The separation of a penicillium fungal extract by thin layer chromatography and HPLC: A comparative study

The separation of a Penicillium fungal extract by two dimensional-multimoda l thin layer chromatography and gradient high performance liquid chromatogr aphy is reported. The fungal extract was resolved on a cyano derivatized si lica gel TLC plate using two dimensional development; first dimension in me thylene chloride: hexane:acetic acid (9:10:1), taken out, dried, turned 90 degrees, and developed in the second dimension in acetonitrile:methanol: wa ter (40:37:32). Over 20 different spots were observed under UV radiation. T hree different mobile phase gradients, acetonitrile, methanol, and tetrahyd rofuran as primary organic solvents were used. Different selectivities; resulted in each gradient. The best result was obt ained using the acetonitrile gradient, 25% to 100% acetonitrile in 30 minut es. Also, it was found that using photo diode array detection in natural pr oducts research is a must when quantitation is required, since the mixture contains compounds of different absorptivities. The results show that chang ing the wave length of detection resulted in different peak heights (areas) .