The analytical method of phenobarbital and mephobarbital in the pharmaceutical preparation were investigated by the partition chromatographic method. The influence of 13 kinds of excipients and various kinds of ingredients such as 8 kinds of antipyretics and 5 kinds of antacids on the partition chromatographic analysis of phenobarbital and mephobarbital in the preparation were investigated. The results were not affected by materials with the exception of caffeine, aspirin and sulpyrin.