TAP CH PHAT TRIEN KH&CN, TAP 12, SO 10 - 2009

KHO ST TINH DU V TRI V L TC, Fortunella japonica, Thumb.

Trnh Hong Hiu, Nguyn Th Tho Trn, L Ngc Thch Trng i hc Khoa hc T nhin, HQG-HCM
(Bi nhn ngy 08 thng 01 nm 2009, hon chnh sa cha ngy 24 thng 08 nm 2009)

TM TT: Tc hay cn gi l qut, hnh, h Cam qut (Rutaceae), c trng rt ph bin Vit Nam. ti ny tin hnh kho st tinh du v tri v l, trn nhiu lnh vc: hiu sut ti u theo cc phng php ly trch, ch s vt l v ho hc, thnh phn ha hc v hot tnh sinh hc. S ly trch tinh du c thc hin theo phng php chng ct hi nc un nng c in v chiu x vi sng. Thnh phn ha hc tinh du c xc nh bng phng php GC-MS cho thy tinh du v tri c cu phn chnh l limonen (92%) v tinh du l cha ch yu cc cu phn chnh l elemol (18%), -eudesmol (16%), epibiciclosesquiphelandren (16%). T kha: Fortunella japonica Thumb.,Tc, chng ct hi nc, chiu x vi sng, hm lng ti u, thnh phn ha hc, ch s l-ha, hot tnh sinh hc, limonen, elemol, eudesmol, epi-biciclosesquiphelandren. 1. T VN Tc l mt loi cy d trng. Tri tc ngoi vic lm cnh trong nhng ngy Tt c truyn, cn c dng thay chanh trong mt s mn n, thc ung v gia v. L c hng v c trng. Hoa trng, p v c mi thm. Tn khoa hc l Fortunella japonica Thumb., h Rutaceae. Tn thng gi: hnh, qut, . [1-6] Mt s tn nc ngoi khc nh: calamondin, kumquat. [8] Cc nghin cu v tinh du l v v tri tc c thc hin kh nhiu trn th gii [1, 8-15] nhng ti Vit Nam th cn hn ch. [4,16] Tuy nhin, tt c ch yu quan tm v mt thnh phn ha hc ca tinh du. Cc nghin cu v hot tnh sinh hc tinh du v tri v l tc c tm thy rt t. [12] 2. THC NGHIM 2.1. Nguyn liu V tri v l cy tc, Fortunella japonica Thumb., c thu hi ti huyn Chu Thnh, tnh Tin Giang. 2.2. Xc nh b phn cha tinh du Thc hin s phi phu hc tuyn tinh du xc nh b phn cha tinh du trn nguyn liu kho st. Gii phu hc c thc hin ti B mn Thc vt, Khoa Sinh hc, Trng i hc Khoa hc T nhin Thnh ph H Ch Minh. 2.3. Ly trch Tinh du c ly trch bng phng php chng ct hi nc un nng c in (NC) hoc chiu x vi sng (CXVS) trong l vi sng gia dng ci tin Sanyo EM-D553N, 2450 MHz, 450 W, vi b Clevenger 2.000 ml (b tch tinh du nh). [17-19] Mi ln th nghim s dng 300 g nguyn liu xay nh v 1.000 ml nc. Tin hnh ly trch trong nhng khong thi gian nht nh. S dng dietil eter ly trch phn tinh du, lm khan nc bng Na2SO4 khan v c quay thu hi dung mi. Cn v tnh hiu sut tinh du thu c.

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Science & Technology Development, Vol 12, No.10- 2009 2.4. Thnh phn ha hc Xc nh bng phng php sc k kh ghp khi ph (GC-MS) trn my HP 5890GC/5972MS, ct mao qun: RTX 5 (30,0 m x 250 m x 0,25 m), nhit injector: 250 o C, tc dng: 3 ml/pht. Chng trnh nhit: nhit u 40 oC, tng 2 oC/pht, n 80 oC, lu 4 pht, tng 4 oC/pht n 180 oC, lu 6 pht, tng 15 oC/pht n 280 oC v lu 2 pht. 2.5. Ch s vt l v ha hc T trng c xc nh bng t trng k thy tinh. Ch s khc x c xc nh trn khc x k WYA-S ABBE. Gc quay cc c ghi trn trin quang k A. Krss Optronic. Cc ch s acid (IA), ester (IE), savon ha (IS) c xc nh theo tiu chun Php. [20] 2.6. Hot tnh khng khun Thc hin ti Phng Vi sinh Thc phm, Khoa Xt nghim, Vin Pasteur Tp H Ch Minh theo phng php khuych tn trn a thch, o dng knh vng v trng to ra bi a giy (D = 6 mm) tm tinh du vi cc nng thay i t 100 (tinh du nguyn cht), n cc nng pha long dn. 3. KT QU V BN LUN 3.1. Gii phu hc tuyn tinh du Tinh du phn b trong v tri v l tc di dng tuyn tit, hnh cu hoc hnh bu dc. 3.2. Ly trch tinh du 3.2.1. Kho st t l nguyn liu - nc Tin hnh kho st t l vi 100 g nguyn liu (v tri hoc l) xay nh, vi th tch nc t 500 n 1.100 ml, un trong 1 gi bng phng php chng ct hi nc un nng c in. Kt qu c ghi nhn trong Hnh 1.
1.20 1.11

1.00 0.93

Hiu sut (%)

0.80 0.77 0.60 0.8 0.81

0.87

0.91

Hiu sut tinh du v tri (%) Hiu sut tinh du l (%)

0.40 0.28 0.20 0.14 0.14 0.14 0.15 0.16 0.2

0.00 400 500 600 700 800 900 1000 1100 1200

Th tch nc (mL)

Hnh 1. Hiu sut tinh du v tri v l tc theo th tch nc chng ct

T Hnh 1 cho thy vi 100 g v tri hoc l tc cn c 1.000 ml nc c th thu c hiu sut cao nht. 3.2.2. Kho st thi gian chng ct theo cc phng php 3.2.2.1. V tri 3.2.2.1.1. Chng ct bi s NC: Tin hnh chng ct t 3 n 9 gi, kt qu c ghi nhn trong Hnh 2.

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TAP CH PHAT TRIEN KH&CN, TAP 12, SO 10 - 2009

1.51 1.5 1.49 1.49

1.48

Hiu sut (%)

1.46

1.45 1.44 1.43

1.42 120 180 240 300 360 420 480 540

Thi gian (pht)

Hnh 2. Hiu sut tinh du v tri tc theo thi gian chng ct bi s un NC

Theo Hnh 2, hiu sut tinh du t gi tr cao nht (1,50%) sau 7 gi chng ct. 3.2.2.1.2. Chng ct di s CXVS Tin hnh chng ct t 20 pht n 60 pht, kt qu c biu din theo Hnh 3. Kt qu t Hnh 3 cho thy, hiu sut tinh du v tri t cao nht 0,88% sau 40 pht. Nh vy s chng ct hi nc di s CXVS trong trng hp tinh du ny khng thch hp (0,88% so vi 1,50%). C l l do thnh phn ha hc ca tinh du cha rt nhiu hp cht khng phn cc, do khng chu nh hng ca vi sng. Bn cnh vic thc hin chng ct tinh du di s CXVS-CN (c nc), chng ti cng thc hin vic kho st di s CXVS khng cho thm nc vo nguyn liu (CXVS-KN). Tuy nhin, trng hp CXVS-KN, hiu sut t cao nht ch l 0,39%, sau 20 pht. iu ny chng t lng nc t nhin c sn trong nguyn liu khng li cun ht tinh du. Do , tinh du ly trch t phng php CXVS-KN khng phn nh t nhin nn khng c s dng lm mu vt cho cc kho st tip theo.
0.90 0.88 0.88 0.86 0.87 0.88 0.88

Hiu sut (%)

0.84 0.82 0.80 0.78 0.76 10 20 30 40 50 60 70 0.77

Thi gian (pht)

Hnh 3. Hiu sut tinh du v tri tc theo thi gian chng ct bi s CXVS c nc (CXVS-CN)

3.2.2.2. L 3.2.2.2.1. Chng ct bi s NC: Tin hnh chng ct t 2 gi n 6 gi, kt qu c ghi nhn trong Hnh 4.

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Science & Technology Development, Vol 12, No.10- 2009

0.58 0.56 0.54 0.55 0.56

Hiu sut (%)

0.50

0.51

0.46

0.42

0.38 100

0.39 120 140 160 180 200 220 240 260 280 300 320 340 360 380

Thi gian (pht)

Hnh 4. Hiu sut tinh du l tc theo thi gian chng ct bi s NC

Hiu sut tinh du t gi tr cao nht (0,56%) sau 5 gi chng ct. 3.2.2.2.2. Chng ct di s CXVS: Tin hnh chng ct t 20 pht n 60 pht, kt qu c ghi nhn trong Hnh 5.
0.50

0.45 0.43 0.4 0.35 0.34 0.30 0.3 0.25 0.43

0.40

Hiu sut (%)

CXVS-CN

0.20 0.18 0.15 10 20 30 40 50 60 70 80

Thi gian (pht)

Hnh 5. Hiu sut tinh du l tc theo thi gian chng ct bi s CXVS-CN

Thc nghim cho thy phng php chng ct hi nc di s CXVS-CN a n kt qu c nhiu u im: rt ngn thi gian ly trch (cao nht 0,43% sau 60 pht), tit kim nng lng. Tuy nhin, hiu sut tinh du thu c (0,43%) thp hn t s NC (0,56%). Thc hin s chng ct tinh du bi phng php CXVS-KN, hiu sut t c cao nht l 0,38% sau 20 pht, thp hn phng php CXVS-CN (0,43%) T y, c th ni rng lng nc t nhin c sn trong nguyn liu khng li cun ht tinh du. Do , chng ti khng tip tc nghin cu trn loi sn phm ny. 3.3. Ch s vt l v ha hc Cc ch s vt l, ha hc ca tinh du v v l tc c thc hin trn cc mu tinh du c ly trch theo NC v CXVS-CN.

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TAP CH PHAT TRIEN KH&CN, TAP 12, SO 10 - 2009 Bng 1. Ch s vt l v ha hc

Nguyn liu V L Phng php NC CXVS-CN NC CXVS-CN
31 d 31 29 nD 29 D

IA 3,23 2,22 4,90 2,78

IS 7,33 10,60 9,10 21,00

IE 4,10 7,78 4,20 18,22

0,8373 0,8375 0,9140 0,9170

1,4686 1,4687 1,4935 1,4942

-89,166 -89,119 -7,890 -1,062

Qua kt qu Bng 1, cho thy cc ch s ha l ca tinh du v tri tc thu c t hai phng php NC v CXVS-CN khng khc nhau nhiu lm. Trong khi i vi tinh du l tc c s khc bit r rt gia hai mu tinh du. 3.4. Thnh phn ha hc Thnh phn ha hc ca tinh du v v l tc thu c theo hai phng php NC v CXVS-CN c xc nh bng phng php GC-MS. Bng 2. Mt s cu phn chnh ca tinh du v v l tc
Stt (*) -Pinen Limonen Epi-biciclosesquiphelandren Elemol Guaien -Eudesmol Tng cng Hp cht hidrocarbon Hp cht oxigen * theo th t kt qu phn tch % GC-MS Cu phn NC 4 8 40 46 51 52 0,47 92,62 0,12 0,24 99,76 97,49 2,27 V CXVS-CN 0,74 90,17 0,10 0,33 99,69 97,00 2,69 NC 7,98 1,45 16,64 17,72 6,57 16,65 99,96 53,93 46,03 L CXVS-CN 3,78 0,71 15,16 18,54 9,07 20,27 99,86 48,21 51,65

T kt qu thnh phn ha hc ghi trong Bng 2, c th l gii c ti sao hiu sut chng ct hi nc tinh du v tri tc di s CXVS thp hn s NC. l v tonh du ny c thnh phn hp cht hidrocarbon rt cao (97%). M hidrocarbon l nhng hp cht km phn cc do t chu nh hng ca vi sng. Thnh phn ha hc ca hai loi tinh du l tc phn nh hai kiu kch hot chng ct hi nc (NC/CXVS) rt r rng. Tinh du thu c di s chiu x vi sng c thnh phn hp cht oxigen cao hn hp cht hidrocarbon (52% / 48%) v tinh du thu c bng s un nng c in th c kt qu ngc li (46% / 54%). Vic ny cng nh hng n cc ch s vt l v ha hc (Bng 1). Tuy nhin t l thnh phn bch phn ca hp cht hidrocarbon v hp cht oxigen gn nh tng ng nhau. Chnh v vy, hiu sut tinh du thu c t s NC v CXVS khng cch bit nhau nhiu nhu trong trng hp v tri.

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Science & Technology Development, Vol 12, No.10- 2009 3.5. Th nghim hot tnh khng vi sinh vt Bng 3. Kt qu th nghim hot tnh khng vi sinh vt ca tinh du v tri v l tc
ng knh vng v khun (mm) Vi sinh vt Tinh du NC Bacilus subtilis Escherichia coli ATCC 25922 Staphylococus aureus ATCC 25923 Pseudomonas aeruginosa ATCC 27853 Candida albicans V L V L V L V L V L 18.5 20 9 6 12 19 6 11.5 12 11.5 Nng tinh du 10 0 CXVS-CN 15 25 6 12.5 11 13 6 7 17 14 NC 6 10.5 6 6 6 12 6 10 8.5 10.5 10 -1 CXVSCN 6 12.5 6 8 6 8 6 6 9 13.5 NC 6 9 6 6 6 8.5 6 6 6 9 10 -2 CXVSCN 6 9 6 6 6 7 6 6 6 10

Hot tnh khng vi khun v vi nm ca tinh du l tc tng i mnh hn tinh du v tc trn cc chng kho st. 4. KT LUN - Ly trch tinh du bng phng php chng ct hi nc di s CXVS a n kt qu c nhiu u im nh: rt ngn thi gian ly trch, tit kim nng lng. Tuy nhin, trong trng hp ly trch tinh du v tri tc v l tc, s CXVS c xem nh khng thch hp v cho hiu sut tinh du thp hn s NC. - Thnh phn ha hc tinh du v tc l d-limonen v thnh phn ha hc tinh du l tc l elemol, -eudesmol, epi-biciclosesquiphelandren. - Cc ch s vt l v ha hc thay i theo s kch hot phng php chng ct hi nc. - Hot tnh khng vi sinh vt cng c s khc bit gia hai s kch hot v ty thuc nguyn liu l l hay v tri tc.

STUDY OF PEEL AND LEAF CALAMONDIN OIL, Fotunella japonica Thumb.

Trinh Hoang Hieu, Nguyen Thi Thao Tran, Le Ngoc Thach University of Science, VNU-HCM ABSTRACT: The essential oils of peel and leaf of Fortunella japonica Thumb. were isolated by using classical heating and microwave irradiation hydrodistillation. The oil yields of the different parts were also studied. The physical and chemical indices of the oils were identified. The chemical composition was examined by GC/MS analysis. The main components of peel oils and the leaf oils are limonene (92,62%) and elemol (17,72%), -eudesmol

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TAP CH PHAT TRIEN KH&CN, TAP 12, SO 10 - 2009 (16,65%), epi-bicyclosesquiphellandrene (16,64%) respectively. The antimicrobial property of these oils was determined. Key words: Limonene, elemol, -eudesmol, epi-bicyclosesquiphellandrene, peel and leaf calamondin oil, Fortunella japonica Thumb.. TI LIU THAM KHO [1]. Gupta, G.N., Gupta, J.C. Indian Perfume 1, 36-38 (1957). [2]. Phm Hong H, Cy thuc Vit Nam, quyn 2, Nh xut bn Tr, Tp H Ch Minh, 424 (1996). [3]. V Vn Chi, T in cy thuc Vit Nam, Nh xut bn Y Hc, H Ni, 171-176 (1996). [4]. Nguyn Mnh Pha, V Ngc L. Journal of Essential Oil Research 8, 415-416 (1996). [5]. L nh Mi. Ti nguyn thc vt c tinh du Vit Nam, Tp 3. Nh xut bn Nng nghip, H Ni, 83-84 (2001). [6]. L Ngc Thch, Tinh du, Nh xut bn i Hc Quc Gia, Tp H Ch Minh, 80106 (2003). [7]. Giovanni Dugo, Angelo Di Giacomo. Citrus: The Genus Citrus, Taylor and Francis, Boca Raton, 1-15 (2003). [8]. S. R Agrawal, R. P. Agrawal, G. N. Gupta. Proceedings of the Institution of Chemists (India) 31, 207-209 (1959). [9]. Richard A. Bernhard, Byron Scubis. Journal of Chromatography 5, 137-146 (1961). [10]. A.Koyasako, R. A. Bernhard. Journal of Food Science 48, 1807-1812 (1983). [11]. Katumi Umano, Yukio Hagi, Tomoko Tamra, Akihiro Shoji, Takayuki Shibamoto. Journal of Agricultural and Food Chemistry, 42, 1888-1890 (1994). [12]. Jamal Yulliasri, Agusta Praptiwi dan Andria. Majalah Farmasi Indonesia 11, 77-85 (2000). [13]. S. P.Yo, C. H. Lin. European Journal of Horticultural Science 69, 117-124 (2004). [14]. Hyang-Sook Choi. Journal of Agricultural and Food Chemistry 53, 1642-1647 (2005). [15]. Mohamed A. Ferhat, Brahim Y. Meklati, Jaqueline Smadja, Farid Chemat. Journal of Chromatography A 1112, 12-126 (2006). [16]. Nguyn Th Thanh Lan, L Ngc Thch, Cao Nh Anh, Trn Hu Anh, on Ngc Nhun, Quang Hin. Tuyn tp. Hi ngh Ha hc Ton quc ln th 4 (Ha hc cc Hp cht Thin nhin v Ha Dc), H Ni 20/10/2003, 224-228 (2003). [17]. Nguyen Duong Thanh Thi, Tran Huu Anh, Le Ngoc Thach. Journal of Essential Oil-Bearing Plants 11, 154-161 (2008). [18]. Nguyn Qunh Trang, L Ngc Thch. Tp ch Pht trin Khoa hc v Cng ngh 11, 67-72 (2008). [19]. Bi Th Thanh Thy, L Ngc Thch, Trn Minh Thng. Tp ch Dc liu 13, 116119 (2008). [20]. AFNOR. Huiles essentielles. Syndicat National des Industries Aromatiques Alimentaires, Paris. 37-156 (1992).

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