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Chemistry Analytical Lab > FT- IR Spectroscopy > Sample preparation for FT- IR

Abo ut t he Lab Circular Dichro ism Ele m e nt al Analysis Fluo re sce nce FT-IR Spe ct ro sco py Bruker Mattso n Mass Spe ct ro m e t ry Nucle ar Magne t ic Re so nance Ram an Spe ct ro sco py UV/Vis Spe ct ro sco py X-Ray Facilit y Co nt act Us De part m e nt Ho m e Lab Ho m e

FT-IR sample preparation

1. LIQUIDS:
Place a small dro p o f the co mpo und o n o ne o f the KBr plates. Place the seco nd plate o n to p and make a quarter turn to o btain a nice even film. Place the plates into the sample ho lder and run a spectrum. If the sample is to o co ncentrated, separate the plates and wipe o ne side clean befo re putting them back to gether. The KBr plates must be tho ro ughly cleaned after this pro cedure to prevent co ntaminatio n o f future samples. Wipe the windo ws with a tissue, then wash several times with methylene chlo ride (o r ano ther so lvent that will take o ff yo u sample), then ethano l. Use the po lishing kit in the lab to po lish the windo w surface. Wear glo ves to prevent fo gging. The cleaned surface sho uld be clear and free fro m scratches.

2. SOLIDS (in solution):

Prepare a co ncentrated so lutio n o f yo ur co mpo und in a suitable so lvent (e.g. CH 2 Cl 2 ). At t e nt io n: Re m e m be r t hat all so lve nt s co nt aining wat e r will e it he r disso lve t he KBr plat e s o r m ake t he m f o g up. T he bro ad wat e r band m ight also m ask im po rt ant bands o f yo ur co m po und. There are several ways to do this depending ho w much material yo u have at hand: either yo u place a small amo unt (2-5 mg) o f yo ur co mpo und directly o n the plates and add o ne dro p o f so lvent, o r yo u disso lve it in a small test tube first and transfer this so lutio n with a pipet o nto the IR plates. Reminder: Obtain a spectrum of solvent as well, or run the pure solvent as a baseline to automatically subtract it. The KBr plates must be tho ro ughly cleaned after this pro cedure to prevent co ntaminatio n o f future samples. Wipe the windo ws with a tissue, then wash several times with yo ur so lvent, then ethano l. Use the po lishing kit in the lab to po lish the windo w surface. Wear glo ves to prevent fo gging. The cleaned surface sho uld be clear and free fro m scratches.

D e p art me nt o f C he mist ry and B io che mist ry The Michael Faraday Laboratories, NIU DeKalb, IL 60115- 2862 (815) 753- 1131

3. SOLIDS (as Nujol mulls)

Alternative metho ds to o btain IR spectra o f so lids are Nujo l (mineral o il) mulls between KBr plates.
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Remember that Nujol by itself shows a characteristic spectrum! Go o d results are o btained by this metho d o nly if the average particle size o f the so lid is so mewhat less than the wavelength o f light the particles are to transmit. Samples sho uld therefo re be gro und in a mo rtar to reduce the average particle size to 1 to 2 micro ns. Abo ut 5 to 10 mg o f finely gro und sample are then placed o nto the face o f a KBr plate, a small dro p o f mineral o il is added and the seco nd windo w is placed o n to p. With a gentle circular and back-and-fo rth rubbing mo tio n o f the two windo ws, evenly distribute the mixture between the plates. The mixture sho uld appear slightly translucent, with no bubbles, when pro perly prepared. Place the sandwiched plates in the spectro meter and o btain a spectrum. Ideally, the stro ngest band sho uld have a transmissio n o b 0 to 10 % and sho uld no t be to tally abso rbing fo r mo re than 20 cm -1 . At t e nt io n: If t he bands are dist o rt e d (sho w f ro nt ing o r t ailing) t he part icle size is t o o gre at and so m e radiat io n incide nt o n t he m ull has be e n scat t e re d o ut o f t he sam ple be am (Christ ianse n scat t e ring). If a be t t e r spe ct rum is re quire d re duce t he part icle size f urt he r. The KBr plates must be tho ro ughly cleaned after this pro cedure to prevent co ntaminatio n o f future samples. Wipe the windo ws with a tissue, then wash several times with methylene chlo ride, then ethano l. Use the po lishing kit in the lab to po lish the windo w surface. Wear glo ves to prevent fo gging. The cleaned surface sho uld be clear and free fro m scratches.

4. KBr pellet s/disks (f or solid samples)

In o rder to prepare a KBr pe lle t , fo llo w the pro cedure given belo w:

Sample/KBr ratio
The co ncentratio n o f the sample in KBr sho uld be in the range o f 0 .2% to 1%. The pellet is much thicker than a liquid film, hence a lo wer co ncentratio n in the sample is required (Beer's Law). To o high a co ncentratio n usually causes difficulties o btaining clear pellets. The IR beam is abso rbed co mpletely, o r scattered fro m the sample which results in very no isy spectra.

Sample preparation
Altho ugh a ho mo geneo us mixture will give the best results, excessive grinding o f the po tassium bro mide is no t required. The finely po wdered po tassium bro mide will abso rb mo re humidity (it is hygro sco pic) fro m the air and therefo re lead to an increased backgro und in certain ranges. Make sure to wo rk fast. Transfer so me KBr o ut o f the o ven (ATTENTION: the o ven is at 10 0 o C - yo u can easily burn yo urself!) into a mo rtar. Add abo ut 1 to 2 % o f yo ur sample, mix and grind to a fine po wder. Fo r very hard samples, add the sample first, grind, add KBr and then grind again. The sample must be very finely gro und as in the Nujo l mulling technique to reduce scattering lo sses and abso rptio n band disto rtio ns. Take two stainless steel disks o ut o f the desiccato r. Place a piece o f the precut cardbo ard (in the tin can next to the o ven) o n to p o f o ne disk and fill the cuto ut ho le with the finely gro und mixture. Put the seco nd stainless steel disk o n to p and transfer the sandwich o nto the pistil in the hydraulic press. With a
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pumping mo vement, mo ve the hydraulic pump handle do wnward. The pistil will start to mo ve upward until it reaches the to p o f the pump chamber. Then, mo ve the pump handle upwards and pump until the pressure reaches 20 ,0 0 0 prf. Leave fo r a few seco nds and with the small lever o n the left side, release the pressure (ho ld until the sample and pistil are all the way do wn). Remo ve the disks and pull apart. Remo ve the film, which sho uld be ho mo geno us and transparent in appearance. Insert into the IR sample ho lder and attach with sco tch tape. Run the spectrum. After use, the mo rtar and pistil sho uld be cleaned with aceto ne and do uble distilled water, and be put back o n to p o f the o ven fo r drying.

Reasons for Cloudy Disks

1. KBr mixture no t gro und eno ugh 2. Sample was no t dry 3. Sample: KBr ratio high 4. Pellet to o thick 5. Bo lts no t tightened sufficiently. 6 . Sample has lo w m.p. In o rder to o btain any meaningful info rmatio n fro m yo ur IR spectrum, yo u need to have a decent spectrum with sharp peaks, i.e., with 'go o d' intensity and go o d reso lutio n. Ideally, the largest peak, which is caused by yo ur co mpo und, sho uld have an intensity o f 2-5 % T . Mo st detecto rs o nly respo nd linearly o ver a certain range (typically o ne magnitude). Keep in mind that a transmissio n o f 5% is equivalent to an abso rptio n A=1.3, which is the upper limit fo r mo st detecto rs. If yo u o bserve a higher intensity, all yo ur impurities will seem to be mo re significant than they really are. The peaks with the high intensity will be cut o ff. Therefo re, it is very impo rtant to prepare the sample pro perly.

Last mo dified by HH 0 5-20 0 7

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