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CC PHNG PHP PHN TCH SC K

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Khoa Cng Ngh Ha Hc B mn Ha Phn Tch

H Cng Nghip TP.HCM

Dr. Nguyen Ngoc Vinh

Chng 4:

phng php GC/MS v GC/IR

4.1. Sc k kh ghp khi ph 1. Nguyn tc 2. iu kin k thut 3. Phn tch sc k ghp khi ph 4.2. Sc k ghp hng ngoi 1. Nguyn tc 2. iu kin k thut 3. Phn tch nh tnh v nh lng

1. Sc k kh ghp khi ph
Phng php ph khi lng (Mass Spectrometry - MS) l mt phng php phn tch cng c quan trng trong phn tch thnh phn, cu trc cht Bt u t cui th k XIX, Goldstein (1886) v Wein (1898) thy rng mt chm tia ion dng c th tch ra khi nhau di tc dng ca mt in trng v t trng 1913, Thomson thy kh neon t nhin gm 2 loi c khi lng nguyn t khc nhau (isotope) l 20 v 22 (g/mol), ng cng s dng my ph khi lng trong phn tch ha hc, xc nh khi lng phn t v nguyn t 1940 MS c s dng trong pht hin du m v trong phn tch hormon/steroid (1950). S kt hp GC/MS thc hin nm 1960 v LC/MS nm 1970. Sau nhiu k thut mi trong MS nhanh chng pht trin nh FAB, TS, t cc, TOF, MALDI C phm vi ng dng rt rng ln trong cc ngnh ha hu c, ha sinh, v c v nguyn t ng v

1.1. Nguyn tc chung

C s ca phng php MS i vi cc hp cht hu c l s bn ph cc phn t trung ha thnh cc ion phn t mang in tch dng, hoc ph v cc mnh ion, cc gc theo s sau bng cc phn t mang nng lng cao:
ABCD + e ABCD+ + 2e > 95%

ABCD2+ + 3e

ABCD-

Nng lng bn ph phn t thnh ion phn t khong 10eV, s ph v ny ph thuc cu to cht, phng php v nng lng bn ph qu trnh ion ha. Ion phn t c s khi (m/e) k hiu l M+ C nhiu phng php ion ha khc nhau nh va chm electron, va chm electron, ion ha photon, ion ha trng, bn ph ion, bn ph nguyn t nhanh Phng php khi ph chnh l phng php nghin cu cc cht bng cch o chnh xc khi lng cht

2. C s k thut ca MS
Thit b khi ph u tin c J. Thomson (Anh) ch to nm 1912 v c hon thin hn vo nm 1932. S chung:
Np mu

Ha kh mu

Ion ha

Phn tch ion theo s khi

detctor

Bm ht

X l s liu

1. Np mu v ha kh mu:

Mu np vo my c th dng kh, lng, rn

Mu sau khi bin thnh dng kh i qua sang bung ion ha qua giao din interface.

2. Ion ha:
cc phn t dng kh phi c ion ha bng cc pp thch hp. Phng php va chm electron (Electron impact - EI): Phng php ph bin, dng kh ca mu i vo bung ion ha, va chm vi dng electron t mt si t (catod) chuyn ng vung gc vi dng phn t mu. p sut bung ion ha t 0,005 torr, to 200 0,25oC.

Cc e va chm vi pt trung ho to cc mnh ion, mnh gc hay pt trung ha nh. Sau cc ion i qua mt in trng 400 4000V tng tc, vn tc ion t l vi khi lng ca chng.

Phng php ion ha ha hc (chemical ionization - CI): Ion ha ha hc l cho dng phn t kh va chm vi mt dng ion dng hoc m bin pt trung ha thnh ion phn t hay ion mnh. Dng ion ny c hnh thnh t pt dng kh H2, CH4, H2O, CH3OH, NH3 qua s ion ha nh bn ph chng bng dng electron mang nng lng cao. Mi phn t dng kh c th to ra cc ion dng khc nhau lm tc nhn trong ion ha ha hc (bng di y) Phn t kh Tc nhn ion dng

H2 CH4 H2O CH3OH i-C4H10 NH3 (CH3)2CO

H3+ CH4+, CH3+, CH5+, C2H5+, C3H7+ H+, (H2O)n H+, (CH3OH)n C4H9+, C3H3+ H+, (NH3)n (CH3)2CO+

Ion ha bng ion dng xy ra theo cc bc sau:

Bc 1: pt trung ha (CH4) va chm vi dng electron mang nng lng cao tr thnh ngun ion

CH4 + e CH4+ + 2e
CH4+ + CH4 CH5+ + CH3+ CH4+ CH3+ + H CH4+ CH2+ + H2 CH3+ + CH4 C2H5+ + H2 CH2+ + CH4 C2H4+ + H2 CH2+ + CH4 C2H3+ + H2 + H

C2H3+ + CH4 C3H5+ + H2

Bc 2: cc ion dng ny va chm vi pt mu dng kh CH5+ + MH CH4 + MH2+ CH3+ + MH CH4 + M+ CH4+ + MH CH4 + MH+ (MH l mu cht)

Phng php ion ha trng (field ionization): s dng in trng mnh lm bt ra e t phn t. Vi b mt KL anod c hnh nhn hay si mng v di chn khng cao (10-6 torr) s pht sinh lc tnh in lm bt e ra khi phn t m khng i hi nng lng qu d. Trong pp ion ha trng, ngun ion c to ra nh mt kim nh c d = vi m lm anod gn ngay trc khe vo bung ion ha, khe vo chnh l catod, cn c khe hi t tp trung ngun ion Phng php ion ha photon: Nhiu qu trnh ion ha i hi nng lng t 10eV tng ng vi cc photon c bc sng khong 83 155nm nm trong vng t ngoi chn khng do c th thc hin qu trnh va chm photon. khi ph t c cng tng t pp va chm electron. Do nng lng nh hn nn ph ny ch yu cho ion phn t v mt s mnh c s khi ln tng t pp ion ha trng. Ngi ta cng s dng ngun laser lm ngun ion ha, l ngun n sc (laser rubi) mang nng lng cao.

3. Tch cc Ion theo s khi:

Cc ion hnh thnh c khi lng m v in tch e , t s z = m/e gi l s khi. Chng s c tch ra khi nhau nh mt nam chm c t trng Ho hoc km theo mt in trng na. Cc loi thit b tch gm c:
Thit b khi ph hi t n hoc cn gi l khi ph hnh qut hay lch t. (single-focusing magnetic deffection hay sector mass analyser)

 Thng thng nht l dng thit b t trng hnh qut (sectorfield spectrometer) tch cc ion theo khi lng Cc ion trc khi ra khi bung ion ha c tng tc nh mt in trng c th U, i qua nam chm hnh ng c t trng H. Cc ion s chuyn ng theo hnh vng cung bn knh r trong t trng ny. Vi r:

Vy vi gi tr U v H nht nh th s khi m/e t l vi bn knh r. T biu thc trn nhn thy cc ion c m/e khc nhau s c tch ra khi nhau do r ca vng cung chuyn ng ca chng khc nhau. My hi t n c phn gii thp (1000 5000)

 Thit b khi ph hi t kp (double-focusing sector spectrometer): my c phn gii cao (10.000 100.000) ngi ta ch to b tch ion gm 1 in trng v mt t trng t cnh nhau. Cc ion trc khi qua t trng hnh qut, s i qua 1 in trng tnh in tch bit nhau mt ln na

 S tandem sector instrument vi EBEb ion optic (Jeol HX110, vi h khi ph hi t 4 ln).

 Thit b khi ph t cc (quadrupole): khi ph t cc s dng 4 thanh trn t song song nhau thnh 1 b. Tng cp i din in tch m hay dng ca ngun in 1 chiu (DC). Ngoi ra th in xoay chiu c s dng cho c hai cp.

C 2 trng u khng lm tng tc dng in tch dng t ngun i ra nhng lm chng dao ng quanh trc trung tm khi chuyn ng v ch cc ion c s khi nht nh mi n b phn thu gp. Tn s v th thay i cc ion c s khi khc nhau ln lt n b phn thu gp.

 Cc ion i vo trng t cc theo hng trc z ng thi dao ng theo hng trc x, y di nh hng ca mt trng in tn s cao. Ch cc dao ng ca cc ion c m/e c bit khng tng ln theo bin dao ng v c th i qua tm t cc dc theo trc. Cc ion khc c bin dao ng tng s va p vo thnh cc in cc trc khi c th vt qua cc in cc i vo detector. Phng trnh tng qut ca ph t cc nh sau:

Trong K: hng s, V: in p tn s cao, r: khong cch 2 in cc i nhau, f: tn s dao ng ion

 phn gii ph k t cc thng thng t t 500 1000, mun nng cao Rs, ni 2 3 b t cc vi nhau nh hnh di (Rs c th t ti 20.000):

4. Detector:

Sau khi ion hnh thnh v tch khi nhau theo s khi, chng cn c pht hin v ghi nhn nhim v ca detector. Cc loi detector trong MS gm c cc Faraday, nhn electron th cp, tm knh nh nhy ion, h thng m ion.
5. phn gii ca MS: phn gii l kh nng tch 2 s khi lin nhau m v m + m. phn gii R ca MS c tnh theo:

Cc my c R < 1000 c coi l phn gii thp, R > 10.000 l c phn gii cao

3. Sc k kh ghp MS
Cc mu cht trong phn tch MS phi nm dng hi do hn hp kh mang v mu cht t GC c th a vo bung ion ha ca MS nhng phi gim tc kh mang t ct i ra khong < 2ml/min

Do th ion ha ca He l 24,58 eV, nn tng nhy ca my thng qua vic tng th ion ha ca bung ion ha ln 70eV, ngi ta phi tch He ra khi dng kh trc khi vo bung ion ha.
Cu to ca GC/MS gm 2 phn l thit b GC v MS c ghp vi nhau thng qua b kt ni (interface). Nhm mc ch loi bt He, N2 gim p sut dng kh mang v phn t mu cht i vo bung ion ha, phn GC dng ct mao qun v phn MS s dng bung ion ha EI hay CI vi b tch t cc v detector khi ph

Turbo pump

Rotary pump

4. Sc k lng hiu qu cao ghp MS

Phng php HPLC c dng tch cc cht lng kh bay hi, khng th tch c bng GC. S kt hp gia LC v MS cng cho nhng thun li tng t GC/MS. Tuy nhin interface y ng vai tr quan trng v gm nhiu loi khc nhau.

B kt ni np cht lng trc tip (direct liquid insertion, DLI)

B kt ni bng chuyn (moving belt interface) B kt ni phun nhit (thermospray) B kt ni bn ph nguyn t nhanh dng lin tc (CF-FBA, Continuous Flow Fast Atom Bombardment) B kt ni phun in