Combinatorial pulsed laser deposition (CPLD) is proving to be an efficient, low-cost tool in synthesis and characterization of thermoelectric composition-spread films. With the help of CPLD, (Ca1-xAx)3Co4O9 composition-spread films were fabricated successfully on TiO2 (100) substrate , and their thermoelectric (TE) properties and structure were evaluated by the multi-channel thermoelectric measurement system and concurrent X-ray analysis respectively. The largest power factor in (Ca1-xSrx)3Co4O9 film library was achieved as 1.67µW/cmK2. The influence of substitution on TE properties was investigated by doping different ions into Ca3Co4O9. By comparing Seebeck coefficients and conductivities of the films, it can be drawn that the substitution ions with smaller size are potential candidates for good TE properties. The dependence of TE properties of (Ca1-xSrx)3Co4O9 films on temperature shows that good TE properties can be achieved when temperature is above 150K.