hpltlc notes

8/13/2019 Hpltlc Notes

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HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHY.

PRINCIPLE:

It involves three steps i.e sample appliation ! hromato"ram #evelopment an#

hromato"ram eval$ation.

%tep&: The samples to 'e hromato"raphe# are applie# to the hromato"ram

la(er .vol$me preision an# e)at positionin" are ens$re# '( the $se o* a s$ita'le

instr$ment.

%TEP+: The solvent ,mo'ile phase- mi"rates the pre#etermine# #istane in the la(er

,stationar( phase- '( apillar( ation. In this proess the samples are separate# into

*rations. A*ter evaporation o* the mo'ile phase the *rations remain store# on the

la(er.

%TEP: The tra/s are sanne# in a #ensitometer 0ith a li"ht 'eam in the visi'le or$ltraviolet arn"e o* the spetr$m.a'sor'ane or *l$orensene is meas$re# '(

#i**$se re*l$tane. Alternativel( it an 'e eval$ate# '( vi#eo tehnolo"(.

The Priniple o* separation is a#sorption. One or more ompo$n#s are spotte# on a

thin la(er o* a#sor'ent oate# on a hromato"raphi plate. The mo'ile phase

,#evelopin" solvent- *lo0s thro$"h 'ea$se o* apillar( ation ,a"ainst "ravitational

*ore-. The ompo$n#s move aor#in" their a**inities to0ar#s the a#sor'ent. The

omponent 0ith more a**init( to0ar#s the stationar( phase travels slo0er. The

omponent 0ith lesser a**init( to0ar#s the stationar( phase travels *aster.

INTRO12CTION:

A2TOMATIC PLATE COATER:

The "lass plates to 'e oate# are onve(e# $n#erneath a hopper *ille# 0ith

the a#sor'ent s$spension.

The la(er thi/ness is #etermine# '( either a *i)e# "ate 0ith pre3set spaers

or '( a "ate 0ith a#4$sta'le #istane.

The plates are move# '( a motori5e# onve(in" s(stem at a $ni*orm *ee#in"

rate o* &6 m7s! to ens$re a $ni*orm la(er.

The A$tomati Plate Coater is availa'le normall( 0ith a *i)e# "ate *or pre3

set la(ers o* 66 an# 866 mirons! an a#4$sta'le "ate *or la(er thi/nesses 69

+mm.



1RYING RAC:

• The 1r(in" Ra/ onsists o* ten in#ivi#$al al$min$m tra(s! +6 ) +6 m!

0hih an 'e sta/e# ;$i/l( an# onvenientl(.

• A tin 'o) *or storin" the tra(s an# t0o 0ire han#les! to move the sta/ 0hile

hot! are s$pplie#.

•

• The 1r(in" Ra/ is onvenient to $se! parti$larl( 0hen TLC plates are

prepare# 0ith the a$tomati plate oater in lar"e r$ns.

•

The 1r(in" Ra/ also omes in han#( *or plates smaller than +6 ) +6 m.

PLATE C2TTER:

2se# to $t HPTLC plates easil( an# more preisel(

C$ts plates 0ith a thi/ness $p to mm

1oes not #ama"e the sensitive la(er

Eas( to han#le. Rea# the re;$ire# si5e *rom the sale #iretl(

Helps savin" osts on pre oate# plates o* hi"h ;$alit( '( preventin" o** $ts.

IMMER%ION 1E<ICE:

For proper e)e$tion o* the #ippin" tehni;$e! the hromato"ram m$st 'e

immerse# an# 0ith#ra0n at a ontrolle# $ni*orm spee#.

e( *eat$res:

2ni*orm vertial spee#! *reel( seleta'le 'et0een 6 mm7s an# 86 mm7s.

Immersion time seleta'le 'et0een & an# = seon#s an# in#e*initel(

The #evie an 'e set to aommo#ate &6 m an# +6 m plate hei"ht

>atter( operate#! in#epen#ent o* po0er s$ppl(.



PLATE HEATER:

The TLC Plate Heater is #esi"ne# *or heatin" TLC plates to a "iven

temperat$re! 0hile ens$rin" homo"eno$s heatin" aross the plate.

The TLC Plate Heater has a CERAN? heatin" s$r*ae 0hih is resistant toall ommon rea"ents an# is easil( leane#.

The +6 ) +6 m heatin" s$r*ae has a "ri# to *ailitate orret positionin" o*

the TLC plate.

Pro"ramme# an# at$al temperat$re are #i"itall( #ispla(e#.

The temperat$re is seleta'le 'et0een +8 an# +66 @C. The plate heater is

protete# *rom overheatin".

%EMI A2TOMATIC %AMPLE APPLICATOR:

The instr$ment is s$ita'le *or ro$tine $se *or me#i$m sample thro$"hp$t. In

ontrast to the A$tomati TLC %ampler ! han"in" the sample *or the

Linomat re;$ires presene o* an operator.

ith the Linomat samples are spra(e# onto the hromato"raphi la(er in

the *orm o* narro0 'an#s.

This tehni;$e allo0s lar"er vol$mes to 'e applie# than '( ontat trans*er

,spottin"-.

1$rin" the spra(in" the solvent o* the sample evaporates almost entirel(

onentratin" the sample into a narro0 'an# o* seleta'le len"th.

%tartin" 5ones spra(e# on as narro0 'an#s ens$re the hi"hest resol$tion

attaina'le 0ith an( "iven Thin3La(er Chromato"raphi s(stem.

MAN2AL %AMPLE APPLICATOR:

The Nanomat serves *or eas( appliation o* samples in the *orm o* spots onto

TLC an# HPTLC la(ers! preisel( positione# an# 0itho$t #ama"e to the

la(er.



The at$al sample #osa"e is per*orme# 0ith 1isposa'le Capillar( Pipettes!

0hih are preisel( "$i#e# '( the Capillar( Hol#er.

The Nanomat is s$ita'le *or

Conventional TLC plates inl$#in" sel*3oate# plates $p to +6 ) +6 m

HPTLC plates &6 ) &6 m an# +6 ) &6 m TLC an# HPTLC sheets $p to +6 ) +6 m.

A2TOMATIC %AMPLE APPLICATOR:

A$tomati sample appliation is a /e( *ator *or pro#$tivit( o* the TLC

la'orator(.

The re;$irements *or an instr$ment servin" this p$rpose! i.e. preision!

ro'$stness #$rin" ro$tine $se an# onvenient han#lin" are *$ll( met '( the

A$tomati TLC %ampler .

The AT% o**ers *$ll( a$tomati sample appliation *or ;$alitative an#

;$antitative anal(ses as 0ell as *or preparative separations. It is s$ite# *or

ro$tine $se an# hi"h sample thro$"hp$t in mass anal(sis.

%amples are either applie# as spots thro$"h ontat trans*er ,6.&98 miro

liter- or as 'an#s or retan"les ,6.8 to B 86 miro liter- $sin" the spra(3on

tehni;$e.

%tartin" 5ones spra(e# on as narro0 'an#s o**er the 'est separation

attaina'le 0ith a "iven hromato"raphi s(stem.

Appliation in the *orm o* retan"les allo0s preise appliation o* lar"e

vol$mes 0itho$t #ama"in" the la(er.

Prior to hromato"raph(! these retan"les are *o$se# into narro0 'an#s

0ith a solvent o* hi"h el$tion stren"th.

The AT% allo0s over spottin"D! i.e. a se;$ential appliation *rom #i**erent

vials onto the same position.

This tehni;$e an 'e $se# e.". in pre3hromato"raphi #erivati5ation

A2TOMATIC 1E<ELOPING CHAM>ER:



The A$tomati 1evelopin" Cham'er o**ers onveniene! sa*et( an#

repro#$i'ilit( *or #evelopments o* TLC7HPTLC plates an# *oils 0ith the

*ormat $p to +6 ) +6 m.

Chromato"ram #evelopment is the most ritial step o* Thin3La(erChromato"raph(. In the A$tomati 1evelopin" Cham'er A1C this step is

*$ll( a$tomati an# in#epen#ent o* environmental e**ets.

The ativit( an# pre3on#itionin" o* the la(er! ham'er sat$ration!

#evelopin" #istane an# *inal #r(in" an 'e pre3set an# a$tomatiall(

monitore# '( the A1C .

T0o mo#es o* operation are possi'le: stan#3alone 0ith inp$t o* parameters

via /e(pa#! or remote operation *rom 0in CAT% 0ith proess monitorin"!

#o$mentation o* operatin" parameters an# reportin".

A onventional T0in Tro$"h Cham'er is $se# *or #evelopment. ith s$h

ham'ers e)istin" anal(tial proe#$res an 'e $se# 0itho$t han"e! 0hile at

the same time! environmental e**ets an# operator errors are eliminate#.

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