Original research articleHPLC-UV method for simultaneous determination of MK-1775 and AZD-7762 in both acetonitrile-aqueous solution and mouse plasma

- HPLC/UV method was developed to analyze and separate MK-1775 and AZD-7762 in ACN-aqueous solution.
- HPLC/UV method was developed to analyze and separate MK-1775 and AZD-7762 in mouse plasma.
- The developed method was accurate, sensitive and precise according to the ICH guidelines.

A sensitive and precise method is described for the simultaneous determination of two small molecule kinase inhibitors: MK-1775 (MK) and AZD-7762 (AZD), in acetonitrile (ACN)-aqueous solution and in mouse plasma. A Nova-Pak C18 reversed phase column (3.9 mm × 150 mm, 4 μm, 60 Å) was utilized in the separation using an isocratic mobile phase of 0.1% v/v triethylamine in phosphate buffer (pH = 7.4): acetonitrile (ACN) (60:40, v/v), at a flow rate of 0.8 mL/min. Detection wavelength was set at 310 nm for both MK and AZD, and 431 nm for the internal standard sunitinib (SUN). The developed method was validated following the ICH guidelines and it was shown to be accurate, precise and linear in the range of 41 ng/mL to 8333 ng/mL for both drugs in the ACN-aqueous solution and from 83 ng/mL to 8333 ng/mL for both drugs in mouse plasma samples. For the first time, the presented data suggest the suitability of this method for the simultaneous separation and quantification of MK and AZD in both ACN aqueous solution as well as in mouse plasma samples.