A single dose of florfenicol (Nuflor®) was administered to eight healthy adult alpacas, at 20mg/kg IM (intramuscular) and 40mg/kg SC (subcutaneous) using a randomized, cross-over design and 28-day washout period. Subsequently, 40mg/kg florfenicol was injected SC every other day for 10 doses to evaluate long-term effects. Maximum plasma florfenicol concentrations (Cmax, measured via high-performance-liquid-chromatography) were achieved rapidly, leading to a higher Cmax of 4.31+/−3.03 μg/ml following administration of 20mg/kg IM than 40mg/kg SC (Cmax: 1.95+/−0.94 μg/ml). Multiple SC dosing at 48hr intervals achieved a Cmax of 4.48+/−1.28 μg/ml at steady state. The area under the curve and terminal elimination half-lives were 51.83+/−11.72μg/ml.h and 17.59+/−11.69 hours after single 20mg/kg IM administration, as well as 99.78+/−23.58μg/ml.h and 99.67+/−59.89 hours following 40mg/kg injection of florfenicol SC, respectively. Florfenicol decreased the following hematological parameters after repeated administration between weeks 0 and 3: total protein (6.38 vs. 5.61 g/dL, P<0.0001), globulin (2.76 vs. 2.16 g/dL, P<0.0003), albumin (3.61 vs. 3.48 g/dL, P=0.0038), white blood cell count (11.89 vs. 9.66 ×10^3/μL, P<0.044), and hematocrit (27.25 vs. 24.88%, P<0.0349). Significant clinical illness was observed in one alpaca. The lowest effective dose of florfenicol should thus be used in alpacas and limited to treatment of highly susceptible pathogens.