Separation and purification of chlorogenic acid from tobacco by-products by polyamide and silicagel column chromatography

Methods for separation of chlorogenic acid (CA) from tobacco by-products were established. The liquid chromatography tandem mass spectrometry (LC-MS) method for identification and analysis of chlorogenic acid from tobacco materials has been developed. CA was isolated by polyamide and further purified by silicagel column chromatography. Results reveal that polyamide is suitable for separation of CA from tobacco extract. After one run treatment with polyamide, the content of chlorogenic acids was 40.3%. The product was further purified using silicagel column chromatography; the content of total chlorogenic acid was increased 2.29-fold from 40.3 to 92.2%. The LC-MS results showed that total chlorogenic acids were made up of four components: 5-CQA, 3-CQA, 4-CQA and cis-5-CQA. The content of 5-CQA was the most (78.2%) and other three chlorogenic acid derivatives were 1.9, 10.1 and 2.0%, respectively. Key words : Tobacco by-products, chlorogenic acid, polyamide, silica gel column, LC-MS.