Introduction: Candidemia is a severe bloodstream infection caused by various Candida species, prevalent in adult intensive care units (ICUs) where patients are often critically ill. Rapid and accurate identification of Candida species is crucial for effective treatment and improved patient outcomes. This study compares the efficacy of Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) and the VITEK-2 Compact system in identifying Candida species from blood cultures in a tertiary care setting in Uttarakhand, India.Methods: A prospective cross-sectional study was conducted in the Microbiology Laboratory of AIIMS Rishikesh from November 2022 to March 2024. The study enrolled ICU patients with sepsis and blood cultures positive for Candida species. The Candida isolates were identified using the VITEK-2 Compact system and MALDI-TOF MS. The results from the two methods were compared to determine concordance and discrepancies in species identification.Results: VITEK-2 and MALDI-TOF identified C. tropicalis and C. albicans as the most common species. VITEK-2 identified 19.4% (n=20) as C. albicans and 35% (n=36) as C. tropicalis, alongside species like C. auris and C. parapsilosis. MALDI-TOF results included 18.4% (n=19) for C. albicans and 34% (n=35) for C. tropicalis, additionally detecting species like C. orthopsilosis and Pichia species. A significant concordance rate of 98.1% (p=0.001) was found between the two methods. However, MALDI-TOF was more precise in identifying less common species and highlighted discrepancies, such as the misidentification of C. orthopsilosis as C. parapsilosis by VITEK-2.Conclusion: The study underscores the importance of accurate species-level identification of Candida in clinical settings. MALDI-TOF offers rapid and reliable identification, which is crucial for managing antifungal therapy and monitoring resistant species. While both methods demonstrated high concordance rates, the complementary use of these technologies, along with potential molecular confirmation, could enhance the precision of Candida species identification, aiding in better infection control and patient care.