Traditional Soxhlet extraction for measuring oil content is time－consuming and consumes a large amount of organic solvents. To solve these problems， seven typical tobacco seed samples， including flue－cured tobacco， sun－cured tobacco， burley tobacco and aromatic tobacco were selected to establish a rapid determination method based on serial petroleum ether Soxhlet extraction. This method extracts more than ten samples by placing multiple samples in different overlapping and tandem sample tubes using a single Soxhlet extractor. Subsequently， the oil content is calculated by the difference in the samples before and after extraction. The establishment of this method provides a new， rapid and accurate analytical approach for measuring oil content in tobacco seeds. This method is also applicable to oilseeds such as dragon fruit seeds， sesame seeds， rapeseeds， and perilla seeds， which indicates its good general applicability.