field (2,4): 1. sun dried; 2. dried in an herbarium oven at 10S°F; 
3. soaked in water: 4. soaked in 95% ethanol; 5. soaked in white 
rum (86 proof, ca. 44% ethanol); 6. soaked in an FAA mixture 
(ethanol 90%: glacial acetic acid 5%: formalin 5%): 7. soaked in 
formalin (100% formalin, ca. 40% formaldehyde) and water ina 
1:1 mixture; 8. soaked in formalin alone. 
The material for treatments 3-8 was stored in the dark for 
four weeks at room temperature, and the leaves were then 
removed from the solutions and air dried. The solutions were 
kept in a refrigerator. 
Two samples of dried leaves from each of the eight treat- 
ments described above were finely macerated, extracted with 
boiling 80% methanol, and the solution concentrated and made 
up to a final volume of 10 ml. The ultra-violet spectrum (220- 
400 nm) of the methanol extract (suitably diluted) was deter- 
mined and the amount of phenolic compounds calculated, 
using caffeic acid (a cinnamic acid) and rutin (quercetin 3-0- 
rhamnoglucoside, a flavonol) as standards. Identical amounts 
of the methanol extracts were also examined by 2-dimensional 
chromatography, using the solvent systems n-butanol-acetic 
acid-water 6:1:2 (BAW) followed by 5% aqueous acetic acid 
(HOAc). 
The solutions in which the leaves had been soaked were 
concentrated to a standard volume and subjected to 2-dimen- 
sional chromatography by the procedure described above. 
The amount of phenolic compounds remaining in the leaf 
material following these various treatments is shown in Table 1. 
TABLE | 
Treatment phenols* mgm/gm dry wt. 
1. Sun dried 12.5 
2. Herbarium dried 11.5 
3. Water 3.0 
4. 95% Ethanol 2.4 
5. White rum 2.4 
6. FAA 0.9 
7. Formalin-water 1:1 1.8 
8. Formalin 1.6 
“as caffeic acid 
259 
