This study evaluated different carbon and nitrogen sources on the growth and production of carbohydrates, protein, lipids, and chlorophyll of Spirulina platensis LEB-52 through an easy successive methodology under aqueous conditions. Spirulina platensis was cultivated at 120 rpm and light intensity of 156 µmol m−2 s−1 in a 500 mL Erlenmeyer flask with a working volume of 250 mL, using Zarrouk's medium. The biomass, carbohydrate, and protein production together with the specific growth rate did not show a significant difference between NaHCO3 and Na2CO3. The salts of urea and ammonium are not an alternative nitrogen sources of low cost for Spirulina platensis cultivation. From the experimental results obtained in this study, a successful estimate of carbohydrate, protein, lipids, and chlorophyll content inside Spirulina platensis was achieved without use advanced analytical techniques, allowing saves resources and time. This method can be extrapolated to other microorganisms and cultivation regimens.