THC HIN KIM TRA MY SC K LNG

Sc k lng hiu nng cao ( HPLC) l mt trong nhng k thut chnh c dng rng ri trong cc phng th nghim phn tch. My sc k lng c pht trin trong ngnh Dc, ha cht , thc phm, m phm, v mi trng. m bo rng d liu thu c t my sc k lng l tin cy, h thng my sc k lng nn c theo di thng xuyn tng thi k. Trong qui trnh ny , vi tiu chun nh gi cho mt h thng HPLC ( bao gm mt bm bn knh, mt b t ng tim mu, mt detector UV-VIS, v mt bung iu khin nhit cho ct) s c tho lun. Bng 1 : Thc hin tiu chun ca HPLC Mdun HPLC Pump Thc hin c tnh chnh xc tc dng chnh xc ca gradient Kim tra p sut Injector ng tuyn tnh Carryover Detector chnh xc bc sng tuyn tnh Nhiu v tri Kt qu mong i 1% ca tc dng ci t 1% tng hp ca gradient Kim tra chc nng valve ng n , khng c r r t bm RSD < 1% R2 0.999 < 1% 2 nm R2 0.999 Noise : 10-5 AU Drift : 10-4 AU/Hr

PUMP : chnh xc tc dng : Mt yu cu quan trng ca Modun Pump l c th duy tr chnh xc ca tc dng pha ng, mc ch iu ny s l cn thit m bo s n nh v lp li tc ng pha phn tch v pha tnh. S chnh xc km s nh hng n thi gian lu ca s chia tch. chnh xc ca t l dng c th tnh n gin bng cn 4 s l. Th d : 0.5 ml/pht , ci t 10 pht , cn trng lng v tnh li trung bnh . 1.0 ml/pht , ci t 5 pht 2.0 ml/pht , ci t 5 pht Sau v th ca lu lng v trng lng tng ng .

 chnh xc Gradient v tuyn tnh : Khi phn tch gradient, kh nng ca pump l phn phi pha ng. S chnh xc v tuyn tnh ca s phn phi gradient dung mi c th kim tra gin tip bng cch quan st s thay i ca hp thu. Kim tra ny c thc hin cho hai knh mt ln. Knh A c dng l dung mi tinh khit chng hn nh methanol, IPA, nc , trong khi knh B l mt dung mi cha mt vt hp th UV chng hn nh 0.5% Acetone trong nc. Khng cho h thng qua ct tch m ch qua cun dy mao qun c p sut vo khong 20 -30 Bar ( 280 psi 420 psi ) 100% B, 0% A trong 10 pht 80% B, 20 % A trong 10 pht 60% B, 40 % A trong 10 pht 40% B, 60 % A trong 10 pht 20% B, 80 % A trong 10 pht 0% B, 80 % A trong 10 pht o bc sng 254 nm Nu hn hp ca 20% A v 80% B l chnh xc th chiu cao hp th ca n s bng 80% ca chiu cao hp th ca 100% B Tip theo x l cho knh C v D cng nh trn. Hnh 1 : tuyn chnh xc ca gradient v tuyn tnh.

Kim tra p sut : Bm LC ph thuc vo chc nng ng n ca check valve v ni dng ng ng n. Kim tra p sut ca bm s hu ch v cht lng kim tra check valve v xc nh c hay khng c r r trong h thng. Trc nht y ng ra ca bm bng mt nt kn v ci t bm t ng ngng ( Shutdown ) p sut 6.000 psi. Bm dung mi methanol 1 ml/pht. p sut trong u bm nhanh v

ng ra b chn li. Khi p sut tng n 3.000 psi, ci t l dng gim xung cn 0.1 ml/pht. Lc ny p sut s tng t t n p sut shutdown, nu check valve c th gi pha ng trong bung bm l mong i bnh thng nh hnh 2. Nu check valve khng hot ng ng n, p sut s dao ng trn 3.000 psi thay v t n p sut t ng ngng. p sut ca bm gim chm t t sau khi t ng ngng.

INJECTOR MODUN ( B phn tim mu ): ng : Kh nng ca van tim mu a cng mt lng mu tim lp li l quyt nh s ng v chnh xc ca din tch nh hay chiu cao nh hu so snh trong nh lng ngoi chun. Nu s thay i ca mu v chun c tim vo ct khng c kim sot cht ch, nguyn tc nh lng ngoi chun l mt so snh ti. Khng th so snh y ngha gia din tch ca mu v chun c th c thc hin. S chnh xc tuyt i ca th tch tim th khng l then cht bng s lng mu ca chun v mu c tim vo. ng ca van tin mu c th biu th bng cch lm t nht 6 ln lp li tim cng mt mu. lch chun tng i ( RSD ) ca din tch ca cc ln tim c tnh ton nh gi s ng. Tuyn tnh : Hu ht van tim mu t ng c kh nng thay i th tch tim khng cn phi thay i vng lp tim. Thay i th tch mu s c a vo vng lp tim mu bi ng tim bi ng tim hay thit b o khc. Tnh ng dng ca vng lp mu v kh nng ca thit b o a nhng s lng khc nhau ca mu trong t l thch ng s nh hng s tuyn tch ca th tch tim. tuyn tnh th quan trng i vi nhng phng php m yu cu dng nhng th tch tim thay i, chng hn nh phng php cao-thp trong nh lng khng tinh khit. 3

Nhim cho mu trc ( Carryover ) : Khng phi nh cung cp no cng thc hin kim tra ny v n rt ph thuc vo phn tch. Mt lng nh ca phn tch c th nhim vo t mu trc i vo mu k tip . S nhim mang sang ny s nh hng n s nh lng chnh xc ca mu tip theo. trnh nhim cho t mu trc, tt c van tim c tip xc vi mu ( vng lp mu, kim tim ,) phi c ra sch sau mi ln tim. Hiu qu ca ra sch c th c nh gi bng cch tim mt mu trng sau khi tim mt mu c nng cao trong phn tch. Din tch ca phn tch tm thy trong mu trng l phn trm ca din tch nng mu c th c dng xc nh cp ca s nhim mang sang.

U D T NGAI KH KIN : chnh xc bc sng . chnh xc bc sng c nh ngha l lch ca bc sng o mt hp thu hay di pht x t bc sng bit ca di. Vn tt , chnh xc v nhy ca php o s l sai st nu chnh xc bc sng c vn . C nhiu cch kim tra chnh xc bc sng ca u d t ngai v kh kin. Kim tra bc sng t bn trong my, vch n deuterium 656 nm, v nhng di hp thu 360, 418, 453, v 536 nm trong knh lc holmium c dng. Vch n D2 v knh homium th d dng, nhng hn ch ch o trong vng kh kin. Kim tra bc sng trong vng t ngai c thc hin bng cch bm y flow cell mt dung dch ca mt cu t bit hp thu v qut ph dung dch cho hp thu cc i v cc tiu. Vi max hay min t ph thu c so snh vi max hay min ca cu t bit xc nh chnh xc ca bc sng. Dung dch ca potassium dichromate trong perchloric acid v holium oxide trong perchloric acid c th c s dng . Tuy nhin , nhng dung dch c tnh acid rt kh lm sch flow cell bng dng chy sau khi o v nhng vt hynh quang t dung dch potassium dichromate. Dung dch pha lang caffeine th d dng chun b vi max 273 nm v 205 nm v min 244 nm. tuyn tnh : Khi phn tch tht s nng c th thay i, do kh nng ca mt u d l p ng tuyn tnh ca nng trong mt vng cho php da trn chnh xc ca din tch v chiu cao nh so snh gia cc chun v cc mu. tuyn tnh ca detector c th kim tra bng cch tim mt dy dung dch chun thay i nng . Nng n t 0 n t nht l 1.0 AU. hp thu vt qu 1.5 AU s nh hng nh sng lc. V th gia nng v p ng tnh tan tuyn tnh da vo h s tng quan ( r2 ). Chun b pha ng acetonitrile: 0.1% acid acetic (13:87), 1ml/pht Bc sng : 273 nm Nhit mi trng ( 15 27oC ). Chun caffeine, cn 75mg pha vo trong bnh nh mc 100 ml (C5), sau pha lang dy nng gc ny: ly 5 ml pha vo 100ml (C1), ly 10ml pha trong 100ml (C2), ly 20ml/100ml (C3), ly 30ml/100ml (C4). Tim mu 10L ca C1, C2, C3, C4, C5 vo my v v th tng ng vi din tch. 4

Nhiu v tri : Khng phi tt c cc nh cung cp thc hin kim tra ny. Vi h thng c c th khng cn ng t s tn hiu trn nhiu nh thit b mi . Mch in t , bm, v nhiu thit b o quang, n yu, o flow cell, nhit khng n nh s gp phn lm nhiu v tri detector. Nhiu cao c th gim nhy ca detector v nh vy nh hng n s nh lng lng phn tch nng thp. Detector tri c th nh hng n s xc nh ng nn v tch phn pick. Nhiu qui trnh nh gi cho nhiu v tri ca detector da trn ASTM ( American Society for Testing and Material) Method E 685. Ngy nay hu ht phn mm sc k u c kh nng tnh tan nhiu v tri ca detector. c bit , detector nn c lm nng trc khi kim tra, v nhng bt n nhit nn c trnh trong khi kim tra. iu kin kim tra ng, methanol c chy qua flow cell l 1ml/pht. p sut vo khang 500 psi phng nga bt kh.