xay dung h© tti6ng«eac c^i s6,«ban ttinh phan tich va phuong phdp Odnh gia nhan dgng ngudn g6c ddu; xdc djnh chi s6 nhan dgng cho nnat s6 ddi tupng ddu fhd Viet Nam CN Phan Nhu Dinh KS Ngo Mai Anh KS[.]
Trang 1xay dung h© tti6ng«eac c^i s6,«ban
ttinh phan tich va phuong phdp Odnh gia nhan dgng ngudn g6c ddu; xdc djnh chi s6 nhan
dgng cho nnat s6 ddi tupng ddu fhd Viet Nam
CN Phan Nhu-Dinh
KS Ngo Mai Anh
KS Nguyin Van Mai
Vien Diu khi Viet Nam
Ngay nay, 6 nhiem moi trw&ng dang tr&
thanh vin de mang tinh toan ciu, trong do
6 nhiem moi trw&ng do diu mo va eae san
phim diu mo la mot trong nhO'ng vin de
dwgv xa hgi quan tam ngay cang nhieu
& Viet Nam, nganh cong nghiep Diu khi
da va dang phat then khong ngirng, ben canh nhO'ng Ig'i ich do dau mo dem lai eOng phai ke den nguy ea gay 6 nhiim moi
trw&ng sinh thai mot each trwe tiep hoae gian tiep tip cac hoat dgng tham do, khai thae, van ehuyen, tang trip va sir dung diu
mo IVIgt trong nhO'ng vin de dang quan tam
ve 6 nhiem moi trw&ng trong nhO'ng nam vwa qua ia sw xuit hien cac vet diu tren bien, tren song hay eae bai bien lien tue bi
6 nhiem b&i eae mang diu troi dat vao biy, nghiem trgng han la eae vu trim diu khong
ro nguon goe gay nhieu kho khan trong eong tae quan ly, ipng eipu, khae phue h$u qua eOng nhw boi thw&ng thiet hai
Khi xay ra sw eo diu tran, mot trong nhwng eong viee can phai khin trwang thwe hien la xae djnh khu vwe bj 6 nhiim, tim ra doi twg'ng va nguon goe diu 6 nhiem Tuy nhien, do sw eo xay ra da lau m&i dwoc phat hien, lue nay nguon va doi twang gai
6 nhiim da each qua xa trong khi cat nguon eo kha nang gay 6 nhiim thi nhiei nen rit kho tim ra doi twg'ng gay 6 nhien Dieu kha pho bien la eae dan vj hay ea nhai gay 6 nhiem diu it tw giae nhan traeh nhi^n
ve minh nen eae nha chipe trach can
Trang 2P E T R O V I E T N A M
• ' # ^ ' '
tien hanh xae minh nguon goe
va dae trwng eua diu 6 nhiem
bing nhO'ng bang ehwng khoa
hge mang tinh thuyet phue
Viec nhan dang nguon goc dau 6 nhiem la rit kho vi thanh
phin ciu thanh eua diu rit
phirc tap, khi tiep xue v&i moi
trw&ng de bj phong hoa va trgn
lin v&i eae thanh phin chit hiru
ea trong tw nhien hoae v&i eae
loai diu khae, dae biet la nai da
tirng xay ra 6 nhiem thw&ng
xuyen nhw: Khu vwe eiu cang,
cipa song cira bien, khu vwe gin
kho ehira diu va ehe bien diu
Vi vay phai lwa ehgn mot he
phwang phap phan tieh thieh
hgp va phai eo doi twg'ng lam
ehi thj de phan tieh so sanh
Hien nay tren the gi&i da eo rit nhieu phwang phap nhan
dang nguon goe diu twang doi
tin eay, mot so phwang phap da
dwg'C xay dwng thanh tieu
ehuin Tuy nhien eae phwang
phap khong phai la phu hg'p eho
tit ea eae loai diu ma irng v&i
mil loai ehi eo the phu hg'p mot
hoae mot vai phwang phap nhan
dang ma thoi Vi du phwang
phap phan tieh Vanadi va Niken
(V, Ni) la rit hieu qua doi v&i eae
loai diu eo ham Iwg'ng V, Ni eao,
nhwng lai kho sir dung de phan
biet eae loai diu tho Viet Nam
v&i nhau do ham Iwg'ng eae kim
loai nay trong hiu hit diu tho
Viet Nam eo gia trj rit thip (xip
xi v&i gi&i han phat hien) hoae
doi v&i phwang phap phan tieh
biomarker eho diu tho Viet Nam
bit bugc phai thwe hien them
bw&e taeh loai parafin trong khi
phin l&n eae loai diu tho khae
CO the ap dung qui trinh phan
tich it phipc tap han, mot so ehi
so thw&ng eo ham Iwg'ng qua
nho dan den do lech ehuin
twang doi eua no trong cae lin
phan tieh lap khong dat, nen ein phai lwa chgn phwang phap va eae ehi so phan tieh phu hg'p va phai CO mot qui trinh phan tieh hg'p ly
Ben canh do viee tim ra miu diu doi chirng de tien hanh phan tieh nhan dang eOng gap nhieu tr& ngai nhw la khong khoanh vung dwg'C doi twg'ng gay 6 nhiim trong hang ngan doi twg'ng nghi ng& nen viee truy tim nguon goe phai thwe hien tren nhieu doi twgng, viee nay se tin rit nhieu th&i gian va ehi phi Do vay viee "Xay dwng he thong eae ehi so, hoan thien qui trinh phan tieh va phwang phap danh gia nhan dang nguon goe diu; xae djnh ehi so nhan dang eho mgt
so doi twg'ng diu tho Viet Nam"
la viee lam mang tinh cip bach
va thiet thwe, eo the ap dung vao thwe tien nhw sau:
+ Co dwgx: ea s& va phwang
phap nhan dang nguon goe diu
on djnh va tin eay
+ He thong eae ehi so nhan
dang lam ea s& doi chirng v&i miu diu 6 nhiim ein xae djnh nguon goe
+ Khoanh vung doi twg'ng
nghi ng& gay 6 nhiem nhanh chong va loai bo cae doi twg'ng
vo can di han ehe s6 mau phan tieh tiet kiem dwg'C th&i gian va chi phi
+ Lam cas&phan biet nhanh
nguon goe diu tran xuit xir tir nw&e ngoai
+ DOng lam tai lieu tham
khao trong eae de tai nghien ewu khoa hge eo lien quan Ngoai ra nghien ciru nay eon mang y nghia l&n trong cong tac quan ly va bao ve moi trwang cho nganh noi rieng va cho quoe gia noi chung
DAU KHi - s o 7/2010 4 7
Trang 3A N T O A N - M O I T R U d N G D A U K H I
'.^::'(i/trfti
'^<um^j^ffA^^rim^^(^^f^^^-^
1 Phu'O'ng phap nhan dang nguon goc dau va he
thong cac chi so nhan dang
Phuang phap nhan dang dau d nhiem dya tren
"Phuang phap tieu ehuin cda Nauy da soat xet de
nhan dang d i u tran" (2002), cd tham khao them tieu
c h u i n cua Hoa Ky ASTM D-5739-95, "Phuang phap
tieu ehuan de nhan dang nguon g i c d i u tran bang
phuang phap sac ky khi va k h i i pho ion ehgn lgc"
(1995) Phuang phap nhan dang nguon gde d i u eua
Nauy (Nordtest) dya tren k i t qua phan tieh d i u d
nhiem va dau doi chu'ng vdi s i c ky khi (GC-FID) va
sac ky khoi pho (GC-MS) nham xac djnh cae dae trung
phan bo n-alkane, hydrocarbon tham da vdng, cac
hgp c h i t danh d i u sinh hgc (biomarker) cdng vdi eae
ehi so nhan dang duge tinh toan dya tren cac dac
trung phan bo nay
Cac bude t i i n hanh phan tieh va xd' ly dd' lieu thyc
nghiem de nhan dang ngudn goe d i u bao g i m cac
bude theo luu do (Hinh 1)
Bwac 1:
Sau khi xd' ly, eae m i u d i u d nhiim va dau d i i
chd-ng duge phan tieh vdi GC-FID nham xae djnh:
+ Dac trung phan b i n-ankanes cda cac mau d i u
(td-CIO d i n C35)
+ Cac chi s i ehpn Ipe eua isoprenoid no mach
nhanh
+ K i i m tra qui luat phong hda d i u (bing each
ehuin hda theo cac hgp chit khdng bj phong hda
hoae b i n vO'ng vdi qua trinh phong hda)
Tai bude nay, d i u d nhiem ed the duge so sanh
mdt each djnh lugng va djnh tinh (nhu tl le eae
iso-prenoid va n-ankan) vdi mau nghi v i n , mau nao khdng
trdng khdp vdi nhau se bj loai bd khdi cae bude phan
tieh k i t i i p Tuy nhien, n i u ed bat cd' dau hieu nghi
v i n nao, qua trinh nhan dien se v i n duge tiip tuc thyc hien vdi cae budc k i i m tra t i i p theo
Bwiyc 2:
Bude t i i p theo la phan tieh mau dau d nhiim va
m i u d i u doi chu'ng vdi s i c ky khi - khoi pho d che do chgn lgc ion (GC/MS-SIM) n h i m xae djnh ham lugng
va sy phan bo cac nhdm chat biomarker va cac hgp
c h i t hydrocaebon tham da vdng (PAH) Cae dU kien ban djnh lugng tren ddng de:
+ Xae djnh bd cac chi s i nhan dang duge tinh toan dya tren eae PAH ehgn Igc n h i m danh gia mu'c dp phong hda va nhan dien ngudn goe d i u d nhiim + Xac djnh bg eae ti le nhan dang duge tinh dua tren cae hgp c h i t biomarker chgn Igc (bao gom cac hgp c h i t hopane va cac triterpanes khac, cac ster-anes thdng thudng va diasterster-anes, triaromatic steroids) dung cho viee nhan dang nguon goc + Thiit lap viec kiem tra sy phong hda danh cho cac nhdm PAH ehgn lgc tren
Bwac 3:
+ Danh gia sy tae dgng eua qua trinh phong hoa (dya tren viec k i i m tra qui luat phong hda eua n-ankane d bude 1 va tham ehi la sy phan bo ban djnh lugng eua eae nhdm PAH d bude 2)
+ Danh gia va loai bd cac ti le nhan dien cd sai so thd do sy sai biet giO'a cae k i t qua phan tich va sy khdng dong n h i t eua mau
+ Phan tieh tuang quan: Xac dinh sy dong dang
v i eae chi so nhan dang cua mau d i u d nhiim va miu dau doi chung ed tinh d i n cac sai lech do phan tich de dua ra cac k i t luan theo eae mu'c: G i i n g , ed the giong, khdng the k i t luan, khdng g i i n g
4 8 DAU KHi - s o 7/2010
Trang 4M a u d l i u tran Oac tru'ng ve c a m q u a n
va tinh chat vat ly
Chiet Lam sach m a u
So sanh vol m a u nghi van
Co khac biet ve Sac do va s y phan bo n-ankane?
Khac biet nay co the
do phong hoa?
K h o n g
K h o n g
Kiem tra m u c do phong hoa
Co SU' khac biet ve phan b6
B i o m a r k e r va PAH '^
D a n h gia do c h i n h xac
cua phu'O'ng phap p h a n tich
Khac biet nay co the
do p h o n g hoa?
Khong
K i e m tra mu'c
dp phong hoa
Tl le nhan d a n g
CO trung k h o p ?
Khac biet nay co the
do phong hoa?
G i o n g (Positive match)
I Mot phan | ' | K h 6 n g
Co the giong (Probably match)
K h o n g the ket luan (Inconclusive)
Khong giong <-(No match)
Khong
Khong
Hinh 1 Ltru do phan tich nhan dang
ion 198pg ng? 70 to 198 30) 0-14 D
300DO
28000
26000
24000
22000
2OO00
18000
16000
14000
12000
10000
BOOO
0000
4000
2000
"""m
100 42 00 43 00 44 00 45 00 46 00 47 00 4fl 00 49 00
Hinh 2 Methyl Dibenzothiophene (m/z 198) trong miu dau thd
II He thong cac chi so nhan dang nguon goc dau
1 Nhan dang phan tieh kiem tra bang GC/FID
Viec su- dung GC/FID nhu la phep kiem tra nhanh de khai quat hda dae tinh toan bd eae m i u d i u
S i c ky d i GC/FID cho ta buc tranh tdng quan v i sy phan bo cae hydrocaebon uu the trong tung
m i u (nhu sy phan giai cda td'ng loai n-ankane va cac isoprenoid ehd yeu) Nd eOng eung c i p thdng tin quan trgng ve md'c do phong hda cda mau d i u tran va eae thdng tin n i u cd ve dae diem rieng va cac thanh phan d nhiem neu cd trong
d i u Ddi vdi cae m i u d i u da bi phan hdy sinh hpe, phan b i n-ankane thudng ed cac pic dae trung uu t h i phan bo d i u dan trong toan vdng s i c do
Tl le giua cac n-ankane va iso-prenoid Pr/C17; Ph/C18 va Pr/Ph duge ddng lam cae chi so nhan dang nguon gde d i u
2 Cae chi so nhan dang eua
PAHs
D i u thd d cac md va vdng khac nhau cd the r i t khac nhau ve phan
b i PAH Mpt vai ti le nhan dang duge su dung de nhan dang nguin
g i c d i u (Radke, 1998; Douglas, 1996; Wang et al 1999 va Stout et al., 2002) Nhdm ti le nhan dang PAH thdng dung nhit la dibenzoth-iophenes va phenanthrenes methyldibenzothiophenes (C1-dibenzothiophenes, vdi dai dp soi
tu C18-C19, m/z 198, Hinh 2) thudng hien dien trong dau tho vdi
Trang 5A N T O A N - M O I T R U d N G D A U K h i
Hinh 3 Retene (m/z 234) trong mau diu thd
1 192 001191 70 10 192 301 O- 4 D
Hinh 4 Methyl Phenanthrene (m/z 192) trong miu diu thd
Chi so nhan dang cua PAHs
1
2
3
4
5
6
C 2 - d b t / C 2 - p h e (%)
C3-dbt/C3-phe (%)
C3-dbt/C3-chr (%)
2 - M P / 1 - M P (%)
4 - M D / 1 - M D (%)
R e t e n e / C 4 - p h e (%)
100 * [ C 2 d i b e n z o t h i o p h e n e s / ( C 2
-d i b e n z o t h i o p h e n e s + C 2 - p h e n a n t h r e n e s l
100 * [ C 3 d i b e n z o t h i o p h e n e s / { C 3 -dibenzottniophenes + C 3 - p h e n a n t h r e n e s ) l
100 * [ C 3 d i b e n z o t h i o p h e n e s / ( C 3
-d i b e n z o t h i o p h e n e s + CS-chrysenes)]
100 * [2-methyi p h e n a n t h r e n e / {2-nnethyl phenanthrene + 1-methyl p h e n a n t h r e n e ) ]
100 ' [4-methyl d i b e n z o t h i o p h e n e / (4-methyl d i b e n z o t h i o p h e n e + 1-methyl
d i b e n z o t h i o p h e n e ) ]
100 * [retene/(retene + C 4 - p h e n a n h r e n e s ) ]
ndng dp dang k i Sy phan bo va ti le giu'a cac d i n g phan C 1
-dibenzothiophene thudng khae biet giua eae loai d i u khac
nhau TI le giua cac dong phan Cl-dibenzothiophenes thudng
it bi b i i n doi do sy b i c hai tu qua trinh phong hda cua eae loai
d i u bj phong hoa nhe hay phong hda trong khoang thdi gian
n g i n Vdi Cl-dibenzothiophenes, eae vi khuan thudng cd
khuynh hudng phan ra chu y i u nhit ddi vdi cac d i n g phan
2-/3-methyldibenzothiophene isomers (Wang and Fingas, 1995)
Do vay ti le 4-MD/1-MD dugc chgn lam chi s i nhan dang d i u
Retene (vdi dai dp sdi td' C19-C22, m/z 234, Hinh 3) la mpt
diterpane tham (1-methyl-7-(1-methylethy phenanthrene) ed ngudn goc tu" nhya (acid abietie), va b i n vO'ng vdi qua trinB phong hda Ti le nhan dang giO'a Retene va tong ham lugng C4-phenanthrenes cd the lai mgt nhan td hOu ich trong viec phan biet cac loai d i u ed dac tinh tuang t y nhau (Stout ef
al., 2002)
Methylphenanthrenes (C1-phenanthrenes, vdi nhiet do sdi trong khoang C18-C19, m/z
192, Hinh 4) ed the cung c i p cac thdng tin hO'u ich trong cac vu d i i u tra tran diu, dac biet trong eae trudng hgp lien quan den nguon nhien lieu a giai doan giu'a cua phan doan ehung c i t vdi thanh phin chu yiu la Cl-phenanthrene Sy phan b i cda cae dong phan Cl-phenanthrene trong d i u thd rit dac trung eho ca tudng da hinh thanh diu va mu'c do trudng thanh nhiet cda d i u md Hen nu'a, d i n g phan 2-Methylphenanthrene cung nhay han vdi qua trinh phan ra sinh hoc trong via ehu'a so vdi eae d i n g phan khac (Radke,
1988 Wang and Fingas, 1995) Do vay ti le 2-MP/1-IV/1P duge chgn dung lam ehi s i nhan dang dau ,, Cac hgp c h i t C3-ehrysene thudng rat it
thay doi trong qua trinh phong hda so vdi 03-dibenzothiophene Do vay ty s i C3-dbt/C3-chr thudng it thay doi trong giai doan dau cua
qua trinh phong hda va thudng giam a giai
doan sau cua qua trinh phong hda nen du'gc lam ehi so de danh gia md'c do phong hioa cua dau Do vay, chi sd C3-dbt/C3-chr thudng duge s u dung de lam "ty s i phong hda" Sy cd mat cua chrysene trong dau c6 the ddng de phan biet eae san pham dau va
d i u thd
Nhu vay, ed t i t ca 6 chi so nhan dang dau
ed ngudn goe PAH duge dinh nghTa d bang tren Tuang t y nhu cae nha dja chat dau khi (Weiss et al., 2000), tat ca cae ti le tren (nhu giO'a pic A va pic B) duge tinh toan theo ti le
% dya tren edng thd'c (1);
100%x[A/(A+B)] 0)
3 Cae ehi so nhan dang eua hgp chat
danh diu sinh hoe biomarker
50 DAU KHI - sO 7/2010
Trang 6Hinh 5 Sic dd ion cua hopanes va cactriterpanes nam vdng
no khac (m/z 191) ciia miu diu thd
Ooi tircyng phan tich va l<y hieu cua cac hopanes va triterpanes
nam vong no khac (m/z191)
TT Kl iiieu Doi tipp-ng phan tfch
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
2 0
27Ts
27Tm
28ab
25nor30ab
29ab
29Ts
3 0 d
29ba
3 0 O
30ab
30ba
31abS
31abR
3 0 G
32abS
32abR
33abS
33abR
34abS
34abR
18a(H)-22,29,30-trisnorhopane 17a(H)-22,29,30-trisnorhopane 17a(H),21 (3(H)-28,30-bisnorhopane 17a(H),21 P(H)-25-norhopane 17a(H),2ip(H)-30-norhopane 18a(H)-30-nomeoHopane 15a-methyl-17a(H)-27-norHopane (diahopane) 17P(H)-21a(H)-30-norHopane (normoretane) 18a(H)-oleanane (not present in or rare In oil from pre-Cretaceous source rocks)
17a(H),2ip(H)-hopane 17p(H)-21a-(H)-hopane (moretane)
1 7 a ( H ) , 2 i p ( H ) , 22S-homohopane
1 7 a ( H ) , 2 i p ( H ) , 22R-homohopane Gammacerane (not abundant in North Sea oils)
1 7 a ( H ) , 2 i p ( H ) , 22S-bishomohopane
1 7 a ( H ) , 2 i p ( H ) , 22R-bishomohopane
1 7 a ( H ) , 2 i p ( H ) , 22S-trishomohopane
1 7 a ( H ) , 2 i p ( H ) , 22R-trishomohopane
1 7a(H),21 P(H), 22S-tetrakishomohopane 17a(H),21 P(H), 22R-tetrakishomohopane
Cae biomarker thudng la cac hgp c h i t hydrocarbon ed d khip mgi
nai va xuit hien mgt each t y nhien, hien dien trong d i u thd va phin
Idn cac san pham d i u md Chung la cac dang hda thach d muc phan
tilp, ed nguon g i c td' cae loai sinh vat
CO Cac biomarker la cac nhdm hgp
c h i t da dang, phu-c tap, tinh dae trung cao va kha b i n vO'ng vdi qua trinh phong hda, do vay chdng la ea sd de tinh cac "chi s i " duge ddng rdng rai trong tim k i i m tham dd d i u khi n h i m dac trung va phan biet eae loai d i u vdi
nhau (Stout etal., 2000) Cac
biomark-er thdng dung nhit dugc eae nha dja hda h&u ea sd' dung la terpanes, ster-anes, mono- va triaromatic steroids (Peters and l\/loldowan,1991, 1993) Td' cae kinh nghiem dja hoa thu dugc trong qua trinh tham dd d i u khi, ngudi ta dua ra mgt bg 18 chi so nhan dang chgn Igc biomarker ddng d i phan biet cac loai d i u cd tinh c h i t
tuang ty nhau (Faksness et al., 2002)
Tieu chi de chgn lya eae ti so nhan dang nay la:
+ Dae trung va da dang
+ B i n vO'ng vdi qua trinh phong hda
+ Do chinh xac/phu'c tap cda phuang phap phan tich
Cae sac do ion vdi ky hieu pie edng nhu ten cda cae doi tugng phan tieh va ky hieu viit t i t eda hopanes va thterpanes nam vdng no khae (m/z 191), steranes thdng thudng, diaster-anes (m/z 217 va 218) va triaromatic steroids (m/z 231) dugc trinh bay trong cac Hinh 5, 6, 7 Cac ti s i nhan dang chpn Ipe cdng dugc dua ra trong cac bang ben dudi Cac ti sd nhan dang nay cua biomarker cung duge tinh toan theo edng thuc (1)
Trang 7A N T O A N - M O I T R U O N G DAU KHI
Ten ty so
%27Ts
"Ts/Tm"
Djnh nghla
100 * [27Ts(191)]/([27Ts(191)] + [27Tm(191)])
Oleanane
Ty s i nay dae trung cho dp trudng thanh cda dau va
tang tu 0 vdi d i u cd dp chin t h i p va ed xu hudng tang eao
d i n 80-90% khi dp chin tang Ty s i nay cdng bj anh
hudng bdi nguon goe tao d i u
Ten ty so
%28ab
Bisnorhopane Dinh nghTa
100 * [28ab(191)]/([28ab(191)] + [30ab(191)])
Ty s i nay la mdt ehi s i dja hoa quan trpng Nd cd t h i
SU' dung lam chi thj eho cac loai vi sinh vat trong mot s i
mdi trudng tao d i u y i m khi Chi s i nay cd the dugc ddng
de danh gia do trudng thanh eda d i u va giam khi do chin
cua dau tang
25-Norhopane Ten ty so
%25nor30ab
(191)
Djnh nghTa
100 * [25nor30ab(191)]/([25nor30ab(191)] +
[30ab(191)])
Day la hgp c h i t Hopane bj loai mdt nhdm methyl C i u
tu' nay dugc tao ra trong eae loai d i u thd bj phan ra sinh
hge Idn Hgp c h i t nay thudng dugc phat hien trong eae
manh ion 191 va 177
Ten ty so
%29ab
Norhopane Djnh nghTa
100 * [29ab(191)]/([29ab(191)] + [30ab(191)])
Ty s i nay dae trung cho ca nguin goe tao d i u eOng
nhu md'c do trudng thanh nhiet eua d i u
Ten ty so
%29Ts
Norneohopane
Djnh nghTa
100 * [29Ts(191)]/([29Ts(191)] + [30ab(191)])
Ty so nay dae trung cho md'c dp trudng thanh nhiet
eda d i u Tuy nhien ty s i nay edng thudng mac sai s i do
kha nang tach giO'a hai c i u td" norneoHopane (29Ts) va
norhopane (29ab)
C30 diahopane Ten ty so
%30d
Djnh nghTa
100 * [30d(191)]/([30d(191)] + [30ab(191)])
Ty so nay dae trung cho md'c dp trudng thanh nhiet
eda d i u Tuy nhien ty so r i t nhd trong cae loai dau cd dp
trudng thanh t h i p eung nhu trung binh va tang manh trong
cae loai d i u co dp trudng thanh eao
Ten ty so
%30O
Djnh nghTa
100 * [30O(191)]/([30O(191)] -i-[30ab(191)])
Sy cd mat eau td- Oleanane eho thiy diu co nguon g i c td' eae loai thyc vat trong dat liin va duge tao ra trong ky Creta thugng mau tre hen Hgp chat thudng ed mat vdi ham lugng kha cao trong cae loai cua Malaysia [3] Ty s i nay cung bj anh hudng do qua trinh trudng thanh nhiet eua dau
Gamacerane Ten ty so
%30G
Djnh nghTa 100*[30G(191)]/([30G(191)] + [30ab(191)])] + [27Tm(191)])
Gamacerane thudng ed mat vdi ham lugng tuang d i i Idn trong eae loai d i u duge tao trong moi trudng sa l i n g qua muoi Ty s i nay thudng Idn khi cae C35 homohopane cOng Idn
Ty so dong phan bishomohopane 6" C20 Ten ty so
%32abS
Djnh nghTa 100*[32abS(191 )]/([32abS(191 )]+[32ab R(191)])
Cae ty s i cua cac d i n g phan isome la cac chi thj cho do trudng thanh eua d i u va thudng dugc sd' dung rgng rai trong dja hoa Ty s i nay thudng tang td" 0 d i n 60% khi do trudng thanh cua dau tang trong khoang dau cda ed'a so dp trudng thanh Chi so nay thudng it b i i n doi trong cac loai
d i u cd dp trudng thanh eao
T i m e
-L m f c
3 SOOO
30000
25000
20000 1SO00
10000 SOOO
°
a n ^
35000
30000
2 5 0 0 0
20000
15000
10000
«
23
66.00
66 00
24
68.00
68 0 0
l o r 21
70 00
l o f i 2
70 00
7 0 0 ( 2 1 6 70 10 2 1 7 3 0 1
li'
72 00 7d 00
S U T U D E N 0
2« 30 3 1 , 3
27 29 •'^
76 00 78 00 eo 00
e 00 121770 ID 21B 30) S U T U O E N D
25
26
72 00 74 00
31
28 ^^33
27
75 00 76 00 SO 00
82 00
62 00
a4oa
S4D0
Hinh 6 Sic do ion cua cac steranes thdng thuang va diasteranes (m/z 217 va 218) cua miu dau tho
52 DAU KHI - so 7/2010
Trang 8P E T R O V I E T N A M
Doi tu'O'ng phan tich va kf hieu cua cac steranes thong thu'O'ng
va diasteranes (m/z 217, 218)
Peak s 6 K i hieu D 6 i tu'O'ng phan tich
23
24
29
30
31
32
33
25
26
27
28
31
32
27clbS
27dbR
28aaR
29aaS
29bbR
29bbS
29aaR
27bbR
27bbS
28bbR
28bbS
29bbR
29bbS
13P (H), 17a(H), 20S - cholestane (diastenane ) 13P(H), 17a(H), 20R - cholestane (diasterane ) 24-Methyl-5a(H), 14a(H), 17a, 20R - cholestane 24-Ethyl-5a(H), 14a(H), 17a, 20S - cholestane 24-Ethyl-5a(H), 14P (H), 17p, 20R - cholestane 24-Ethyl-5a(H), 14P (H), 17P, 20S - cholestane 24-Ethyl-5a(H), 14a(H), 17a, 20R - cholestane 5a (H), 14p(H), 17P(H), 20R - cholestane 5a (H), 14P(H), 17P(H), 20S - cholestane 24-Methyl-5a(H), 14p (H), 17P, 20R - cholestane 24-Methyl-5a(H), 14p (H), 17p, 20S - cholestane 24-Ethyl-5a(H), 14P (H), 17P, 20R - cholestane 24-Ethyl-5a(H), 14p (H), 17p, 20S - cholestane
Diasterane/sterane Ten ty s6
%27dia
Djnh nghTa
100 * ([27dbS(217)] + [27dbR(217)])/([27dbS(217)] + [27dbR(217)] + [27bbR(217)] + [27bbS(217)]
Ty so dong phan sterane 6" C20
Ten ty so
%29aaS
Djnh nghTa
100 * [29aaS(217)]/([29aaS(217)] + [29aaR(217)])
Ty so dong phan sterane 6" 014 va 017
Ten ty so
%29bb
Djnh nghTa
100*([29bbR(217)]+[29bbS(217)])/([29bbS(217)]+[29bbR (217)]+[29aaS(217)]+[29aaR(217)]
Oac ty so phan bo cua cac ho-p chat bb sterane
Ten ty so
%27bbSTER
%28bbSTER
%29bbSTER
Djnh nghTa
100 * [27bb(S+R)(218)]/([27bb(S+R)(218)] + [28bb(S+R)(218)]
+ [29bb(S+R)(218)])
100 * [28bb(S+R)(218)]/([27bb(S+R)(218)] + [28bb(S+R)(218)]
+ [29bb(S+R)(218)])
100 * [29bb(S+R)(218)]/([27bb(S+R)(218)] + [28bb(S+R)(218)]
+ [29bb(S+R)(218)])
Ohi so nhan dang vd'i steranes thong thu'O'ng va diasteranes (m/z
217 va 218) STT
1
2
3
4
5
6
Ky hieu
%27dia
%29aaS
%29bb
%27bbSTER
%28bbSTER
%29bbSTER
Djnh nghTa
100 * ([27dbS(217)]+[27dbR(217)])/ ([27dbS(217)]
+[27dbR(217)] + [27bbR(217)] + [27bbS(217)])
100 * [29aaS(217)]/([29aaS(217)] + [29aaR(217)])
100 * ([29bbR(217)]+[29bbS(217)])/ ([29bbS(217)] + [29bbR(217)] + [29aaS(217)] + [29aaR(217)])
100 * [27bb(S+R)(218)]/([27bb(S+R)(218)] + f28bb(S+R)(218)l+[29bb(S+R)(218)]))
100 * [28bb(S+R)(218)]/([27bb(S+R)(218)] + [28bb(S+R)(218)]+[29bb(S+R)(218)]))
100 * [29bb(S+R)(218)]/([27bb(S+R)(218)] + [28bb(S+R)(218)]+[29bb(S+R)(218)]))
Ty le tuang d i i giua diaster-ane va sterdiaster-ane thudng dugc sd' dung nham xac djnh loai nguon
d i u Tuy nhien chi so nay cOng thudng bj anh hudng do qua trinh trudng thanh nhiet, ty s i nay d i i vdi cae d i u ed dp trudng thanh eao Ty s i nay thudng eao khi dau duge hinh thanh trong da nguin Silic va thudng t h i p khi hinh thanh tu da me la carbonat
Ty s i d i n g phan 20S/20R C29 sterane la chi thj eho md'c
dp trudng thanh cda dau Gia trj nay biin doi tu 0 (dii vdi d i u chua trudng thanh) d i n khoang 55% d i i vdi d i u trudng thanh
Ty le nay eOng phu thupc nhieu bdi qua trinh phan ra sinh hpe do
d i n g phan 20R ed xu hudng mat nhanh han so vdi d i n g phan 20S(Philpand Gilbert, 1986)
Ty s i nay eOng dae trung cho mue dp trudng thanh cua d i u Ty
s i nay cung biin doi tu 0% d i i vdi cae dau chua trudng thanh va
d i n 50-55% doi vdi d i u trudng thanh trung binh va d i n 60% doi vdi eae loai d i u ed dp trudng thanh cao
Phan bd cac hgp chit C27, C28, C29 sterane dugc sd' dung rdng rai trong viec xac djnh ngudn tao d i u
Ngoai ra eae ehi so nhan dang eda nhdm cadinane va bieadinane (m/z 369) cOng la chi
s i dac trung eda d i u it bj tac dpng eda qua trinh phong hda Cac chi s i nhan dang eda nhdm bieardinane (m/z 369) Vdi ky hieu W la trans-trans-trans-bicadinane, T la eis-eis-eis-bieadinane, X la C29 pentaeyelie triterpane, va C30H la C30 Hopane (Yi Zhou va edng sy) Hinh 8 ehi ra vj tri tren s i c do va
ki hieu eua eae cau tu dac trung bieadinane
DAU KHi - sO 7/2010 53
Trang 9A N T O A N - M O I T R U O N G DAU KHI
saoD
3400
2600
» ( 2 3 0 70 ID 231 30) SUTUDENX)
34
> ,»P
35
I l l I I I ,
^!^'" •- ' M J V - W L ,
.IW
39
40
I
-„^jJ ^jw Vv;»Ww^il v^•^« -vvv
5Z.») M.C» M.OO Se.OO 6093 6I.CO 64 00 66 OO 66X 70.00 72M 7< OO 76,00 7800 80.00
H/'n/j 7 sac dd ion ciia cac steroid 3 vdng tham (m/z 231) ciia miu diu thd
Oac Steroid 3 vong t h e m (m/z 231): Doi tu'ang phan tich va ky hieu
Peak so
34
35
36
37
38
39
40
Ky hieu C20TA C21TA SC26TA RC26TA + SC27TA SC28TA RC27TA RC28TA
Doi tu'ang phan tich C20 - triaromatic steroid hydrocarbon C21 - triaromatic steroid hydrocarbon C26, 20S - tharomatic steroid hydrocarbon C26, 20R- + C27, 208 - tharomatic steroid
hydrocarbon C28, 203 - triaromatic steroid hydrocarbon
027, 20R - triaromatic steroid hydrocarbon C28, 20R, triaromatic steroid hydrocarbon
STT
17
18
19
Ti so nhan
Ky hieu
%TA21
%TA26
%TA27
dang cho Triaromatic Steroids (m/z 231)
Djnh nghTa
100 * [C21TA(231)]/([C21TA(231)] + [RC28TA(231)])
100 * [SC26TA(231)]/( [SC26TA(231)] + [SC28TA(231)])
100 * [RC27TA(231)]/([RC27TA(231)] + [RC28TA(231)]) Oac chi so nhan dang nhom bieadinane (m/z 369)
STT
1
2
3
4
Ty so
W/(T + W) T/(X + T) C30H/(X + C30)
C30H/(T + C30H)
Djnh nghTa
100 * (W(369))/(T(369) + W(369))
100 * (T(369))/(T(369) + X(369))
100 * (C30H(369))/(X(369) + C30H(369))
100 * (C30H(369))/(T(369) + C30H(369))
Hinh 8 Sac dd
ion nhdm
biead-inane (m/z 369)
va ciu til' nhan
dang
il )
L Ja4.v.AiiAJU]uLlU,
III Oac qui trinh phan tich
1 Qui trinh xw ly mau
Can khoang 0,1g mau dau trar hoae m i u d i u thd hda tan tronj 20ml dung mdi dichloromethane (DCM) Hoa tan m i u bang each ch( vao b i n sieu am 15 phdt de dau tai hoan toan vao dung mdi, lay ra ch vao td lanh va d i qua dem
Gan p h i n dung dieh de loai b eat va eae tap chat ca hoc, tii hanh nhoi ept d i lam sach mil bang 5g silicagel da dugc hoat hoi qua dem d 300°C chuyin toan bp dung djch len d i u ept va ru'a giai vdi 3 l i n the tich cot (45ml) bing hon hgp n-Hexane: DCIVI (1:2) Lay khoang 1/4 p h i n dung djch vu'a duge rCpa giai va cd dac din 1ml de phan tieh GC/FID
Phan cdn lai duoi dung moi
b i n g ddng N2 d i n the tich khoang 1ml Sau dd t i i n hanh loai bd toan
bd DCM ed trong mau bang each them vao dung djch 3ml n-Hexane rdi eho bay hai Lap lai viec them n-hecxan 3 l i n de loai hoan toan DCM
P h i n dung djch nay se dugc tach cac hgp chat hydrocarbon no
va hydrocarbon tham tren ept dxit j nhdm vdi cac dung mdi ru'a giai j tuang d'ng la n-Hexane cho phan
no va DCM eho phan tham: Cho toan bd m i u da loai dung mdi DCM len edt dxit nhdm (5g) ru'a giai vai
3 lan t h i tich cdt (45ml) dung moi n-hexane
Phan vd'a ru'a giai (hydrocar-bon no) dugc dem di loai bd dung mdi n-hexane b i n g each cho bay hai tdi khi cdn khoang 1ml, sau d6 them vao 3ml Isooctane Tien hanh bay hai va them Isooctane 3 lan nO'a loai bd hoan toan n-hexane
De loai bd hoan toan n-alkane trong phan no, dung 0,5g ray phan td' 5A da chiit b i n g dung moi
1^ 54 OAU KHi - SO 7/2010
Trang 10<^ ^
toluene va hoat hda d 500°C cho vao dung djch De
hdn hgp qua dem, ddng xyring hut p h i n dung djch va
tiin hanh phan tieh Biomarker p h i n no tren thiit bj
GC/MS
Phan cdn lai tren edt (hydrocarbon tham) d u g c
ru'a giai bang 45ml DCM, toan bg p h i n rd'a giai nay
duge ed dae d i n 0,5ml de phan tich hydrocarbon
tham tren t h i i t bj sac ky khi ghep khoi pho
(GC/MS)
2 Qui trinh phan tieh phan n-alkanes (C10 - C35),
Pristane va Phytane tren thiet bj sae ky khi
Thanh p h i n dau sau khi duge lam sach d qui trinh
xd' ly mau se dugc phan tieh tren may sac ky khi
(GC/FID)
Sac ky khi dugc t i i n hanh tren thiit bj HP-6890
vdi eae d i i u kien phan tieh nhu sau:
cot mao quSn
Khi mang
Bu6nq bcym mau~
Chu'ong trinh nhl§t do
Oku d6
Sac ky khi HP-6890
HP-1 0.25 nm film X 60m x 0.32mm ID Nhigt ap t6i da 320°C
Helium, luu lirynq 1,5 ml/phut Buong bgm m i u splitysplitless Nhi^t dp khcfi diem: 50 C giO 3 phut Gia nhi^t 15°C/phut den 120°C, Gia nhi^t 4°C/phut den 290°C, giu- 50 phut
FID 370"C
Ap suat khi n6n: 400ml/phut
Ap suat khi H2: 40ml/phut LU'U lu'p'ng khi b6 sung: 40ml/phut Phuang phap nay cho phep phan tich
hydrocar-bon td' n-CIOden n-C35
Viee phan tieh tren GC ehi eho thay duge cac
dac d i i m nhan dang han che khi m i u d i u da bj
phong hda manh hoae khi loai dau trong cac m i u r i t
giing nhau
De cd dugc bd'c tranh toan canh cua md-c dp
phong hda, ed the thyc hien mpt budc k i i m tra dan
gian bing each lay tich phan (hoae do chiiu cao) cda
n-ankane trong s i c ky do GC - FID va ve bieu do so
sanh cac m i u sau khi da ehuin hda vdi eae hgp c h i t khdng bj phong hda Sy phan bo n-ankane trong cac
m i u eOng phan anh thdng tin v i kha nang tai phan bo paraffin nhu mgt p h i n cua qua trinh phong hda (Strom
- Kristiansen et al., 1997)
Day phan b i n-alkane duge b i i u d i i n tren sac do duge ehuan hda theo edng thd'c sau:
C,(ch) = C,(dt) * 2/[C29(dt) + C3o(dt)]
Trong dd:
Cx (ch): Ham lugng c i u td' X trong day n-alkane
da duge ehuin hda
Cx (dt): Ham lugng e i u td" X trong day n-alkane tinh theo dien tieh sac do
3 Qui trinh phan tieh hydrocarbon thom tren GC/MS
Thanh phin hydrocarbon tham sau khi duge chiit
d khau xu' ly m i u duge phan tich tren thiit bj GC/MS vdi cac d i i u kien phan tich nhu sau:
Thi^t bi
c o t Khi mang Bcym mSu Chu'O'ng trinh nhi0t cJO NSnq iLFp'nq electron Thd'i qian logi b6 dunq moi
C h ^ dp cua kh6i ph6
GC Agilent Technology 6890N Supeico EQUITY ^" -5 0,25 m film x 60m x 0,25mm ID NhiOt dO t6i da 350°C
He Luu lu'O'nq: Iml/phut
BO bom mSu SPLIT/SPLITLESS SOCC
Nhi^t dO kh&\ di^m: 50''C, giO 3 phut
Gia nhi^t: 4''C/phut d^n 290°C qii> tronq 40 phut
70 Ev TCf 14 phut
C h ^ dO chgn Ioc ion (SIM)
4 Qui trinh phan tieh biomarker no tren GC/MS
Dieu kien phan tich phan Biomarker No tren thiit
bi GC/MS nhu sau:
Thi^t bi
c o t Khi mang Born m i u Chu-ong trinh nhi$t dO NSnq lu'p'ng electron ThO-i qian lo?! bO dunq mOi
Ch6 dO cua khdi p h i
GC Agilent Technology 6890N Supeico EQUITY ' " -1 0,25 m x 60m x 0,25mm Nhi^t dO t6i da 350°C
He
L i / u l u p p q : 1 ml/phut
BO b a m mSu SPLIT/SPLITLESS 300"C Nhi$t dO khd'i di6m: SO'-'C, giO 3 phut Gia nhi0t:50 - 290''C tOc dO 4"C/phut
GiO 6r nhi§t dO 290''C tronq 50 phut
70 Ev Ti> 8 phut Ch6 dO chpn lpc Ion (SIM)
DAU KHi - sO 7/2010 55