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HPLC and CE Principle and Practice

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peak

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gain

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HPLC

Problem

2. mobile ptrase

qdt: iltgr: g ]u?ltgr: ulJ tl1 lJ 1'l lnilu

tilotpump

pump

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till.t Uqryv{nil : o0n

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tlJ ,i,

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2.4

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lu n]ou purge il:vruru

prime ivlJU

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pump

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3.1

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3.2

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mobile phase

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column [tOvfl0nUUnnU[Ul

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lritiouno''uhlfi pu-p

rIryur

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Audv

filtJ guard column

4.2 guard column 14:OflOnilUqqnU

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guard column I

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ohase

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nJfiuurnJn':kirrn)

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S.:

fivlolornrg'lu

A l1'{lJt [OV

purnp (retention time 0v

OFlttUU random)

lrir,'los ornrfloon 0"rnf? pump

lril

doliltufl1i

mobile pnus. 5.4

/t4 r

-i-s

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'Aiar OUl.lU.lllJO:

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cty

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mobile phase riou trlrn

rfr

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lao

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lil (retention time ovtiounldrtrnto.t ia

ddo

d15 vrq olurFt on lJuu

0'tu'lu lJtnrnu n?'r

ud,

n?rlJqlonnooilu)

- 3/57 -

ctgr

ida-tA

d]:

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column mUqU?U (qâ&#x201A;ŹUHQulO{fl OnilUm Yl eAlaaul

5.5 u?a10{

dcgas

o,ngt

lS column oven YllSOflO

fl {Uu 0v

Problem

ttio

check valve

nJduu seal 1JO{ pu-p

tiir j'-jU'-jtJL-

mobile

phase [1Jfl5t]J?l[fl:0{ HPLC Fld}J rr-robile -

[1{ retention time

l: lJ:ilOn:ldlilUO'1

9lT?0d0Ufl

od'ru

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I on

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frqrrj?rursufi#ounl niu fie t0 ul rr?rud

ovrflu roo ul rflud'u Aurdr

[000'1.191?OUl.1[lJU l:

l0 l[0U 1:100

rIryrn

cnrfiel

nr:ufr^ln

s.o d'rvirnynrucrosei?oeirs

hiarrur:

0 zudrr

6. peak

hirrun

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ro.:,du

ruh.r ri tirril u 1il

lpi

til OVAlUgl?0U1.:91?U mobile ohase

qe/

(retention time ovnioul

lTfl 0n]J u

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Jfi â&#x201A;Źruei'?ri r ny n

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9l:

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(rflurua1# retention time rilduu)

lilt ll lfliuU'Ut,L-----'\,tril il -'-rt*l

6.2 suard column H:OnOAilU9lU

qffo +.2

Norrnal

r lilll illr

^'HIUVUL-'r Problem

peak

tnnnt: split

z.t finr:rlurlioufi

guard column

ttio

noCIlJu z.z

I q{u +.2 i I

pi'rriravarutoid'lodxkiarrt:n2oru | rJfunJ6ou6'rlirovnrulnil tirrfluhJ'ld'

9 v

lqY

l'Ulflunu mobilc phase

lfinvaf

u I e) un?OU'l{Fl?U mobile phase

lr{t.

ILJIUU-lr Problem

peak U tarl Fl.1[[Flt:UIt:fl

doul

d14

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mq0o{qadl:tl'l9l:$1U

9l?1 peak }J tail

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vrnqo0na9l

fit ltju rld9li?tlnq0tndt:

11:0 tu

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' ^ AO{ tSflOAtJUSU9l0U ISU

column frpril:vrnvr

.i ar

n:ilm c1

analyte

g.)cty

rljudl:lJ5vn ouvrlljutud til

tryfl 0

nilu

odtY

S1lfl deactivated column IilUAU 8.3

to{

mobile phase

uhitrltvaru

rJfu pu tos mobile phase'lrrj

dr:il:ynoufirflurua nr:ld ovvirl# 8.4 analyte r4rrJg^fi?orn'u active sites

flonlJu

eJO.l

peak fi

pH firirn.r

nlrudlJiltn TlJrndu

[9llJ ion-pair reagent tfi

ar:

modifi er yu^nfi tfl uruadarrLr:o

:yt1.1u'i9{

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nr:ufflt

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rJanrr 9. peak tfludulJ'lgl:

d6lvl

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9.2 mobilc phase

[1J0UU[1-Jl-llJ tatl

qdo s.t

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AdIAI

[dOlJdnlil Ufll:1JU[lJ0u aal<ldl

9.3 guard column

tOnO RilUlJnl:ilU[1JOU

: ? 0dOUfl l5 t9l: UIJ mobile Phase

q&o q.z

11:OS',l19

10. peak

[[Un0lnnU

d6lul

14 ld

[il91

(fronting peaks)

0. 1

q{o

udt::un?u [u9l?ou'l.t ffdnutuvrflu'hri uto

10.2 peak

baseline

s.r

rfi urJ : v fi vrfr n ruuio rr]fi uu

tioun#r

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peat

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fidnutuvrioul qw-u orlog i

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ft?tlldrlJl5 a'lu nt:#nrfrofl

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yfrqfi{:

Problem

I peak liio

dil rfryu^fl du nirr

nJd

ut ,oJ

d a

rrunzuo

{

--^ [1JUflOn]JU olnaonilusuq lilil1? c18

irl

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d-1510

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q{o

cyano phase

rilurlu

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o.+ d'r

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r{rriuril

ut o{ pT? o d r s'l il ar il r : r} t'l d:J

:-unJ fi uu

d'rlir

nvnr

u'lr ri #rrilu hl 16'

b{ov nrufirodxdtu mobile

mobile phase

phase

riount:nnaoil 1# Flush FtoolJuryuq:J 'fird'ru ethylacetate Yu^q HPLC

dtu'lu

2oo ml

'luo'qr:rnr:'lr1a

[n:91 2.5

ml/min qrildhu solvcnt su^ofi

rirrjru

fl01.1

t. ilaru peat fid'nueuvruu

'hirrvoru (rounded peaks)

t t.

t rirq.:n'jl

linear dynamic range tlo'1

o fl

il?ilriu

ei'ro

detector eranl

t t.Z

d'lrir gain UO{ detector srrtnu'hi d4

1.3

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lJ?n10{dl:ilqq[11fl 0niluil0]u?u at4

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[lJ

5157 -

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u $o n r11r

uru n {

nr:uffh

drilql

rJn;vr 11

laaA

d I

rilfi uu grrtrL[t:.tllo{ buffer, pH

lnnilnn:ul5v1{?l.lflonilufl u |

inilsvnou

[u Fl']ou'ln tcl

d I

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dfl

lfl

taaa

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riluqiuHQlJl0{n0ailu

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d{[nu a

recorder 12. baseline

drift (baseline

qqos iu

r"r, 1r^

cr /ur

[il

odil

idr

nn!ils?ltilu

do49.

[ilauulilu{[anuounvll

)

si'seir

u 0 .1 fl

ilJu{

lilnuuryuq10{noa}Ju

[9] )

t t.S d'{fir time constant 1Jo{ detector u$o al4

mobile phase

10 { 0 ifl il : vft 0u11 0q tu9l?0u-l.l 0vry1unl9l

r9ltirl:

:tc{ruto{

t1q nr

[1'l baseline

4'A

uio

ru^

ti-. constant

al JA ' illqfu?lQlJfl {t{n cycle) UoUn:

ovfi r

tov rnobile

q l^r-r o I n odtL dt

phase [[0& 010

cry

[Sqiln:il[[antUaUU

il il n lJ^lr o i c{ I : n v j

tqduo

n:v 11u9r09rsun1: fi fl[14lJO.1[[dnttAenlS U1

iliorirfi

t9lp.lR14qnq9r

fl ?u qlJ qfu

pirafl

ia -i

[lJu{ tu

I ufi ou

rrlr dcrector

l: cvnlun{

[v{0a9 qiufi Qil1 0{fl

qY t?l

;

Fl{vl

Normal

lr'l$lrto.t detector triu uv detector fifinrrru ctY t4 a "[rar liiont] ['f {1u lu:vilu indirect 6l

photometric rnode Problem

i 2.2 rr,obile

rrsu rfr o td

uriYu e

tlun:tfi fi tflu

pnase'hi rual

baseline rinovfi

d'nuorsi.rffu

lri

'ld solvent sfre uPLc

dr:rnfifildt{iu

tn;91

buffer rtflv aclditive c{ol

AA*

iln?1xJU5q1]5q{

cycle)

riouldsru

degas mobile phase

1#fi'rry6rdurutffo degas mobile phase 'lu I

:Y1'1?1.: [5.1'lU I

2.3 detector cell

dnil:nufofi l'lolornrg

Flush detector cell d'lu methanol solvent

a{ ia OU 11}J 9l?1tJ

It:

tlio

{ fl { n

l4rntrtavotfl detector

cett rTlu

Hxo, (r?rrLl#Hct) 12.4 outlet line 1Jo{ detector qoroi'u

n?lild'ulu

detector cell {'1

detector cell window baseline

rflurrqt#

tlnn#l? a.rrua1rl

hi6uu

to1d.11'lqqglU outlet line 1J0{ detector

oon

trjduu outlet line 'Iuri nJduu detector cell window'luri

12.5 flt:zudlJ mobile phas"

n]:

dr1#

'hi6 riodsr:r

dou

Yal

Fl?1}J 0{ fl

AU14

o.l

lJ 0

mobile phase 14ioogl:lfllt

t1.1RIUOUU

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5 1 fr

tiln

nr:u#l,u

d]rfi9l

rlcymr

tz.o nod'ru#uhq cquilibrution dt Jeutqilrv

Flush fl00uU9l'lU solvent

?luil?lu[[:

odxOrrfi ofi nr: nJduu mobile phase ru^el

rJrunax ttnuiu flow

hiri

ln rirJ:vrror 0-20 rvirqo.rr]?rurpr:

mobile phase Tu^Fr

udl

fl0nlJu noufl-l:?19lflou 12.7 mobile ptrase 'hiavorpr adoia; 1.1

O [9r

fl 1: lfi]J

UtJ 0 1n

rforfiolJdRt?\

lil

fl [Un

9l: ?0dOUnlS [Fl5Uu mobile ohase

9l'l

J4t

2.8 udlT 11d1il t:0u9l9r91 0u tun0nilu[1Ju

'l# guard column

r?n ru

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11t57 -

sp:t$d 2 - sp:tsd tz r{lu{oryaornn#rdo Guide to successful operation of your LC j A - .z-.---.---q. i u r"r < d lildilo{dllaYAlU gt't:t{fr 2 baseline'lritiuU }oufrOttnqtnu?tilonnill6iull'lin1:

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rfiqo rnfi ?'lo'r ornrfl fi $o

tfi o

ki 1 rtti'l

n'lrlriu rflu nu d

pump'lrri

Tl 1 {'l U

{ pump

alu

? 1 Yl 1 { 1 lJ [ 9l 11 {

dO.l pump

414'

check valvc. plunger seal,

1450 tlJ

plunger "frqrt

drld

d:yrruiru 'l# a.nu, lnri

rot,r",rt

#r rfl u

u gradient

run t tu

iA4 fr fr I fr or rH E 0r n n ?r L il

fi n'l r ru piu'hi

rfl u a l rJ n

st^

ff q fi

rilfi uu hJ

rff o dn : rei

u u o{

mobilc nhase tilfi uuttilo r oojie)

qrfl il[9rtJ119n:t iyt 7 1o 1.3

ra444

lJ:lJtA:nqaH:Ofl

?-UJ

o q9,

'hi

equilibrate

solvent

to/o

n'i

fr fr n

il;ru

r t ru r r: I

rJ'1 n

qJoril?lrn : ryo l'i

^g) 1.4 0fuhfllJ140n tilfli11 qg

0% I

Qil fl fl

n { q [u ?i

-lT,r [

[?q a

n'i

t:.t u

: vo.r'i

llu

d-t

n oii'ru

q9,)

{u cr o r 6'r o d x

r r r ulil

ann

solvent it fu a r

ohase d t

rnu til 111tl{floa}Ju tu'

h I

: fi fi e H io

Y qY "tS

9Ylr

[1'N1U10nfl?Ouli]J1fl a

un?t

rr o a''li

et 1

d'l il il

^ { [r-l9 r

tir

n'i r

11 n r

atrLr : o fi e'ld

AYq

t 1.t

d? u mobi le

il ry

1.4.1 ldcolumn heater fioruunfi 5 qq

1r 1 u.]

.1s)

u |]l

"c rrfro

^9 ofll?f nruilo.t rYvd

1.4.2 9l:?0dOU?l solvent zudlJttlnU9 gt

1.4.3 utuln:

0{1,1

: ofl

jj /tneo t? tud0tuTlilofiJilnu

0nilu

y d 4 q 9u . flfl -11^l[t?nno]Jn{11 l.t:o tnnn]J n ta

1.5 nl:zuCllJ solvent []J9

?0d o

u'i r il il

H r tn^q o r fl

n'l :

qqIeuIuoIntfl drJ solvent hi

r.t

r?wt io 1 il

lnufio p!

dil

standard

[0.1

1.6

ilnrrilu[lJOuvl

re^il

oi r 1#dr.:

o.z dril

solvent'hi

td9l $ ?1il

nJ fi

u fl ry

iu H r &r n

a r rL r : o

p.r

rfi q r

od'l u,

{'j'r fi n r :

rflr

rou r rd

dr ri'u 1 d'

mobile phase r5o guard column

- 23t57 -

lln

6U 1'l

q 0 1n

duluri

run d'au column

tir hi

Fl'111

phase a

hifio?l h{qrfiqrr:rs fr z 4s z.z

fl:?odoulqu r o.

fi'l retention time

r'rdlr solvent 4rAd

: srfl lquidmobile

il:yrurru

u rdo u fi

o d'ru

er.:'i r o'r o rfi m

?o fi n r :

ttio

filter

r-J

pr o

rnnr

u rtl o u

1u:yul

2. retention time

nt:rrfi'h

flt[1491

116U141

'lunr:fiPt

2.1 pump flow rate

Yl tdl

rlnavfl:.:[ilaBu[[un{

:: ^"[ulr t rurnlLruH5onqa r -x

96rgtA

slu taqtu l.t u {fl a oqt?a1

,il

rilnuurrljR{

[11 U Vl I

uhrdA

2.3 :vuuu.1 trJrdfru: 1.1:o

2.4

equilibrate

?11.1lfl

dUn

15 fl 1:

m A

dOU U U dl

rnr

qrio t.+

^9J 2.2 aatr flN}j 0.1 tlJ Fl.lll qq

tis

dsr

UAd

-]v

:'l fl o'u o I solvent h?fi n I r rL

pi'.r ni r

d o u fl r :

qdo r.r

drAd

q{o

iln1:!u[lJ0uvr

t.o

flonilu 2.5

u tA tdoxJdfily,l

n OaIJU

(qcg rfi uq ar ar rTfi

2.6

nll

degas

t'tio

dou

laavd

lJ 5 y

dil t n1il

u o s no n

lJu

rs tn fi )

sparge

mobile phase'hitrrurvau

mobile ptrase'Iil degas rfo sparge dru helium

r'hJ equilibrate

ii u

-'l

heliurn sparge ri tb1dfl : rd?u

lJ o

ttdr

rto.r : s iq o d r'l #n r : li

solvent

nJ 6

uu'1

z.t flnr:rJurtlou'lu

rnâ&#x201A;Źtlll mobile ptrase -'luri lqu'l#ar:rnfififiqarnrnd rray

mobile phase

solvcnt ryfiq HPLC rn:Fl

"; lirnrrruavotel fitter riodlutrali5Q mobile filtcr 'irl sonicate 1r,

phase

*,rric acid ,-tudruri.t

loudr

n#.r trny

a1:Jd?U methanol

Flush trOY equilibrateSsUllr

:.i

2.8 inlet filter Uo'l mobile phase

r?o filter'lu:vltu

? 0 cf o

:'t r o yir : y r u'i r d'u fi o q 1 n

arjol

fly0'19114:0lilnuu

o r o r{o s

dr { 1 #

tilil

lirnrtrruavo-re filter fioqjluzuroruTT! mobile phase loudr

qqei'u

fitter 'hJ sonicate

e N Nitric acid srrrud'rurir

nr:% rrnv

flrild1u methanol ororlosnJduu nit fioqjlu filter'Iuri 2.9

<,Lao

lJnl: :?cillJ tll: uulJ

u ntting

v{{

u'rqsy'

q dril u n r : ir

dru

q or 1 n

#n ry u

rqV

al4t./u

[l.tr[uu (:u?i0ut tl.rrluuu]n[nu til) f]'lunfl{:10q

n-J6uu fittins ttoy ferrule 3. rctention time rqrd4uda fr't t1.{ }J ?'l

rilduutflu

3.1 mobile pnure

fi { l1il nfl :

fl4fqrrAg,J

w q9l

r[9t []J [ryfl ]yrfifi9l o.: q

ftfl 9to{

9,'!

'lrri

tn5uru mobile phase 1rru

' : ouo9r5-tc{?uvl

,",

fildhi

[14

^d tlJQnno{cf 1145u?rnlT

lletdou#u 3.2 pump flow rate

? 0 rf o

u fl I : ri'wit dsr : r n I UAA

ril

rilnuurrljRi

r14

- 24/57

u'r3

dil nil

nt:

yl9 do

:'lr J

l,J

u u c]

solvent

hifi

r ru

ilcgrr

nr:rttl'lt

d't[14E

3.3 pump r4rsrufieilnel^ o q 9v 111

lilOA:

tflt:

Ft:xrdoulau

[140 tlJ

q9)

tTfl:VU0fl9l?{1JU19l 5 ml ?1.110{:Udl:nvnlul]00fl

0]fl detector

flflno.l

nrn mobile pnur.

lorul# flow rate srtrLif;nqaoufil'ff

rYd

ou].:uou 4 u'lyt

rirursu flow rate fild'o?.r

lo*

Ot/

rate

ltetfiri't{#u traql'ir pu,np dtlrufiqilns]^o?s d

[1.r8vren:'r{vr

3.4 qiu?r u

t i ^g) O{ ur lilnsvl

qrio t.a

QrJ14

riln uu otu1.t fl lJvr tu{'lu qqu

3.5 n{nlqfu?rQtJ11

column heater 3.6

oiq t

fifl

t?l

Fro

tnt9)

lilQnilo{

Aavq

xulall:oTua10'1

?1dOU1U1n"d : OTU n1

{fl AO}JU

-/tnt9) floailu tllflnflo{ q

3.7

mobile phase

Atn r4u

lT solvent n lilil?n0nUtdu

fi stabilizer oqjlu A 115O

stabilizer rrJd t,ru an

ril

dtAd

3.8 lJnl:ilur1JOUlr

q{o

r.o

dou'jrfi nr:nJduurnJ nsfi fluidic system

floauu 3.9 gradient delay volume U0.1

fluidic system

[]J

nt 5 tfi il q il

0flnoq

nfzu aru

ar:

u'j r fl n r : uJ d u u r ril

volume

- 25t51 -

'lril

o.r fl I

riohj

rryu

run gradient rfl u d'u dr

o r o v r{o r ri r u I ru gradient delay

flr:rifi s gr.fdr.rto{ peak fiorJnfi flaur t. 'hirJ:rng

peat<

nr:rri1'lzu

fl't[14E

t.t ilqnrnrfl

ddd

9ln9n:t{il

injector

t|ovgr]:lnn

niu fiqfiq vial, 'hifi vial j

- a

' oH tu9r" rItl.'l u 1ilfl { il91,

l^utl4t

1J:tJt9r:Ft?0U'l{ [U

;;

nr:finhi

tfiurvlo,

Normal pea!<s

d d

Qfler0{, [1:Jqqqqnu

{lu

rlu 7

1.2 mobile pttut"'hi'lun

ilio'iilnrlou No peaks

t0 rl,t 9/at

tu [9t[!Fl pump ?t5o pn-p finrJnfi qfinr:r.rfi t lq fi 'lilrfi

I ilo z

o'ldrst^rL mobile phase

Mobile phase'l

inlet filter

mobile phase m6o filter lu:vt-luqfloi'u

1J0.1

v,l

jie

1.3

n{nl

t4'

detector [lJ

quat:'livt

4 1J0 2.8

9t: ? 0d0uI

ravn{nr lil nfl n o {

I qY

qndo.: rryu etlruurr

fl0il dt

sensrtrr,lty, autozero 9)

rlJU9tU

r.+

lrild'rjfu

zero detector baseline

'"rofi

9l'1

detector

9rnfl151.1vr i I

detector

t.S

ilcyurorfl

{oo

ISU detector flow cell, power supply,'l'10:'lllff r dty

tljU9lU 1.6

nt:

arr

[90]J9l O:v?1',1]{

detector fl UnOilYl?[9lO: lnr tlJ[14 tJ

atl

lJ 9r o I 0.1 d] u diu fu 1 fil I tn v fl 1 T 9l { Fl'l I A- - 2t- qY o i ' [u9l1rlfi u {11Qnno.1 u 0{fl l:rryoxJgro t1'r og'ct

d o il n 1 : lso

lv d]J jq

1.7 mobile pttate

ttr

fi1dhi

dot

t9l:UlJ mobile ohase yq49luaa<a

fi I

frfl9r0.1

1,i 1J

lJ fU

-1

U 1\

: 0 il

[?1]J

n U n ?',I ru 9l U

llt

anns fl0u1Jo{gr?0u'l{ otA 1

?OU'1.1[do]Jdnlv'l

d0u dn

y't lJ

?ou1.1

addutJ

?1?U

d0 u ?r il1: ra 5 uil

dvrql

[9]:urua?out.l n4]J

- 26t57 -

u'1.1

O',i 0

tfl

] fl fl 1 T

uful.t1 ddrA

2. peak IJlJU

lFl[nUn?lil filFl 11 ?.1

, I

JUr.,]r-_JL__J

61fl9tO.1

loop 1u injector

llr

dou

sample toop fi 1d

o r od'osn-J fi

1r ritfJ u

fllJ

t9l?][1.1:JtS

1JU

Qnfl0'1

;;

9/r

tyu fl?]}JUl?

gldU sensitivity, autozero

-l

luugru

Smaller than expected peaks

2.4

9r:?r fl ou[[Rv9t ifi't [14OnR0.:

>l cl

I qy

z.: d.rrir detector'Iil c|flgro{

;;;-\-

Normal peaks

l-- lu=

[!nuuu:]J19r:mq9l

2.1 ilTrJ19r:ilqq t:J

2.21Ul911O.l sample

:lt l,l ll iltt ti l_

(qiU[f,U sensrtrvrty)

.ll :l |

nr:r[fi'h

dr[14E

h 'tn9/ rLU t9l1j5U

)

zeroYl

91.1

zero dctector baseline

detector rY 4i

jtl

2.5 n'r5 rToufl

v 1'l'11.1

uad

detector fl iln O!?'l?[FlO h'

tu[14

2.6

: ? t fl o u n'l t t1

no10

i fl

r 619/

t6l o Outv lU9nt

10{nt:[YolJ9lO tfi

-i

d frjooâ&#x201A;Źu I

I a

[a s n']

tfi USnfl nnon

tV d:J

drard

ilnr:ilu[lJou11

detector flor.r, cell

firn'lrruayorn6'{fi 2.6.

I Itl'lU?lfi0OlJUFl?U

z.o.z

union

lun:tfid,uu *n leul#

frn|rdof 1# nusn

rftuti

mobile ptlur"

rrrild'?u methanol (dr

methanol d-tlJt:CIp.tdurcirfi'u solvent d,uu .yy Y

drflu

*n ld'l srrru

9r?uutofln:{ a

d u

z.o.: lun:tfrfitnu run lnuld mobile ptrase vfror'hifirTr 1# nurn d?uar: 2oru : vH'irn THF rrnydr'ludsr: 5

0/5

0 (dr dr :

dil,u o.t THF

r.r

rn y

lir dar

rriru

au,,?l

ru'r, o ^

-ih

ftu solvent lttfiu run tsl) srrrud'ru 100 % THF z.o.+

2.7 detector lamp "irqor

tirtirvrliliyrr t#

et:?0do1l lamp energy diagnostics

lru h{nlfiuu 4-Y od]ul5

u?1

-27t51-

"n"rgy tamo

tv ,

lJ 5

nusn

druar:ovnrufirr: rdu

leulilfiil#r

ri r

n'j r rn ar qirfi

detector o

tfi u u riu

r?ru

lrri odrsl:fintru detector ur{

u lamp energy

rnvA a I l9t tlJ olJ

v n'1aq u oua

{

flr:rrri'h

ilryrtorfl

Z.S r

tlil

j;! injector

qmmt:1.1?t 8 trCI3Fl't5-t.11'l 9

t4 rd

vral, [uU vial

A9zuq

tio)a

ou'.1tu E|lttl.t u.1il fl {aq.

,lt-,rrrUrr,,,trf -, 4Ahl

tylu.l?,1o,

qnd'or,

nltaq til

rfi rufi

qeroiu

riJ

Ftu @r4d

2.9 a'l

rut

lnttn:

oonturixrr:nro{1fl : ru fi

n'nusuv broad

?1 1-l

[0001.19t?ou].1

tfl1l 3. peak fi

aot

o ut.1 lJ fi ?'t lJ

[11 d4

YYvl tJ

:ltt

a n rJ?

3.1 1j5il191:V]AAH:O Fr'lltJ[lJ

n9t o9t

0.19t? 0 U1.1

^h';qtud illnlnu til vn t?tn0Rilu

FlFlFt U

: fi fi e H f

?0u].11JO.ltUilAA

oaqn

r r ru rc?il

{u I o.r pi'r o d r.r d'l o y

9) ctt)

. -ia moblle phase 01 [T solvent yltJ9l?11J[[:.lUOun?l 0V

a r il r : u

'hi equilibrate

? [u n']5

fi q'l d'l

:.i ru r n

n'i

o.i

il;il

rsr

: so

l'i

tY 19J

n0n3JU It9]fi'l tS solvent ?l]Jfi11]Jt[:.1U-1flfl?t 0y a t r t : o fi sr'16'1:.i u I n

n'ir

% I o{

il?ilrfl

roldt lu

fl0nuu . Time

do td

3.2 tubing

Normalpeaks

Fl:?0dOU tubing. fitting [tOv fcm,le

tSlJUUlFl

[duzu]fluufl alnnlu tu

fild

l4r t].J

n'^

,lâ&#x201A;Źl

?l il,I l\r i I-.--JVLJ\ LJ \-J

QflFtO'1 1.1:Ofl',l:9191 ln rd

9,/

tubing [ilFr ?'t:o IT firting

;; Broad peaks (earlv eluters only)

[t0Y femrle'hirurLryaru

-.:

u i

3.3 tun1: 09l9tuu1.1d?u yl

drqooi'ufi tubing l#nifi

in-line filter ^^=^ 1150 guard

9uaAAv?

4udA

column H:OflOOIJU 14:0

uulrri

fi'lO9l9lUll in-line filter ttiO guard column U5OgrOn'Uu q yv

h{o r wirn

rril

rqilf

onJ 6uu

1r ri

trr fir n o

:.+ fleyurorfl injeuor

iJi

9rmFn5]{m 8 ttnyfll:1.111 9

rl,Jad4

ruu ?'rn1:?ryuH:ou9l aoddv4

flqnu [uilq9qflauH:0 "frqo injector port dl

Qord'u

[uuau 3.5

tJUlAlO{ sample

loop

lu injector'hi

:'l 0 dou sample toop fi 1# or opfo.rnJd

uu'191?1tl{}Jt&du

qnno{

28t5'7

uu 1r rirfl

g.O

qrdlt

ri'wir detector time

constant o

4. peak Yl{H:JFlilOflUftJV broad

ntsr[d'h

dr[1.14

6U141

[?tu

hlqnd'os

drArd

ainl

I ufil:lJU[1J

0U11 guard

9vdA

CI1{nOAilUillO guard column

Aqd

tdvd4

column 14:0fl0OuU

IUO UUfl

4.2 nOnilUld0lJdnlvl

: ? I do u il : q

rfl qru tAuil :

v fr n fr'n rrnr

OnilU

: O guard column

v fr

fr n r vr cr o':

q Y ti

fl o d'L

l6t

u01J:u tl4tllnuun ontuu

flr

n'i r rn a ryi

t14:J

Aarrcl

;# Normal Peaks

fi50lnqs0.1?1.: [u

ftodlJu fi o ri'r o tj r.r J q

4.3 guard column j

rtdn {'jriliyl4 ttfi

ldolJdfl'r1\

colr,-n T'm

4.4 solvent

il 6'o s fi guard column d,r'ld' peut

tryRYAlU

lJ:

4.5 1U-r9r145oSUqUOn /t0'

: n #o o n'i A

n guard column 1#nJfi ou guura

nuotuti'?odt ld':u mobile phase rr?o1# solr ent tifi n?Iu

il1flfl?1 mobile phase

lrri

Broad peaks (all peaks)

9r?out{rufl?1uil:.1

q 0'l

rJ n

I mobile

phase

m?o

fi o pi'r o eir.:'l

9.)

191:l]U 0UA.1 4aud

9l: ?0 floulJu19t14: OTU fllJOnflOAlJU

flonlJu ttJ0nfl0{ q qs)

^9) 4.6 0â&#x201A;ŹlrllRtJUOn qU td

rllauu

^ 1'lR]Jfl

OnilU

[1J

ryntd

4.7 :yUUU{ t}JldctU: 4

lT column heater [n 0n?ilfl]JOtU qq{

ul0

rfi

1'l:OUi Iu equilibrate

111.1

#ar

yns

r.r

: v u u tsu column

It0u detector finrrrurAfiUT [[OU equilibrate ?1'l{lflfi I

oul {[1^lu{1,{o t9luqq' Otndnl]y

tnlJ

nl:fl{nl 1

iqy

tT [Y1] mobi le ohase

ad9)

detector [[nySUft detector []JU9lU

u n : tfi fi run gradient

rio r 1 #n

: v u u n d'rL r{r 4

'jdutvtl 0vaqnxout i9l?gto [lJ eouilibrate noum

lu n : tfi fi 1 d ion-pairing reagent o v'ldtta solvent'iiu ru u ds lu

dr:avnrur{ru{u

fl 1 T equilibrate n o d'lL

druru

s mlr,l

I ml/min rflueiu 4.8

daad

n?tilt:? tun]:v,turl

f4r9)

tilflfl9ro{

- 29t57 -

r t to v rJ?ru r

u : un?trut: ? tu n]:T'tu1.,,1

100 ml

tf niu

fidsr:rnr:'lrn

U6U?,I

5. peak fi d'nu arv rr Y

fioutfl u

dl4

nr:Itfi'lt

dlrlJE

peak SOllflU 14:0}J [1]

t fi nr: qorri'ulr.r ritufi

iluiaa@J

q 9)y r

dtatd

?l al It I Ill ?lI lrI t il 11 lt illl l- JILJL'Jr-_J t.--

5.2 uil 1: 1 Jutil ou?] guard

dvJ

;;

5.3

n rn

rSonon:1il

r I f l 11 1r fl v o ra11 io nl 6 ou 1u ri

iluda

Rlin0R:JUfi I 0 suard column ldv6a

- tA flonilutflolJflnlvi

d o u il r y fr

fi n r rnr r o.r

u0tJ: 1J tl.trllnuufi

01nnr: rnqryo.r'ir.:1u

NormalPeaks

guard column

[lJOUUfl OnilUH : O suard column

column 1'15On0AilU

t14tJ

#rqqd'ufi in-line filtcr u?o

guard column 14t0

ftoouu I

yuda9ltdcll

0lqAnUil tubing t11[lJRUu

o d'rL

rf tlr qi r n'i r

n ar

/cl

nlJu tl.lu

nonruu

:l-l_{-L_l\iL I r I

Double/shoulder Peal<s

fiqei'rodr{lerukielosfi guard colum" drld peak rlnd

5.4 guard column j

[tdq{'jt il iU t4 I tn^q o I n guard column

[doildn]Tr

ou guoro

"olumn'lrri o q il3ilrn : fi fi n il ?o a q n rr il ulrL du uor o{r o dr s dru

t^dda

5^5 ]J:lJ1$15?]AqH:O n

1#nJ

rr il r{il slu t ot d'r o cirr

fifinlltr:l#oun'jr

mobile phur" 6't1# solvent

n'ir

unrfiuhJ firldnod'rurf

nrtl r : cr fi q'l d'kir

'1il equilibrarc

n0nlJU [[9lCI] tS solvcnt ?lXJfl']'lut[:{}J1flfl?l 0v d1il

,e) qy

ofi

0% I o{ il;ursr : ryo.r'ir

a'l6"irir rnn'ir I % t o Irj?l

rsr: vo I i rI

floailu 6.

peak'hjaruilrfl : drur#r

r^iaa

6.1 1J5tu19r5TQ911450

fi?lu

(fronting peak)

xr r n

uhi

9Yul

[1]J1JUlJ 0 iFl? 0U]'1

rn^u'lil

d r1 #n o d'ru

il?xr r fl : fi fi

v ?o n n

tY qY

n'ir

r{rL du

I pi'r o dr I dr u

fifinrtrLtt:srioan'jr tv

mobile phur. d'i1# solvent

dril r : 0 fi q'i6'1riru

o{ il;rL

: vo

n0nLU [[n0] tS solvent ?1ilfl?1ilU:.1]J1flfl11

equilibrate

il r : 0 fr q'l d''hi

rL r

t'ir

n'i

%'u

il3il

: rio.r'i

r.r

noailu

ffi Nonnal Peaks

6.2 solvent

fildnvnru

Ulnn'j'1mobile phase

lJ:lJtgl5llUOUn

dr.rnodlLrjuio

guard column

dtat

Aul

column 1'l:0fl0ClJU 6.4

- tA n0nlJuldOildnlr' ar16l

01nn15lfl9tSO{?1'l IU

noalJu

- 30ls'7 -

I mobile phase r io fi n pi'r o dt'r 1 u

n?I

6.3 xJnl5 lutilo1.J11 guard Tfr

pi'rodttfin r"rilrr:.1 l

Fronting peaks

AyAlSgl?0U1{9t?U mobile phasc 145O [U solvent YIU xJ [

[: { #o u n'i Ac)

fdud4

[il nuun oniluH 1o suard colurnn a

do lt lJ : y dm - qY ri

5 fl 'l 1^'t I o { fl o n

uolJSlJ tl4rilnuuFronilu

t14 xJ

u fi 1 gn fl ? 1 [n fu c?l

ilryrr

nr:rri1'ln

dlrHq 6.5 guard column j

s'qeTrodrllqulileio{fi

[doudRtl.i'l

rldq

i'jtil

tirld

guard column

iUil rtfr n o t n guard column 1 #rrJ

peak rJnfi

u guuro

"olu-n'lril 7. peak

lilaruursr:druvd'{

7. 1

}Jn]:ilutilout't

guard

4ud

(tailing pcak)

column 14:OflOnlJU

lr ?l I,I It [ [ ll <l I I

dl4t4

7.2

c{nlv'l

ar16l

0'lnnt:tnqs0.11l.1tu

lr |lil llil nI |lt IUUL--JLJ T11

;;

l_-]Uuu!_J

ldadA

dOU U : y dnI n IV'l I

U0u: U

qS)

t1'l[1J RUU

On f l OA

?gt

il U n Ifl I n ?Itn ilcl1

rlOnilU

Il'J lJ

n0nlJu 7.3 guard column

tdolJdnlil

lr ?l Lirt

I O guard column

lilOUU nOO ]J 1-111 T 0 suard column

ri'r o dr I I q

da {'j I il

Normat Peaks

3l ot lill lil |l il

fl00ilu[d0u

YvdA

A1{nOnilU

il elo { fi g*ard column tir'lei' peat

[U t4 I lfl'Fr 0 I

r-l n

fl guard column 1 #nJ fi u u gr.uro

column'luri ll

L-

;;

Fronting peaks

f .+

ilErirorfl

a L a

injector

911191-t:1{yl 8 tlnyq151.1yl 9

aze

lYu ?]n?:?ryilr:ou9l vddqa

nonu r1jilqqqnnuH:o . ^9^-^ T l!91 rnlcctor port ^^^QFl9llJ <t

[uu9]u Z.S 6'snir detector time

6'qnirlyri

t4r9)

constant []JQnflO.1 8. uogr peak [[UU 'hitruaru

a.t d'lrir detector 'lil

9t5?0

vtcty9)

flOullnyn{fl I tH0nfl

9)l

fiflgro{ [ryu fl']'luul? fl0U sensrtrvrty. autozero 4e)

tljU9rU Normal peaks

8.2 recorder input voltage

rJir rirn rrlsir.r

frn

#lylft r1n ri

1ilqnd'os 8.3

raAda

lJ:lJ'rgr:ilqqil:O 9tYul

n?1il['uutul0nn?oul{ ilma

Tailing peaks

aVtloct/ud

ru]nlflu [u 111n4flOnilu

tA'joug9J9)ut9)

il :

Iu equilibrate

]J 11 lJ u

{

9l ?

0uInn?u

[S solvent yl]Jfl?1]J[[:.1X]OUflll 0Y rn

n'ir

tY etl)

i il?ru

rsr

: ryo

dki

ru r

n'i

rir lu

[[A0l [5 so]vent ll]Jn?lU[[:.llJlnn?]

a r il r : n fi q'l

nonlJu

t q t

fll

dril r : 0 fi q'ld'hiru nOnruU

- 31t57 -

a q il : o a q n

mobile ohase

lnt

il;ru

rio.:'ix 1 u

ilqur 9. pcak

nn'l#r

nr:trfi'ln

d,UlJE

(negative peak)

e.t kiarui{'qiyreu -,1,a/

ntou

[fl peak

o I

o1 0 in0lJ?\ ?[9]

a a r u iY q!

6U'r ru rqt 0

il fl 0 n

lqY

a v{ ? rel 0

}'l

I Q

f,i el

o.l

naun']u yl

9.2

uld glifll

polarity

d'wir polaritv

lrri

a^d

detector

OnOilY{ ?1910:

'hirururvaru

-Iido

Negadve peaks (all Peaks)

10. peak U'1.1

n#lrir

(negative peak)

l0.l rilu

)^ i peak frtnrrzuu

'lu:vriu'uol

peak

t, ?l |, I !t rl I I il lr

ifl

mobile phase cl

1.i

1t [14

tj:t-ttilnuu condition [1]fll5 run

separation

d'llJ15 ilItUfl 0Ofl

to.z 1ud'rodr{f,

dr5il:vnoilfifinir nr

rJurflou'lu

0lfl

. -.-a

peak Yuglu

peak ttO.t analyte

d o u'jr pcak n d'u fi'r ri'u

r n pi"l o

dx v i oa

lJuLr-L_JL-Jt-

;;

rioun'jr mobile

Normal Peaks

rl ,t |l l, Iil :t ?l ililil

ion-pair

v19/

RYO-lUgl ?OU1{9t?U

phasc

solvent

- tilrurornrirodr{l#ilfurilfi

uu condition

lunr:

nln v

- tltlro tnar: rlu rtl ou1il solvcnt 1#rrj fi uu solvent

l-uL-L,uJ'*

luri

;;

Negadve Peaks (one or more)

aty

10.3 mobile phase

nYAlU9ll0U'1.:91 ?U mobile phase

qflndutra{'ldxrrn

ilfunJfiuu

condition

'lunr:

,un 1# peak

vfinfi

f{l}J1: filtufl OOf,i0lfl pcak 1O$ analyte n J fi

r r ru u r r

du fi

ldl dfi ni r q.r n'j r uv-cut off 'u 0.1

mobile phase trjd uuryfi 10.4 autoinjector

qfi

ornrgr{r'hJ'}u:vutl

- 32t57 -

tiu

qto{

mobile phase

jie)

9lflgn:t{il

9 u0

Av4Ylaultz)

I 5 t.r

fr 6 tJan I {t(i ru q iun I il fl t o n I no I ulj : lJ I cu lilQ

flqxr 1. 5311. peak zuOS analyte

nn0

{

nr:Irfi'h

flrtfiq fie

r.r larirlu:uuil{'anl:

dorynhignd'o.r

: l a ri r r r a v rJ :-u r t fi 1 #q

sample table

calibration table

refcrcnce peak

peak window t6'{

peak threshold

retention time

Ju 1

idou riu ilioqir

tln#u

1.1.2

run standard

luri

rlo.r d'.r fi

{r.r tfi u hJ)

qzuafild'

9tmgr't:'lnil 4

1.2 retention time

rjl TlJOUU[[Un{

z.'hirJ:rnn

peat<

q{o t.t

r'lcirirlu :vuud'qnr:

riorynhiqnrlor 9l?19|t:1{vl 4

2.2 retention time ,it'

lilauurlila i :. rJ:rnn p"'L

fikinr:fi

osstU [fltru1 t9ltlfl:u (ghost

:.t finr:ilurtJoulu

rniut

mobile phase

solvent

peak)

lrri

mobile phase

rfirr

HPLC rn:Fl mobile phase lotuti'l

qrf

sonicate

adrarcl

H:Oilnl:ilu[lJOU : vfi

3.3

rdul

?l{nl:

tglT

1'l

rfi u 1 [v{.1

#rhjnl

9t: ?0fl

vr?udou?5fl 1:[9]:uugl?ou].luu

[Fr:u]J9r?0uln lil]J

ydd

ouflnlnro{91'?od1{

durot

unrT

run

n1{n

c{-r rJ r

flflun9r

ri"t o d r': rfi o 1 #a r :

vnou

fl elute

oonil1l.til9r ru

6r nq{

?u solvent

e.fi'u o

qfinr:xfi q {o

3.4 :vUUO'l{[Xilq9l

ghost peak Itda{'j-1

Aa4vrJ

0&tlfl elute OOfllJl

n9.: rrnv

standard q9,, tr'q s. q r iltuH]tfl9r0tfl9r?ouli t14[tfl [u [0u

ar:rJ:vnou'lu

?o u-t{ nq q fl 0 u

Ululu

a5?0c{ou]nuflt:fin

u}J9l?0u].:

utiotYA 9r

Nitric acid srrrud'lurir

rudlu methanol

Flush t[03 equilibrateT

rrnv

lirnrrtavoter filter fioqjlutrnIIIQ filter'hl

:.2 d'rotixtfiorLanrn

lpruldar:rnfififiqflinrv{fi

33157 -

dd ?1il n ?'l}J

[

.-C_ { }J1 nt?{ oryv fl

u n o d'lr rf 1 du r u o o n 1 u : v fi

a.:

'j'1.1

: yl fl 'l 5 run

ilcgrr

nr:rrd'lci

fllr1.1E

:.s nr:dx

9,,yrdr

nli

sample loop

sample loop 0t'lU mobile phase TY11?1{fl1:qnt[9]Ov

'hirfro.rrnro

Fl5.:

a i

c'vd

try[uilaqTflvo'r9l

:.0 rfirufiq'lildvorer

dt.: injection port

:.2 finr:rJuttJoufi

injector

rr]duu seal [[03 filter

3.8

inlector [tOu sample loop

9)v(

drArd

nr{flon3Ju

ilnr?ilurlj0u11

rrlnuunonruu

floatJu

jm d A v I o e, lv '[d 'oigronu ldu ryi010 91 0.1u: u condition'lu solvent fi 'trifi

3.9 lJ stabilizer 0Q

mobile phase

t1J

nr: run'Irri

145O

stabilizer lljouu dn111 +.

r1:rnn p"ut riounitti

nrqfiqlulnirltlelrn:tL

vt4

a{o r.:

I n?0U1{[do]Jdnl]^l

+.2 qtgtfio resolution

d o il il : v fi u fr n r vl

1r er

a9)

f t{r #o un'i

o { o o d'u

r rn

u qiu

o rL

l14t1lRUUflOnilXl

jo9u, 4.3 mobile phasc 11lS

tll

rsr?urL mobile phase Yo4dY^44 '111

c|flFro.1

5.1

nl:ri1u?6tt peak

height

r$o

| [U 1'l

l','|

5 0 :J

fl il n 1 ]

I U t]\ ]J I U O 1 o tfl 9l 0 I fl fl I :

navfloulJo{a?ou'1.1

5. qâ&#x201A;ŹU[dU accuracy

tru

5. 14r

peak area IU

flflp{o.l

? 0 do

u n r : 1 ri

ni r r tn v rJ

sample amount

scale factor

intemal standard amount

retention time

t r r11

o ,

n o{o

I d't

i',rv

5.1.2 0lnUU run standard [14]J qt{nm l9l s.

3'[u n : tfr fr : v ti u o'q n r : rio

amount

qn: ru{1u

finilniq l#nJ:^uurfiuu

t't

response

a componcnt

tos

peak

rilnir

1JO{ standard tO.l

- tlrr.r u'j r response q n r{0.:

[

[e{q

{'j I il iU 1'l'}]J I 0 I n

quantitation

- t{rvr u'ir rcsponse hi qn do.r integration u6o chromatography

- 34t57

tdq{'j1il

iut't

til

-Ifl

nr:rrii'lr

drrl4E

lJiu141 ut4

q{o:.2

5.2 9l?OU]{[dO]Jdn]il v

4dtdt

14501JflT:1JU[110U r4

:v11 ?1{fl 'l:

tgr

[11

: u}J9r?oul{ d

-la

5.4

^Yd tuqruilQilqluu

F11.1

nr:rn;urud'rodrshl

ll?UdoUnt:

dorrYYc3l

: U]Jg]?OU]{ ITU fi2lrJ[UulJu

[9r

: 0.1

[1JU

glu

Qflelo{ 5.s

Seal ltCIYlfl U9l?oU'l{

5.3 lJnt::s[14U1JO{

nr:tn6uil

: UtJ

[9r

091

i.lY

' : r d?U Tt tT ITU

; U

lil U- nllnvfl

'.l: [0 0

dty

hiqnd'o'r

standard

dounl:

li lU

intemal

0'1.1

tljilau

[nEuil intemal standard 1uil s.o

ilrgrrortlnrsfi q ('lu oiq

drld

nxeo-toop manual injector o{ il?r:

nlOll'l lT external

tvirzu

standard)

dtld

5 loop

uii'hJlu

1#1aar:ovotu'fi'lu?u

toop riou d o g d'{ fi el

partiat-loop manual injector

fi#oun'jr

plol fi q ar : 1u rJ?ru r sr :

50% 1Jo{il;}J1Fri sample loop

v qr)

fl1tT syring-type mauual injcctor 49d; ^61 - [llftufl [ufl tTqanyno{iln?tiln{l] - ldturqto{ loop qnd'0.:

a d 4 - 6lt[Tyu9ru0{t1ilq90fl

ao.1

l1ilqflfly0191

loading port llJS?SlJ

ln ki' <

y q9

fi1 [T autoinjector

lry1U191110{ loop

Qfi9loi

a d d I - Q9 lsryuqloi[tilqqQnnor - lililorn'rfl tu[1]Jq9l h

I d

vial lJFl?OUl.lOU[l.''lU.lltO {

rd .9 "i - l4[]Jufl1::?ryil

hi'jr

tdt

manual u?o autoinj ector ov d'o

rduyrxnr:'l?to

- 3s/5',7 -

equilibrate Jq u

?t.t manual injector

purge autoinjector

lilusirrrurj{fifl

s vi

i'Ifi

n'l:rrfih

fl'l[1.jq

tU 1.1I

qqtdU precision

n]:d]u?fl,l

6.1

peak

q{o

s.r

q{o

s.o

height lt6o peak u."u 'hj

qnr{or

flqmrornnr:fiq (lu

o.z

oiq

n:filll

[Y extemal

standard)

O.: flcymrorfl injector [sl.J

t[AyAl:t{11

Q?]Fn:"r{11 4

[rAVn]:l{?]

91119n1'1.111

Ai'-<a4a

t'lo ?: ?ryufi: ou9rgr9tflu ddvAo

[1LA9qqflUH5

OY']:"91 -dt9)

injector port Q0au tUU9lU 6.4

utA 9|

?0 u]'l !d0

o{o :.2

f{n lv'l

4draro 4dtAtcl

?i:0iln1:iluru0u lu :

rdul

v l.t

?1.1fl1: tgt: ulJ9l?OU'1{

o.s ln : rLrlsr rrns:Jfr fl cuil'l lS1-! retention time ri

rilauulruni

tl il:oau:li

peak [.l9lUfl9l 6.6 response

detector

crl)

fll:lnil 7 ilâ&#x201A;Źufilnr:luir1l{

fild'orn

dFlO.1

pump

1ifu14'1 1.

pump

.ihidxru s[9ruvtTl9l l,t

nru[tnU front panel lru

lixrurryuri'u

n1:rrfl'lu

er1r14E ln rl/tt4

1.1 trJ torduuilnn trl lr

Vt 19,

dou n1: rfi uu ar8'ly,l

d o 1r reir rfi

1.2 trJrJfl:v[[d [1^lTllvl rgt1lduuilon tyl

"[crll

u il d'n'l

d A gt d u r rfr I u fr rei r rfi

{s r u'[

fi r r r u r rae q'i r refi r fi u u

rJnhlr,^'l1uri 1.3 fuse

1]9

n]fiuu nrse'[mri

- 36157 -

1 or o

ri r q rl n : srfl

tiy

rJ a''n 1 v'l

d'n

lvl

v'l

fi r fi r nj 1 o'j r

q rJ n

: afl il fi

r hi'nj

fr u u rd'r

tir

fi il iu u

rfi u L

ilryrr 2. solvent

hilrn

nr:rrfi'Izu

dt[1.19t

2.1 fuse 1l9l

njfiuu fus" tnri

2.2pump hjrfrorLsioriu

pump controller

dltu

dOU nl : [5O]J

to v fl

9l,: 9nt[1.1 u.1

[ lCI

?1.1

o.: IEEE-48

pump controller

kilq u hido.: ld

controller fi il I u

tu IEEE-48 8)

[ [6] U

UISOIJF| O

pump control ter t r fl et {'j 1 pump

r ( #r ru o o e o r u rfi o ru si o 1 u t

y a9l

B (6 1

6nnfl tit p.rnp ntilt;fi

oon olnfutflqcfiytli pn-p ri r I r u

pump f]U pump controllcr

qadA'

il 9l d?1'l T pump

ru v

fi q rJ n :

il{c{o.1lil9td??tsog)

limit

n9/

1J0.1

pump tX({n?1

iqY

fl'tluflu11til{1u

oto.t

Aqgt

o qy

nlfl ?'lfi?'llJ9l Ul1 tU{lu 01q in?10 111{lU

z.+ #.rdfl : rnr:'hi

tol p"*p 1nill#qnrlo.r

6'lnir to* pressure lirnit

2.3 flSn'l low pressure

U ?11

ll4 pump

(ovelol

nqnl:

) 14

tlIUOfl;tn1: lltA

drl

pump [UUfluU 2.5 pressure transducer 0

'Iil

4 id . uqq'6r tu 9nill.i u{' }'llu u fl u u

1.15

2.6

ilt

prerrrr.e transduccr 1#ril u qu ri'lu I

lilld'prime iltt

2.7 draw-off valve 4o,"s

tHArllUflUU q

prlme

1'10?1J

fi dn : r nr

tndr?

cioilHioril6uu

0vr:91

â&#x201A;Źu v

Pressure transducer

ilru'lrri

14tl1l draw-off valve

1ri'rnju

H:O:?ry}J

flJfiuu seal1o.1 draw-off val,r"

2.8 solvent'Iu pump head

'hi a r ru i : n p.r a ru uir riu 1 d'

lnOfl tT solvent

?0d o

il'i I solvent dl

q9i

lJ'l

htri

r{rrYu 1 6'u io

d I u vt9) yldltJl:ntiflil[11flU t9l

gtA purge pump 9l?U solvent 11tl1lJledlJ 2.9 check valve

l4io outlet

dtu

inlet

anil:nrio

dou'jtfi

rfi

ariol

fl e o19rl.t : otil a uu

t1.1

inlet

uio

outlet 0 tfl

1il-l

dr.: li rn r rrL

dr.rrufiqilnfi 2.10 plunger seal

rqot

ififi

solvent

frdruoonvrrldrurfrl

a9.r'

v fl

[fl n 0 :

seal'it!pt

-37t57 -

O U

11'ln9l I U t4 0.1

qv .tfnjduu rj

seat

lrri

pump head

Niorfie

pump hcad [ ldn i ?1 piunger

1lfu111

nr:rui'h

fr][1491

d mobile pnur. ri'.r o qj1 u : v d'il rd u rrYu u ?o pi r n'jr

z.tt fiornrg'lu pu*p

head (tqy'ttv pump 9l?1lJ

pump

ri'uq.r;

tubing fimr.:uh pump 14o?u

q'fl

99 4 oO

cl I t 11 U

1f n : r

U fi

{

To ujd

[

[?'l u.1

Q.l fi ?1

pump

ro uioqnri'u 1#rr1ul#trrju

uu"lr ri

degas mobile phase

1il6 1#

aegus

'luri

filter fioqjlutler mobile phase dnil:n 1#ooo filter'lrldt

fi?'ltJcjvola]nudrlil

sonicate

N Nitric acid qtile{?u

rii : rrir [tayfltilel?u

methanol

ln^nfll:5vtHUllo'l mobile phase 1Jâ&#x201A;ŹtJUoqjlu pu*p head 1# prime pump tubing

Aydyr.?--q9) 11 11 1 n t1J

1 pump lJlllJ1fl tdU ft 1qil Ufl

o rq,i

A1{

n 1 U tU U

g) oY rj

n r19/

lnu til}lnt lH mobile phase t1r0!1!l ll'ilURUU lii tio

t14lJ11il1JU 191ril

2.12 n1YU1{

fi 'lv

ilt (pump dampener)

',l

r?o

O U

l"d rubing

il]y du

9t:?0cfoufi-l::-?6il rrjdeJu oumn damoener u$o gi'rn:o.t rdu': t?ix

d'rn: orrfiul'lu:vuu

r: n'r'rud'ud{"f qq

2.8

?^rior X1A1[Uff?U111111#

solvent ruarLtr?rriu 6

r6o

p'lush dr s r r d: tail

0u tfi[1Jduu

?o q rJ n : aipr a

ar:

tYr ti'r

s rnru

fl iy u

t?flJ

qiln:niruarudr:1u:vuu

fiilryrrlunr:dr{'ru d

fla9l0s]n

2.14}JO[FIO:11O{ pump

u-rlel AqflOS1.1

2.15 circuit board "frqn 3.

o,Lai

ilnl::-)ryilil

pump head

3.1 pump plunger seal

dfl

njfiau seat'lrri

fl:ou 4t

3.2 plunger dnfl5OU

.lioil 1.riotil6 uu plunger'lu ri

3.3 pump head 140llJ

ilryudngf ido{c.ro{ pump head t#rnju loruovdorur;u q y

trty

tl"l[tUU[yl'lrlflU ilqyilUtlO? seal Ovqnt191 tq [[nyOyAO{

l4rrat4t

lil?luuttuu0urflu tu

- 381s7 -

n1:rrtl'h 3.4 inlet ?1?o outlet check

r 4u hlrnju

valve 14d?u

rrsi:

titting

itodrl#rnju

v femrle'j r r

urfr u'lr-J

I r io nj u rn^u'hJ n ?o r

^o-^__- 4 t4 r

q r9,/ o ri Ft1..t : 0 tlJ rljCIuu tu ru nttitl0r

s'l: 4.

<'i'a.i

ilnl::?ryilil

draw-off

valve

s. n

dq:rnr: lHaHiod'srrv

&ar^

! udt:

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ilfl

4.1 draw-off valve

l#rnju

draw-off valve

14tlxJ

1'10?tJ

14:OSt:91

ttiduu

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nJfiuu inlet line

'lrri

rit mobile

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5.1

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seal lJOi drar,v-off

phasc

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u u lH *;

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{

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prime flru1uri

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'hirfio.rno e4 q

prime ilru1r :

dt 5.5 inlet

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ljio

outlet check

dfliltnu$olixru

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u'iI hifi

v'l o s

lil

mobile phase

I fl ?l

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degas

r?o

d'lu helium

sparge

dV O-l9l check valve

dO U

rdrjilv -l'l lJ fl I : n : O.: solvent

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t^lailfl9r

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dtL tfr O tl

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: r ar

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solvennriovioal

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dt:CIyntuo9lfl'u

lirntrruavotFl filrer fioqjlurroruit0 mobile

filter'lil

sonicate

gvrAi'o

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s.z

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rfo

nlush fir

tdd

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o{ hinJfioulrri

0iln:ilrududt: [u:vuu <q4 rJ u[u].t

?t1,1'lu

- 39t57 -

phase

Nitric acid srrild'ruu't

Inodr

n9.r rrov

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nfru aru a r

riu

tir rir

w r fl q1 u r

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dt[1.rE

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:ou

lfl

t3[11ufl

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l5: ?ry]J1'li

pump head

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neaa'hi ar u

flu

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lu pu*p

rL'r

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rbd,t-laa!

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d,ru',rauusluriot

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UFI

nanuttuu 8,3 indicator rod

9,/44

(ftlu)

U9l

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manual injector

rlcunr a,i,<.i

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nt:trfftn

fltlfi91 1.1 injector port

rjfur?onjfi uu seal'lm

rqfl

1.2 syringe needle g"ug"

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ector port

sea

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41157 -

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1'150

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backflush injector

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injector

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sample loop

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port 5.3 al

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injector

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d-li qQdnu potl tlJ0{O1fl

gnment

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injector port

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crt)

9ll:l{lYl 9 lJCUfllnl:[U{lu autoinjector

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l. autoinjecto.'hjrit'ttu Jq ufi

d'qailr[0u front panel

hidr.rruniuflu

ntflrffh

cl'ilHq 1.1

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d.: r u 1 d':L r rfi

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do ufl 't: rfi uu ar u'lrnl

vi r r r

d o 1l e{r rH u u rl d'n

nlfruu fuse

- 42157

u fi u{r rfr u u

r re{r

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luri

fi I u il n'n

ri r q il n : a:'t v'ht r fi

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fi il iy u

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rlsurir tn ro

2. autoinjector [lJ?11illJ

s i-

lFlu111''l9r

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nr:uiT1zu

cTtlfi91

t6orL

2.1 autoinjector

fl : ? 0 dO U fl I : [50

]J 9l O

gioii'l controller

y ?l ? I

{

autoiniector flU controller il q9t

rrovfl'r:a inrl[]1u{uoi IEEE-488 (01 t.ir IEEE-488)

lirrru

retdtu160ileio vv

oon 0tn#urtiqA?md autoinjector 6nnfs d1 autoinjector

d'lUlTCllll{lU ..

control ler

149,/s

4s4

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ctY

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fitting Hn?ilil?o[ldu

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lnu tu 3.2 valve seal "frqot

rilduu

3.3 seal pack

trJfiuu seal pack

"f t:"or

seat

nJ6uu fluid pack

3.4 fluid pack "irqn

4o@A-qdd

d4dq

3.5 rlruanlJUqH'r

purge autoinlec,tor lfl 0fl I 0A yu^q4dq9r't.{da

0lu{rnqil6u11 ton 3.6 syringe

qqnuH:0

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t}1[ilnuu[1ilqn[[nv

seal pack

trJfruu syringe

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tllu

tflu

fitting fln?ilH?ottdu

: ? t do U fitting

njduu fluid valve

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trJ o'

y fermle

tlJ

4.2 fluid valve "ftqrt

d'rodx'hir,ir.nu

3. lJnt:5?rylJ

tor

a^oq'l

dAA

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tu ' Isllul.l:0 t]J' ot llJflnaoi t14]J5ulrfl I

3Jfl

?t:o txl glr

2.3

autoi

- 43t57 -

Uu needlewash pump

nJ 6 u

u 1 H ri tir

"f r

;

q t

9) 1.,1

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nr:u#lru

cTil149l I

needlewash

I r q needleu'ash ldo

p1.:

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n'jr

qjri r

autoinj ector

'lunn'rq

autoinjector s. flcyurnr:finpi'rodrs

Isu

5.1 injection valve

purge autoinjection

"ftlot

lnrda

tliti.r

e]?our{ tilQnaa H:0 gr"l:r.r peat fiorrJnrr

5.2

ddua

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il cgr r1#qio

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sample looP

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u r.ptum

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u o 1 n 1 fl a n

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5.10

nl: align :vlldrril

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5.1 1

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alig'n'lilrj

nJfiuu injector seal

- 44t57 -

1Y lu

Yq--^l

fl1 tilQnflo{ t1'l1J:uttfl[14]J

zuoila19l

injector seal "irqol

riu s 1 { a 1 T

n1:uiT1r 6.

1ilfi solvent lnor.iru

injector

m In

9y'

lavio:vu'jri

6.1 tlJ t9lFlOilO:VH?li

injector

fil pu-p 'lt{ solvent lmne.iruki

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autoinjector

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4 t4r

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lJfll; 7.

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purge automJector u

qyt

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11910

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loop QOflU

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pump rrhq injector outlet tubing ta

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t .q.z fladum

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- 45/51 -

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tonfrr50l.los Andrea Weston, Phyllis R. Brown. 1997 . HPLC and CE Principie and Practicc. W.B. Saunders company USA,

Calilbmia. 280 p. Guide to successful operation of your LC system. 1994.Waters. 34 Maple Street, Milford,

revision 1.5194.

- 57t57 -

MA 01757, 022318TP,