Determination by liquid chromatography with fluorescence detection of total 7-ethyl-10-hydroxy-camptothecin (SN-38) in beagle dog plasma after intravenous administration of liposome-based SN-38 (LE-SN38).
An HPLC- fluorescence method to quantitate total 7-ethyl-10-hydroxy-camptothecin (SN-38) in beagle dog plasma spiked with liposome based formulation of SN-38 (LE-SN38) and using camptothecin (CPT) as the internal standard (I.S.) was developed and validated to support pharmacokinetics/toxicokinetics studies. Sample preparation was done by protein precipitation using acetonitrile with 0.5% acetic acid. The supernatant was evaporated, and reconstituted in acetonitrile-20 mM ammonium acetate, pH 3.5 (20:80, v/v). When injected onto a Zorbax SB-C(18) HPLC column SN-38 as well as I.S. were detected by fluorescence using an excitation at 368 nm and emission at 515 nm. The SN-38 concentrations in samples were calculated from a standard curve of peak area ratios of SN-38 to the I.S. using weighted linear regression. The sensitivity limit for SN-38 was 1.00 ng/ml in beagle dog plasma with a precision (expressed as relative standard deviation) of 12.4% and an accuracy (expressed as analytical recovery) of 104%. The assay was linear within the standard curve range of 1-750 ng/ml. Acceptable precision and accuracy were also obtained for concentrations over the balance of the standard curve range from between-run and within-run calculations.
Guo
W
W
Pharmacokinetics, Metabolism and Bioanalytical Department, Research and Development, NeoPharm, 1850 Lakeside Drive, Waukegan, IL 60085, USA.
Ahmad
A
A
Khan
S
S
Dahhani
F
F
Wang
Y F
YF
Ahmad
I
I
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000008:administration & dosage* / Q000097:blood* / Q000493:pharmacokinetics; D002166:Camptothecin / Q000008:administration & dosage* / Q000031:analogs & derivatives* / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004285:Dogs; D004791:Enzyme Inhibitors / Q000008:administration & dosage* / Q000097:blood* / Q000493:pharmacokinetics; D000077146:Irinotecan; D008081:Liposomes; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
D000972:Antineoplastic Agents, Phytogenic; D004791:Enzyme Inhibitors; D008081:Liposomes; D000077146:Irinotecan; D002166:Camptothecin