Daptomycin (DPT) is an important antimicrobial agent used in clinical practice because it is very active against several types of medicinally challenging Gram-positive bacteria, such as methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococci. In addition to concerns about the quality of the analytical methods used in the QC of drugs, there is also concern about the impact of these methods on the environment. The trend toward sustainable consumption is increasingly evident and has forced the pharmaceutical industry to reduce the generation of toxic waste. In this context, IR spectrophotometry stands out because it does not use organic solvents and, although it is formally accepted for the identification of individual compounds, also allows the quantification of substances. Therefore, the aim of this work was to develop and validate a green analytical method for the analysis of DPT in a lyophilized powder for injection by FTIR spectrophotometry. The method involved absorbance measurements in the spectral region of 1700-1600 cm-1. The method was properly validated and found to be linear, precise, accurate, selective, and robust for the concentration range between 0.2 and 0.6 mg/150 mg. The validated method was able to quantify DPT powder for injection and can be used as an environmentally friendly alternative for routine analysis in QC.