Briefly, plasma aliquots (500 l) were stirred and pinoline harmine and soma centrifuged (650 RCF, 10 minutes, 4 ° C) after pinoline harmine and soma adding 80 l of internal standard (3.4 mcg / ml of chloroquine) and 5 ml methyl t-butyl-ether. After centrifugation, the analytes were extracted by acidification with 1.2 ml of hydrochloric acid at 5% and back-extracted with 1.2 ml of 1 N sodium hydroxide and 5 ml of methyl t-butyl ether . The supernatants were evaporated to dryness with nitrogen gas (37 ° C) and reconstituted in 250 l of mobile phase and 200 aliquots were injected into the HPLC (Hitachi High Technologies, San Jose, CA) . The conditions of high-performance liquid chromatography were identical to those published previously, 3 with the only difference is that our composition of the mobile phase was 32/68 (v / v) acetonitrile/50 mM potassium phosphate buffer containing 0, 2% triethylamine (pH 4.8). In our conditions, the retention times of chloroquine and OSI-774 was 2 and 18 minutes respectively. erlotinib concentrations were calculated using a standard curve (range 15.3 to 4103.3 ng / mL). Samples with concentrations that were away pinoline harmine and soma from the standard curve were re-analyzed by dilution of plasma prior to extraction blank, and the final concentration values were determined after the application of the dilution factor. intra-assay reproducibility (CV% = 1.1 to 10.0) and accuracy (range pinoline harmine and soma of 94.6 to 107.3) was determined by performing three of the seven measures same levels in the same day. inter-assay reproducibility (CV% = 4.5 to 11.2) and accuracy (range 96.3 to 104.5) was determined by testing the same seven standards in triplicate on three days of Statistics and data analysis software NONMEM (version V4;. GloboMax LLC, Hanover, MD) was used for pharmacokinetic analysis.Characteristics of the dose, the concentration of erlotinib and the patient responds the same time.
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