TP CH KHOA HC V CNG NGH, I HC NNG - S 3(44).

2011

NGHIN CU CHIT TCH V XC INH THNH PHN HA HC CA HP CHT POLYPHENOL NHM TANIN T V KEO L TRM
A STUDY ON THE EXTRACTION AND DETERMINATION OF CHEMICAL CONSTITUENTS OF POLYPHENOL TANNIN FROM ACACIA AURICULIFORMIS BARK L T Hi, Nguyn Th Lan Anh, Lu V Dim Hng
Trng i hc S phm, i hc Nng
TM TT nh hng ca cc yu t n qu trnh chit tch polyphenol nhm tanin trong v keo l trm c nghin cu. Cc nhm chc trong tanin c xc nh bng phng php quang ph hng ngoi (IR). Thnh phn ha hc v cu to ca mt s cu t chnh trong tanin c phn tch bng phng php sc k lng cao p kt ni khi ph (HPLC-MS). Kt qu nghin cu tm c iu kin ti u cho qu trnh chit tch polyphenol nhm tanin trong v keo l trm: kch thc nguyn liu dng bt, t l nc: etanol 50 : 50, nhit 0 80 C, thi gian chit 75 pht, t l rn: lng 2g : 70ml. v hiu sut tch tanin l 21,60%. so vi nguyn liu v kh. Polyphenol trong v keo l trm c cha tanin ngng t v tanin thy phn. ABSTRACT The influence of some factors on the extraction yield of polyphenol tannin from Acacia Auriculiformis bark was investigated. The function groups of tannin were determined by the infrared spectroscopy (IR). The chemical components and the structure of main compounds of tannin were determined by high pressure liquid chromatography with mass spectroscopy (HPLC-MS). The optimum conditions for the extraction of polyphenol tannin from Acacia Auriculiformis bark were powder, size of bark, ratio of water: ethanol (%V) 50 : 50, temperature: o 80 C, extraction time 75 mins, ratio of material : solvent 2g : 70 ml and the yield of tannin extraction was 21.60%. The polyphenol tannin from Acacia Auriculiformis bark was hydrolysable - condensed type.

1. t vn Ngy nay, cc vt liu kt dnh mi c ngun gc t thc vt ang c cc nh khoa hc quan tm nghin cu [1]. c bit, i vi ngnh sn xut vn g nhn to th nhu cu s dng cc cht kt dnh l rt ln. Trong qu trnh sn xut vn g nhn to, ngi ta thng s dng cc loi keo nh phenol formaldehyde (PF), phenolresorcin formaldehyde (PRF),... Tuy nhin, vic s dng cc loi keo dn bt ngun t cc ha cht ca cng nghip du m thng c gi thnh t v gy c hi vi mi trng. Do vy, xu hng nghin cu tm cc cht khng c hi thay th mt phn hoc ton b nguyn liu gc du m bng cc nguyn liu ti to c ngun gc thc vt l cng ngh hp dn v mt kinh t v mi trng ang c cc nh khoa hc trn th
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gii quan tm. Mt trong s l cc hp cht polyphenol nhm tanin c tch ra t cc loi thc vt nh keo l trm, keo en, keo tai tng, c, thng c s dng cho tng hp keo polyphenol formaldehyde [2, 3, 4, 5]. nc ta cy keo l trm hay cn gi l Trm bng vng (Acacia auriculiformis) c trng vi mt din tch ln trn khp mi min t quc nhm phc v cho cng nghip g, cng nghip sn xut bt giy. T , mt lng rt ln phn v cha polyphenol b b i hoc lm ci t. Do vy, vic nghin cu chit tch v xc nh thnh phn ha hc ca hp cht polyphenol nhm tanin t v keo l trm s c ngha quan trng v mt khoa hc v thc tin trong vic nghin cu tng hp mt loi keo dn g thn thin mi trng. 2. Phng php thc nghim 2.1. Nguyn liu Nguyn liu l v keo l trm cc khu rng Qung Nam Nng v c ra sch, thi nh bng dao kim loi khng g, sy 50oC n kh. 2.2. Nghin cu nh hng cc yu t n qu trnh chit tch polyphenol nhm tanin Cc yu t nh hng n qu trnh chit tch polyphenol nhm tanin nh: kch thc nguyn liu, t l dung mi nguyn liu, thi gian, nhit , t l dung mi nc etanol. nh lng tanin thu c trong dch chit bng phng php Lowenthal: oxi ho bng KMnO4 vi cht ch th inigocacmin [6]. 2.3. nh tnh phn bit polyphenol nhm tanin ngng t v tanin thy phn Dng phng php Stiasny xc nh s c mt ca polyphenol nhm tanin ngng t v tanin thy phn c trong dch chit v keo l trm [7]. 2.4. Xc nh thnh phn ha hc v cu trc ca cc hp cht polyphenol nhm tanin Thnh phn ha hc v cu trc ca cc hp cht polyphenol nhm tanin trong v keo l trm c xc nh bng phng php quang ph hng ngoi (IR), sc k lng cao p kt ni khi ph (HPLC-MS) ti Vin Ha hc cc hp cht thin nhin Vin Khoa hc v Cng ngh Vit Nam. 3. Kt qu v tho lun 3.1. nh tnh polyphenol nhm tanin Dch chit v keo l trm cho phn ng dng tnh vi dung dch FeCl3 5% (c mu xanh en) nn trong dch chit c mt polyphenol nhm tanin. Tin hnh phn ng Stiasny ca dch chit vi HCHO trong mi trng axit HCl thy c xut hin nhiu kt ta vn mu gch chng t c polyphenol nhm tanin ngng t.
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Dch lc sau khi loi b kt ta cho mu xanh ru khi cho vo dung dch CH3COONa d vi vi git dung dch FeCl3, chng t c polyphenol nhm tanin thy phn. 3.2. nh hng ca cc yu t n qu trnh chit tch polyphenol nhm tanin t v keo l trm 3.2.1. nh hng ca thi gian nh hng ca thi gian n qu trnh chit tch tanin c tin hnh nh sau: Ly 2g v keo l trm thi nh c kch thc 3 5 cm cho vo 100ml nc un nhit 70oC trong cc khong thi gian khc nhau. Kt qu thu c trnh by hnh 1.
8.5 8 7.5 X(%) 7 6.5 6 5.5 5 4.5

30 45 60 75 90 105 Hnh 1 cho thy, khi thi gian t(min) un cng tng th lng tanin tch ra cng nhiu v n 75 pht th lng Hnh 1. nh hng ca thi gian n hiu sut chit tch tanin tch ra t gi tr n nh. Nh vy thi gian tch ti u l 75 pht.

3.2.2. nh hng ca t l cht rn v dung mi chit nh hng ca t l cht rn v dung mi n hiu sut chit tch tanin c thc hin nh 3.2.1 trong thi gian 75 pht v th tch ca dung mi t 30ml n 80ml. Kt qu thu c trnh by hnh 2. Hnh 2 cho thy, t l cht rn v dung mi ti u cho qu trnh chit tch tanin t v keo l trm l 70ml cho 2g v keo l trm. Nguyn nhn l do khi lng dung mi tng ln th kh nng tip xc vi nguyn liu cng ln v lng tanin tch ra cng nhiu, nhng dng lng dung mi qu ln th tanin cng khng tch thm v lc tanin c trong v keo l trm hu nh c tch hon ton.
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8.5 8 7.5 X(%) 7 6.5 6 5.5 5 30 40 50 60 V(ml) 70 80

Hnh 2. nh hng ca t l cht rn v dung mi n hiu sut chit tch tanin

TP CH KHOA HC V CNG NGH, I HC NNG - S 3(44).2011

3.2.3. nh hng ca nhit nh hng ca nhit n hiu sut chit tch tanin c tin hnh nh 3.2.2 ti cc nhit khc nhau: 40, 50, 60, 70, 80, 90oC. Kt qu thu c trnh by hnh 3. Nh vy, nhit tng th hiu sut chit tch tanin tng v t ti u ti 80oC. iu ny c th gii thch l do khi nhit tng th thun li cho vic ph hy mng t bo thc vt v tng ha tan tanin, dn n tng hiu qu chit tch.
12.5 11.5 10.5 9.5 X(%) 8.5 7.5 6.5 5.5 4.5 40 50 60 70 80 90 nhit (oC)

Hnh 3. nh hng ca nhit n hiu sut chit tch tanin

3.2.4. nh hng ca kch thc v keo l trm Kt qu nghin cu nh hng ca kch thc v keo l trm n hiu sut chit tch tanin c trnh by hnh 4. Kt qu cho thy, khi kch thc nguyn liu cng nh th hiu sut chit tch tanin tng; nguyn nhn l do tng kh nng tip xc gia nguyn liu v dung mi, to iu kin thun li cho qu trnh tch tanin t v keo l trm.
20 18 16 X(%) 14 12 10 8
3c m 11, 5c m 23m m da ng bo t 5c m 3-

2-

kich thuoc

Hnh 4. nh hng ca kch thc nguyn liu n hiu sut chit tch tanin

3.2.5. nh hng ca t l nc : etanol nh hng ca t l nc: etanol c tin hnh nh sau: Ly 5g v keo vo 70ml dung mi vi t l nc/etanol khc nhau un 80oC trong thi gian 75 pht. Kt qu thu c trnh by hnh 5. T hnh 5 cho thy, t l % nc : etanol ti u l 50 : 50. Sau t l ny th hiu qu chit tch b gim i; nguyn nhn c th l do tanin tan tt trong etanol hn nc nn khi tng th tch etanol th tanin tch ra cng tng. Tuy nhin, trong v keo c mt s hp cht cng tan tt trong etanol nn tch ra cng vi qu trnh tch tanin, do khi lng etanol qu ln th cc cht ny tch ra cng nhiu v gim hiu qu ca qu trnh tch tanin.
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0 80 /2 0

60 /4 0

50 /5 0

40 /6 0

10 0/ 0

3.3. Tch tanin rn v xc nh ch s Stiasny

V/V (%)

Hnh 5. nh hng ca t l nc: etanol n

hiu sut chit tch tanin Sau khi x l v trm bng dung mi chit th ngoi tanin ra cn c mt s tp cht khc phi loi b. loi b tp cht, dch chit s c x l qua cloroform. Tch tng cloroform th thu c dch chit tanin. C cn dch chit 70oC thu c tanin dng bt.

Ch s Stiasny ca tanin rn c xc nh nh [7] v c gi tr l 69,05. 3.4. Phn tch sn phm tanin 3.4.1. Ph hng ngoi ca mu tanin rn Ph hng ngoi mu tanin tch t dung mi ru- nc c trnh by hnh 6.

Hnh 6. Ph hng ngoi ca tanin tch ra t dung mi ru nc

Ph cho thy trong tanin v keo l trm c cha cc nhm chc c trng nh: -OH (3380 cm-1), C=O (1610cm-1), C=C thm (1515 cm-1, 1448 cm-1), = C O C (1204 cm-1), - C O C (1095 cm-1), CH benzen th para (845 cm-1), CH thm (722 cm-1). 3.4.2. Phn tch sc k lng kt ni khi ph (HPLC-MS) Tin hnh phn tch HPLC-MS mu tanin trong dung mi metanol-nc ti
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00 /1 0

20 /8 0

Nh vy, iu kin ti u cho qu trnh chit tch tanin t v keo l trm l: nguyn liu dng bt, t l nc: etanol 50 : 50 ti nhit 800C, thi gian ti u l 75 pht, t l rn: lng 2g : 70ml. Vi iu kin ny th lng tanin tch c l 21,60%. so vi nguyn liu v kh.

22 21 20 X(%) 19 18 17 16 15 14

TP CH KHOA HC V CNG NGH, I HC NNG - S 3(44).2011

Vin Ha hc cc hp cht thin nhin Vin KH&CN Vit Nam thu c kt qu trnh by hnh 7.

Hnh 7. Ph sc k HPLC-MS ca tanin v keo l trm

T kt qu ph HPLC-MS, kt hp vi mt s d liu v ph chun ca mt s hp cht tanin cho php d on s c mt mt s hp cht polyphenol nhm tanin tch ra t v keo l trm c trnh by bng 1.
Bng 1. Cc hp cht polyphenol nhm tanin trong v keo l trm

Cu t 1- Rettime: 4,4 min M = 472 CTPT: C22H16O12 3 - 0 - galoyl epigallo catechin

OH HO

Cng thc cu to
OH OH O

OH OH O OH O

OH
OH

2- Rettime: 4,9 min

OH

M = 414 CTPT: C21H18O9 3-0-pyrocatechin-gallo catechin

HO O

O OH

OH

OH OH

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3- - Rettime: 20,5 min M = 442 Cng thc phn t : C22H18O10 Epicatechin gallate(ECG)
OH O HO O

OH OH

O OH

OH OH

4- Rettime: 24,3 min M = 576 Cng thc phn t : C30H24O12 Proanthocyanidin

5- Rettime: 25,4 min M = 578 Cng thc phn t : C30H26O12 Epicatechin-(4 ->8)-epicatechin

4. Kt lun - V keo l trm c cha hai loi polyphenol nhm tanin: Tanin thy phn v tanin ngng t. Ch s Stiasny trong tanin rn tch ra l 69,09. - iu kin ti u cho qu trnh chit tch tanin t v keo l trm l: kch thc nguyn liu dng bt, t l nc: etanol (%V) 50 : 50 ti nhit 800C, thi gian ti u l 75 pht, t l rn: lng 2g : 70ml. Vi iu kin ny th lng tanin tch c l 21,60%. so vi nguyn liu v kh. - Kt qu phn tch ph IR v HPLC-MS cho thy tanin tch t v keo l trm c cc dao ng chnh: -OH, C=O, C=C thm, = C O C, - C O C, CH benzen th
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para, CH thm v gm cc hp cht polyphenol thuc nhm tanin thy phn v tanin ngng t. - V keo l trm l ngun nguyn liu t nhin chit tch polyphenol nhm tanin ng dng trong cng nghip tng hp keo, lm cht thuc da... TI LIU THAM KHO [1] K. Van Langenberg, W. Grigsby and G. Ryan, Green Adhesives (2010): Options for the Australian industry summary of recent research into green adhesives from renewable materials, Forest & Wood Products, Australia. [2] Mohd. Nor b. Mohd. Yusoff, L.T. Chew (1998), Mangrove tannin as an adhesive for wood-based panels, Journal of Tropical Forest Science 1(2): 97-102. [3] M.T.Paridah & O. C. Musgrave (2006), Alkaline treatment of sulfited tannin-based adhesive from mangrove to increase bond integrity of beech slips, Journal of Tropical Forest Science 18(2): 137-145. [4] rika da S. Ferreira (2008), Use of Tannin from Pinus oocarpa Bark for Manufacture of Plywood, Proceedings of the 51st International Convention of Society of Wood Science and Technology, November 10-12, 2008 Concepcin, CHILE, Paper WS-22 [5] G. Kay Nemlu, G. Rsel .Olakoulu (2005), Effects of Mimosa Bark Usage on Some Properties of Particleboard, Turk J Agric For 29, 227-230. [6] Ann E. Hagerman (1998), Tannin Chemistry, Department of Chemistry and Biochemistry, Miami University, USA. [7] Roland E. Vetter (1996), Mangrove bark: A renewable resin source for wood adhesives, Acta Amazonica 25(1/2), 69-72. (BBT nhn bi: 06/05/2011, phn bin xong: 10/05/2011)

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