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then another bowl was turned over to cover the first
one. The enfleurage bowls were left overnight. The
following morning, a new set of fresh M. alba flowers
(100 g) replaced the old ones and the same process was
repeated for 14 consecutive days. At the end of the
extraction process, the buffalo fat was saturated with
fragrance; it was then scraped out and transferred to a
clean bottle. Thereafter, 100 ml of cooled ethanol was
added to the bottle, left overnight, then filtered to sepa-
rate the ethanol extract from the fat. The aforemen-
tioned method was repeated until there was no fra-
grance left in the buffalo fat. The combined ethanol
filtrate was then evaporated using a rotary evaporator.
The concentrated extract was then centrifuged and the
Figure 1 Processed buffalo fat for enfleurage
upper layer was kept for analysis of its chemical compo-
sition.

Preparing buffalo fat for the enfleurage method
The buffalo fat was cleaned and unwanted parts Extraction of M. alba oil by steam distillation
such as blood vessel and fascia were removed. Then The basic steam distillation set was connected and
the fat was sliced into small pieces; 100 g of sliced fat 300 g of fresh M. alba flowers was extracted with 600 ml
was weighed and transferred to a glass bottle. Dichloro- distilled water for two hours. The final extract was
methane (Sigma, lab grade) (66.7 ml) was added to the cooled and centrifuged to separate the fragrant oil;
bottle to attain a fat to solvent ratio of 3:2 weight/vol- then the oil was kept for the analysis of its chemical
ume. Then the bottle was tightly closed and kept at composition.
ambient temperature for 10 days. The dichloromethane
layer containing the extracted fat was separated from Extraction of M. alba oil by hexane
the fat by filtration. The filtrate was transferred to an A total of 200 grams of fresh M. alba flowers was
aluminum tray and exposed to the sun during daytime weighed and transferred to a brown bottle; 1,000 ml of
and dried overnight in a fume hood. Extracted buffalo hexane was added, then the bottle was left for extrac-
fat appeared to be clear white (Figure 1); it was scraped tion for one month; and the sample was shaken daily.
out and kept in a clean bottle. Ethanol was added to the After one month, the extract was filtered and the hex-
extracted fat in a ratio of 1:1 w/v and the fat was kept ane (solvent) removed by rotary evaporator. The oil in
for seven days prior to use. Finally, ethanol was poured the hexane extract was extracted again by partition
out of the bottle and the extracted fat was transferred to with cooled ethanol. The ethanol layer was then col-
a 250 ml beaker and placed superficially above a water lected and quickly evaporated using a rotary evapora-
bath at 50˚C until the fat began to aggregate into semi- tor, followed by centrifugation, and the oil was kept for
solid form. Extracted fat from the final process was the analysis of its chemical composition.
kept for the enfleurage method.
Analysis of chemical composition of oil samples

Extraction of M. alba oil by the enfleurage method using gas chromatography-mass spectrometry
A total of 100 g of extracted fat from the above- (GC-MS)
mentioned method was weighed and then smeared onto Oil samples from the three extraction methods were
two glass bowls (20 centimeters in diameter); 100 g of analyzed for their chemical composition using an Agilent
fresh M. alba flowers were put into the first bowl and Technologies 6890 N GC-MS with a 5973 innert quadru-