Anemia is prevalent in pregnant ladies. It is also common in college girls and other age groups of female. Anemia has been found to be associated with low heart rate variability (HRV) indicating increased cardiac risk. Medical students are under stress, which may affect both the hemoglobin level and HRV. Thus, we aimed to assess the hemoglobin, iron status, and HRV. A cross-sectional comparative study was conducted on 40 unmarried female medical students of age 18-30 years. They were divided into three groups based on hemoglobin (Hb) level following WHO guidelines: normal Hb group (Hb≥12g/dl), mild anemia (Hb<11.9-10.0g/dl) and moderate anemia (Hb 9.9-7.0g/dl). Their mean corpuscular volume, RBC counts and ferritin levels were measured. Short-term heart rate variability and cardio-respiratory variables were recorded in all subjects after 15 min of supine rest and compared among the groups. The relationship between HRV and blood parameters was studied using Spearman’s Rank correlation. Kruskal Wallis followed by multiple comparisons was applied to compare the variables of blood and HRV among the groups. According to Hb level, 17.5% students were moderately anemic, 50% were mild anemic and 32.5% were with normal Hb level. They were comparable in terms of their age, BMI, diastolic blood pressure and heart rate. However, systolic blood pressure [92(90.0-110.0) mmHg vs. 110(108.5-112) mmHg, p=0.022] and respiratory rate [18(16-18) per minute vs. 20(19-22) per minute p= 0.001] were lower in moderate anemic group compared to normal group. Also systolic blood pressure [92(90.0-110.0) mmHg vs. 110(102-110) mmHg, p= 0.042] was lower in moderate anemic group than mild anemic group. The Hb level, RBC count, MCV, and ferritin levels { [9.2 (9.0-9.8) g/dl vs. 11.0 (10.17-11.45) g/dl vs. 12.4(12.0-12.65) g/dl, p= 0.001], [3.7(3.5-3.7) per mm3 vs. 4.0(3.9-4.1) per mm3 vs. 4.2(4.11-4.36) per mm3, p= 0.001], [77.94(77.29-82.97) fl vs. 85(78.76-88.7) fl vs. 90.06(86.33-92.54) fl, p= 0.01] and [7.9(6.5-19.4) ng/ml vs. 18.85(15.85-38.0) ng/ml vs. 28.9(17.3-44.7) ng/ml p=0.01]} respectively were lower in moderate anemic group than mild anemic and normal groups. However, there were no significant differences in HRV parameters among the groups, and no significant correlations were obtained between HRV and blood parameters. Moderately anemic female medical students had lower systolic BP and respiratory rate compared to mildly anemic female medical students. However, there were no significant differences in HRV parameters among the groups. It indicates mild and moderate anemia may not have effect on cardiac autonomic modulation in this female age group.