PJB-2018-1808
Qualitative and quantitative analysis of ergot alkaloids produced by Aspergillus niger through surface culture fermentation process
Memuna Ghafoor Shahid, Shahjahan Baig, Muhammad Saleem, Rabia Arif, Gul Zareen Ghafoor and Ayesha Liaqat
Abstract
The present study deals with the qualitative and quantitative analysis of ergot alkaloids produced by Aspergillus niger using surface culture fermentation technique. Two different fermentation media (A and B) were prepared for the production of extracellular and intracellular extracts of Aspergillus niger. The fermentation media A and B were further modified as A1, A2, A3, A4, B1, B2, B3 and B4 respectively. After maintaining the pH of the media at 5.0 using 0.1N HCl and 0.1N NaOH, these flasks were incubated at 25ºC for 10 days. After 10 days of incubation, the ergot alkaloids determination was done using spectrophotometry and thin layer chromatography (TLC) techniques. Maximum ergot alkaloids yield was obtained from extracellular and intracellular extracts of fermentation medium A4 (3.04 0.1 mg/ml) and A2 (0.93 0.01 mg/ml) respectively. Likewise, maximum ergot alkaloids yield was achieved from extracellular (2.45 0.03 mg/ml) and intracellular (1.33 0.01 mg/ml) extracts of fermentation medium B3. For thin layer chromatographic studies, various mobile phases were screened and TLC plates of mobile phase G (chloroform 140: propanediol 55: water 15) revealed Van Urk reagent positive spots that produced remarkable pinkish purple color on silica gel plates. Maximum Rf value was measured from extracellular extracts of sample A3EE (Rf= 0.95) and intracellular extract of B3EE (Rf = 1.21) in mobile phase G which confirmed the presence of ergocryptine and ergotamine in the samples. After modification in the mobile phase G, maximum Rfvalue was obtained from extracellular extract of sample A3EE (Rf= 1.32) intracellular extract of B4ME (Rf =1.12) that also indicated the presence of ergocryptine and ergotamine in the extracts of Aspergillus niger
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