Spectrophotometer Modern

8/20/2019 Spectrophotometer Modern

1/21

ASSIGNMENT

SPECTROPHOTOMETER MODERN

Class : 2B

By :

Afifah Rizannah 12002

Firdanatul Lailia 12030

Istiana Maida Maria 12041

Muh.Harist Nur Afif 12049

Risa !urnia 120"#

$iti $yarifah 120#"

A!A%&MI FARMA$I HAN' ()AH *A!AR(A


2/21

CHAPTER 1

INTRODUCTION

A. Backgroun

The spectrophotometer has well been called the workhorse of the modern

laboratory. In particular, ultraviolet and visible spectrophotometry is the method of

choice in most laboratories concerned with the identification and measurement of

organic and inorganic compounds in a wide range of products and processes - in

nucleic acids and proteins, foodstuffs, pharmaceuticals and fertilisers, in mineral oils

and in paint. In every branch of molecular biology, medicine and the life sciences, the

spectrophotometer is an essential aid to both research and routine control.

Modern spectrophotometers are quick, accurate and reliable and make only

small demands on the time and skills of the operator. However, the user who wants to

optimise the functions of his instrument and to be able to monitor its performance in

critical areas will need to understand the elementary physics of the absorption process

as well as the basic elements of spectrophotometer design.

B. Purpose

To know identification of spectrophotometer, Identification Instrument of

spectrophotometer, Instrument Component of spectrophotometer, and Procedure by

spectrophotometer

CHAPTER II


3/21

DISCUSSION

A. D!"#n#$#on

A s+,-tr+ht/,t,r is an instru/,nt fr /,asurin th, trans/ittan-, r

asran-, f a sa/+l, as a fun-tin f a,l,nth. th, instru/,nt /ay , /anual r

r,-rdin.$+,-tr+ht/,t,r -an , us,d d,t,r/in, th, -n-,ntratin f a slutin

thruh th, asr+tin int,nsity at -,rtain a,l,nths. %,+,ndin n th, ran, f

a,l,nth f liht sur-, it -an , -lassifi,d int t diff,r,nt ty+,s:

)56isil, s+,-tr+ht/,t,r: us,s liht ,r th, ultrail,t ran, 718 6 400 n/

and isil, ran, 7400 6 #00 n/ f ,l,-tr/an,ti- radiatin s+,-tru/.

IR s+,-tr+ht/,t,r: us,s liht ,r th, infrar,d ran, 7#00 6 1000 n/ f

,l,-tr/an,ti- radiatin s+,-tru/.

In isil, s+,-tr+ht/,try th, asr+tin r th, trans/issin f a -,rtain

sustan-, -an , d,t,r/in,d y th, s,r,d -lr. Fr instan-, a slutin sa/+l,

that asrs liht ,r all isil, ran,s 7i.,. trans/its nn, f isil, a,l,nths

a++,ars la- in th,ry. ;n th, th,r hand if all isil, a,l,nths ar, trans/itt,d

7i.,. asrs nthin th, slutin sa/+l, a++,ars hit,. If a slutin sa/+l, asrs

r,d liht 7


4/21

B. I!n$#"#ca$#on Ins$ru(!n$

(h,r, ar, /any diff,r,nt /d,ls f s+,-tr+ht/,t,r. But all

s+,-tr+ht/,t,r r in a si/ilar fashin. All s+,-tr+ht/,t,r shin, a ,a/ f liht n a sa/+l,. (h, /l,-ul,s in th, sa/+l, int,ra-t ith th, liht a,s and ,ith,r

asr liht ,n,ry r r,fl,-t 7r trans/it it. (h, s+,-tr+ht/,t,r /,asur,s th,

a/unt f asran-, r trans/ittan-, f liht y a sa/+l,. By -/+arin th,

asran-, data t standards f nn -n-,ntratin th, a/unt f /l,-ul,s in th,

sa/+l, is d,t,r/in,d. $/, s+,-tr+ht/,t,r ar, ,ry s+histi-at,d and ,=+,nsi,.

$/, us, nly n, ty+, f liht and nt th,rs. $/, i, diital dis+lays r r,aduts

hil, th,rs r h,d u+ t -/+ut,r that ath,r analyz, and +rint ut th, data

-ll,-t,d. All th, s+,-tr+ht/,t,r ha, s/, -//n f,atur,s. (h, /st asi-

s+,-tr+ht/,t,r has a la/+ a +ris/ that -han, th, -lur f liht ,/itt,d a

sa/+l, hld,r and a dis+lay. !ns r uttns t -alirat, th, s+,-tr+ht/,t,r

all th, /a-hin, t n hi-h /l,-ul, is t , /,asur,d.

(h, /ini/u/ r,>uir,/,nts f an instru/,nt t study asr+tin s+,-tra 7a

s+,-tr+ht/,t,r ar, shn ,l:

1. A sur-, f radiatin f a++r+riat, a,l,nths.

2. A /,ans f islatin liht f a sinl, a,l,nth and ,ttin it t th, sa/+l,

6 /n-hr/atr and +ti-al ,/,try.

3. A /,ans f intrdu-in th, t,st sa/+l, int th, liht ,a/ 6 sa/+l, handlin.

4. A /,ans f d,t,-tin and /,asurin th, liht int,nsity.

Figure 1. Basic structure of spectrophotometers


5/21

A /d,rn s+,-tr+ht/,t,r -nsists f ar, ar,as r s,-tins: radiatin

sur-, /n-hr/atr +ht/,t,r sa/+l, ar,a and d,t,-tr ar,a. A

s+,-tr+ht/,t,r -ntains tunst,n la/+ 7hit, liht a d,ut,riu/ la/+ 7)5 liht

r th. A tunst,n la/+ +rdu-,s hit, liht in th, isil, a,l,nths. A

s+,-tr+ht/,t,r ith a tunst,n la/+ is -all,d a ?5I$@ s+,-tr+ht/,t,r ,-aus,

it +rdu-,s liht in th, isil, s+,-tru/. A d,ut,riu/ la/+ +rdu-,s liht in th,

ultrail,t 7)5 +art f th, s+,-tru/. In a 5I$ s+,-tr+ht/,t,r h,n th, hit,

liht th, +ris/ it is s+lit int th, -lurs f th, rain. (h, a,l,nth -ntrl

rtat,s th, +ris/ dir,-tin diff,r,nt -lur f liht tards th, sa/+l,. (h,

a,l,nths a liht +rdu-,d y th, tunst,n la/+ a ran, fr/ 30 n/ 7+ur+l, liht

t #00 n/ 7r,d liht. All th, th,rs -lurs f th, rain ha, a,l,nth in

,t,,n th,s, alu,s. By turnin th, a,l,nth -ntrl ns any a,l,nth f

isil, liht ,t,,n 30 n/ and #00 n/ -an int,ra-t ith th, sa/+l,. (h, /l,-ul,s

in th, sa/+l, ,ith,r asr r trans/it th, liht ,n,ry f n, a,l,nth r anth,r.

(h, d,t,-tr s,ns,s th, a/unt f liht ,in trans/itt,d y th, sa/+l, and r,+rts.

(hat alu, dir,-tly 7 trans/ittan-, r -n,rts it t th, a/unt f liht asr,d in

asran-, unit.

A s+,-tr+ht/,t,r in ,n,ral -nsists f t d,i-,s a s+,-tr/,t,r and a +ht/,t,r. A s+,-tr/,t,r is a d,i-, that +rdu-,s ty+i-ally dis+,rs,s and

/,asur,s liht. A +ht/,t,r indi-at,s th, +ht,l,-tri- d,t,-tr that /,asur,s th,

int,nsity f liht.

S&!c$ro(!$!r: It +rdu-,s a d,sir,d ran, f a,l,nth f liht. First a

-lli/atr 7l,ns trans/its a straiht ,a/ f liht 7+htns that +ass,s thruh a

/n-hr/atr 7+ris/ t s+lit it int s,,ral -/+n,nt a,l,nths 7s+,-tru/.

(h,n a a,l,nth s,l,-tr 7slit trans/its nly th, d,sir,d a,l,nths as shn

in Fiur, 1.

P'o$o(!$!r: Aft,r th, d,sir,d ran, f a,l,nth f liht +ass,s thruh th,

slutin f a sa/+l, in -u,tt, th, +ht/,t,r d,t,-ts th, a/unt f +htns that

is asr,d and th,n s,nds a sinal t a alan/,t,r r a diital dis+lay as

illustrat,d in Fiur, 1.

C. Ins$ru(!n$al Co(&on!n$


6/21

1. $ur-,

(h, r,>uir,/,nts ar, that th, sur-, shuld , stal, durin th,

/,asur,/,nt +,rid i.,. that th, int,nsity f ,/itt,d radiatin shuld nt

flu-tuat, and that th,r, shuld , ad,>uat, int,nsity ,r as lar, a a,l,nth

r,in as +ssil,.)ltrail,t liht is ,n,rally d,ri,d fr/ a d,ut,riu/ ar- that

+rid,s ,/issin f hih int,nsity and ad,>uat, -ntinuity in th, 190 6 380 n/

ran,. A >uartz r sili-a ,n,l+, is n,-,ssary nt nly ,-aus, f th, h,at

,n,rat,d ut als t trans/it th, shrt,r a,l,nths f th, ultrail,t radiatin.

(h, li/itin fa-tr is nr/ally th, l,r li/it fat/s+h,ri- trans/issin at

aut 190 n/.

5isil, liht is nr/ally su++li,d y a tunst,n la/+ r in /d,rn

syst,/s y a tunst,n6hal,n 7alsd,s-ri,d as>uartz6idin, la/+ hi-h

has hih,r r,lati, ut+ut in th, -rss6

,r r,in 7320 6 380 n/. (h, ln

a,l,nth li/itis usually th, -ut6ff f

th, lass r >uartz ,n,l+, nr/ally

,ll ,ynd th, us,ful isil, li/it at

900 n/.

In /st /d,rn

s+,-tr+ht/,t,rs th,

+,r su++ly arran,/,nts in-ludin

any n,-,ssaryFigure 2. UV/Vis Light Sources

start6u+ s,>u,n-,s fr ar- la/+s as ,ll as th, -rss6,r ,t,,n sur-,s at th,

a++r+riat, a,l,nth ar, aut/ati- /,-hani-al s,>u,n-,s. La/+ sar, usually

su++li,d n +r,6s,t f-us /unts r in-r+rat, si/+l, adCust/,nt /,-hanis/s

fr ,asy r,+la-,/,nt.

2. Mn-hr/atr

(h, fun-tin f a /n-hr/atr is t +rdu-, a ,a/ f /n-hr/ati-

7sinl, a,l,nth radiatin that -an , s,l,-t,d fr/ a id, ran, f

a,l,nths. (h, ,ss,ntial -/+n,nts ar, ,ntran-, slit -lli/atin d,i-, 7t

+rdu-, +arall,l liht a a,l,nth s,l,-tin r dis+,rsin syst,/ f-usin l,ns

r /irrr and an ,=it slit.


7/21

( asi- /,thds f a,l,nth s,l,-tin /ay , nt,d filt,rs and a

dis+,rsin syst,/ 7,.. a +ris/ r diffra-tin ratin.

Dart f Mn-hr/atr :

a. Filt,r

Filt,rs f -lur,d lass r ,latin, ar, th, si/+l,st fr/ f s,l,-tin ut th,y

ar, s,,r,ly li/it,d in us,fuln,ss ,-aus, th,y ar, r,stri-t,d t th, isil, r,in

and th,y ha, id, s+,-tral and idths. (y+i-al and idths ar, rar,ly ,tt,r

than 30 6 40 n/. Int,rf,r,n-, filt,rs ,ss,ntially a sustrat, 7lass nr/ally ut

/ay , sili-a n hi-h /at,rials f diff,r,nt r,fra-ti, indi-,s ha, ,,n

d,+sit,d -an , -nstru-t,d ith andidths f th, rd,r f 10 n/ r l,ss.

H,,r th,

-/+arati,ly id, andidth 6 and th,r,fr, li/it,d r,slutin 6 f filt,rst,th,r ith th,ir inaility t +rid, a -ntinuus s+,-tru/ 7,=-,+t in s+,-ial

fr/ su-h as ,d, filt,rs /a, th,/ ina++r+riat, fr us, in rutin, laratry

s+,-tr+ht/,try in s+it, f l -st and t,-hni-al si/+li-ity.

. Dris/s

A +ris/ f suital, /at,rial and ,/,try ill +rid, a -ntinuus s+,-tru/

in hi-h th, -/+n,nt a,l,nths ar, s,+arat,d in s+a-,. It is usual t i/+r,

th, d,finitin f th, liht ,t,,n th, sur-, and th, +ris/ y usin an ,ntran-,

slit 7t d,fin, th, in-id,nt ,a/ and a -lli/atr 7t +rdu-, a +arall,l ,a/ at

th, +ris/. Aft,r dis+,rsin th, s+,-tru/ is f-us,d at th, ,=it slit hi-h /ay ,

s-ann,d a-rss th, ,a/ t islat, th, r,>uir,d a,l,nth. In +ra-ti-, th, +ris/

is nr/ally rtat,d t -aus, th, s+,-tru/ t /, a-rss th, ,=it slit. R,fl,-tin

-/+n,nts i., /irrrs inst,ad f l,ns,s ar, d,siral, in )5 syst,/s fr th

,ffi-i,n-y and -st -nsid,ratins.

Dris/ /n-hr/atrs ith and idths in th, )5E5is f 1 n/ r ,tt,r ar,

a-hi,,d ithut r,at diffi-ulty and s +,rfr/an-, is r,atly i/+r,d

-/+ar,d ith filt,r6as,d d,sins. H,,r th,r, ar, draa-s ass-iat,d ith

usin +ris/s: th,ir nn6lin,ar dis+,rsin th, t,/+,ratur, r,lat,d -hara-t,risti-s f

th, -//nly us,d +ris/ /at,rials and th, -/+li-at,d +ris/ dri, /,-hanis/

n,-,ssary t +rid, a -n,ni,nt a,l,nth -ntrl and r,adut.

Prism monochromators with bandwidths in the UV/Vis of 1 nm or better

are achieved without great difficulty and so performance is greatly improved

compared with filter-based designs. However, there are drawbacks associated


8/21

with using prisms: (1) their

non-linear dispersion, (2) the

temperature related

characteristics of thecommonly used prism

materials and (3) the

complicated prism drive

mechanism necessary to

provide a convenient

wavelength control and

readout.

-. %iffra-tin 'ratin

'ratins +rid, an alt,rnati, /,ans f +rdu-in /n-hr/ati- liht. A

diffra-tin ratin -nsists f a s,ri,s f +arall,l r,s 7lin,s n a r,fl,-tin

surfa-, that is +rdu-,d y tain a r,+li-a fr/ a /ast,r -ar,fully +r,+ar,d usin

a /a-hin, r in-r,asinly fr/ n, hi-h is hlra+hi-ally ,n,rat,d. (h,

r,s -an , -nsid,r,d as s,+arat, /irrrs fr/ hi-h th, r,fl,-t,d liht

int,ra-ts ith liht r,fl,-t,d fr/ n,ihurin r,s t +rdu-, int,rf,r,n-,

and s t s,l,-t +r,f,r,ntially th, a,l,nth that is r,fl,-t,d h,n th, anl, f

th, ratin t th, in-id,nt ,a/ is -han,d.

A/n th, adanta,s that ratins ff,r 7-/+ar,d t +ris/s ar, ,tt,r

r,slutin lin,ar dis+,rsin and th,r,fr, -nstant andidth and si/+l,r

/,-hani-al d,sin fr a,l,nth.Figure 4. Diffraction Grating Monochromator


9/21

( additinal -hara-t,risti-s f ratins /ay

, nt,d:

1. If a,l,nthλ

is r,fl,-t,d fr a i,nanl, θ th,n λE2 λE3 and s n ar, als

r,fl,-t,d at that anl,. (h,s, ,rla++in

s+,-tra nn as s,-nd and third rd,rs

,t- -an , r,/,d ith filt,rs r ith a

+r,6/n-hr/atr.

Figure 5. Structure Schematic relationship

of energy distribution and grating line density

2. ar,ful s,l,-tin f th, laz, anl, 7th, anl, at hi-h th, r, is -ut

ill +,a th, ,n,ry at th, a,l,nth f th, laz, ty+i-ally 20 n/ fr

instru/,nts f th, ind und,r dis-ussin.

3. Bth th, ,n,ry and th, r,slutin f a ratin ar, dir,-tly +r+rtinalt th, nu/,r f lin,s.

Fr /a=i/u/ ,ffi-i,n-y th, lin, s,+aratin shuld , as -ls, as

+ssil, t n, a,l,nth and fr )5E5is ratins th, lin, d,nsity is

ty+i-ally1200 +,r //.

'ratins ha, th, fllin adanta,s ,r +ris/s:

7a B,tt,r r,slutin and ,n,ry transf,r.

7 Lin,ar dis+,rsin and th,r,fr, -nstant andidth.

7- L,ss -/+li-at,d a,l,nth dri, /,-hanis/ is r,>uir,d.

7d $tray liht is li/it,d t i/+,rf,-tins at th, ratin surfa-,.

d. ;+ti-al ,/,try

As all asr+tin /,asur,/,nts ar, rati d,+,nd,nt 7IEI it is n,-,ssary t

r,-rd a r,f,r,n-, slutin ,fr, rinin th, sa/+l, und,r t,st int th, liht

+ath. (h,s, /,asur,/,nts ar, dn, usin a -u,tt, 7/at-h,d if +ssil, t that

-ntainin th, t,st sa/+l, in th, liht +ath fill,d ith th, a++r+riat, sl,nt.


10/21

(h, r,f,r,n-, int,nsity 7I ari,s ith a,l,nth in a -/+li-at,d /ulti6

fun-tin ay 7du, /ainly t sur-, ,n,ry /n-hr/atr trans/issin slit

idth and d,t,-tr r,s+ns, s it is ,ss,ntial h,n /,asurin asr+tin t r,6

/,asur, th, r,f,r,n-, fr ,a-h dis-r,t, a,l,nth at hi-h /,asur,/,nt is t ,

/ad,. All /d,rn instru/,nts ar, /i-r+r-,ssr as,d and ha, th, fa-ility t

str, a as,lin, that is 100( r 0 A s,t at ,a-h a,l,nth in th, ran,

,r-/in this r,>uir,/,nt. (hishas all,d sinl, ,a/ s+,-tr+ht/,t,rs t

-/+,t, n +,rfr/an-, ith th, /r, ,=+,nsi, dul, ,a/instru/,nts.

Additinal adanta,s f /i-r+r-,ssr handlin f th, d,t,-tr ut+ut ar, th,

aility t intrdu-, -/+n,nt fa-trs 7,.. -n-,ntratin r /lar asr+tin

data and t +r,s,nt r,sults in alt,rnati, fr/ats ithut additinal

/anual-al-ulatin. An i/+rtant -nsid,ratin in s/, laratri,s is th, aility

t int,rfa-, ith +,rsnal -/+ut,rs s that r,sults -an , in-r+rat,d int a

laratry infr/atin /ana,/,nt syst,/ 7LIM$ r transf,rr,d t disfr

ar-hiin r data /ani+ulatin +ur+s,s.

1. $inl, ,a/ +ti-s

(h, d,,l+/,nt f th, /i-r+r-,ssr has /ad, it +ssil, t a-hi,,

,=-,ll,nt r,sults usin a sinl, ,a/ -nfiuratin h,n -/+ar,d t a dul,

,a/ -nfiuratin this r,sults in r,at,r +ti-al and /,-hani-al si/+li-ity.

(h, +r-,ss f -/+arisn ,t,,n r,f,r,n-, and sa/+l, -,lls -an ,

a-hi,,d ith sinl, ,a/instru/,ntatin y f,,din th, +st d,t,-tr sinal

t a /i-r+r-,ssr hi-h str,s th, r,f,r,n-, data fr sutra-tinfr/ th,

sa/+l, sinal +rir t +rintin r dis+layin th, r,f,r,n-, -rr,-t,d r,sult 7th,

as,lin,. $inal l,,ls -an ,-/+ar,d ,t,,n diff,r,nt sa/+l,s at n,

a,l,nth at a s,ri,s f +r,d,t,r/in,d a,l,nths r if a,l,nth dri,is

+rid,d a -/+l,t, asr+tin s+,-tru/ -an , tain,d.

Figure 6. Single beam spectophotometer

2. $+lit 7r,f,r,n-, ,a/ +ti-s


11/21

Gith th, intrdu-tin f =,nn flash la/+s int s+,-tr+ht/,t,rs

th, s+lit ,a/ -nfiuratin has ,-/, n,-,ssary this is ,-aus, th, hih

int,nsity flash,s fr/ th, =,nn +uls, la/+ ar, nt alays f ,>ual

/anitud,. (hus a++r=. #0 f th, ,n,ry fr/ th, /n-hr/atr is +ass,d

thruh th, sa/+l, ith th, r,st in t a s,+arat, f,,da-d,t,-tr

,nalin a /,ans f tain int a--unt dr+sEains in ,n,ry ia a f,,da-

ain l+ in th, d,t,-tr ,l,-trni-s. (his stailiz,s th, syst,/ and th,r, ar,

n lar, ,=tra -st ,l,/,nts inl,d.

3. %ul, ,a/ +ti-s

(raditinally th, +r,f,rr,d t,-hni>u, as dul,6,a/ ,/,try in th,

sa/+l, handlin ar,a. %ul,6,a/+,ratin is a-hi,,d y a ti/,6sharin

syst,/ in hi-h th, liht +ath is dir,-t,d 7y rtatin s,-tinal /irrr r

si/ilard,i-, alt,rnat,ly thruh th, sa/+l, and th, r,f,r,n-, -,ll. (h,

a,l,nth d,+,nd,nt fun-tins f th, instru/,nt ar,sinifi-antly r,du-,d t

i, /u-h i/+r,d +,ratin -hara-t,risti-s y a f,,da- syst,/ in th,

r,f,r,n-, -hann,l thatadCusts th, d,t,-tr ain t -/+,nsat, fr sur-, and

d,t,-tr ariatins.

Figure 7. Double beam spectrophotometer

%iff,r,n-, f sinl, ,a/ s+,-tr+ht/,t,r ith dul, ,a/ s+,-tr+ht/,t,r

ar, :

$inl, ,a/ %ul, ,a/

1. %,t,r/inatin f th, asr+tin

s+,-tra /anually s ast,ful f

1. %,t,r/inatin f th, asr+tin

s+,-tra aut/ati-ally thus sain


12/21

ti/,.

2. h,a+,r +ri-,

3. 'd fr >ualitati, analysis

ti/,.

2. (h, +ri-, is /r, ,=+,nsi,.

3. 'd fr >uantitati, analysis

,-aus, it is /r, a--urat,.

4. %id, array +ti-s

A furth +ti-al -nfiuratin is th, did, array h,r, liht is

/n-hr/at,d aft,r +assin thruh th, sa/+l, hi-h /,ans that n sa/+l,

-/+art/,nt lid is n,-,ssary. (h, th,r /aCr diff,r,n-, is that th, dis+,rsi,

,l,/,nt 7ratinis fi=,d and d,s nt /, as in /r, -n,ntinal syst,/s.

3. $a/+lin handlin

In +ra-ti-, y far th, r,at,r +art f all /,asur,/,nts ill , /ad, n

sa/+l,s in slutin. 5a+urs and slids -an , a--//dat,d ut /st

instru/,nts ar, d,sin,d ith a standard -,ll

7r -u,tt, as th, nr/al sa/+l, -ntain,r.

(h, d,sin -nstru-tin and /at,rial f th,

-u,tt, ar, all i/+rtant t a--urat,

/,asur,/,nts as ar, +,ratr +ra-ti-, and

sa/+l, +r,+aratin.

Figure 8. Sample Solution in cuvette

u,tt,s ar, ty+i-ally /ad, f lass r )5 rad, sili-a 7a--rdin t th,

a,l,nth ran, f int,r,st ar, fus,d rath,r than -,/,nt,d 7t r,sist th, a-tin

f s/, sl,nts and ha, th, fllin -hara-t,risti-s:

1. ;+ti-al inds 7th, sid,s thruh hi-h th, ,a/ +ass,s ar, hihly

+lish,d +arall,l and flat


13/21

2. &ntran-, and ,=it surfa-,s ar, ,=a-tly +arall,l and rthnal

3. Liht +ath 7distan-, ,t,,n inn,r surfa-,s f inds is tihtly

-ntrll,d.

(h, hld,r that l-at,s th, -u,tt, in th, liht ,a/ /ust ,nsur, +r,-is,

and r,+rdu-il, l-atin ith r,s+,-t t th, ,a/.(h, /st -//nly us,d

-u,tt, has a liht +athl,nth f 10 // ut ln,r r shrt,r +athl,nths ar,

us,ful if -n-,ntratin r asran-, fall utsid, nr/al ran,s ithut furth,r

+r-,ssin 6 ,.. sl,nt ,=tra-tin r dilutin. Mi-r-,lls ar, +arti-ularly us,ful

h,r, sa/+l, lu/,s ar, r,stri-t,d: as -,lls fl -,lls and dis+sal, -,lls ar,

all aailal, t ,=t,nd th, us,fuln,ss f th, t,-hni>u,.

%is+sal, +lasti- -u,tt,s ar, us,d t aid t,dius ashin +r-,dur,s

+arti-ularly in hih,r thruh+ut a++li-atinssu-h as . Mr, r,-,ntly n,

+lasti- /at,rials that trans/it dn int )5 ha, ,-/, aailal,.

Asr+tin s+,-tr+ht/,try is a +,rful aid th t th, id,ntifi-atin

f th, -/+n,nts f an unnn 7>ualitati,analysis and t th, /,asur,/,nt

f th, a/unts f indiidual -/+n,nts +r,s,nt 7>uantitati, analysis.

In>ualitati, r it is usually n,-,ssary t /,asur, asr+tin ,r a ran, f

a,l,nths. $/, fr/ f a,l,nths-annin /,-hanis/ and a /,ans f

-/+arin th, unnn slutin ith a r,f,r,n-, ar, d,siral, f,atur,s

fs+,-tr+ht/,t,rs t , us,d fr s+,-tru/ +lttin. H,,r in /any

laratri,s th, r,>uir,/,nt is t /,asur, lar,nu/,rs f si/ilar sa/+l,s at n,

r a f, a,l,nths nly 6 ,.. ld sa/+l,s in a usy +athly laratry.

(h,n,,d t +lt a -/+l,t, asr+tin s+,-tru/ is rar,. In /st +ra-ti-al

situatins th,r,fr, sinl, ,a/ instru/,ntatinis +r,f,rr,d fr s+,,d

-n,ni,n-, and si/+li-ity f +,ratin and -ns,>u,ntly r,du-,d li,lihd f

+,ratr ,rrr.Instru/,nt r,>uir,/,nts ar, usually l,ss -/+l,= and th,r,fr, l,ss,=+,nsi,.

(h, t,-hni>u, /ay , assist,d y a /,ans f /anually intrdu-in a

standard r r,f,r,n-, slutin int th, liht +ath ats,t int,rals t -h,- 0 ( and

100 ( s,ttins. In su-h -as,s a--ura-y ill , ,nhan-,d if th, r,f,r,n-, and

sa/+l,slutins ar, -ntain,d in -u,tt,s ith -ls,ly /at-hin trans/issin

-hara-t,risti-s at th, a,l,nth f int,r,st.

4. %,t,-tr


14/21

;f th, fur +rin-i+al ty+,s f d,t,-trs fund in s+,-tr+ht/,t,rs n,

th, +ht-ndu-ti, -,ll 7ty+i-ally a lihts,nsiti, lay,r ,.. s,l,niu/ n a /,tal

sustrat, is s s,,r,ly r,stri-t,d in th a,l,nth r,s+ns, and s,nsitiitythat

it is al/st n,,r fund in instru/,nts f th, -lass und,r -nsid,ratin. (h, /st

-//nly ,n-unt,r,d d,t,-trsar, th, +ht/ulti+li,r th, sili-n did, and th,

did, array.

Dart f %,t,-tr:

a. Dht/ulti+li,r

Dht/ulti+li,rs ha, an int,rnal a/+lifi-atin that i,s th,/ r,at

s,nsitiity and a id, s+,-tral ran,. Liht -aus,s ,/issin f ,l,-trns fr/ a

+ht-athd, hi-h a--,l,rat, +ast a s,ri,s f dynd,s /aintain,d at

+rr,ssi,ly in-r,asin +t,ntials. &l,-trns striin th, first dynd, r,l,as, a

s,-ndary ,/issin that is strn,r than th, riinal ,a/ and s n thruh th,

dynd, -hain t +rdu-, a -as-ad, ,ff,-t. (h, ,l,-trn d,nsity r,l,as,d y th,

final dynd, t th, and, -an , /any rd,rs f /anitud, r,at,r than that fr/

th, -athd, ut it r,/ains +r+rtinal t th, int,nsity f th, in-id,nt radiatin.

( ty+,s f +ht/ulti+li,r ,/,try /ay , nt,d: sid, ind and ,nd

ind. In /st -as,s th, ,nd ind +,r/its th, ,=+sur, t th, ,a/ f a

lar,r -athd, ar,a and is th, +r,f,rr,d fr/ fr hih,r s+,-ifi-atin instru/,nts .

A sinl, +ht/ulti+li,r ill +rid, ad,>uat, s,nsitiity ,r th, hl, )5E5is

r,in ut at s/, -nsid,ral, -st th f th, -/+n,nt its,lf and th,

n,-,ssary +,r su++ly and -ntrl -ir-uitry.

. $ili-n did,

$ili-n did, d,t,-trs ha, +,rfr/an-, -hara-t,risti-s hi-h 7h,n th,

d,i-, is int,rat,d ith an +,ratinala/+lifi,r -/+ar, ith ths, f a

+ht/ulti+li,r ut hain a id,r a,l,nth ran, ut l,ss s,nsitiity. (h,y

ar,/,-hani-ally rust 7,in slid6stat, d,i-,s and ,l,-trni- ,n,fits in-lud,

r,du-,d +,r su++ly and -ntrl -ir-uitr,>uir,/,nts.

-. %id, array

A did, array is an ass,/ly f indiidual d,t,-tr ,l,/,nts in lin,ar r

/atri= fr/ hi-h in a s+,-tr+ht/,t,r -an, /unt,d s that th, -/+l,t,

s+,-tru/ is f-us,d n t an array f a++r+riat, siz,. N a,l,nth

-han,/,-hanis/ is r,>uir,d and ut+ut +r,s,ntatin is irtually instantan,us.


15/21

R,slutin h,,r is li/it,d y th, +hysi-alsiz, f indiidual d,t,-tr ,l,/,nts

7ty+i-ally 2 n/ and a,l,nth ran, is usually li/it,d t aut 1 -ta, 72006

400n/ 3006"00n/ 4006800n/. (his is ,-aus, in a did, array instru/,nt it is

diffi-ult t intrdu-, filt,rs t/itiat, th, ,ff,-ts f stray liht fr/ th, s,-nd

rd,r s+,-tru/.

. M,asurin syst,/

(h, +ri/ary fun-tin f a s+,-tr+ht/,t,r ,nds ith th, +risin f a

sinal 7nr/ally an ,l,-tri-al lta, that is+r+rtinal t th, asr+tin y a

sa/+l, at a i,n a,l,nth. (h, sinal handlin and /,asurin syst,/s -an ,

assi/+l, as an a/+lifi,r and a /,t,r r as ,larat, as a +,rsnal -/+ut,r and

+rint,r d,+,ndin n th, a++li-atin.

In si/+l,st fr/ a /,t,r ill s,r, ,ith,r t indi-at, th, aslut, alu, f

th, ut+ut sinal r in s/, instan-,s th,null +int in a a-6ff -ir-uit. %iital

r,aduts 7L&% r L% ar, faur,d fr -larity and la- f a/iuity and /ay,

lin,d ith a /i-r+r-,ssr su-h that r,adut is in any +r,f,rr,d t,r/s 6 dir,-tly

in -n-,ntratin units fr ,=a/+l,.hart 7r th,r r,-rd,rs -an , us,d ith

instru/,nts ,>ui++,d ith a,l,nth s-annin syst,/s t +rid, dir,-tlyan

asr+tin s+,-tru/. (h,y ar, als us,ful in th, study f r,a-tin rat,s h,r, th,

r,>uir,/,nt /ay , t +ltasr+tin aainst ti/, at a fi=,d a,l,nth.A l- diara/ f th, +st d,t,-tr ,l,-trni- handlin and f th,

int,rat,d ut+ut and dri, syst,/s f a /d,rns+histi-at,d sinl, ,a/

s+,-tr+ht/,t,r all -ntrll,d ia a sinl, /i-r+r-,ssr is shn in.

;n-, th, +,ratr has d,fin,d th, +ara/,t,rs 7,..a,l,nth ut+ut

/d, and r,l,ant -/+utin fa-trs th,syst,/ ill ,nsur, th, -rr,-t and

+ti/u/ -/inatin fall th, arial,s aailal,. $,l,-tin f sur-, and

d,t,-trar, aut/ati-ally d,t,r/in,d any filt,rs 7,.. rd,rsu++r,ssin filt,rs r

th,r -/+n,nts ill , intrdu-,dint th, +ti-al train at a++r+riat, +ints and

sa/+l, andr,f,r,n-, -,lls -rr,-tly /ana,d in th, sa/+l, ar,a. ;ut+utin th,

r,>uir,d t,r/s 7trans/ittan-, asran-, -n-,ntratin ,t-. ill , +r,s,nt,d

and th, r,l,ant sa/+l,id,ntifi,d.

$,-ndary rutin,s su-h as a,l,nth -aliratin and th,rs,lf6t,sts /ay

, aailal, n d,/and and int,rfa-in ith,=t,rnal -/+ut,rs r th,r

instru/,ntatin 6 ,.. aut/ati-sa/+lin d,i-,s 6 is ,asy.

D. Proc!ur!

$+,-tr+ht/,t,r +,ratins uid,


16/21

1. h,- t , sur, th, -,ll hld,r is ,/+ty.

2. (urn n th, s+,-tr+ht/,t,r usin th, +,r sit-h n th, l,ft sid, f th,

a- f th, instru/,nt.

3. All th, s+,-tr+ht/,t,r t ar/ u+ fr 30 /inut,s.

4. Dr,ss AE(E t s,l,-t th, asran-, r trans/ittan-, /d,.

. $,t th, a,l,nth t th, d,sir,d alu,. Hldin th, ,y dn ill -aus, th,

a,l,nth t -han, /r, >ui-ly.

". Ins,rt th, aln int th, -,ll hld,r ith th, -l,ar alls f th, -u,tt, t th,

frnt and a- f th, instru/,nt. ls, th, sa/+l, -/+art/,nt. (h, lan is

us,d t z,r asran-, r,adins du, t i/+,rf,-tins in th, -u,tt, and fr

th, asran-, f liht y th, /,diu/ r -arri,r slutins th, +r-,dur, ar,

usin ill -ntain s+,-ifi- instru-tins aut th, -/+sitin f th, lan.

#. Dr,ss 0 AB$E100( t lan th, instru/,nt.

8. R,/, th, lan fr/ th, -,ll hrd,r ins,rt th, sa/+l, and -ls, th, sa/+l,

-/+art,/,nt.

9. R,-rd th, instru/,nt r,adin.

10. R,/, th, sa/+l, fr/ th, sa/+l, -/+art/,nt.

CHAPTER III

CONC+USION


17/21

A s+,-tr+ht/,t,r is an instru/,nt fr /,asurin th, trans/ittan-, r

asran-, f a sa/+l, as a fun-tin f a,l,nth. th, instru/,nt /ay , /anual r

r,-rdin.$+,-tr+ht/,t,r -an , us,d d,t,r/in, th, -n-,ntratin f a slutin

thruh th, asr+tin int,nsity at -,rtain a,l,nths. %,+,ndin n th, ran, f

a,l,nth f liht sur-, it -an , -lassifi,d int t diff,r,nt ty+,s:

U,-#s#)l! s&!c$ro&'o$o(!$!r: us,s liht ,r th, ultrail,t ran, 718 6 400

n/ and isil, ran, 7400 6 #00 n/ f ,l,-tr/an,ti- radiatin s+,-tru/.

IR s&!c$ro&'o$o(!$!r: us,s liht ,r th, infrar,d ran, 7#00 6 1000 n/ f

,l,-tr/an,ti- radiatin s+,-tru/.

ATTACHMENT

T*&! o" S&!c$ro&'o$o(!$!r


18/21

Attachment 1. Spectrophotometer Modern




19/21



20/21


Attachment 6 . Spectrophotometer Modern


21/21

DA/TAR PUSTA0A

Dr. Ambasta, B.K, ______“Chemistry for Engineers”

http://chemwii.!c"a#is.e"!/$hysica%_Chemistry/Kinetics/&eaction_&

ates/E'perimenta%_Determination_of_Kinetcs/(pectrophotometry

)ytochemica% *echni+!es, raaman, ha% -

www.rg!.ac.!/0/1213213spectrophotometer45.p"f

www.microeg!i"e.com/e+!!ipment/spectrophotometer

(pectro4E"!cationa%4Boo%et436.p"f
http://chemwiki.ucdavis.edu/Physical_Chemistry/Kinetics/Reaction_Rates/Experimental_Determination_of_Kinetcs/Spectrophotometryhttp://chemwiki.ucdavis.edu/Physical_Chemistry/Kinetics/Reaction_Rates/Experimental_Determination_of_Kinetcs/Spectrophotometryhttp://www.microeguide.com/equuipment/spectrophotometerhttp://www.microeguide.com/equuipment/spectrophotometerhttp://chemwiki.ucdavis.edu/Physical_Chemistry/Kinetics/Reaction_Rates/Experimental_Determination_of_Kinetcs/Spectrophotometryhttp://chemwiki.ucdavis.edu/Physical_Chemistry/Kinetics/Reaction_Rates/Experimental_Determination_of_Kinetcs/Spectrophotometry

Documents

Spectrophotometer Modern