Dioscorea spp. tubers are known commonly as camote de cerro to the state of Jalisco, México. Besides its use as food, it contains diosgenin, a raw material used widely for synthesising steroid hormones. The purpose of the present research was to determine diosgenin contents by gas chromatography-mass spectrometry (GC-MS), as well as physical and chemical characteristics of sixty Dioscorea spp. accessions cultivated in mesh shade coming from eleven localities in the state of Jalisco. Extraction protocol for diosgenin was maceration combined with ethanol 80%. Significant statistical differences for diosgenin content were found between accessions, finding levels between 0.02 and 0.16 mg kg−1 in dry basis. Length, weight tuber, proportion pulp/skin, percentage of pulp and percentage of skin showed significant differences. Chemical composition based in fresh weight presented in moisture percentage a range of 71.93–83.26%, crude protein 1.42–1.74%, ashes 0.80–1.16%, lipid 0.13–0.17%, crude fibre 3.80–4.02% and total carbohydrates 13.97–25.44%. Results show that tubers from Jalisco can be an important source of nutrients for consumers.