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using the exact procedure as in the study of 30 g of repeated until there was no fragrance left in the buf-
animal fat. falo the fat. The combined ethanol filtrate was then
The percentage of purified fat from the three evaporated using a rotary evaporator. The concen-
samples from each extraction period was calculated trated extract was then centrifuged and the upper
from day 1 to day 30 on the same basis as that of the layer was kept for the analysis of its chemical compo-
previous study. sition.
The duration that resulted in the highest
Result and Discussion
average percentage of extracted fat would chosen as
the optimum extracting period. All extracted fat
Preliminary extraction study using
samples were soaked with 95 percent ethanol for pre-
dichloromethane
servative purposes and kept for further study.
4. Essential oil extraction by enfleurage method According to the study, the extracted pig fat
using purified animal fat appeared as a liquid in the dichloromethane layer
The optimum animal fat, the extraction ratio and (Figure 2A). After dichloromethane was completely
extraction duration determined by the previous study evaporated, extracted pig fat appeared as a white
was used to prepare extracted fat for the enfleurage viscous semi-solid (Figure 2B). Extracted bovine fat
method. Purified fat (100 g) was weighed and smeared appeared as a white solid stain interspersed with
onto two glass bowls (20 centimeters in diameter); yellow oil droplets (Figure 3A) and a yellow-white solid
100 g of fresh Michelia alba flower was put into the with intermittent yellow droplets (Figure 3B) after the
first bowl and then another bowl was overturned to solvent was removed. Extracted buffalo fat appeared
serve as a lid covering the first bowel. The enfleurage as a thick, white homogenous semi-solid before the
bowl set was left overnight. On the following morn- solvent was evaporated (Figure 4A) and a white ho-
ing, a new set of 100 g of fresh M. alba flowers re- mogenous solid with wax-like texture after the sol-
placed the old one and the same process was re- vent was evaporated (Figure 4B). The details of the
peated for 14 consecutive days. At the end of extrac- three extracted fats are provided in Table 1.
tion process, the buffalo fat saturated with fragrance According to the overall physical properties of
was then scraped out and transferred to a clean bottle; extracted fat, buffalo fat was the most satisfactory
100 ml of cooled ethanol was added to the bottle, left one because of its texture and solidity. Buffalo fat
overnight, and then filtered to separate the ethanol was therefore selected for further studies regarding
extract from the fat. The aforementioned method was the optimum extraction ratio and optimum extraction

A B

Fig. 2A Extracted pig fat in dichloromethane layer
2B Purified pig fat