Previous studies have suggested that growth-hormone-releasing factor (GRF) enhanced growth and advanced puberty onset along with hormonal changes in buffalo heifers (Bubalus bubalis). However, it is not known to what extent exogenous GRF could influence blood metabolites and minerals to bring about puberty in buffalo heifers. Therefore, we planned to investigate the effect of exogenous bovine GRF (bGRF) on blood metabolites and minerals in buffalo heifers during a 3-month pre-treatment period, 9-month treatment period and 1-month post-treatment period. Six buffalo heifers were treated intravenously with bGRF (10 mug per 100 kg body weight) at 15-day interval for 9 months. Another six buffalo heifers of weight- and age-matched received requisite amount of vehicle (0.9% NaCl solution) during the same period. Exogenous bGRF enhanced (p < 0.01) plasma non-esterified fatty acids (NEFA) concentrations in treatment group when compared with control group during the treatment and post-treatment period, while plasma alpha-amino nitrogen (AAN) concentrations showed a decreasing trend (p < 0.05) in the treatment group when compared with the control group during the treatment and post-treatment periods. The plasma inorganic phosphorus (Pi) was found to be higher (p < 0.05) in the treatment group animals in comparison with the levels recorded in the control group animals during the treatment as well as post-treatment periods. However, there was no change (p > 0.05) in plasma glucose and calcium concentrations between the two groups. Plasma NEFA was found to be positively correlated with plasma growth hormone (GH); however, it was only significant for the treatment group (r = + 0.76; p < 0.05). Plasma AAN in the treatment group exhibited negative correlation with plasma GH (r = 0.72; p < 0.05), while plasma AAN and GH were recorded to be positively correlated in the control group (r = 0.47; p < 0.05). The present findings suggest that exogenous bGRF induces GH release that increases plasma NEFA and Pi and decreases AAN concentrations, which probably help to reach a certain physiological state that initiates events necessary for bringing about puberty in buffalo heifers.