of CHCl; and O,1-N HCl were added, and the organic layer was 
discarded after shaking. The aqueous layer was washed with 
CHCI and was then made alkaline (pH = 9.0) with solid Na2CO3. 
The liberated organic bases were extracted with CHCl. The dried 
extract was dissolved in a suitable amount of mixture MeOH / 
CHC} (1:1) and submitted to TLC, GC and GC-MS analyses. 
Thin-layer Chromatography (TLC) 
Alkaloid constituents were separated on Silica Gel G TLC 
ready made plates (Merck No. 5748) with methanol-ammonia 
(99:1) as solvent. Alkaloids were located with Dragendorff’s 
reagent and tryptamines with Erlich’s reagent. 
Gas Chromatography (GLC) 
Gas chromatographic analysis was performed with two 
commerical apparatus (Pye Unical Model 104 and Schimadzu 
Model GC mini-1), equipped with hydrogen flame ionization 
detection systems. 
The stationary phases used were: 
1) 3% SE—30 ultraphase (1,50 m X 2.0 mm glass tube); 
2) 3% OV—17 ultraphase (1,50 m X 2.0 mm glass tube); 
3) SE—30 capillary WCOT column (25 m X 0.2 mm glass tube). 
The columns were operated with temperature programming 
from 150° to 280° at 5°/min. rate. The injector block and the 
detector chamber were kept at 300°. For capillary column, a 
“solvent free” injection device was used (van den Berg and Cox 
1972). 
The amount of alkaloids in mg/100 g plant material and the 
percentage of each alkaloid in the alkaloid mixture was 
determined with the capillary column using DMT asa standard. 
Gas Chromatography- Mass Spectrometry (GC-MS) 
The principles of the technique have been described earlier 
(Holmstedt and Lindgren 1967). The mass spectrometry work 
was carried out with an LKB 2091 gas chromatograph-mass 
spectrometer. The ion source was at 270°, the electron energy 
was 70 e V and the electron ionization current 50 yA, 
respectively. The separations were made on columns consisting 
of 3% SE-30 and 3% OV-17 (2 m X 2.0 mm glass tube) with 
temperature programming. 
220 
