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The product was then extracted with DCM (3x20ml), dried over MgSO 4 and concentrated.
The upper layer (organic) was dried over MgSO 4 , and the products were obtained by evaporating solvents.
N medium was composed of 0.2 g K 2 HPO 4 , 0.8 g KH 2 PO 4 , 0.2 g MgSO 4 , 0.1 g CaSO 4 *H 2 O, 0.0033 g NaMoO 4 , and 0.005 g FeSO 4 *H 2 O per liter of distilled water, with 1 000 mg/L nicotine.
In addition, NaCl, KCl, Na 2 SO 4 , ZnSO 4 , MgSO 4 , ZnCl 2 , ammonium acetate, Na 2 HPO 4 , n-butyl alcohol, MIBC, CH 2 Cl 2, CCl 4 , cyclohexane and acetone were prepared immediately before each experiment.
Usually, inorganic salt such as ZnSO 4 , MgSO 4 and ZnCl 2 was applied for protein precipitation.10 ml milk mixed with 12.6 mg precipitants (e.g.