Traditional Chinese Veterinary Medicine practitioners commonly prescribe herbal formulas. These plant-based medicines are highly complex with a number of different components sensitive to variable growing environments and processing. The development and optimization of analytical methods to identify ingredients and to assure consistent levels of biologically active molecules in these plant-based formulas is critical. A high pressure liquid chromatography (HPLC)-based method for reliable, cost efficient and consistent analysis of the Chinese herbal medicine Da Huo Luo Dan used in the integrative treatment of paresis or paralysis was developed. The objective of the analysis was to evaluate the levels of the clinically useful neuroexcitatory alkaloids brucine and strychnine, compounds that are also associated with toxicity at high levels. Initial screening using a gas chromatography-mass spectrometry method (GC-MS) at an analytical laboratory was performed to identify the organic plant compounds consistent with the constituent herbs of the formula along with the alkaloids of interest. The HPLC method was then performed for quantification. The samples contained 128 mg/kg as-fed (AF) brucine and 196 mg/ kg AF strychnine in one batch lot, and 143 mg/kg AF brucine and 202 mg/kg AF strychnine in another. This is equivalent to a strychnine dose of 0.02 mg/kg body weight in dogs when the herbal formula is administered at recommended levels, and is well below the suggested toxic dose in dogs of 0.45-0.75 mg/kg body weight.