[Objectives] To establish a method for the determination of heavy metals and pesticide residues in Dendrobium officinale produced in Guangxi and its culture substrate. [Methods] The contents of five heavy metals(Pb, Cd, As, Hg and Cu) were determined by the atomic absorption spectrophotometric(AAS) method; residues of 9 organochlorine pesticides(α-BHC, β-BHC, γ-BHC, δ-BHC, p,p′-DDT, p,p′-DDE, p,p′-DDD, o,p′-DDT and pentachloronitrobenzene) were determined by gas chromatography(GC). [Results] The contents of five heavy metals in the medicinal materials of D. officinale produced in Guangxi were 0.96, 0.21, 0.021 mg/kg, and not detected, and 0.35 mg/kg. The contents of five heavy metals in the culture substrate were 0.03, 0.19, 0.21, 0.004 and 1.80 mg/kg, respectively; the content of 9 organochlorine pesticide residues in the sample was not detected. [Conclusions] The detection results of heavy metal content and organochlorine pesticide residues in D. officinale produced in Guangxi and its substrate are consistent with the limit requirements of the Chinese Pharmacopoeia. This experiment is expected to provide reliable data reference for ensuring the safety of the drug resources of D. officinale in Guangxi.