XC NH ACETALDEHYDE TRONG ETHANOL NHIN LIU BNG

HPLC VI U D IN HA
Chu trnh hot ng in phn ca acetaldehyde v sn phm dn xut vi
dinitrophenylhydrazine 2,4 (DNPHi) c nghin cu mt in cc carbon
thy tinh. Nghin cu ny c s dng ti u ha cc iu kin th nghim tt
nht xc nh chng bng phng php tch sc k lng hiu nng cao (HPLC)
cng vi u d in ha. Acetaldehyd 2 ,4-dinitrophenylhydrazone (ADNPH)
c rav tch ra bng ct pha o C18, di iu kin ng th vi pha ng
cha hn hp 2 cht methanol/LiCl(aq) nng l 1.10-3 M (80:20 v/v), tc
dng chy 1,0 mL min-1.iu kin ti u u d in ha ca
ADNPH l 1,0 V so vi Ag / AgCl nh mt in cc tham kho. Phng php
c xut n gin, nhanh chng (phn tch thi gian 7 pht) v nhy(gii hn
pht hin l 3.80 L-1) ti mt t l tn hiu: nhiul 3:1. N cng rt chn lc
v c lp li.[ lch chun 8,2% 0,36 (n = 5).ng cong phn tch ca
ADNPH tuyn tnh trn phm vi 3 - 300 mg L -1cho mi ln bm mu(20 v
lp li ca php phn tch l >99%.
GII THIU
Ti Brazil, vic s dng ethanol nh mt nhin liu t hoc mt ph gia
oxy ha xng du tng ln ng k t nm 1975 l kt qu ca Chng trnh
Ru Quc gia a ra nh l mt chin lc tr nn c lp hn v du nhp
khu.Hin nay, sn xut ethanol nhin liu hng nm ca Brazil l khong 14 t
m3/nm, n thuc loi mt nh sn xut ln nht th gii.
S c mt ca cht gy nhim v c v hu c trong nhin liu ethanol
c kim sot mt cch cht ch bi mt c quan qun l lin bang (Agncia
Nacional do Petroleo) theo gii hn cho thnh phn ca tp cht.Mc d, nhng
cht gy nhim c coi l l mt sn phm th cp, nhung chng c nh hng

quan trng c bn khi ng c hin i c s dng, c pht trin vi cng

nghcao, v do i hi nhin liu phi c tinh khit cao. Khi vic sdng
nhin liu ethanol c hn ch, th phng php phn tch c sn s nhanh, chn
lc v nhy nh lng cht nhim hu c tr nn kh khn.
Acetaldehyd l tp cht quan trng nht c thy trong nhin liu ethanol
cc mc khc nhau ca nng , m ph thuc vo qui trnh sn xut, s tch ly
v cc yu t khc.
Mc d tm quan trng ca n c bit n nhiu, nhng c ch l mt vi
bi vit trong cc ti liu c lin quan n cc ngh thay thphng php xc
nh v nh lng ca n, m chng rt quan trng i vi mc ch kim sot
cht lng, chng hn nh phi bit thnh phn ha hc c th.
Hu ht cc phng php HPLC lin quan n acetaldehyd u c da
trn mt phn ng vi dinitrophenylhydrazine 2,4 (DNPHi) ci thin
tnh nhy ca sphn tch cc hp cht cacbonyl. Mc ch ny c thit lp bi
Kissinger et al, pht hin nhng hp cht ny vi u d in ha.Tuy nhin,c
mt phng php HPLC xc nh acetaldehyde trong ethanol nhin liu bng cch
s dng u d in ha c m t.Chng ti bo co v s pht trin ca mt
phng php n gin cho nh lng acetaldehyde trong etanol nhin liu da
trn phng php HPLC vi u d in ha.
THC NGHIM
Thuc th
Tt c cc ho cht s dng ca loi thuc th ACS hoc

tt hn.

Acetaldehyd l t Merck (Darmstadt, c). Tt c cc dung mi c s dng, nh

ethanol, methanol v acetonitril, phi c cp t Mallinckrodt HPLC (Xalostoc,
Mexico). Nc kh khong c thu thp t mt H thng nc Milli-Q
(Millipore, CA, USA). 2,4-dinitrophenylhydrazine (DNPHi) t Merck (Darmstadt,
c) c tinh ch bng ba ln recrystallizations lien tip tmethanol.H 3PO4 l
t Aldrich Chemical Co (Milwaukee, M). LiCl v LiOH, thu c t Aldrich

Chemical Co (Milwaukee, M), c s dng h tr in phn trong nc

kh khong.
Dn xut acetaldehyd-2 ,4-dinitrophenylhydrazone (ADNPH)
ADNPH c s dng nh l mt cht chun c chun b t phn
ngc bit n nhiu

ca cc hp cht cacbonyl vi 2,4-dinitro-

phenylhydrazine (DNPHi), thu c nh m t. DNPHi (0,4 g; ca. 2 mmol) c

ha tan trong axit photphoric m c (2 mL) vnc kh khong (3 ml). vi s
ha tan ny acetaldehyde chun (1g) c ha tan trong methanol (15ml) c
thm vo. Sn phm phn c tch ra thng qua lc v c lm tinh khit (hai
ln) bng recrystallization t ethanol tinh khit.
Dung dch chun c chun b bng cch ha tan tng trng lng (1g L -1)
ca ADNPH tinh khit trong acetonitrile. Thnh phn ca dung dch ny sau
c pha long vi hn hp ethanol v nc thch hp(95:5 v/v) ti thiu ha
nh hng nn, cng nh lm dung dch chun cho mc ch hiu chun, chun b
cho dy nng t 3 300 mg L1.
Dn xut mu
Mt dung dch 0.4% ca DNPHi c chun b bng cch ha tan DNPHi (0.4g)
bng acetonitrile (100ml). Trong mt erlen cha 0.9ml ca dung dch DNPHi,
4.0ml mu (c pha long trc) v 50H3PO4 1.0 mol/L c gii thiu,
lin tc. Sau dung dch ny c em khuy nhit phng trong 20min.
dn xut mu c lc qua b lc Millipore 0.45 v sau
c tim vo h thng HPLC.
Cc iu kin sc k
Mt b my HPLCProstar Varian c s dng di iu kin ng th vi mt
pha ng cha hn hp methanol/LiCl(aq)(1,0.10-3M 80:20 v/v) Mt van tim mu
Rheodyne 7725 vi vng lp mu 20 uL c s dng, mt ct pha o vi ct
nhi Shimadzu Shim C18 (150 x 6mm i.d. 5um) c p dng hu ht cho cc php

o vi mt ct bo vShimadzu Shim C18.

u d in ha (Varian ProStar 370) da trn nguyn l wall-jet, bao gm
mt in cc hot ng cacbon thy tinh, in cc so snh bc bc clorua v mt
in cc i khng c s dng. Trc khi s dng dung mi pha ng phi c
lc qua b lc 0.45um (Millipore) v c kh vi kh nit tinh khit cao. u d
in ha c ci t 1.0 V v tc dng l 1.0uL/min.
Cc iu kin in phn
Php o in phn bng cch s dng potentiostat / galvanostat (EG & G
Princeton Nghin cu ng dng Corp (PARC), Model 173)
Mt pin in th cha ba ien cc c s dng mt in cc carbon thy
tinh (Methrom, din tch = 0,0314 cm 2) nh in cc hot ng, mt si dy bch
kim nhin cc i khng v mt in cc calomel bo ha nh in cc so
snh(Methrom).in cc hot ng c nh bng gia th nghim vi 0.05um
bn nhm, v ra sch vi nc kh khong.
Cc thng s my in th c cn bng trong 15s v qut vi tc
50mL/s, dung dch ca LiCl (1 mol/L), v LiOH(1 mol/L) cng nh
methanol/LiCl(aq)(1 mol/L,50:50 v/v) v acetonitrile/ LiCl(aq) (1 mol/L, 50:50 v/v)
c s dng h tr in phn. Tt c cc dung dch c lm sch vi nit tinh
khit cao trong 10min trc khi tin hnh ghi in th v dng nit tip tc i qua
dung dch trong sut qu trnh o.
Phn tch nh lng
S chuyn i hm lng acetaldehyt trong etanol nhin liu thnh dng dn
xut (ADNPH) t c bng cch s dng mt lng d ln DNPHi 4-6. ng
cong tiu chun cho ADNPH t c bi s hi quy tuyn tnh, gin din tch
peak t l vi nng . Cc h s tng quan gn nh ng nht.

Phn tch mu etanol nhin liu

Cc mu etanol nhin liu thng mi c sn khc nhau c thu thp xung
quanh Araraquara, So Paulo, Brazil.Nhng mu ny c lu tr trong cc bnh
thy tinh Pyrex nhit kim sot 20oC trong tt c cc php o.thnh phn
acetaldehyt trong nhng mu ny c dn xut v phn tch c tin hnh trn
h thng HPLC.
KT QU V THO LUN
Phn tch HPLC cho cht chun ADNPH
Khng c vn v s hp th trn b mt in cc c tm thy trong
khi o acetaldehyt cho s pht sinh oxy ha v s gim thy ng lc in th
c m bo. c c php o phn tch cho cc in cc hot ng trn cc
h thngHPLC, n cn thit phi nh bng b mt ln u mt cch cn thn vi
0.05 um bn nhm v trc mi in th khc nhau n cn thit l mt in ha
sch chng trnh c t 1,0 v -1,0 V trong 20 giy.
nh hng ca in th ln thy ng lc in th c ghi li cho cht
chun ADNPH 50 mg/ L cc in th lm vic khc nhau s dng mt pha ng
cha methanol/LiCl(aq)(1,0.10-3M 80:20 v/v) c ch ra trong hnh 1.Tuy nhin,
ng cong qu trnh oxy ha thu c cho ADNPH theo cc iu kin th nghim
m t trn, c th hin trong Hnh 1a.Mc d mt tn hiu nh c quan st
thy 0,8 V,nhng gi tr ti a ca peak hin ti c quan st thy mt in th
cao hn 1,0 V.in th lm vic ti u choqu trnh oxy ha ca ADNPH c la
chn vo khong 1,0 V.Ti in th cao hn 1,2 V thy c s gia tng ca nh
hng nn do lm tng n. S gim thy ng lc in th tng ng (hnh
1) ch ra khng c tn hiu ng k di mc gi tr -0.7 V, nhng tn hiu ln
nht nhn c in th cao hn -0.9Vni c hai nhm nitro c th c gim.
chn u d in th ti u, l mong mun ti a ha phn ng hin
thi v ng thi ti thiu ha nh hng nn.Trong trng hp ca ADNPH, chai
in th-0,8 V hoc 1,0 V u c th c s dng t thy ng lc hc in th.

Ni v s xem xt ny, sc k cho cht chun ADNPH 50 mg/ L c ghi li khi s

dng mt ct pha o (Shimadzu Shim-pack C18) tin hnh di iu kin ng
th, s dng pha ng lmethanol/LiCl(aq)(1,0.10-3M 80:20 v/v) ti tc dng l
1mL/min. Sc ph thu c khi in th c mt gim mt s gp phn cao ca nh
hng nn do c s can thip ca oxy ha tan khng c loi b trong thi gian
di iu kin desaeration.Nh vy, ti u ha thng qua kh nng oxy ha
c nghin cu.
Hnh 2 cho thy s phn tch sc k in hnh cho cht chun ADNPH vi
u d in ha. Nh vy cho thy s phn tch sc k cho ADNPH t tt iu
kin th nghim ti u, v s ra gii ca hp cht nghin cu hon thnh trong
thi gian t hn 7min trong mt qu trnh chy sc k. S xc nh peak sc k da
vo thi gian lu tr (5.8 min) v c xc nhn bi xc thc dung dch chun
ca ADNPH.
ng chun phn tch ca u d in ha, da trn din tch peak t l vi
nng , c u tin cho nhng peak cao caADNPH do kh nng ng c trn
b mt in cc, chng lm sai lch kt qu nh lng.Cc ng cong phn tch
cho ADNPH c xy dng bi biu ca din tch peak vi nng , v mi
quan h tuyn tnh t c t 3300mg/L vi cc thng s nh sau: din
tch=1.52 x 105+ 0.1 x 105[ADNPH] vi r2=0.9998. Gii hn pht hin (LOD) xc
nh nng c tim thp nht c thc hin t l tn hiu: nhiu bng 3:1
l 3.8ug/L.
Xc nh acetaldehyt trong mu etanol nhin liu
S dng iu kin th nghim tt nht nh nghin cu trn, nhiu mu
etanol nhin liu t nhiu ngun khc nhau c x l bng DNHPi, chng c
phn tch bng HPLC vi u d in ha c iu chnh 1.0V t c php
tch tt nht. Bn hnh 3 ch ra mt php tch sc k tiu biu t c t mt
loi mu etanol nhin liu thng mi. S c mt DNHPi nng tng i cao
khng gy ra bt nh hng r rng no ln tn hiu ca sn phm ADNPH c

to ra sau phn ng, chng c tch t mi phn ng khc. Ngoi ra nhng

peak tng ng vi ADNPH,c mt thm mt vi peak khc v xc nh hu ht
chng. Nhng peak xut hin thm c cho l s c mt cu cc hp cht
cacbonyl. Thm vo , phng php ny c kim tra cho nhiu mu etanol
nhin liu, kt qu nh lng c lin quan c lit k bng 1.thng thng
thnh phn acetaldehyt trong etanol nhin liu kh cao, v im c gi tr nng
cao ca acetaldehyt trong nhin liu thay i nng t 83 343mg/L . Nhng
pht hin phn nh nhng kh khn lin quan vi mt hn hp nn.
Hi quy c nh gi cho ADNPH bng cch s dng cc dung dch dn
xut chun ca ADNPH mc khong 25% php o v thc hin nm ln phn
tch sau mi ln b sung (Bng 1). Nng ca ADNPH c tnh ton da vo
din tch peak. N c tnh ton da trn phng trnh hi quy tuyn tnh ca
phng php thm chun.
lp li ca phng php ny c xc nh bng cch tin hnh nm
th nghim trn cng mt mu hn hai ngy trong 1 bn. Gi tr ca lch chun
thp (8,2% 0,36) v h s bin i l 3,2%. Tuy nhin, phng php xut
cho thy chn lc v nhy xc nh chng.
KT LUN
Mt phung php sc k lng hiu nng cao vi u d in ha c ti u
ha xc nh acetaldehyt trong mu etanol nhin liu nghin cu thnh cng.
c bit u d in ha th hin nhy hn v tnh chn lc cao hn, iu rt
hu ch khi phn tch mu tht, bi v chng lm gim nh hng nn ,do ci
thin vic nh lng acetaldehyde.V vy, pht hin ny chng ti a ra mt
phng php nhanh chng, ng tin cy v n gin, cho nh lngv xc nh
chng trong ethanol nhin liu.