Abstract: The physico-chemical and microbiological quality of honey samples obtained from the tropical rainforest agro-ecological zone of Nigeria were investigated. There is paucity of information on the biochemical characteristics of honey produced from the rain forest zone of Nigeria; hence, the need to provide a database on the characterization of honey produced in this zone. A total of eighteen honey samples collected from this zone were analyzed for their chemical properties including moisture, ash, pH, glucose and fructose content, while the microbial population including total plate count (TPC), total coliform and the levels of spore-forming bacteria and fungi were determined. Data obtained were analyzed statistically by Analysis of Variance (ANOVA), while sample means were separated with the aid of Least Significant Difference (LSD) at P ≤ 0.05 using Statistical Analysis System (SAS) package. The mean moisture content values ranged from 12.32 – 34.02 %, while the mean ash content ranged from 0.74 – 1.22 g 100 g-1. The mean pH value of 2.6 obtained from honey samples collected in Akwa Ibom was significantly very low when compared with that of 4.3 and 3.2 obtained from Edo and Bayelsa States honey samples respectively. Glucose and fructose mean values detected in honey samples also showed significant values. Four mineral elements: Potassium (K), Magnesium (Mg), Calcium (Ca) and Iron (Fe) were detected, but with levels varying from one place to another. Potassium (K) was the most abundant element with a mean value of 8.06 ppm obtained from Akwa Ibom honey samples. Results of microbiological characteristics showed that microbial profile were low for all microorganisms detected in honey samples produced from the tropical rainforest agro-ecological zone of Nigeria.