A collaborative study regarding analysis of minor alkaloids in tobacco was initiated by TSRC, and the results were reported in 2005. In this study, a mixed standard solution of 4 minor alkaloids (nornicotine, anabasine, anatabine, and myosmine) was used. When we verified the collaborative study, it was clear that the level of recovery of myosmine (Laboratory Fortified Blanks: LFB) was extremely low (around 30%). The preparation of the standard solution should be modified for the accurate quantification of myosmine.

When we performed individual analyses of the minor alkaloid standard reagents, we found that myosmine was also contained in the nornicotine standard reagent we used. We tested nornicotine standard reagents from several different manufacturers and found that they all contained myosmine.

When preparing the standard solutions above, using the improved TSRC collaborative method (ITCM), the recovery rate of nornicotine, anabasine, anatabine and myosmine in LFB were 100.3%, 100.2%, 96.8% and 103.4%, respectively; 84.8%, 99.3%, 95.7% and 105.3%, respectively, in the Laboratory Fortified Matrix (LFM) of burley; and 82.7%, 99.2%, 95.0% and 106.5%, respectively, in the LFM of flue-cured.