Eluent

column was clamped in a vertical potion to the stand. 8mL of eluent was added to the column and the silica gel was mixed with the remainder of the eluent. The mixture was carefully poured into the column. More eluent was added to the beaker until all the silica could be poured into the column. The eluent was drained and a 1cm layer of sand was added to the top of the silica. More eluent was added and drained until it was level with the top of the sand.

Extraction of pigments
3-4 baby spinach leaves were gathered and placed into a mortar with a small amount of sand. 8mL of methanol was measured out and added to the mortar. A stemmed funnel lined with a filter was clamped and placed into a 125mL separatory funnel (with stopcock closed). The spinach was crushed with a pestle until it turned a dark green
1mL of the concentrated solution was saved for TLC. The stopcock was opened and the eluent was continuously added to the top of the column until the pigment separated into three separately coloured layers. The eluent was collected at the bottom and reused to flush the column. When the first band arrived at the bottom of the column, a clean and labelled 20mL beaker collected the pigment. This was repeated for the two other layers. Once all layers were collected in 20mL beakers, they were placed on a hot plate on high for 2 minutes to concentrate.

Thin Layer Chromatography
Three TLC plates were prepared and marked with C, Y, B and G. Each of the concentrated pigments was dabbed onto their respective rows on each of the three TLC plates using a plastic pipet tip. 3 TLC chambers were prepared using 7mL of each different mobile phase (9:1, 8:2 and 7:3 pet ether: acetone solution). The TLC’s were performed and Rf values were measured.

All materials were disposed of properly and all glassware was cleaned following correct laboratory