Chemistry Division, CSIR-Central Drug Research Institute, Lucknow 226 031, India.
Prem Man Singh
PM
Chauhan
Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research (NIPER), Raebareli 229 010, India.
Nagarjun
N
Rangaraj
Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research (NIPER), Raebareli 229 010, India.
Kalpesh
K
Vaghasiya
Pharmacokinetics & Metabolism Division, CSIR-Central Drug Research Institute, Jankipuram Extension, Sitapur Road, Lucknow 226 031, India Academy of Scientific and Innovative Research, New Delhi, India j_lal@cdri.res.in.
Jawahar
J
Lal
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006224:Cricetinae; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D008647:Mesocricetus; D052999:Quinazolinones / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014344:Trypanocidal Agents / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics
D052999:Quinazolinones; D014344:Trypanocidal Agents