TAP CH PHAT TRIEN KH&CN, TAP 12, SO 07 - 2009

NGHIN CU S DNG THIT B SOXHLET-VI SNG LY TRCH MT S HP CHT THIN NHIN

Phm Thnh Lc, L Ngc Thch Trng i hc Khoa hc T nhin, HQG-HCM
(Bi nhn ngy 02 thng 11 n m 2008, hon chnh sa cha ngy 10 thng 03 n m 2009)

TM TT: L vi sng gia dng c chuyn i thnh thit b ly trch Soxhlet-Vi sng, s dng vo vic ly trch hp cht thin nhin. Vic nghin cu s dng c thc hin trn: cafein t bp tr, Camellia sinensis L.; - steviosid t c ngt, Stevia rebaudiana (Bert.) Hemsl; - artermisinin t hoa thanh hao hoa vng, Artemisia annua L.; - rutin t hoa he, Sophora japonica L.; - tinh du tri i hi, Illicium verum Hook. f.; - tinh du ht th l, Anethum graveolens L.. Cc s ly trch ni trn cng c thc hin song song trn h thng Soxhlet un nng truyn thng. S so snh hai phng php kch hot c thc hin da trn thi gian, hiu sut v phm cht ca sn phm ly trch. T kha: H thng ly trch Soxhlet-Vi sng, ly trch hp cht thin nhin, Camellia sinensis, Stevia rebaudiana, Artemisia annua, Sophora japonica, Illicium verum, Anethum graveolens. 1. T VN L vi sng do Percy Spencer pht minh u tin nm 1947.[1] Tuy nhin, mi n nm 1978 Michael J. Collin mi thit k l vi sng u tin p dng cho phng th nghim phn tch.[2] Sau hng lot thit b vi sng c pht minh phc v vo nghin cu cng nh phc v sn xut cng nghip.[3-9] Vic p dng nng lng vi sng h tr thc hin phn ng ha hc v ly trch hp cht thin nhin hin ang rt c quan tm.[10] Cc thit b vi sng chuyn dng rt t tin nn vic trang b cc loi thit b ny khng n gin i vi cc phng th nghim Vit Nam. Trong iu kin , l vi sng gia dng tr thnh la chn u tin v chi ph trang b v chuyn i cng nng thp. Hin nay nhiu phng th nghim Vit Nam bt u a l vi sng vo phc v cho nghin cu, tuy nhin ch mt vi phng th nghim tham gia ci tin l vi sng gia dng thnh nhng thit b chuyn dng phc v cho nhng mc ch nghin cu chuyn ngnh. Trn c s nghin cu chuyn i l vi sng gia dng thnh thit b ly trch hp cht thin nhin v thc hin tng hp hu c[11] nhm nghin cu v Ha hc Xanh tip tc nghin cu chuyn i l vi sng gia dng thnh thit b Soxhlet-chiu x vi sng. Bi bo ny trnh by kt qu thu c khi s dng thit b ni trn vo vic ly trch mt s nguyn liu thc vt xc nh kh nng v tm cch ci tin hot ng ca thit b ny. 2. THC NGHIM 2.1. Nguyn liu Vic ly trch c thc hin trn cc i tng thu mua ti nhng a phng nht nh: - Bp Tr long (Cng ty Tr Tm Chu, Bo Lc, Lm ng). - Phn trn mt t ca cy C ngt (Lm ng) - Hoa Thanh hao hoa vng (Trung Quc) - Hoa He (Hip Thnh Dc Hng, Qun 6, Tp H Ch Minh). - Tri i hi (Cao Bng). - Ht Th l (Thi Bnh).

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Science & Technology Development, Vol 12, No.07 - 2009 2.2. Ly trch: Ly trch kit trong nhng iu kin (lng nguyn liu, th tch dung mi) nh nhau, song song trn hai thit b ly trch Soxhlet-un nng c in (A) (Ph lc 1) v Soxhlet-chiu x vi sng (B) (Ph lc 2). 2.2.1. Thc nghim ly trch trn thit b ly trch Soxhlet-chiu x vi sng: Cc mu cy c khi lng (20 g) v 500 mL dung mi c np vo h thng (B). u tin, un bnh cha dung mi cho n khi si. Hi dung mi ngng t ri xung phn cha nguyn liu. Khi ng s chiu x vi sng. Ngng chiu x khi nhn thy dung mi bt u mun si. Lp i lp li thao tc trn cho n khi dung mi bt u quay v bnh cha dung mi th khng chiu x na. Sau , ch n khi dung mi ngng t, trong ln k tip, ri xung phn cha nguyn liu. Lp li cc thao tc nh trn cho n khi s trch kit c xc nh. Thi gian chiu x chung l tng s cc khong thi gian chiu x ngt khong. Sau khi thu hi dung mi, tt c cc loi cao (cao 1) thu t hai phng php ly trch A v B c x l hon ton ging nhau c c sn phm th (cao 2) t yu cu phn tch. Vic iu ch cao 2 t cao 1 c thc hin theo cc quy trnh ring: 2.2.2. Ly trch cafein t l tr: [12] Cao 1 c ho tan hon ton vo 100 mL nc ct, un si nh, thm t t 1 g Ca(OH)2, tip tc un si nh trong 1 pht. ngui n nhit phng, lc qua Bchner, c dng 1 g bt tr lc. Dung dch qua lc, c ly trch bng CHCl3 (7 x 20 mL). Ra dung dch ly trch vi 15 mL nc ct. Nc ra c ly trch li bng CHCl3 (3 x 15 mL). Gp tt c cc dung dch CHCl3 li, lm khan. Lc, c quay thu hi dung mi, xc nh khi lng cao 2. 2.2.3. Ly trch steviosid t phn trn mt t ca cy c ngt: [13] Cao 1 c ho tan vo 50 mL nc ct, thm 50 mL dung dch Na2SO4 bo ha, lc u. Ly trch bng n-butanol (10 x 50 mL), dung dch ly trch c ra bng nc ct (3 x 20 mL). Nc ra c ly trch li bng n-butanol (3 x 20 mL). Gom tt c dung dch ly trch li, lm khan. Lc, thu hi dung mi, xc nh khi lng cao 2. 2.2.4. Ly trch artermisinin t hoa cy thanh hao hoa vng: [14] Cao 1 c ho tan vo 20 mL eter du ho, ly trch bng dung dch etanol : nc c t l 1 :1 (10 x 20 mL). Thu c dung dch mu trng sa. C quay, xc nh khi lng cao 2. 2.2.5. Ly trch rutin t hoa ho: [15] Cao 1 c ho tan vo 200 mL etanol nng, un nh n khi dung dch bt u kt tinh. ngui n nhit phng. yn 24 gi rutin kt tinh hon ton. Lc qua Bchner, sn phm th c ra bng eter du ho lnh cho n khi nc ra khng cn mu xanh th dng li. kh t nhin n khi lng khng i, xc nh khi lng cao 2. 2.2.6. Ly trch tinh du t ht th l v t tri i hi: [16,17,18] Cao 1 (nha du) c ho tan vo etanol. Lm lnh trong 2 gi. Lc nhanh qua h thng phiu lc lnh. C quay thu hi dung mi. Xc nh khi lng cao 2 (tinh du). 2.3. Xc nh hm lng cu phn chnh Hm lng cc cu phn chnh (%) trong cao 2 c cung cp t kt qu ca cc phng php phn tch HPLC/UV, HPLC/MS/MS, GC/MS. 2.3.1. GC/MS: Trn my QP2010. Ct mao qun MDN-5S1: 0.25 mm x 30 m x 0,25 m. Chng trnh nhit: Nhit u 60 oC gi 4 pht, tng 2 oC/pht cho n 80 oC sau tng 5 o C/pht n 180 oC gi 6 pht, tng 10 oC n 200 oC gi 5 pht. 2.3.2. HPLC/UV: - nh lng cafein: Trn my Shimadzu LC 10A. di sng 276 nm. Dung mi acetonitril:H2O (0.05 % H3PO4) l 15:85. Ct C18(250 mm x 4.6 mm). Tc dng: 0.8 ml/pht.

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TAP CH PHAT TRIEN KH&CN, TAP 12, SO 07 - 2009 - nh lng rutin: Trn my Shimadzu LC 1100. di sng 254 nm. Dung mi MeOH:H2O l 4:6. Ct C18 (250 mm x 2.0 mm). Tc dng: 0.2 ml/pht. 2.3.3. HPLC/MS/MS: - nh lng steviosid: Trn my Themo Quantum Access. p sut u ct: 10 bar. Dung mi acetonitril (0.1 % HCOOH):H2O (0.1 % HCOOH) l 65:35. Ct C18 (250 mm x 4.6 mm). Tc dng: 400 l/pht. nh lng theo ion c trng c m/z= 664.3-665.3. - nh lng artermisinin: Trn my Themo Quantum Access. p sut u ct: 10 bar. Dung mi MeOH (0.1 % HCOOH):H2O(0.1 % HCOOH) l 9:1. Ct C18 (250 mm x 4.6 mm). Tc dng: 500 l/pht. nh lng theo ion c trng m/z= 246.4-247.4 2.4. Hiu sut cc ln ly trch c xc nh nh sau:
Hieu suat cao 1 = Khoi lng cao 1 Khoi lng nguyen lieu Khoi lng cao 2 Khoi lng nguyen lieu x 100

Hieu suat cao 2 =

x 100

Khoi lng cau phan chnh = Khoi lng cao 2 x Ham lng cau phan chnh (*) (*) Khi lng cu phn chnh l khi lng l thuyt ca cu phn chnh trong cao 2 da theo kt qu phn tch.
3. KT QU V THO LUN 3.1. Xc nh cng sut chiu x Cng sut chiu x thch hp cho mi nguyn liu v dung mi tng ng l cng sut tha iu kin: Cho hiu sut ly trch cao, dung mi ly trch ti bu ly trch khng b un si qu nhanh khi chiu x. tm cng sut chiu x thch hp cho tng loi nguyn liu v dung mi, u tin thi gian ly trch c c nh l 30 pht. Chiu x vi sng cc cng sut khc nhau t thp n cao. Ngng chiu x, thu hi dung mi, cn cao 1, xc nh hiu sut. Bng 1. Kt qu kho st hiu sut ly trch theo cng sut chiu x, trong 30 pht.
Hiu sut (%) Cng sut (W) Bp tr H2O 10.36 13.02 16.65 19.19 19.95 20.72 C ngt H2O 8.06 13.43 15.65 19.70 20.36 20.74 Hoa he EtOH 15.42 19.54 21.31 23.56 (*) Hoa thanh hao hoa vng Eter du ha 60-80 oC 1.81 2.19 2.93 (*) Tri i hi Eter du ha 30-60 oC 10.27 11.20 (*) Ht th l Eter du ha 30-60 oC 5.76 6.05 (*) -

80 150 300 450 750 900

(*): Khng kho st tip v dung mi si qu nhanh khi ti n hnh chiu x.

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Science & Technology Development, Vol 12, No.07 - 2009 V th biu din v chn cng sut ph hp.

th 1. Hiu sut cao 1 theo cng sut chiu x

3.2. So snh kt qu ly trch trn hai thit b ly trch Soxhlet-un nng c in v Soxhlet-chiu x vi sng Chng ti thc hin vic ly trch kit cc loi nguyn liu nu trn vi 2 phng php (A) v (B). Kt qu ghi trong Bng 2.

th 2. So snh thi gian v khi lng cu phn chnh khi p dng phng php ly trch (A) v (B)

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TAP CH PHAT TRIEN KH&CN, TAP 12, SO 07 - 2009

th 3. So snh thi gian v kh i lng cu phn chnh khi p dng phng php (A) v (B) ly trch steviosid v artermisinin (khng th y r th 2)

Kt qu th 2 v 3 cho thy khi ly trch kit cc hp cht thin nhin bng thit b ly trch Soxhlet-chiu x vi sng kt qu thu c nhanh 12-16 ln hn so vi ly trch kit bng thit b ly trch Soxhlet-un nng c in. Khi lng cao 1 v cao 2 thu c bng thit b ly trch Soxhlet-chiu x vi sng cng cao hn, iu chng t h thng ny ly trch c nhiu sn phm hn phng php Soxhlet un nng c in khi p dng trch kit trn cng khi lng nguyn liu v dung mi. Trong trng hp artermisinin, hm l ng cu phn chnh trong phng php (B) cao hn phng php (A) v hiu sut cao 2 phng php (A) li thp hn phng php (B). iu ny c th gii thch l do thi gian ly trch artermisinin bng phng php (A) kh di (1.560 pht), m artermisinin d b phn hy. Qua ta thy thm mt u im ca h thng (B) l thi gian thc hin ly trch nhanh nn t gy phn hy hot cht. Tuy nhin nhn chung, c hai phng php u trch kit sn phm nh mong mun, nn hiu sut chung gn bng nhau. 4. KT LUN u im ca thit b ly trch Soxhlet-chiu x vi sng: nhanh, an ton i vi nhng cht d b phn hy do thi gian cp nhit rt ngn, tit kim nng lng. Khuyt im ca thit b ly trch Soxhlet-chiu x vi sng l cha t ng ha c qu trnh ly trch, tt c mi hot ng iu khin u thc hin bng tay. Lng mu np vo gii hn. 5. PHNG HNG PHT TRIN Thng qua thc nghim, thit b ly trch Soxhlet-chiu x vi sng cho thy cn mt s im cn khc phc vic ly trch mi ny c hon thin hn: - Lp t thm mt thit b kim sot nhit n nh nhit ni v tr t nguyn liu. - Lp t thm mt cm bin kim sot mc dung mi ni v tr t nguyn liu.

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Science & Technology Development, Vol 12, No.07 - 2009 - Lp t thm mt b phn theo di nng dung dch ly trch chy ra t bu t nguyn liu nhm xc nh thi im hon thnh qu trnh ly trch kit. - Mt mch in t kt hp cc b phn ni trn v ni vo b phn iu khin thi gian chiu x ca l vi sng. Nu thc hin c cc ci tin trn, thit b ly trch Soxhlet-chiu x vi sng mi s hot ng hon ton t ng trong sut qu trnh vn hnh.

USING MICROWAVE-SOXHLET EXTRACTOR TO EXTRACT SOME NATURAL PRODUCTS

Pham Thanh Loc, Le Ngoc Thach University of Science, VNU-HCM ABSTRACT: Domestic microwave oven was modified into Soxhlet-Microwave Extractor for extracting of natural products. The study of its extracted capacity was realized on: - caffein from Camellia sinensis L.; - stevioside from Stevia rebaudiana (Bert.) Hemsl.; artermisinin from Artemisia annua L.; - rutine from Sophora japonica L.; - fruit oil from Illicium verum Hook. f.; - seed oil from Anethum graveolens L.. These extracted results were compared with the Soxhlet-conventional heating on the time, yield and quality. Keywords: Soxhlet-Microwave Extractor, natural product extraction, Camellia sinensis, Stevia rebaudiana, Artemisia annua, Sophora japonica, Illicium verum, Anethum graveolens. TI LIU THAM KHO [1]. Mike Taylor, Bhuwan Singh Atri, Sonal Minhas. Developments in Microwave Chemistry, Evalueserve 5-18 (2005). [2]. Michael J. Collins, Dennis P. Manchester. United States Patent 4835354 (1988). [3]. C. Oliver Kappe. Angew. Chem. Int. Ed. 43, 6250-6284 (2004). [4]. J. L. Luque-Garca, M. D. Luque de Castro. Anal. Chem. 73, 5903-5908 (2001). [5]. J. Parera, F. J. Santos, M. T. Galceran. Journal of Chromatography A 1046, 19-26 (2004). [6]. J. R. Jocelyn Par, Michel Sigouin, Jacques Lapointe. United States Patent 5002784 (1991). [7]. M. Letellier, H. Budzinski. Analusis 27, 259-271 (1999). [8]. V. Fernndez-Prez, L. E. Garca-Ayuso, M. D. Luque de Castro. Analyst 125, 317322 (2000). [9]. Xiaolan Zhu, Qingde Su, Jibao Cai, Jun Yang. Analytica Chimica Acta 579, 88-94 (2006). [10]. F. Chemat, M. -E. Lucchesi, Chapter 22: Microwave-assisted Extraction of Essential Oils, A. Loupy, Ed.. Microwaves in Organic Synthesis, VCH, Weinheim, 959-983 (2006). [11]. L Ngc Thch. Nghin cu chuyn i l vi sng gia dng thnh thit b ly trch hp cht thin nhin v thc hin tng hp hu c. Tuyn tp Hi tho Sng to khoa

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TAP CH PHAT TRIEN KH&CN, TAP 12, SO 07 - 2009 hc vi s nghip cng nghip ha, hin i ha t nc. Nng, 19-23/07/2006, 204-212 (2006). [12]. Md. Mumin Abdul, Farida Akhter Kazi, Md. Zainal Abedin, Md. Zakir Hossain. Malaysian Journal of Chemistry 8(1), 45-51 (2006). [13]. Georgia J. Persons, United States Patent 3723410 (1973). [14]. 14. A. R. Bilia, P. Melillo de Malgalhaes, M. C. Bergonzi, F. F. Vincieri, Phytomedicine 13, 487-493 (2006) [15]. Farouk S. Elferaly, Hala N. ElSohly. United States Patent 4952603 (1980) [16]. Nguyn Th Hoi, L Ngc Thch. Bo co Khoa hc, Ngy Ha hc Tp HCM ln 5, 11/10/2007, 78-83 (2007). [17]. Wang Qin, Jiang Lin, Wen Qibiao. Wuhan University Journal of Natural Sciences 12(2), 529-534 (2007). [18]. Nguyn Th Kim Phng, L Ngc Thch. Tuyn tp cc cng trnh, Hi ngh Khoa hc v Cng ngh Ha hc Hu c Ton quc ln th 4, H Ni 20/10/2007, 485-489 (2007). PH LC

Ph lc 1. Thit b Soxhlet- un nng c in (A)

Ph lc 2. Thit b Soxhlet-chiu x vi sng (B)

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Science & Technology Development, Vol 12, No.07 - 2009 Bng 2. So snh kt qu ly trch theo hai phng php (A) v (B)
Nguyn liu Cu phn chnh Phng php Thi gian (pht) 1670 137 2340 150 1380 112 1560 118 1500 95 1260 85 Khi lng cao 1 (g) 8.2516 8.3925 6.4654 6.7951 8.2085 8.7970 0.8742 1.1098 2.9199 3.1406 1.9537 2.1051 Hiu sut cao 1 (%) Khi lng cao 2 (g) 0.2477 0.2629 1.3673 1.5344 3.3142 3.5073 0.3514 0.4280 2.3876 2.3960 1.1692 1.2969 Hiu sut cao 2 (%) Hm lng cu phn chnh (%)* 84.97 83.78 16.05 14.16 99.45 93.69 0.65 0.84 94.10 94.10 32.93 29.39 Khi lng cu phn chnh (g) 0.2105 0.2203 0.0157 0.0174 3.2959 3.2860 0.0023 0.0036 2.2467 2.2546 0.3850 0.3812

Bp tr lng

A Cafein B A Steviosid B A

41.72 42.00 33.76 35.49 44.18 47.18 4.41 5.62 16.68 17.90 10.20 11.02

1.25 1.33 7.01 8.01 17.84 18.81 1.77 2.17 13.64 13.66 6.14 6.79

C ngt

Hoa he

Rutin B A B A (E)-Anetol B A Apiol B

Hoa thanh hao hoa Artermisinin vng Tri i hi

Ht th l

* Theo kt qu % ca GC-MS, HPLC-MS, hoc HPLC-UV

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