Background: Poisoning cases are among the most frequent medico-legal emergencies presenting to hospitals. In addition to medical management, healthcare professionals must fulfill crucial legal responsibilities including proper documentation, sample preservation, and communication with law enforcement. Failure in these aspects can compromise judicial proceedings. This study aimed to assess the knowledge, attitude, and practices (KAP) of healthcare professionals regarding medicolegal handling of poisoning cases in tertiary care centres of Madhya Pradesh. Methods: A cross-sectional, questionnaire-based KAP study was designed using a validated tool approved by the Institutional Ethics Committee, NSCB Medical College, Jabalpur. 250 healthcare professionals (interns, postgraduate residents, medical officers, consultants, and nursing staff) were included in the survey. The instrument comprised 20 knowledge multiple-choice questions, 10 attitude statements on a five-point Likert scale, and 10 practice items. Descriptive statistics and group comparisons were performed by designation. Results: The mean age of participants was 35.3 ± 9.8 years; most interns were aged 25–26 years. Males comprised 55%. Designations were interns (40%), PG residents (30%), medical officers (15%), consultants (10%), and nursing staff (5%). Mean knowledge score was 13.8 ± 3.2 (out of 20). Consultants and medical officers scored higher (16.5 ± 2.1 and 15.1 ± 2.7) than interns (12.6 ± 3.1) and nurses (10.4 ± 3.4). Attitude scores reflected strong agreement that medicolegal documentation improves transparency and that training should be mandatory (overall mean = 4.02 ± 0.48). Practice indicators showed that 72% had handled poisoning cases personally, 68% routinely intimated police, and 60% consistently preserved biological samples before treatment. Conclusion: The study demonstrates good overall attitude but only moderate knowledge and inconsistent practices among healthcare professionals, particularly interns and nurses. Structured medicolegal training, clear hospital protocols, and reinforcement of the chain of custody and documentation procedures are essential to improve the quality of medicolegal management of poisoning cases.