Phng php ph khi lng

Phng php ph khi lng

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Phng php ph khi l mt k thut dng o c t l khi lng-trn-in tch ca

ion; dng thit b chuyn dng l khi ph k. K thut ny c nhiu ng dng, bao gm:

M hnh c bn ca mt khi ph k.
Xc nh cc hp cht cha bit bng cch da vo khi lng ca phn t hp
cht hay tng phn tch ring ca n
Xc nh kt cu cht ng v ca cc thnh phn trong hp cht
Xc nh cu trc ca mt hp cht bng cch quan st tng phn tch ring
ca n
nh lng lng hp cht trong mt mu dng cc phng php khc
(phng php ph khi vn khng phi l nh lng)
Nghin cu c s ca ha hc ion th kh (ngnh ha hc v ion v cht trung
tnh trong chn khng)
Xc nh cc thuc tnh vt l, ha hc hay ngay c sinh hc ca hp cht vi
nhiu hng tip cn khc nhau.

Mt khi ph k l mt thit b dng cho phng php ph khi, cho ra ph khi lng
ca mt mu tm ra thnh phn ca n. C th ion ha mu v tch cc ion ca n vi
cc khi lng khc nhau v lu li thng tin da vo vic o c cng dng ion.
Mt khi ph k thng thng gm 3 phn: phn ngun ion, phn phn tch khi lng,
v phn o c.

V d v cch hot ng

Cc ha cht khc nhau th c khi lng phn t khc nhau. Da vo , khi ph k

s xc nh cht ha hc no c nm trong mu. V d, mui NaCl hp th nng lng

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Phng php ph khi lng

(nng lng hp th ty theo ngun ion, v d MALDI nng lng l tia laser) tch
ra thnh cc phn t tch in, gi l ion), trong giai on u ca phng php ph
khi. Cc ion Na+, Cl- c trng lng nguyn t khc bit. Do chng tch in, ngha
l ng i ca chng c th c iu khin bng in trng hoc t trng. Cc ion
c a vo bung gia tc v i qua mt khe vo ming kim loi. Mt t trng c
a vo bung . T trng s tc ng vo mi ion vi cng mt lc v lm trch
hng chng v pha u o. Ion nh hn s b lnh nhiu hn ion nng v theo nh
lut chuyn ng ca Newton gia tc t l nghch vi khi lng ca phn t. u o s
xc nh xem ion b lnh bao nhiu, v t gi tr o ny, t l khi lng-trn-in tch
ca ion c th c tnh ton. T , c th xc inh c thnh phn ha hc ca mt
mu gc. Trn thc t th hai ion Na+ v Cl- s khng c o trong cng mt ln, v
cc my o ch c th nhn ra ion in tch dng hoc in tch m nn nu my khi
ph k c iu chnh o cc ion in tch dng th ch c ion Na+ l c nhn ra
bi my. .Mt trong nhng tnh nng ln ca khi ph lng l c th tm thy cu to
khng gian ca phn t v d phn t C7H14O2 c th l acid hoc ester ... V kh nng
pht hin ra hp cht vi nhy cc cao t 10-6 dn 10-12 gram. Di y l mt khi
ph (electrospray)ca phn t Kaempferol-rhamnose-rhamnose-glucose(m/z 741) trong
loi c thaliana, phn tch vi 5.10-6L (nu dng my MALDI th ch cn 0,5.10-6L).

ng dng sinh hc

Khi ph ca protein

Protein v cc phn mnh peptit

Protein m cc nh nghin cu sinh hc quan tm thng l s kt hp phc tp ca

nhiu protein v phn t khc nhau, cng tn ti trong mt mi trng sinh hc. iu
ny t ra hai vn chnh. Th nht, hai k thut ion ha dng cho cc phn t ln ch
lm vic tt khi m hn hp t cc thnh phn c cu to gn ging, trong khi trong cc
mu sinh hc, cc protein khc nhau thng l c lng khc bit nhau ln. Nu hn
hp c ion ha dng phng php phun ion hay MALDI, th nhng protein dng m
d tha nhiu c xu hng gim tn hiu so vi nhng ci t d tha hn. Vn th
hai, quang ph khi t hn hp phc tp l rt kh nghin cu do c qu nhiu thnh
phn phc hp. l v vi tc ng ca enzym, mt protein to ra hng lot sn phm
peptit.

gii quyt vn ny, hai phng php c s dng rng ri phn mnh protein,
hay cc sn phm peptit t s tc ng ca enzym. Phng php u tin s phn mnh
ton b protein v c gi l in chuyn gel hai chiu (2-DE: two-dimensional gel
electrophoresis). Phng php th hai, ghi sc lng hiu sut cao (HPLC) c dng
vi cc phn mnh peptit sau khi protein phn tch bi tc ng ca enzym. Trong mt
s tnh hung, c th cn phi kt hp c hai phng php.

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Phng php ph khi lng

Cc vt gel c xc nh trn 2D Gel thng l thuc v mt protein. Nu cn bit nh

danh ca protein , th c th xem xt vt gel . Khi peptit kt qu t tc ng ca
enzym ln protein c th c xc nh bng khi ph dng ly du khi peptit. Nu
thng tin ny khng cho php xc nh danh tnh ca protein mt cch chnh xc, cc
peptit ca n c th xem l thuc v o ph khi tandem.

Vic xc nh c tnh ca hn hp protein dng HPLC/MS cn c gi l shotgun

proteomics v mudpit. Mt hn hp l kt qu ca s tc ng ca enzym ln hn hp
protein s c phn mnh theo mt hay hai bc bng ghi sc lng. Cht tch ra t
giai on ghi sc c th hoc l trc tip a vo my o ph khi thng qua ion ha
phun in t (ESI), hay tch ra thnh mt lot cc vt nh s dng sau ny trong phn
tch khi bng MALDI.

Xc nh Protein

C 2 cch chnh trong khi ph xc nh protein.

Ly du khi peptit (PMF) dng khi ca cc peptit phn gii lm u vo

tm kim trong CSDL ca cc khi bit trc t danh sch cc protein
bit. Nu mt chui protein trong danh sch tham kho trng khp vi gi tr
th nghim th c l do tin rng protein c tn ti trong mu gc.
Tandem MS ang tr thnh mt phng php th nghim ph bin xc nh
protein. Phn ly do va chm (CID) c dng trong cc ng dng chnh
khi to mt tp cc phn mnh t mt ion peptit c th. Qu trnh phn tch
ch yu da vo cc ch phm phn tch b gy lin kt peptit. V s n
gin ca vic phn tch ny, n c th dng khi ca cc phn mnh quan st
c so trng CSDL ca cc khi bit vi mt hay nhiu chui peptit.

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