Paper Chromatography Ultraviolet Spectrophotometer

In the development of disease resistant varieties, secondary alkaloids are often a concern for flue-cured tobacco breeders. A rapid color test developed by Glock and Wright (1963) is used to quantify nornicotine in flue-cured tobacco. The method involves color development from the reaction of nornicotine and 1,3-indanedione. This test is sensitive enough to detect microgram levels of nornicotine. The method is used quantitatively with a spectrophotometer because color development is linear from 5.0 to lOOpg/ml of nornicotine reacted with 1,3-indanedione. 

A 100- to 1000-mg sample of ground air-dried flue-cured tobacco is weighed into a 125-ml Erlenmeyer flask. Ten ml of 2 N NaOH and 50 ml of chloroform are added to the flask. The flask is shaken vigorously for 30min and then 2g of analytical grade celite are added to absorb water. The extraction process is completed by filtering the tobacco extract through two layers of Whatman 40 paper. 

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New techniques and instruments, such as partition chromatography on paper strips and the photoelectric ultraviolet spectrophotometer, stimulate the development of biochemistry after World War II. New... 

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