Ba d£mg sinh hgc vd cdc chdt co hogt tinh sinh hgc
DISTRIBUTION FATTY ACIDS OF EACH LIPID CLASS FROM WILD SEED OIL - BURSERACEAE FAMILY
Nguyen Thi Nguyet', Doan Lan FbuongS Vu Thi OanhS Nguyen Quoc BinhS Vu Ha Nguyet H a n J , Ngnyen Van Tuyen AnhS Pham Thi Hong Minh', Nguyen Thi NgaS Pham Quoc Long*
' Institute of natural product chemistry - Viet Nam academy of science and technology
^ Vietnam National Museum of Nature - Viet Nam academy of science and technology
^ Hanoi Pedagogical University 2 ABSTRACT
The Burseraceae is the large family that have used in biblical times In incense, in medicine and for embalming.
Therefore tree filled with many beneficial effects in prevention and treatment disease, as well as economy, livelihood In China. Canarium album isbasicalfy used for oil, like European olive, hence it is also called Chinese Olive. Thefhdt, mil, seed, and root o/Canarium have a long history of usage as traditional medicines for the treatment of swollen and sore throat, excessive thirst, hematemesis due to cough, laclllary dysentery, epilepsy, puftfer poisoning, and alcoholism. The dried fruits are also reported tohave anti-virus, anti-bacterial, anti-inflammation and detoxification activities. The lipid of this one have studied and showed distribution of fatty acid of each lipid class such as: polar lipid (PL), Sterol (Sl), Diacylglycerol (DG), Triacylglycerol (TG), Free fatty acid (FFA) and Hidrocarbon+wax (HW). There 16:0, 18:2n-6, I8:ln-9 and 18:0 acids were principal component from total lipid, beside some fatty acids such as 14:0,15:0, 24:0 were contained with small content
Keywords: Seed oil. Canarium album Lour. (BH60B), Canarium album Lour. (BH60^, Canarlttm toHnense (BH76;, Canarium tramdenum (BH85), Polar lipid (PL), Sterol (ST), Free fetly acid (FFA), Diacylglycerol (DG). Triacylglycerol (TG), Hidrocarbon+wax (HW), Total lipid CTL).
L INTRODUCTION
Burseraceae is family of flowering plants in the order S^indales, composed of about 17-18 genera of resinous trees and shrubs. They are native primarily to tropical America, but a few species occur in A£rica and Asia. Fresh green rhombic has alcohol detox effect and treatment of fish crab poisoning. Ripe fiuit have a sedative effect, cure epilepsy. The oleo-gum resin from several species of the genus Boswellia, caUed fianldncense, was used in biblical times in incense, in medicine, and for embalming. Therefore tree fiUed with many beneficial effects in prevention and treatment disease, as well as economy, UveUhood [1^].
H. MATERIALS AND METHODS 2.1. Materials
The samples were collected at some places such as: Son La, Me Linh (Ha noi), Krong Bong (Dak Lak), in April-July for Canarium album Lour., Canarium toldnensis and August-October
f01 Canarium tramdenum 2014. The science names were appraised by Dr. Nguyen Quoc Bmh - VAST, The samples were collected fbm fi^sh and old seed and stored at -4°C in our laboratory. All data about information of sample were reported in table 1.
No 1 2 3 4 IX
Table 1. Information about the samples of Burseraceae family Science name Canarium allmm Lour.
Canarium album Lour.
Canarium tokinense Canarium tramdenum Methods
Symbol BH60 BH60B BH76 BH8S
Place SocLa DSkLik _ MeLU SouU
2.2.1. Lipid extraction
Total lipid was extracted by BUgh and T)y&i method [3j. The seeds (lOg) were crushed and Upidj was extracted at 4*C for 24h by stirring with a mixture of chloroform/meflianol (1:2. v/v). AfteTi then, the nuxture was filtered and divided two
H$i thdo khoa hgc ky m$m 40 nSm thdnh ldp Vign Hdn l&m KH&CN VigtNam phases by added IV chloroform and IV H2O. After
isWitig (24h). lower phases were collected and aqtreous phases were extracted by IV chloroform again. All organic phases were removed solvents by roto-evaporated under reduced pressure. Total lipid was stored at -20°C in pure chloroform. Total lipid content of Madhuca elHptica (Pierre ex Duibard) H. J. Lam was determined gravimetrically [41.
2.2.2. Composition and content of lipid classes from total lipid
Total Upid compositions were analyzed by one dimensioi^ diin layer chromatography (TLC) usmg precoated silica gel plates (10 cm x 10 cm) Sorbfil PTLC-AF-V (Sorbfil, Krasnodar, Russia).
The plates were first developed to their fUll lengtti with n-hexan/EtzO/AcOH (70/20/1, by vol) and finally to 25% length with CHCl3/MeOH/C6Hs/NH40H (65/30/5/10. by vol).
After drying stream of air, plates were sprayed witti 10% H2S04/MeOH and heated at 240"C for lOmins. The chromatograms were scanned using an image scanner (Epson Perfection 2400 PHOTO) in a grayscale mode. The percentages of Upid contents were based on band intensity usmg an image analysis program (Sorbfil TLC Videodensitometer, Krasnodar, Russia). Peak areas and Upid class percentages were calculated according to [5,6].
, 2.2.3. Separation Upid classes
Using thin layer chromatography (TLC) to separated phosphoUpid classes from polar lipid with system solvents CHCb/MeOH/NHj/CeHe = 70/30/5/10 (v/v/v/v). Each class was compared with standard in the TLC with the system solvents and the specific spray reagent u s ^ earUer were
^employed. [5]
X2.4. Methods to identified compositions and content of fatty acid
To identified composition and content of fetty acid, total lipid was given mefliyl ester of fetty acid, after then analysis by GC-MS Shnnadzu QP 2010wifli column LBX(30mx0.25(imx0.25^m).
Temperature of detector and injector were 250"^
and 200°C (respectively). Temperature program increase 2''C, from 160°C to 260°C, hold 20mins.
Total time for one sample is 70mins. Fatty acid Empositions were identified by mass spectrum KdoMipared widi standard spectrum. [7]
m . RESULTS AND DISSCUSTION 3.1. Content of totalUpid
Content of total Upid was identified by percent ratio weigjit of total lipid compared with weight of firesh seeds. There results were reported in table 2.
Table 2. Information about sample - Burseraceae family No
1 2 3 4
Samples Canarium album Lour.
Canarium allmm Lour.
Canarium lohnense Canarium tramdenum
Symbol BH«0 BH60B BH76 BH8S
Content 2.5 2.6 7.9 13 Contents of total lipid were showed that specific difference divided into two groups BH60, BH60B (lower 3%) and BH76, BH85 (higher 7%).
There results were similar about shape and size of genus. The Canarium album Zour.genus 03H60 and BH60B) were eUipse shape and there kernel were smaller than Canarium tokinense (BH76) and Canarium tramdenum (BH85).
With Canarium album iour. genus were coUected at two place and different weadier (Son La and Dak Lak) but content of total Upids were the same. Thus, anyplace and any weadier don't effect to content of total Upid of wild seeds Canarium album Lour, genus.
32. Composition and content of Upid classes from total lipid
Table 3. Composition and content of lipid classes from total lipid PL
ST FFA DG TG HW
BH60 5.78 6.43 3.98 75.4
-
8.41 BHeo
5.24 5.38 4.04 74.67
-
10.67 BH76
7.67 9.12 5.78 6.74 63.78 6.91
BH8S 9.34 10.56 6.67 67.12
-
6.31 The composition of the Upids of plant tissues have been reviewed [8]. Those plant tissues diat serve as major food materials have received most study obviously. Triacylglycerol tend to be the most abundant class of storage lipid in tissues that are rich in lipids. On the other hand, diere are exceptions and jojoba oil consists mainly of wax estHs.
r
199Ba dgng sinh hgc vd cdc chdt cd hogt tinh sinh hgc
Total Upid of aU san^ile contained basic Upid classes such as polar Upid (PL), sterol (ST), free fatty acid (FFA), Triacylglycerol (TG) and Hidrocarbon + wax (HW), beside Diacylglycerol Q3G) class was only appeared in BH76 sample (6.74%).
3.3. Composition and content of fatty acids from total Upid
Con^sition and content of fritty acids from wild seed oil - Burseraceae femily were reported table 4.
Table 4. Composition and content of fatty acid from total Qpid
^ ^
12:0 a-14:0 14:0 a-15:0 15:0 a-16:0 16:ln-9 16:0 17:ln 17:0 18:2n-6 18:ln-9 18:ln-7 18:0 I9:ln-9 20:0 20:3n-9 22:0 22:ln-9 23:0 24:0 26:0 SFA USFA
Content (•/.) BH60B
0.09 0.18
-
0.13 0.11 0.02 0.18 27 0.46 33.64 25.33 0.33 10.84 0.07 0.38 0.04 0.23 0.8 0.17
-
39.61
.
60J9 BH60
- -
0.24 0.15 0.35
- -
18.69
- -
43.04 32.19 5.16
- - - - - - -
0.18 24.77
.
75J3 BH76
0.05 0.15 0.03 0.07 0.12 0.09 0.28 24.51 0.16 0.08 42.84 24.01 0.1 7.1 0.01
.
0.05
.
0.06
-
0.11 0.18 32.61 6739
BH85 0.07 0.05
-
0.03
-
0.12 0.34 27.04 0.11
-
32.7 32 0.15 7.04 0.09
- - -
0.12
-
0.14 34.81
_
65.19 The results showed that abundant appearance of fatty acids from Cu to C24 aU sanqiles and only present super-long fatty acid 26:0 with smaU content (0.18%) in Canarium tokinensis. Main fatty acids were 16:0, 18:2n-6, 18:ln-9 and 18:0 vdiich were principal conqxnient of plant seed.
Specially, Unoleic (18:2n-Q was in^rtant fatty acid forbiosyndietic process mto PUFAs. Content of this one was the sunilar some other seed oil
(SFA: TottI saturated &tty Bcidi, USFA: Total '•'llt?^^l^•^^^ fitty acids)
sunflower, oUve ...[9]. Beside, oniega9 acid (18: ln-9) was high content, too.
Total saturated fiitty acids (SFA) and total unsaturated fatty acids (USFA) don't diffoence with BH76 and BH85. However, BH60 and BH60B were genus together, this cUfference was very much. Thus the place and weadier don't effect to total Upid but effect to quaUty of lipid.
3.4. Composition and content of fatty acids from each lipid classes
Distribution &tty acids in Upid classes were reported in table 5. Cou^are with total saturated
&tty acids (SFA) and total unsaturated fatty acids were described in chart 1-4
Chart 1. Total saturated fatty acids and total unsaturated fatty acids of BH60B The sample BH60B contained five basic Upid classes such as: PL, ST, FFA, TG, and HW.^
Distribution of fatty acids were the same and different, too. Present of 16:0. 18:2n-6, 18:ln-9 and 18:0 acids with hig^ content in aU class was the simUarly. However, 19:ln-9 acid (0.07%)]
were contained in total Upid but only presentation m HW class. Other exan:q>le, 15:0 acid was contained in total Upid and ST class but not appeared other classes. Beside, &tty acids 17:0,:
20:3n-9 exits m PL classes.
Chart 2. Total saturated fatty acids and total unsaturated fatty acids of BH60
lipid classes ds from each ttjraci
•s
conte
a o
5. Co Table BH8S BH76 BH60 BH«OB Sample
t
H
s
0 H
1
£
g
W O
g 1
^
E-
FFA
g 1
si
[;
i
s
1
0.1 1.37 1.23 1 12:0 0.73 1.35 1.55
•
•
•
•
•
•
1 a-14:0 0.35 1 0.6 0.28 5.83 0.11 3.05 5.4, 1.92 7.2 0.24 4.86 2.13 4.91
n
5.29 2.94 6.4 12.27
•
1 14fl 1.81 0.39 1.21 0.89 1.03 0.81
•
§
0.18 1 3.82 1.76 1.58 4.01 2.77
•
2.52 4.14 3.14
•
3.41
•
•
•
1 15:0
'
1.14
'
1.08
•
1.12
•
•
•
•
•
2.6 1 11-16:0 0.23 1
•
8.41 1.49 2.64 1.37 3.38
•
2.54
•
•
3.65 6.04
•
•
1 16:lii-7 26.91 1 36.49 35.07 41.88 27.88 33.57 29.19 47.73 40.37 22.94 26.13 30.01 S
46.43 27.35 32.03 26.35 33.6 54.02 28.9 19.79 1 16:0 0.81 1 17:ln 0.17 1
•
0.77
•
0.44 1.51
•
1.08
•
•
•
0.92 17:0 28.66 1 16.81 22.52 9.94 28.77 31.47 26.53 13.38
•1
45.71 31.36 39.78 21.58 14.55
s.
19.3 21.68 7.34
S
28.8 12.81
1
34.86 1 36.04 32.9 17.86 36.53 18.68 24.43 17.6
1
24.81 14.29 20.27 19.3 14.59 20.69
^
32.91 19.72 13.65 33.23 37.5
^ i
0.56
cl
7.2 10.06
<i
10.35 6.49 7.95 8.1 10.08 5.74 7.55 7.81
2
11.49 3.87 13.62 9.77 9.77 12.89 9.07 12.59 18:0 3.88
?
0.26 1
•
0.22 0.53 0.43
•
0.72
•
0.5
•
•
•
0.93 4.2
Bl
0.24 1
•
•
'
0.68 1.35 0.71
•
1.2
•
•
•
•
•
•
0.76 22:0 12.91 6.69 1.42 23:0 0.26 1
•
0.72
'
2.84 1.98
•
•
•
0.94
•
1.06 24:0 0.74
1
35.67 1 47.15 44.58 63.79 34.7
"^
1
^
s66.73 28J12
S
1
59.12 70.86
2
56.05 41.21
? s
79.18 37.97 39.12 SFA
<^
52.85 55.42
3
53.6
1
w
71.08
a
60.05 40.88
rf
68.78 43J)5 58.79 46.01 20.82
s
60.88
1
o
Q
+ 1
I I
Ba d^g sinh hgc vd cdc chdt cd hogt tinh sinh hgc
Similarly BH60B, the sanqile BH60 contained five basic Upid classes. Total saturated fatty acids were higher than total unsaturated &tty acid in FFA, HW and ST class. Contrary, in TG and PL class, total saturated &tty acid was lower than total unsaturated &tty acids class. Beside, &tty acids such as: 15:0,16:ln-7 and 24:0 were contained in HW class and not ^^>eared in other classes.
Chart 3. Total saturated Tatty adds and total unsaturated fatty acids of BH76 The san^le BH76 super-long fatty acids as 23:0 and 26:0 were only present at FFA class but not appeared other class. Total unsaturated fatty acid of TG, ST and PL were higher than total saturated fatty acids.
60 • 4 0 • 20 • 0 •
.« ^ ^
HW TO FFA ST PL
in
• SFA - SUSFA
Chart 4. Total Total saturated fatty acids and total unsaturated fatty adds of BH85 Compared with oflier san^les, total saturated
&tty acid of TG class was higher than total unsaturated &tty acid in sanqile BH85. There were 22:0 and 24:0 acids which existed in polar Upid and not present of other classes.
ACKNOWLEDGMENTS: This work was supported by cooperation-project Germany and
Vietnam. Project code: 44/2014/HD-NDT.
REFERENCES
1. L. Watson va M.J. DaUwitz, Cdc hp thyc v$t c6 hoa (1992)
2. Burseraceae, Encyclopsedia Britannica
Lie, 2015.
http://www.britannica.com/plant/Burseraceae.
3. Extraction of Lipids in solution Ity the method of Bligh & Dyer. A rapid method for total lipid extraction and purification.
4. Hamoutene D, Puestow T, MiUer-Banoub J, Wareham V (2008) Main lipid classes in some of deep-sea corals in the Newfoundland and Labrador region (Nothwest Atlantic Ocean). Coral Reefs 27:237-246.
5. Hamoutene D, Puestow T, MiUer-Banoub J, Wareham V (2008) " Main lipid classes in some species of deep sea corals in the Newfoundland and Labrador re^on"(Northwest Atlantic Ocean).
Coral Reefe 27:237-246.
6. Khotunchenko SV, KuUkova IV (2000)"
Lipids of different parts of the lamina ofLaminaria japonica Aresch" Bot Mar 43:87-141.
7. Audrey imbs "Fatty acid, Upid class, and phospholipid molecular species composition of the soft coralXenia.^" No.6. Volume 50. ISSN 0024- 4201.2015
8. Harwood, JJC (1980) Plant acyl lipids:
structure, distribution and analysis. In: The Biochemistry of Plant. Vol 4. Lipid: Stmcture and Funtion, pp. 1-55 (ed. PK. Stumpf, Academy Press, New York).
9. Sheppard, A, J., Iverson, J.L. and Weihauch, J.L (1978) Composition of selected dietary fats, oils, margarines and butter. In handbook of Upid reseach. Vol. 1 Fatty acids and Glycerides, pp. 341-379 (ed. A. Kukis, Plenum Press, New York).
Corresponding author. Doan Lan Phuong
Institute of natural product chemisby - Viet Nam academy of science and technology Email: dnanlanphuong75@p[mail cnm
Telephone number: 0904221818
