Tgp chi phdn tich Hda, Ly vd Sinh hgc - Tap 19, So 2/2014
TONG HOP, NGHIEN CUtJ VA THAM DO HOAT TINH SINH HOC CilA PHtrc CHAT HON HOP EUROPI
AXIT L-GLUTAMIC, O-PHENANTROLIN
DintdasognlO-9-2013
Le Hiru Thiing, Nguyin Thi Hoai Anh, Ngo Thi Hoa Khoa Hda hgc, Tru&ng Dgi hgc Suphgm Thdi Nguyen
SUMMARY
SYNTHESIS STUDY AND APPROACH ON BIOACTIVITY OF THE MIXED LIGAND COMPLEX OF EUROPIUM WITH L-GLUTAMIC ACID O-
PHENANTROLINE
The mixed Hgand complex of europium with L-glutamic acid and o-phenanthroline was synthesized in ethanol aqueous solution. The structure of the complex has been recognized by the elemental analysis, IR spectra and thermal analysis methods. The complex have the formula Eu(Glu)iPhenCli.3H20. The antibacterial activity of complex was tested in vitro against three Gram-positiveamd, three Gram-negative bacterial and one type of fungus. The results of antibacterial activity test shows that this complex exhibit excellent antibacterial ability against Stcphylococcus aureus. Bacillus subtilis, Lactobacillus fermentum, Salmonea enterica, Escherichia coli and Candida albicans.
Keyword: mixed Hgand complex. L-glutamic acid, o-phenanthroline. europium.
1. MCi D A U tham do hoat tinh sinh hpc ciia phiic chit hfln Hien nay, phiic chit cua cac nguyen tfl dit hpp Eu (HI) axit L-glutamic, o-phenantrolin.
hiim (NTDH) vdi cac aminoaxit 4rpc quan 2 THUC NGHIEM tam nghien cuu Wong ddi rpng rai do chiing cd ^.1. Tdng hop phiic chit
hoattinhsinhhpc[l,2.3,4.5,6,7].Tuynhien Hda tan axit L-glutamie (Giu) va o- phiic chit hdn hpp tao nen giiia NIDH, phenanUolin (phen) (theo tile moi Giu: phen = aminoaxit va phdi hi huu CP ttung hda edn it 3 ^ i)taigdungmdinudc-etanol(l: l).Dun dupc nghi&i Cliu, dac biet la hoat tmh sinh hpc ^^ ^^ ^^ ^^ ^^^ ^^ ^ ^ ^ ^ ciia chiingTrong bai bao nay, chiing tdi ttinh Them tir tir dung djch EuCh ttxing etanol vao
hdn hpp tten (theo ti le moi EuCls: Glu = 1:3), dimg dung dich NaOH loang dieu chinh pH din 6,5 dung hfli luu ttxxig 6 gid. Cd each diily hdn hpp phan ling den khi xuit him vang be mat. Di ngudi cac tinh thi phiic se tach ra. Lpc nia phuc chit bing axeton, bao quan ttong binh hiit am[8].
2,2. Cac phuong phap n ^ e n cuu Ham luong (%) Eu ttong phiic chat dupc xac dinh bing phuong phap chuan dp Complexon, dung djch chuan dp DTPA chi thj asenazo (E) 0,1 %i dung djch dem cd pH
= 42.
Ham lupng (%) C, N ttong phiic chit duoc xac djnh bing phuang phap khfli pho plasma cao tin cam ling (ICPMS) tten may phan tich nguyen tfl Truspec - CNS Leco (My).
Phfl hap thu hdng ngoai aia cac phdi tu va phiic chit ghi tien may Mangna IR 760
Sepcttometer ESP Nicinet (My). Cac miu dupc ttpn diu, n ^ i n nhd va ep vien vdi KBr,
^ phd ttang vimg tin sd tir 400+4000 cm"'.
(Jian do phan tich nhiet ciia phiic ehit 4ipc ghi ttea may DTG - 60H Shimazu (Nhat). Tic dp nang nhiet ciia mau la 10°C/phut, ttaig khflng khi, khoang nhiet (K tir 30 - 800''C.
Thu boat tinh khang sinh cua cac phfli tii, mufli ciia Eu va phiic chit dupc thuc hioi ted Vien Hda hpc - Vien Han lam Khoa hpc va Cdng nghe Viet Nam, theo ihuong phap khuich tan tren thach, mdi ttircmg Mueller - Hinton vdi 7 chung khuan bd sung dung djch vi smh vat kiem djnh nflng dp 5.10' CFU/ml, u d 37°C.
3. KET QUA VA THAO LUAN 3.1. Ket qua phan tich thanh phan phiic chat
Kit qua phan tich thanh phin phiic chat dupc chiradbang 1.
Bdng 1. Hdm lugng (%) cdc nguyen td Eu, C, N cua phuc chdt
Cdng thuc gia thiet
Eu(Glu)3phenCl3.3H20
Eu LT 16,27
TN 16,21
C LT 34,72
TN 34,05
N LT 7,50
TN 6,98 LT: li thuyet; TN: thuc nghiem
Ket qua d bang 1 cho thay ham lupng nguyen td (Eu. C, N) theo thuc nghiem kha phii hpp vdi ket qua tinh theo eflng thiic gia fliiet (H thuyit). Trong cflng thirc gia thiet ciia phiic chat sfl phan tii nudc xac djnh bing phucmg phap phan tich
nhiet.
3.2. Nghien cihi phiic chat bang phirong phap phan tich nhiet
Gian dfl nhiet va kit qua phan tich gian do nhiet ciia phu:c chit chi ra d hinh 1 va bang 2.
Hinhl. Gidn do phdn tich nhiet ciia phuc chdt
Bdng 2. Ket qua phdn tich gidn d6 nhiet cika phuc chat
Phuc chit
Eu(Gln)3phenCl3.3H20 Nhiet dp pic
CO 125,03
482,24 Hieu
ling nhiet Thu nhiet
Tda nhiet
Dp giam khdi lupng(%) LT 5,787
80,003 TN 6,289 37,867 30,642 4,916 79,714
Dudoan cau tli tach
ra hoac phan huy
3H2O
chay va phan hiiy phiic chat
Dudoan san pham cudi cimg
EU2O3
(-) Khong xac dinh ; LT:
Nghien cuu gian do nhiet cua phuc chat, cho thay tren dudng DTA cd mot hieu ung thu nhiet tai 125,03°C va mpt hieu umg tda nhiet tai 482,24°C.
Qua tinh loan do giam khoi luong phuc ch4t tren dudng TGA (hinh 1) cho thay d hieu ung thu nhiet cd xap xi ba phan tu nudc tach ra. Nhiet do tach cac phan tii nudc thap va thupc khoang nhiet dp tach nudc ket tinh cua cac hpp chat, chung td nudc cd trong phuc chat la nudc ket tinh.
li thuyet; TN: thuc nghiem
Tai hieu ung tda nhiet (482.24°C) ung vdi qua trinh chay va phan huy phuc chit.
Nhiet dp phan huy thap, chiing td phuc chat kem ben nhiet.
Ci nhiet dp cao hon nhiet dp ciia hieu iing tda nhiet, dp giam khdi lupng ciia phuc chit khdng dang ke, cd the d day da cd su hin thanh ueropi oxit (EU2O3).
3.3. Nghien cuu phirc chat b^ng phuong phap pho h i p thu hong ngoai
; !
^1"
.I'll
'ri^jli
Hinh 2. ?/id' /ia> * « hdng ngoai ciia axil Hmh 3. Phi hdp thu hong ngogi cia
L-glutamic o-phenantrolin Phd hip thu bflng ngoai ciia axit L- cac dai hap thu dac tnmg ttong cac pho
glutamic, o-phenanttolin va phiic chit hdngngoaiduatlieo tai lieu [8].
dupc chi ra d cac hinh 2. 3, 4. Su quy ket
Bdng 3. Cdc tdn so hdp thu dgc trung (cm'') ciia axit L-glutamic, o-phenantrolin vd phitc chdt
Hpp chit Axit L-glutamic
o-pbenantrohn Eu(Glu)3phenCl33H20
^,°"'
3524,14
"2"'
30743
2977,25
^™;
2742,04
2891,81 yT' 1512,60
1558,10 V/™"
1259,43
1309,61 v^-c
164238 1634,77
^c=*
1584,01 1609,58 (-) Khflng xac djnh
Trong phfl hdng ngoai ciia axit L-glutamic, dai hip thu d cac tin sd 3074,20 cm'';
2742,04 cm"' quy ket tuong irng cho dao ddng hda trj bat ddi xiing va ddi xiing
ciia nhdm "NH3. Cac dai hip thu d cac tin sfl 1512,60 cm'' va 1259,43 cm'' quy ket lin lupt cho dao dpng hda ttj bat dfli xiing va dfli xiing ciia nhdm COO'.
2000
*(cn>-1) EuCglu)3pfianCI3 3H20 KBr
Binh 4. Pho hdp thu hong ngogi ctia phuc
Trong phd hdng ngoai ciia o- phenantrolin, cac dai hap thu d cac tan sd 1642,38 an"'; 1584,01 cm'^ quy ket lan lugt cho dao dpng hda trj ciia C = C v a C = N.
So sanh pho hdng ngoai ciia phuc chat (hinh 4) vdi pho hdng ngoai ciia axit L- glutamic d trang thai tu do (hinh 2) cho thay cac dai hap thu 3074,20 cm"' va 2742,04 cm"' dac trung cho dao ddng hda tri bat ddi xung ( vJ"') va ddi xung (Vj'^"') ciia nhdm NH3* tren pho axit L- glutamic tu do dich chuyen \ e cac vdng tan sd tuong ung 2977,25 cm'';
2891.81 cm"'. Dieu nay chung td axit L- glutaraic d5 phdi tri vdi ion Eu^~ qua nhdm amin. Cac dai hap thu d 1512,60 cm''; 1259,43 cm'' dac trung cho dao ddng hda tri bit ddi xung ( v ^ ) va ddi xung {v^^'^ ) ciia nhdm COO' dich chuyen
len cac viing tan sd cao hon tuong ung 1558,10 c m ' ' v a 1309,61 cm"\
chung td axit L-glutamic cung da phdi tri vdi ion Eu^^ qua nguyen tu oxi cua nhdm cacboxyl.
Ngoai ra, tren phd ciia phuc chat cdn xuat hien dai hap thu d tan sd 3524,24 cm"' dac trung cho dao dpng hda tri ciia nhdm OH' (v"^^ ) ciia nudc. Dieu nay chung td trong phuc chat cd chua nudc va hoan toan phu hop vdi ket qua nghien cuu phuc chat bang phuong phap phan tich nhiet d phan tren.
So sanh phd hdng ngoai ciia phuc chit (hinh 4 ) vdi hdng ngoai ciia o- phenantrolin d trang thai tu do (hinh 3) cho thay cac dai hap thu d 1642,38 cm' ' dac trung cho dao dong hoa tri ciia C = C (v*^^*^) va d 1584,01 cm'' dac tnmg cho dao ddng hoa tri cua C = N
(v<:'") tten phfl o-phenanttolin tit do K^t q"* dem thu vdi 4 mlu gdm: 2 mau dich chuyin vi cac viing t i n s d tuong P ^ i tu (axit L-glutamic, o- img 1634 77 cm'' va 1609,58 cm'' phenantrolin), 1 man mudi kim loai Diiu nay chiing td o-phenanttolin da (EuClj) va I mau phuc chit tren 3 ddng phdi tri vdi Eu'* qua hai nguyen hi vi khuan gram (+); 3 ddng vi khuan gram (-) va I ddng nam dupc chi ra 6 3.4. Thu hoat tinh Hiang khuin, khang ^ ^ S ^•
nam ciia cac phoi tv, muoi ldm loai va phuc chat
Bdng4Kit qud thii hogt tinh Ididng khudn, khdng nam cua axit L-glutamic, o-phenantrolin, mudi europi vd phiic chdt
Ten chiing vi sinh vat kiem dinh
Gram Gram
(- )
Nam
Staphylpcoccus aureus Bacillus subtilis Lactobacillus fermentum
Salmonea enterica Escherichia coli Pseudomonas aeruginosa
Candida albicans
Ten mau
L-glutamie EuCl3 o-phenanttolin Phiic chat GiatrjIC5o(/^/m/)
>128
>I28
>128
>128
>I28
>128
>128
>128
>128
>128
>I28
>128
>128
>128
6,67 20,12 113,78 11,00 17,19 96,43
20,40
23,48 75,59 92,40 71,53 20,37
>128
101,81
* Tir cac gia ttj ICjo pgtml trong bang 4 cd the so bd nhan xet:
Hai dung djch axit L-glutamic va mudi EuCl3 cd ndng dp gay chet 50% (IC50) vupt qua ndng dp thu cue dai nen cd thi coi la khdng cd kha nang khang khuin va khang nim, o-phenanfrolin cd kha nang khang dupe 7 chimg khuin va nim dem thii.
Phiic chat cd kha nang khang vdi loai vi khuan Staphylococcus aureus (IC50' 23,A%ijglml), Bacillus subtilis (IC50 • 75,59/g/m/), Lactobacillus fermentim (IC50 92,40/g/m/), Sahnone enterica(IC5o 71,53 i^lnd\
Escherichia colt
(.lCn,= 20,37 f^/ml) vi Candida albican
^•"^^ (IC50 = 1 0 1 , 8 8 / ^ / m / ) . Kha nang khang khuan ciia phuc chat doi vdi vi khuan Lactobacillus fermentum tdt hon so vdi o- ''^ phenantrolin, 5 loai vi khuan va nam cdn
^"^^ lai thi ySu hon o-phenantrolin.
4. K E T LUAN
Da tdng hpp duoc phuc ran ciia Eu^"^ vdi axit
L-glutamic va o-phenantrolin.
^ija^ Bang cac phuong phap: phan tich nguyen td, phan tich nhiet v^ quang ph6 hap thu hong ngoai cd the k^t luan:
— — r • Phuc chat co thanh phan antrolin Eu(Glu)3phenCl3.3H20.
L • Mdi phan tu axit L - glutamic chi^m ' 2 vi tri phdi tri trong phiic chat, lien ket vdi 7 lion Eu^^ qua nguyen tii nito cua nhom lamin (-NH2) va qua nguyen hi oxi ciia ciia ]nhdm cacboxyl (-C00"). Phan hi 0-
"phenantrolin cung chiem 2 vi tri phdi tri Trong phiic chat, lien ket vdi Eu^^ qua 2 -nguyen tu nito ciia di vong.
*K6t qua thii hoat tinh khang khuin, khang T i l m cho thiy phiic chit cd kha nang khang
duac 6/7 chiing khuin va nam dem thu.
78
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