To determine the accuracy of the light hydrocarbon parameter obtained from the headspace gas, crude oils and associated gases from six wells in the western part of the Qaidam Basin were collected. The composition and carbon isotopes of individual light hydrocarbons from headspace gas and natural gas were analyzed and compared. The results show that the content of C₅—C₇ light hydrocarbons obtained from headspace gas is higher than that in natural gas, but the relative contents of each light hydrocarbon obtained from two methods are similar. It is found that light hydrocarbon parameters calculated from headspace gas and natural gas share similar results in studying the genesis type, generation temperature and other aspects, but get different results in evaluating the maturity by using heptane and isoheptane values. Besides, the carbon isotopes of individual light hydrocarbons in headspace gas are greater than those in natural gas, among which cyclo-alkanes have the smallest carbon isotope difference. Particularly, the carbon isotopes of methylcyclohexane (δ¹³C_MCC6) in the headspace gas are close to those in natural gas. Therefore, the main parameters of light hydrocarbons and δ¹³ C_MCC6 can be accurately obtained from the headspace gas of crude oil. The research results provide a basis for enriching the application of light hydrocarbon geochemistry in oil and gas accumulation.