Abstract: Background and Objective: Jojoba is an oilseed plant with cosmetics, pharmaceuticals and other industrial applications. Once the oil is extracted, the remaining cake has shown good potential as animal feed supplement but there are certain toxicity issues with the leftover feed cake due to the presence of simmondsin. This study was aimed to develop precise, faster and efficient High Performance Liquid Chromatographic (HPLC) method for the quantification of simmondsin in different accessions of jojoba and to find out best accession with low simmondsin and high oil content among them. Materials and Methods: Seed samples were collected from Association of Rajasthan for Jojoba Plantation and Research Project (AJORP), Rajasthan, India. Oil and simmondsin were removed by repeated extraction with water at 90°C for 1.5 h from ground jojoba seeds. Quantitative analysis of simmondsin was done by HPLC method. The separation was achieved by reverse phase C18 column with a mobile phase consisting of water-methanol (80/20; v/v), at a flow rate of 0.75 mL min–1. The resultant chromatograms were analyzed by CLASS-VP V6.14 SP1 software. The correlation between concentration and peak area was calculated by Excel 2010 software. Results: A great diversity was observed between the simmondsin content of jojoba accessions. Oil yield were ranged from 39.0 to 45.0%. The simmondsin concentration varied from 2.6-4.2 g per 100 g seeds. On the basis of above results, the best accession under study was "48-25" which had nearly 44.7% oil content and a low content of 2.61 g simmondsin per 100 g seeds. Conclusion: It is concluded that planting accessions with high oil content and low simmondsin will be beneficial for both farmers and the end users of the remaining feed cake.