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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-
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