ผลและการสนทนาความมั่นคงทางกายภาพ และเคมีทำให้เปอร์เซ็นต์ของสารสกัดจากน้ำมันหอมระเหย pupae ไหมไทยพื้นเมือง (Keawsakolพันธุ์ Nangnoi ศรีสะเกษ Nangleung สำโรง และ Noneruesee) (รูปที่ 1 และ 2) โดย soxhletและวิธี maceration สกัด 24-29% และ 4-7% ตามลำดับ สกัด soxhletวิธีให้ผลผลิตสูงกว่าที่ได้รับจากวิธีการสกัด maceration (p0.05)รูปที่ 1 Keawsakol, Nangnoi Si Sa Ket สำโรง Nangleung และน้ำมันวิธี Maceration pupae Nonerueseeรูปที่ 2 Keawsakol, Nangnoi Si Sa Ket สำโรง Nangleung และน้ำมันวิธี Soxhlet pupae NonerueseeA1 A2 E1 E2เรื่องของกรดไขมันกรดไขมันถูกแยกออกจากน้ำมัน pupae ไหม โดยสกัด soxhlet และ macerationวิธี องค์ประกอบกรดไขมันของน้ำมันในรูป 3A, 3B, C 3, 3D และ 3E ได้ค่อนข้างกำหนดโดยก๊าซ Chromatography (GC) กำหนดให้น้ำมันดักแด้ไหมประกอบด้วยทั้งกรดไขมันอิ่มตัว และในระดับที่สมกัน กรดไขมันในระดับที่สม alpha linolenic กรดและกรด linoleicแก้ว S akol0 0 0 1.4413.6660.2815.717.260.7232.065.6838.8218.05ภาษาอังกฤษ 4.68 0.7142.186.3527.3118.370.445.35010203040506070กรด palmiticกรด stearicกรด Arachidicกรด PalmitoleicOleic กรดกรด linoleicAlpha linolenic กรดน้ำมันอโวคาโดสกัด Soxhletแยก macerationนางน้อย0.7215.717.2613.661.4460.280.346.630.5421.636.8544.7619.255.7818.80.522.457.6544.82010203040506070Palmitic acidStearic acidArachidic acidPalmitoleic acidOleic acidLinoleic acidAlpha linolenic acidAvocado oilSoxhlet extractionMaceration extractionNang L eung0 0 081.4413.6660.280.7215.717.2643.037.6527.810.65.1515.7646.721.770.445.1817.91010203040506070Palmitic acidStearic acidArachidic acidPalmitoleic acidOleic acidLinoleic acidAlpha linolenic acidAvocado oilSoxhlet extractionMaceration extractionS um Rong1.4413.6660.287.260.7215.7134.826.6430.70.667.0619.830.2940.348.5526.110.645.3418.70.32010203040506070Palmitic acidStearic acidArachidic acidPalmitoleic acidOleic acidLin oleic acidAlpha linolenic acidAvocado oilSoxhlet extractionMaceration extractionNone Ruesee1.4413.6660.287.260.7215.7132.46.0433.70.37 0.686.0120.842.317.1530.170.23 0.545.2119.86010203040506070Palmitic acidStearic acidArachidic acidPalmitoleic acidOleic acidLinoleic acidAlpha linolenic acidAvocado oilSoxhlet extractionMaceration extractionFigure 3 Percentage composition of fatty acid identified in native Thai silkworm oil extractedby soxhlet and maceration methods: A. Keawsakol oil, B.Nangnoi oil, C. Nangleung oil,D. Somrong oil, and E. Noneruesee oilA BC DEwere essential fatty acids found in maceration extracts more than soxhlet extracts. Because themaceration extraction was not treated by heat thus the chemical compounds were not destroyed andstable more than those extracted by soxhlet extraction method. The silkworm pupa oils obtained frommaceration extraction of Nonruesee variety contained the highest oleic acid 30.17% as maincomponent but less than that of avocado oil and 42.31 % alpha linolenic acid. The oil from soxhletextraction of Khewsakol variety contained highest oleic acid 38.82% as a component and 32.06 %alpha linolenic acid but it was more than avocado oil. These two types of fatty acid were essential forhuman bodies. It composed of phospholipids which were cell structure and could reduce cholesterolin blood. The lack of fatty acid would affect on growth rate and skin ulcer. Human bodies could notproduce this linolenic acid and therefore it had to be acquired from food (Sappayatosok, 1988).Silkworm pupa contained oil more than that in soybean which comprised only 18.7 % (Manit et al,2000). The silkworm pupa would be a good source of functional fatty acid -linolenic acid. Thus,making it to be potentially useful for food and cosmetic industryLinoleic acid assayThe percentages of linoleic acid from Keawsakol, Nangnoi Si Sa Ket, Nangleung, Somrong,and Noneruesee oils extracted by soxhlet extraction were 2.19, 1.60, 5.89, 5.27 and 4.63,respectively, whereas those of Keawsakol, Nangnoi Si Sa Ket, Nangleung, Somrong, and Nonerueseeoils extracted by maceration extraction were 2.10, 2.25, 2.57, 2.02 and 2.50, respectively. The soxhletmethod appeared to give higher contents of linoleic acid than the maceration method. However, theamounts of linoleic acid in oils were found in trace, which agreed with that previously reported(Kotake et al., 2002). The lipids extracted from several silkworm pupae mainly consisted oftriacylglycerol, phosphatidylethanolamine, and phosphatidylcoline (Kotake et al., 2002). Althoughlinoleic acid is heat labile, the higher amount of linoleic acid was found in oil extracted by the soxhletextraction method than by the maceration extraction method. This may be due to the higher yields ofthe oil by the soxhlet extraction method.Free radical scavenging activityThe free radical scavenging activities of the oils were assayed by DPPH method (Jung, et al.,2006). The SC50 values (mg/ml) of the native Thai silkworm pupae oil were shown in Figure 4. TheSC50 (mg/ml) of the Keawsakol, Nangnoi Sa Sa Ket, Nangleung, Somrong, and Noneruesee oilextracted by soxhlet extraction method were 14.81 ± 0.30, 18.25 ± 0.09, 19.40 ± 0.22, 18.22 ± 0.08and 17.01 ± 0.17, respectively, and for the Keawsakol, Nangnoi Sa Sa Ket, Nangleung, Somrong, andNoneruesee oil extracted by maceration methods were 17.96 ± 0.30, 18.77 ± 0.23, 18.22 ±0.13, 12.51± 0.04, and 10.08 ± 0.14, respectively. This study indicated that Noneruesee oil extracted by
maceration method showed higher DPPH scavenging activity than those from other native Thai
Figure 4 SC50 (mg/ml) values by DPPH scavenging assay of oil from various Thai native silkworm
pupae prepared by the soxhlet and maceration process comparing to the standard
antioxidants (Vitamin C, vitamin E and BHT)
silkworm pupae oils (p<0.05). However, the SC50 of the native Thai silkworm pupae oils were higher
than those of standard vitamin C, vitamin E and BHT (0.42±0.04, 0.54±0.09 and 0.53 ± 0.02 mg/ml,
respectively) (p<0.05). The native Thai silkworm pupae oil contained antioxidant, tocopherol (Kotake
et al., 2002). Some carotenoids such as lutein and nioxantine which were more potent antioxidants
than linoleic acid were also found in the oils (Kotake et al., 2002).In addition, phospholipids and
tocopherol existing in the oils may also play an important role in protecting the lipids especially
linoleic acid against oxidation.(Eiichi et al., 2002).
Tyrosinase inhibition activity
The tyrosinase inhibition activities of oils were assayed by the modified dopachrome method
using tyrosine as a substrate (Long et al., 2000). The IC50(mg/ml) of the Noneruesee oil extracted by
soxhlet method and Nangleung, Somrong,and Noneruesee oils extracted by maceration method were
7.36 ± 0.20, 31.56 ± 0.49, 16.01± 0.07, and 26.02± 0.07, respectively (Figure 5), however, none of
these was detected in Keawsakol, Nangnoi Si Sa Ket, Nangleung, Sumrong extracted by soxhlet
method, and Keawsakol and Nangnoi Si Sa Ket extracted by maceration method.
0.42 0.54 0.53
14.81
18.25 19.4 18.22
17.69 17.01 18.77 18.22
12.51
10.08
0 5
10
15
20
25
30
35
40
Standrad
vitamin C
Standrad
vitamin E
Standrad
BHT
Keawsakol
Nangnoi
Nangleung
Samrong
Noneruesee
SC50 (mg/ml) Hot prosess
Cold process
Soxhlet method
Maceration method
SC 50 (mg/ml)
7.36
0.36 0.15 NA NA NA NA NA
26.02
16.01
31.56
NA
0 5
10
15
20
25
30
35
40
Standrad
vitamin C
Standrad
kojic acid
Keawsakol
Nangnoi
Nangleung
Samrong
Noneruesee
IC50 (mg/ml) Hot process
Cold process
Figure 5 IC50 (mg/ml) values by tyrosinase inhibition activity assay of oil from various Thai native
silkworm pupae prepared by the soxhlet and maceration process comparing to the
standard antioxidants (Vitamin C and kojic acid)
Most oils extracted by the maceration method gave higher tyrosinase inhibition activity than
those by the soxhlet method except for the oil from Noneruesee by the soxhlet method which gave
the highest of this activity (p<0.05). However, the IC50 values of the native Thai silkworm pupae oil
were higher than that of standard vitamin C and kojic acid (0.36 and 0.15, respectively) (p<0.05).
In summary, most oil from the soxhlet method gave lower free radical scavenging and no
tyrosinase inhibition activity except oil from Noneruesee. Three oils of the five samples from the
maceration method have this activity. The bioactive compounds in oil especially linoleic acid, which
are responsible for free radical scavenging activity, may be heat labile. The oil from Noneruesee by
the maceration method showed the highest free radical activity at the SC50 value of 10.08 ± 0.14
mg/ml. The oil from Noneruesee by the soxhlet method gave the highest tyrosinase inhibition activity
at the IC50 value of 7.36 ± 0.20.
Conclusion and Recommendation
This present study compared the extraction efficiency and the two biological activities (free
radical scavenging and tyrosinase inhibition) of oil from the five Thai native silk pupae by soxhlet and
maceration methods. The results suggest the possible application of oil from Noneruesee prepared
by the maceration methods in food and cosmetics products since their free radical scavenging,
tyrosinase inhibition activities and fatty acid profile have been demonstrated.
Acknowledgements
This work was financially supported by Thailand Toray Science Foundation (TTSF). The
authors would like to thank the staffs of Kasetsart Agricultural and Agro-Industrial Product
Improvement institute (KAPI), Kasetsart University, Kasetsart University Research and Development
Soxhlet method
Maceration method
IC 50 (mg/ml)
Institute (KURDI), the Queen Sirikit Institute of Sericulture in the North-eastern region of Thailand,
NPRDC-IST and,Chaing Mai University.
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