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Chapter 9

TABLE OF CONTENTS
Nitrogen
Page
Section 1: Introduction to Nitrogen...................................................................................................... 3
Section 2: Glossary............................................................................................................................. 3
Section 3: Safety and Hygiene............................................................................................................ 3-4
Section 4: Sampling............................................................................................................................ 4
Section 5: Sample reser!ation......................................................................................................... 4
Section ": Sample #ontainers............................................................................................................ 4
$ui% &.1................................................................................................................................................ 4
Section ': Summary of (et)ods......................................................................................................... 4-5
Section *: +mmonia Nitrogen ,etermination...................................................................................... 5
Section &: -.uipment and /eagents................................................................................................... 5-"
Section 10: ,istillation rocedure....................................................................................................... "-'
Section 11: +mmonia Nitrogen ,etermination.................................................................................... '
Section 12: -.uipment and /eagents................................................................................................. '
Section 13: #olorimetric rocedure.................................................................................................... '-*
Section 14: reparing #ali1ration #ur!e............................................................................................ *
Section 15: #alculation....................................................................................................................... *
Section 1": Interferences.................................................................................................................... &
Section 1': Sensiti!ity........................................................................................................................ &
Section 1*: +mmonia Nitrogen ,etermination.................................................................................... &
Section 1&: -.uipment and /eagents................................................................................................. &
Section 20: 2itrimetric rocedure........................................................................................................ &-10
Section 21: #alculation....................................................................................................................... 10
Section 22: Interferences.................................................................................................................... 10
Section 23: +mmonia Nitrogen ,etermination.................................................................................... 10
Section 24: -.uipment and /eagents................................................................................................. 10
Section 25: -lectrode rocedure........................................................................................................ 10
Section 2": reparing #alculation #ur!e............................................................................................ 10
Section 2': Interferences.................................................................................................................... 10
Section 2*: Sensiti!ity........................................................................................................................ 11-12
#)apter & - 1
Section 2&: Interpretation of /esults................................................................................................... 12
$ui% &.2................................................................................................................................................ 12
Section 30: 2otal 34elda)l Nitrogen ,etermination.............................................................................. 12
Section 31: -.uipment and /eagents................................................................................................. 12-13
Section 32: 34elda)l ,igestion rocedure........................................................................................... 13-14
Section 33: #alculations..................................................................................................................... 14
Section 34: 5rganic Nitrogen ,etermination...................................................................................... 14
Section 35: Interferences.................................................................................................................... 14
Section 3": Interpretation of /esults................................................................................................... 14
Section 3': $+6$#............................................................................................................................. 15
$ui% &.3................................................................................................................................................ 1"
+ns7ers to $ui%%es.............................................................................................................................. 1'-1*
+ppendi8 +: /eferences
+ppendi8 9: /eagent reparation
+ppendi8 #: Sample 9enc) S)eets
+ppendi8 ,: reparation of a #alculation #ur!e
+ppendi8 -: (et)ods #)ec:ist +mmonia Nessleri%ation
+ppendi8 ;: (et)ods #)ec:list ,istillation
+ppendi8 G: (et)ods #)ec:list -lectrode
#)apter & - 2
Chapter 9
NITROGEN
Section 1: INTRODUCTION TO NITROGEN
Nitrogen compounds are of interest to 7aste7ater treatment plant operators 1ecause of t)e importance
of nitrogen in t)e life cycles of plants and animals. Nitrogen is a nutrient and occurs in many forms
including ammonia< organic< nitrate and nitrite eac) of 7)ic) may 1e tested for in a !ariety of 7ays. /a7
7aste7ater nitrogen is normally present in t)e organic nitrogen and ammonia forms< 7it) small .uantities
of t)e nitrite and nitrate forms. ,epending on t)e amount of nitrification 7)ic) occurs 7it)in t)e plant<
t)e effluent may contain eit)er ammonia or nitrate nitrogen. =nder normal circumstances< t)e nitrite
form of nitrogen 7ill not 1e present in large .uantities due to its rapid o8idation or con!ersion to nitrate.
2)e presence of large concentrations of ammonia in a stream or la:e can create a large o8ygen
demand. 2)is demand is caused 1y t)e con!ersion of ammonia to nitrate. Hig) concentrations of nitrate
in 7aste7ater treatment plant effluent can cause algae to gro7 in large .uantities. ,ead and decaying
algae can cause o8ygen depletion pro1lems 7)ic) in turn can :ill fis) and ot)er a.uatic organisms in
streams. ;or t)is reason< testing for nitrogen in t)e plant effluent is critical.
Section : GLOSSAR!
B"an#: + preliminary analysis omitting only t)e sample to pro!ide an un1iased reference point or
1aseline for comparison. 2)e 9lan: is usually run on distilled 7ater.
Co"ori$etric %ea&'re$ent: 2)e met)od for measuring un:no7n concentrations of analytes in a
sample 1y measuring t)e sample>s color intensity. 2)e color of t)e sample after adding specific
c)emicals ?reagents@ is compared 7it) colors of :no7n concentrations.
Nitri(ication: +n aero1ic process in 7)ic) 1acteria c)ange t)e ammonia and organic nitrogen in
7aste7ater into o8idi%ed nitrogen ?usually nitrate@.
N'trient: +ny su1stance used 1y li!ing t)ings t)at promotes gro7t). 2)e term is generally applied to
nitrogen and p)osp)orus in 7ater< 1ut is also applied to ot)er essential and trace elements.
O)i*ation: 58idation is t)e addition of o8ygen< remo!al of )ydrogen< or t)e remo!al of electrons from
an element or compound. In 7aste7ater treatment< organic material is o8idi%ed to more sta1le
su1stances. In nitrogen monitoring< ammonia is o8idi%ed to nitrite and t)en nitrate depending on !arious
factors suc) as 7aste7ater temperature< contact time 7it) microorganisms and t)e amount of o8ygen
a!aila1le.
Titrate: 2o titrate a sample< a c)emical solution of :no7n strengt) is added on a drop 1y drop 1asis until
a color c)ange< precipitate or pH c)ange in t)e sample is o1ser!ed ?end-point@. 2itration is t)e process
of adding t)e c)emical solution to completion of t)e reaction as signaled 1y t)e end point.
Section +: SAFET! AND ,!GIENE
A)ene!er samples of 7aste7ater are )andled< it is !ery important t)at operators 7as) )is or )er )ands
1efore eating or smo:ing. A)ile some la1oratory c)emicals are not dangerous< many of t)em are
poisonous or )armful to s:in and clot)ing. /u11er glo!es and safety glasses s)ould 1e used. It is
important to 7as) t)oroug)ly 7it) soap and 7ater after )andling la1oratory c)emicals< especially if
c)emicals come into contact 7it) t)e s:in. 3eep 1enc) areas free of clutter and clean 1enc) surfaces
7it) disinfectant after testing.
#)apter & - 3
/ead t)e la1els carefully and :no7 7)at to do in case of an accidental spill. +l7ays clean up spills
.uic:ly and in t)e safest possi1le manner using disposa1le rags or to7els.
+cids and 1ases can 1e corrosi!e. #are s)ould 1e ta:en 7)en )andling t)em. Ne!er add 7ater to acid.
+l7ays add acid to 7ater !ery slo7ly 1ecause acids and 1ases can generate )eat 7)en mi8ed 7it)
7ater.
Some nitrogen testing in!ol!es )eating samples. 2o a!oid in4uries caused 1y )eat< use glo!es or tongs
to )andle )ot or potentially )ot glass7are. Ne!er assume t)at a 1ea:er or flas: is cool.
Section -: SA%PLING
Samples used for t)e determination of nitrogen can 1e eit)er gra1 or composite. 2)e type of sample
used 7ill depend on t)e monitoring re.uirements< plant operating procedures and t)e testing and sample
storage capa1ilities of indi!idual plants. Samples s)ould 1e collected from 7ell-mi8ed areas in t)e
process flo7 so t)at t)ey are representati!e of t)e total flo7.
Section .: SA%PLE PRESER/ATION
(ost relia1le results are o1tained using fres)ly collected samples. Ho7e!er< if samples must 1e )eld<
residual c)lorine s)ould 1e remo!ed 1y adding Sodium t)iosulfate immediately after collection. +cidify
t)e sample to less t)an pH 2 7it) sulfuric acid and cool to 4B#. +cidified and cooled samples may 1e
)eld for up to 2* days. If t)e test is run immediately< no preser!ati!e is re.uired.
Section 0: SA%PLE CONTAINERS
Special sampling de!ices and storage containers are not necessary for nitrogen testing. Sampling
de!ices s)ould dra7 from 7ell-mi8ed areas of tan:s or pipes< 1e made of resistant materials t)at 7ill not
rust or corrode< 1e capa1le of ta:ing samples t)at are proportional to t)e plant>s flo7< and 1e easily and
t)oroug)ly cleaned.
Storage containers s)ould 1e made of corrosion resistant material 7)ic) can stand repeated
refrigeration. 2)ese containers s)ould )a!e lea:-proof tops. #ontainers s)ould 1e 7as)ed 7it) soap
and 7ater and rinsed 7ell 7it) distilled 7ater.
1'i2 931
1.A)at are t)e four forms of nitrogen found in 7aste7aterC
2.A)at occurs 7)en e8cessi!e effluent ammonia is disc)arged into recei!ing 7atersC
3.If nitrogen testing is not going to 1e run immediately< )o7 s)ould samples 1e preser!edC
Section 4: SU%%AR! OF %ET,ODS
A$$onia Nitrogen
+mmonia nitrogen can 1e determined 1y se!eral different met)ods. Selection of specific met)ods must
1e 1ased upon t)e concentration le!el and t)e amount and types of interferences present.
In drin:ing 7ater< surface 7aters and some )ig)ly purified 7aste7ater effluents< it may 1e possi1le to
determine ammonia concentrations directly 1y colorimetric met)ods. Ho7e!er< t)e appro!ed met)od for
7aste7ater is preliminary distillation of t)e ammonia into an acid a1sor1ing solution for colorimetric<
titrimetric or specific ion electrode determination.
#)apter & - 4
If t)e preliminary distillation step is omitted< comparison data must 1e a!aila1le in t)e la1oratory
indicating no need for t)is step.
Tota" 56e"*ah" Nitrogen
2otal 34elda)l Nitrogen ?23N@ is an analysis to determine 1ot) t)e organic nitrogen and t)e ammonia
nitrogen. 2)e analysis in!ol!es a preliminary digestion to con!ert t)e organic nitrogen to ammonia< t)en
distillation of t)e total ammonia into an acid a1sor1ing solution and determination of t)e ammonia 1y an
appropriate met)od.
Organic Nitrogen
5rganic nitrogen can 1e determined on t)e sample left after distillation in t)e ammonia test. 2)e left
o!er sample is digested< as in t)e 23N procedure< and t)e con!erted ammonia distilled for analysis.
Deter$ination 78 Di((erence
If eit)er t)e organic nitrogen or 23N is determined< t)e ot)er parameter can 1e determined 1y difference
or addition 7it) t)e ammonia !alue.
23N D 5rganic N E +mmonia N
5rganic N D 23N - +mmonia N
Nitrate&9Nitrite&
2)e procedures for nitrate and nitrite analysis are not included in t)is manual. #onsult t)e 1*t)t) edition
of Standard (et)ods for t)is information.
Section :: A%%ONIA NITROGEN DETER%INATION
DISTILLATION STEP
A)en ammonia gas is dissol!ed in 7ater< it 7ill react 7it) t)e 7ater to form some ammonium ions.
,epending upon t)e pH of t)e solution< t)e ratio of ammonia to ammonium 7ill !ary. +t a )ig)er pH<
t)ere is more ammonia. +t a lo7er pH< t)ere is more ammonium.
In t)e distillation procedure< t)e sample pH is raised to &.5 and t)e ammonia gas formed is remo!ed 1y
distillation. 2)e ammonia gas is t)en a1sor1ed in a acid solution 7)ere it is con!erted 1ac: to
ammonium. 2)e distillation remo!es t)e ammonia from t)e sample and lea!es su1stances 7)ic) may
interfere 7it) t)e analysis 1e)ind.
Section 9: E1UIP%ENT AND REAGENTS
E1UIP%ENT
1. ,istillation apparatus consisting of:
500 or *00 mF 34elda)l flas:s
#onnecting 1ul1s
Gertical condensers
Hot plates
300 mF recei!ing 1ea:ers or flas:s
#)apter & - 5
2. pH meter
REAGENTS
1. +mmonia free distilled 7ater
2. 9orate 1uffer solution
3. Sodium )ydro8ide solution " N
4. Sodium t)iosulfate solution
5. Sodium )ydro8ide 1 N
". Sulfuric acid 1 N
'. 9oric acid a1sor1ing solution
*. Sulfuric acid a1sor1ing solution 0.04 N
See +ppendi8 9 for t)e procedures for preparation of t)e reagents used in t)is met)od.
Section 1;: DISTILLATION PROCEDURE
1. +dd 500 mF of ammonia free distilled 7ater and 20 mF 1orate 1uffer to a distillation flas: and ad4ust
t)e pH to &.5 7it) "N Na5H solution. +dd a fe7 glass 1eads or 1oiling c)ips.
2. +ttac) t)e flas: to t)e distillation unit and distill until t)e distillate s)o7s no trace of ammonia.
3. Fea!e t)e distillation unit assem1led until ready to attac) t)e distillation flas: containing t)e sample.
4. (easure out 500 mF of dec)lorinated sample or an ali.uot of sample diluted to 500 mF 7it)
ammonia free distilled 7ater.
NOTE: /emo!e residual c)lorine 1y adding sufficient Sodium t)iosulfate solution to 4ust neutrali%e t)e
residual c)lorine in t)e sample. =se 1 mF to remo!e 1 mg6F residual c)lorine in a 500 mF sample. If
necessary< neutrali%e t)e sample to appro8imately pH ' 7it) eit)er t)e 1 N Sodium )ydro8ide or 1 N
Sulfuric acid solution.
5. +dd 25 mF 9orate 1uffer solution to t)e solution.
". +d4ust sample pH to &.5 7it) " N Sodium )ydro8ide.
'. lace sample in distillation flas: and add se!eral glass 1eads.
*. (easure 50 mF of 9oric acid a1sor1ing solution into 300 mF recei!ing 1ea:er or flas:.
NOTE: =se 0.04 N Sulfuric acid as t)e a1sor1ing solution for samples to 1e tested 1y t)e ammonia
specific ion electrode met)od.
&. lace sample flas: on t)e distillation apparatus and ad4ust )eat to pro!ide a distillation rate of " to
10 mF per minute and collect at least 200 mF of distillate.
NOTE: 9e sure t)e tip of t)e condenser is 1elo7 t)e surface of t)e a1sor1ing solution in t)e recei!ing
1ea:er or flas:.
#)apter & - "
10. Fo7er t)e recei!ing flas: until t)e condenser is free of a1sor1ing solution and allo7 t)e distillation to
continue 2 to 3 minutes longer.
11. ,ilute to 500 mF 7it) ammonia free distilled 7ater.
12. ,etermine t)e ammonia concentration of t)e distillate.
Section 11: A%%ONIA NITROGEN DETER%INATION
B! COLORI%ETRIC %ET,OD
2)e addition of Nessler reagent to a sample of distillate 7ill produce a color 7)ic) ranges from pale
yello7 to 1ro7n depending upon t)e amount of ammonia present. 2)e pale yello7 color 7ill 1e present if
t)e ammonia nitrogen le!el in a 50 mF sample is 20 to 50 micrograms. 2)e 7a!elengt) at 7)ic) t)e
measurement is made is dependent on t)e concentration le!el e8pected.
Section 1: E1UIP%ENT AND REAGENTS
E1UIP%ENT
1. Spectrop)otometer 7it) lig)t pat) of 1 cm or more or ;ilter p)otometer 7it) lig)t pat) of 1 cm and
!iolet filter 7it) ma8imum transmittance 400 to 425 nm
2. pH meter 7it) )ig) pH electrode
3. 9ea:ers
4. 125 mF -rlenmeyer flas:
5. #u!ets< matc)ed set
REAGENTS
1. Stoc: +mmonium solution
2. Standard +mmonium solution
3. Nessler reagent
See +ppendi8 9 for t)e procedures for preparation of t)e reagents used in t)is met)od.
Section 1+: COLORI%ETRIC PROCEDURE
1. ipette 50 mF of distillate or a suita1le ali.uot diluted to 50 mF 7it) ammonia free distilled 7ater
into a 125 mF -rlenmeyer flas:.
2. Neutrali%e t)e 1oric acid a1sor1ing solution 1y:
a. +dding 2 mF Nessler reagent or
1. +dding Sodium )ydro8ide to pH near ' follo7ed 1y t)e addition of 1 mF of Nessler reagent
3. #ap t)e -rlenmeyer flas: 7it) a clean ru11er stopper and mi8 t)oroug)ly.
4. +llo7 10 minutes for color de!elopment. ?Sample< 1lan:< and standards must 1e maintained at t)e
same temperature and color de!elopment time.@
#)apter & - '
NOTE: If ammonia nitrogen is e8tremely lo7< allo7 30 minute de!elopment.
5. (easure percent transmittance of t)e sample using a distilled 7ater 1lan: as reference ?H2 D 100@ at
t)e selected 7a!elengt). ?Since t)e 7a!elengt) used is dependent on t)e ammonia concentration<
t)e 7a!elengt) must 1e determined e8perimentally and t)en remain constant for standards and
samples.@
Concentration <$g9L= >a?e"ength <n$=
0.4 to 5.0 mg6F 400 to 425
approac)ing 10 mg6F 450 to 500
NOTE: =sing a 5 cm cell can e8tend t)e concentration range to include 0.1 mg6F to 1.2 mg6F.
". /ead t)e nitrogen concentration in ug from t)e cali1ration cur!e and calculate t)e ammonia
nitrogen using t)e formula in Section 12?d@.
Section 1-: PREPARING CALIBRATION CUR/E
Standards must 1e treated in t)e same manner as samples. 2)e !olumes of standard solution selected
must 1e placed in t)e distillation assem1ly and distilled. 2)e distillate is diluted to 500 mF and a 50 mF
ali.uot is used for t)e preparation of t)e standard cur!e.
,epending on t)e range of ammonia nitrogen to 1e determined< select a series of standard ammonia
solution !olumes. ;or samples 7it) a concentration in t)e range of 0.4 to 5.0 mg6F< prepare standard
solutions 1y pipetting appropriate !olumes of 0.5 to 250 microgram650 mF ali.uot into distillation flas:s<
and dilute to 500 mF 7it) ammonia free 7ater.
lace 500 mF of ammonia free distilled 7ater in a distillation flas:< proceed 7it) t)e distillation and
colorimetric procedures.
lot t)e percent transmittance of t)e standards at 425 nm !ersus t)e micrograms of nitrogen on standard
grap) paper.
Section 1.: CALCULATION
2o determine t)e amount of ammonia nitrogen< use t)e follo7ing formula:
mg6F ammonia N D +6mF sample 8 96#
+ D micrograms of N from cali1ration cur!e
9 D Golume of distillate collected ?500 mF@
# D Golume of distillate used for color de!elopment
mF sample D mF of original sample placed in distillation flas:
Section 10: INTERFERENCES
#ertain organic compounds ?including urea and cyanates@ 7ill )ydroly%e during distillation at a &.5 pH.
A)ile t)e )ydrolysis is relati!ely small ?5 to 'H@< t)e result of t)e analysis can 1e increased slig)tly.
/esidual c)lorine 7ill cause interference and must 1e remo!ed 1efore t)e ammonia determination.
#)apter & - *
Section 14: SENSITI/IT!
=tili%ing carefully prepared Nessler reagent< concentrations of as little as 1 g650 mF may 1e detected.
/esults at t)is le!el are e8tremely erratic.
Section 1:: A%%ONIA NITROGEN DETER%INATION
B! TITRI%ETRIC %ET,OD
2)e titrimetric procedure for t)e determination of ammonia nitrogen can 1e used only for samples 7)ic)
)a!e 1een treated 1y t)e preliminary distillation into 1oric acid a1sor1ing solution. In t)is procedure< t)e
ammonium concentration of t)e 1oric acid solution is titrated 7it) a strong acid titrant to t)e pale
la!ender end-point of met)yl red-met)ylene 1lue indicator.
Section 19: E1UIP%ENT AND REAGENTS
E1UIP%ENT
1. ,istillation apparatus
2. 50 mF 9urette
REAGENTS
1. Indicating 9oric acid solution
2. (et)yl red-(et)ylene 1lue mi8ed indicator
3. Sulfuric acid standard titrant 0.02 N
See +ppendi8 9 for t)e procedures for preparation of t)e reagents used in t)is met)od.
Section ;: TITRI%ETRIC PROCEDURE
1. ,istill t)e sample as descri1ed in Section 11 using indicating 9oric acid solution as t)e a1sor1ent for
t)e distillate.
2. Select an appropriate ali.uot of distillate 1ased upon e8pected ammonia concentration.
e)pecte* a$$onia $L *i&ti""ate
nitrogen concentration to '&e
5 to 10 mg6F
250
10 to 20 mg6F
100
20 to 50 mg6F
50
50 to 100 mg6F 25

3. 2itrate t)e sample 7it) 0.02 N standard sulfuric acid titrant until t)e indicator turns a pale la!ender.
4. /epeat t)e entire procedure using an ammonia free distilled 7ater 1lan:.
#)apter & - &
Section 1: CALCULATION
2o determine t)e concentration of ammonia nitrogen< use t)e follo7ing formula:
mg6F +mmonia N D ??+ - 9@ 8 2*0@6mF sample
+ D !olume of sulfuric acid titrated for sample ?mF@
9 D !olume of sulfuric acid titrated for 1lan: ?mF@
Section : INTERFERENCES
Golatile al:aline su1stances may cause !ariations in t)e test results.
NOTE: 2)e titration met)od is 1est used for ammonia nitrogen concentrations in e8cess of 5 mg6F.
Section +: A%%ONIA NITROGEN DETER%INATION
B! ION SELECTI/E ELECTRODE %ET,OD
2)e ammonia electrode uses a )ydrop)o1ic ?7ater repelling@ gas permea1le mem1rane to separate a
sample solution from t)e electrode internal solution. ,issol!ed ammonia in t)e sample 7ill pass t)roug)
t)e mem1rane until t)e partial pressure of ammonia is e.uali%ed. 2)e ammonia gas reacts 7it) t)e
internal filling solution creating an electrical current 7)ic) 7ill 1e proportional to t)e ammonia nitrogen
concentration.
Section -: E1UIP%ENT AND REAGENTS
E1UIP%ENT
1. +mmonia electrode
2. pH meter capa1le of reading relati!e milli!olt potential or Specific ion meter
3. (agnetic stirring plate and stirring 1ars
REAGENTS
1. Sodium )ydro8ide 10 N
2. Standard ammonium solution
3. -lectrode filling solution
4. 9uffer pH 4.0
5. 9uffer pH '.0
". +mmonia free distilled 7ater
See +ppendi8 9 for t)e procedures for preparation of t)e reagents used in t)is met)od.
Section .: ELECTRODE PROCEDURE
Since t)e step 1y step procedure 7ill 1e dependent on t)e meter and electrode system c)osen for t)e
determination< it is not possi1le to pro!ide a detailed procedure. 2)is discussion 7ill 1e limited to general
terms.
#)apter & - 10
2)e electrode and mem1rane must 1e tested daily to ensure proper operation. 2)e met)od for t)is
usually in!ol!es determination of t)e slope of t)e cali1ration cur!e. ?2)e slope 7ill normally 1e 5* E6- 1
m! at 25B#.@
2)e samples and standards must 1e at t)e same temperature. + !ariation of 1B# 7ill result in a 2H error
in t)e measurement.
Samples and standards must 1e ad4usted to a pH of 11 to 14 4ust prior to analysis. =sually adding 1 mF
of 10 N Na5H to a 50 or 100 mF sample is sufficient.
Samples and standards must 1e stirred constantly during t)e analysis. + magnetic stirring 1ar in t)e
sample placed on a stirring plate 7ill pro!ide constant stirring.
+mmonia determinations may 1e made directly on t)e distillate or may 1e made 1y a :no7n addition
met)od.
Section 0: PREPARING CALIBRATION CUR/E
If t)e milli!olt scale of a pH meter is used< a cali1ration cur!e s)ould 1e prepared using a standard
ammonium solution. 2)e cur!e is prepared 1y measuring t)e milli!olt reading for samples containing
0.1< 1< 10< 100< and 1000 mg6F. Since t)e electrode measures relati!e milli!olt production< t)e normal
procedure is to set t)e milli!olt reading for t)e 10 mg6F concentration to %ero and determine t)e
remaining standard milli!olt production relati!e to t)is !alue. 2)e results are t)en plotted on semi-log
grap) paper.
2)e standard reference or cali1ration cur!e s)ould 1e c)ec:ed daily 1y testing a standard ammonium
solution 7)ic) )as 1een processed in t)e same manner as t)e samples to 1e tested.
Section 4: INTERFERENCES
Golatile amines 7ill interfere 7it) operation of t)e electrode. Hig) concentrations of ionic su1stances can
also interfere due to t)eir effects on t)e solu1ility of ammonia.
+mmonia 7ill form comple8es 7it) certain metallic ions causing lo7er test results. 2)e most li:ely
interfering metallic ion is t)e mercury ion 7)ic) is not remo!ed as )ydro8ide at )ig) pH. If mercury is
suspected< it s)ould 1e remo!ed 1y comple8ing 7it) iodide.
Aater !apor may act as an interference if it passes t)roug) t)e mem1rane and dilutes t)e internal filling
solution. Normally t)is situation is eliminated 1y ad4usting t)e pH 7it) 10 N sodium )ydro8ide.
2)e presence of 7etting agents suc) as t)ose found in commercial detergents 7ill destroy t)e
)ydrop)o1ic c)aracter of t)e mem1rane. 2)is allo7s 7ater to pass t)roug) t)e mem1rane diluting t)e
internal filling solution.
Section :: SENSITI/IT!
2)e specific ion electrode met)od )as accepta1le accuracy to as lo7 as 1 mg6F ammonia nitrogen.
9elo7 t)is le!el< t)e diffusion of ammonia t)roug) t)e mem1rane 1ecomes !ery slo7 and re.uires
special precautions and procedures.
/esults can 1e reproduced to 7it)in E6- 2H if t)e meter is recali1rated )ourly during use. /eproduci1ility
of results is dependent on temperature !ariations< meter drift< and electronic noise.
/esults are affected greatly 1y temperature !ariations. + 1B# temperature c)ange may produce a E6- 2H
error in measurement. 2)e )eating effects of a magnetic stirrer may 1e sufficient to affect test results.
#)apter & - 11
2o a!oid t)is< t)e 1ea:er s)ould 1e insulated from contact 7it) t)e stirrer 1y means of a t)in plate of
Styrofoam.
Section 9: INTERPRETATION OF RESULTS
Since t)e ammonia form of nitrogen can 1e modified 1y 1iological acti!ity< it is possi1le to )a!e !arying
concentration ranges for ammonia nitrogen at eac) stage of treatment. ;or e8ample< t)e effluent from
an acti!ated sludge plant may contain )ig) le!els of ammonia nitrogen if t)e process is operated in suc)
a 7ay as to retard t)e gro7t) of nitrogen related 1acteria or t)e effluent may contain !ery lo7 le!els of
ammonia nitrogen if t)e process contains a large num1er of nitrifying 1acteria.
Influent concentrations for ammonia nitrogen 7ill normally fall in t)e range of 0.01 to 50 mg6F depending
on t)e c)aracteristics of contri1utors to t)e system.
1'i2 93
1. ,escri1e t)e distillation apparatus for t)e ammonia determination.
2. A)at t7o c)emicals are accepta1le for use as a1sor1ents for t)e distilled sampleC
3. A)at 7a!elengt)?s@ can 1e used to measure ammonia in t)e colorimetric procedureC
4. A)at s)ould 1e added to t)e a1sor1ing solution for t)e titrimetric end-point in t)e
ammonia procedureC
5. A)at instruments are accepta1le for use in t)e electrode procedure for ammoniaC
Section +;: TOTAL 5@ELDA,L NITROGEN DETER%INATION
2otal 34elda)l Nitrogen ?23N@ is t)e sum of t)e organic nitrogen and t)e ammonia nitrogen forms in a
sample. In t)e presence of sulfuric acid< potassium sulfate< and a mercuric sulfate catalyst< t)e nitrogen
7)ic) is part of organic matter is con!erted to ammonia 1y sodium t)iosulfate and t)en distilled from
al:aline solution as descri1ed in t)e ammonia nitrogen discussion.
;or 23N ?or organic@ !alues less t)an 5 mg6F< t)e colorimetric met)od s)ould 1e used. Ho7e!er< for
samples of un:no7n concentrations or concentrations greater t)an 5 mg6F< t)e titrimetric met)od s)ould
1e used.
Section +1: E1UIP%ENT AND REAGENTS
E1UIP%ENT
1. 34elda)l digestion apparatus
2. Heating unit capa1le of )eating a 250 mF sample from 25B# to 1oiling 7it)in 5 minutes
3. ;ume )ood or e8)aust fume e4ector
4. -.uipment for distillation and measurement of ammonia nitrogen
REAGENTS
1. 34elda)l digestion reagent consisting of:
#)apter & - 12
#oncentrated Sulfuric acid
otassium Sulfate
(ercuric 58ide
2. )enolp)t)alein indicator
3. Sodium )ydro8ide - Sodium t)iosulfate reagent
4. 9orate 1uffer solution
5. Sodium )ydro8ide " N
". /eagents for distillation and measurement of +mmonia nitrogen
See +ppendi8 9 for t)e procedures for preparation of t)e reagents used in t)is met)od.
Section +: 5@ELDA,L DIGESTION PROCEDURE
1. Select an appropriate !olume of sample to 1e placed in t)e 500 to *00 mF 34elda)l digestion flas:.
e)pecte* T5N $L o( &a$p"e
concentration <$g9L= to '&e
0 to 1 500 ?use *00 mF flas:@
1 to 10 250
10 to 20 100
20 to 50 50
50 to 00 25
,ilute if necessary to 300 mF 7it) ammonia free distilled 7ater.
2. #+/-;=FFI add 50 mF of digestion reagent. If large amounts of organic matter are present< an
additional 50 mF of reagent must 1e added per gram of organic matter. ?2)is may 1e estimated
from !olatile solids information@.
3. (i8 t)oroug)ly. ?Incomplete mi8ing can cause 1umping during t)e digestion and may result in
glass7are 1rea:ing or loss of sample@.
4. +dd se!eral glass 1eads of 1oiling c)ips to t)e flas:.
5. lace flas: on digestion apparatus and )eat to 1oiling and continue 1oiling until you see t)e
formation of dense 7)ite fumes ?S5
2
@.
". #ontinue to digest t)e sample for 30 minutes more. +s t)e digestion continues< colored or tur1id
samples 7ill turn clear or stra7 colored.
'. #ool t)e flas: and dilute t)e sample 7it) 300 mF of ammonia free distilled 7ater. (i8.
*. +dd 0.5 mF p)enolp)t)alein indicator.
&. 2ilt t)e digestion flas: and #+/-;=FFI add a sufficient amount of sodium )ydro8ide - t)iosulfate
reagent to form an al:aline layer ?pin: %one@ in t)e 1ottom of t)e flas:. =sually 50 mF of reagent is
needed for e!ery 50 mF of digestion reagent used.
#)apter & - 13
10. #onnect t)e flas: to t)e distillation apparatus< mi8 t)oroug)ly and distill 200 mF of distillate into a
1oric acid a1sor1ing solution. ?See Section 11 for t)e distillation procedure.@
11. ,etermine 2otal 34elda)l Nitrogen as ammonia using one of t)e met)ods outlined in Sections 12<
13< and 14.
NOTE: ;or t)e spectrop)otometric met)od< t)e 1lan: and standards must 1e treated 1y t)e digestion
procedure.@
Section ++: CALCULATIONS
#alculate t)e ammonia measured as 23N using t)e procedures found in Sections 12< 13< and 14.
Section +-: ORGANIC NITROGEN DETER%INATION
Since 23N represents t)e sum of t)e organic and ammonia nitrogen concentrations< t)e amount of
organic nitrogen normally can 1e determined 1y su1traction.
Organic nitrogen A T5N B A$$onia nitrogen
If t)e a!aila1le e.uipment and6or la1oratory space is limited< t)e 23N procedure can 1e performed using
t)e solution left in t)e distillation flas: after t)e determination of ammonia nitrogen.
NOTE: 2)e solution s)ould 1e diluted to appro8imately 300 mF 1efore t)e digestion is performed.
2)e results of t)is test 7ill 1e t)e organic nitrogen in t)e sample. 2)e 23N concentration can t)en 1e
found 1y adding t)e ammonia and organic nitrogen concentrations.
T5N A A$$onia nitrogen C Organic nitrogen
Section +.: INTERFERENCES
2)e actual determination of 23N and organic nitrogen is 1ased upon t)e analysis of t)e con!erted
ammonia. 2)erefore< t)e primary interest s)ould focus on t)e sensiti!ity limits and interferences noted
for t)ese tests.
2)e 23N and organic nitrogen procedure as outlined does not reco!er suc) organic nitrogen forms as
a%ide< a%o< )ydro%ones< nitrile< semicar1o%ones< and o8imes.
Section +0: INTERPRETATION OF RESULTS
+s outlined in t)e discussion of normal ranges for ammonia concentrations< t)e concentrations of 23N
and organic nitrogen 7ill 1e dependent on t)e type of facility and t)e le!els at 7)ic) it is operated.
Normal influent 23N !alues 7ill 1e in t)e range of 0.01 to "0 mg6F. 2)e !alues for organic nitrogen 7ill
1e in t)e range of 0.01 to 50 mg6F.
Section +4: 1A91C
+ $uality +ssurance6$uality #ontrol program is re.uired 1y t)e N,-S permit. $uality +ssurance ?$+@
is a set of operating principles t)at must 1e follo7ed during sample collection and analysis. Fa1 1enc)
s)eets must 1e maintained t)at document 7)en t)e sample 7as collected< )o7 it 7as preser!ed< and
7)at results 7ere o1tained.
$uality #ontrol ?$#@ includes any testing 7)ic) is done to pro!e t)at t)e results are relia1le. 5ne of
e!ery ten samples analy%ed s)ould 1e a $# c)ec:. 2)is may include duplicate samples< spi:e samples<
reagent 1lan: analyses and :no7n $# samples o1tained from outside sources.
#)apter & - 14
,uplicate sample analysis in!ol!es analy%ing t)e same sample t7ice and comparing t)e results. 2)e
closer t)e results< t)e more accurate t)e analysis. /esults s)ould not differ 1y more t)an 10H. Spi:e
sample analysis in!ol!es adding :no7n amounts of analyte to a sample and calculating t)e percent
reco!ery. 2)ese are discussed furt)er in #)apter 10.
In Nitrogen analysis< a distilled 7ater 1lan: must 1e run 7it) e!ery 1atc) of samples tested. It is used to
s)o7 t)at t)e glass7are and reagents are not contaminated.
Spi:e samples are run 1y adding standard to a sample. ;or e8ample< a 100 mF -ffluent sample is
spi:ed 7it) 1 mF of t)e 100 mg6F +mmonia nitrogen standard yielding t)e follo7ing results:
-ffluent sample result: 0.1'4 mg6F
Spi:ed -ffluent sample result: 1.21 mg6F
#onc. of 1 mF of 100 mg6F standard in 100 mF: 1.00 mg6F
-8pected spi:e result D sample result E standard conc.
D 0.1'4 E 1.00
D 1.1'4
ercent reco!ery D spi:e result di!ided 1y e8pected result 8 spi:e !olume o!er original !olume 8 100H
D 1.21 di!ided 1y 1.1'4 8 1016100 8 100H
D 104H
$uality #ontrol samples 7it) :no7n concentrations of +mmonia< 5rganic or 2otal Nitrogen can 1e
purc)ased from c)emical supply companies. 2)e results from duplicate< spi:e and outside $# samples
s)ould 1e recorded in a $# note1oo:.
Sample 1enc) s)eets are included in +ppendi8 #.
1'i2 93+
1. A)y is a digestion procedure necessary in t)e analysis of 23N or 5rganic nitrogenC
2. ;or 7)at purpose is t)e mercuric sulfate used in t)e digestion reagent for t)e 23N
procedureC
3. A)at are t)e 7)ite fumes generated near t)e end of t)e digestion in t)e 23N
procedureC
4. A)y is it a good practice to mi8 all solutions t)oroug)ly and use glass 1eads during t)e
23N procedureC
#)apter & - 15
An&Der& to 1'i22e&
1'i2 931
1. A)at are t)e four forms of nitrogen found in 7aste7aterC
The (o'r (or$& o( nitrogen (o'n* in Da&teDater are Organic nitrogenE A$$onia nitrogenE
Nitrate nitrogen an* Nitrite nitrogen3
2. A)at occurs 7)en e8cessi!e effluent ammonia is disc)arged into recei!ing 7atersC
The con?er&ion o( a$$onia to nitrate create& a "arge o)8gen *e$an*3 Thi& re*'ce& the
a$o'nt o( o)8gen a?ai"a7"e to aF'atic organi&$ in the recei?ing &trea$3
3. If nitrogen testing is not going to 1e run immediately< )o7 s)ould samples 1e preser!edC
Aci*i(8 the &a$p"e to "e&& than p, Dith &'"('ric aci* an* coo" to -GC3
1'i2 93
1. ,escri1e t)e distillation apparatus for t)e ammonia determination.
A .;; or :;; $L 56e"*ah" ("a&# connecte* to a ?ertica" con*en&er 78 $ean& o( a connecting
7'"7 an* a hot p"ate or ("a$e capa7"e o( 7eing reg'"ate* to a""oD the proper ?o"'$e per
$in'te to 7e *i&ti""e*3
2. A)at t7o c)emicals are accepta1le for use as a1sor1ents for t)e distilled sampleC
The tDo che$ica"& that are accepta7"e (or '&e a& a7&or7ent& (or the *i&ti""e* &a$p"e are
Boric aci* or ;3- N S'"('ric aci* &o"'tion&3
3. A)at 7a!elengt)?s@ can 1e used to measure ammonia in t)e colorimetric procedureC
Thi& $'&t 7e *eter$ine* e)peri$enta""8E 7't &ho'"* range 7etDeen -;; an* .;; nano$eter&
*epen*ing o( the concentration o( a$$onia pre&ent in the &a$p"e3
4. A)at s)ould 1e added to t)e a1sor1ing solution for t)e titrimetric end-point in t)e ammonia
procedureC
%eth8" re* B %eth8"ene 7"'e $i)e* in*icator &o"'tion
5. A)at instruments are accepta1le for use in t)e electrode procedure for ammoniaC
p, $eter capa7"e o( rea*ing re"ati?e $i""i?o"t potentia" or a &peci(ic ion $eter
1'i2 93+
1. A)y is a digestion procedure necessary in t)e analysis of 23N or 5rganic nitrogenC
A *ige&tion proce*'re i& nece&&ar8 in the ana"8&i& o( T5N or Organic nitrogen 7eca'&e it i&
'&e* to tran&(or$ the Organic nitrogen to the A$$onia (or$ Dhich can then 7e $ea&'re*3
2. ;or 7)at purpose is t)e mercuric sulfate used in t)e digestion reagent for t)e 23N procedureC
#)apter & - 1"
It i& a cata"8&t3
3. A)at are t)e 7)ite fumes generated near t)e end of t)e digestion in t)e 23N procedureC
SO

4. A)y is it a good practice to mi8 all solutions t)oroug)ly and use glass 1eads during t)e 23N
procedureC
To pre?ent "oca" heating an* 7'$ping *'ring the *i&ti""ation proce*'re3
#)apter & - 1'
APPENDIH A
Re(erence&
Standard (et)ods for t)e -8amination of Aater and Aaste7ater< 1*t)t) -dition< +AA+< +H+< A#;J
Aater ollution #ontrol ;ederation< Aas)ington< ,#< 1&&2.
(et)ods for #)emical +nalysis of Aater and Aastes< =.S. -+ - "0064-'&-020< (arc) 1&'&.
+ ;ield Study rogram< 5peration of Aaste7ater 2reatment lants< 3erri< 3ennet) et al< =ni!ersity of
#alifornia< Sacramento.
NOTES:
#)apter & 6 +ppendi8 + 6 age 1
APPENDIH B
Reagent Preparation
REAGENTS FOR PRELI%INAR! DISTILLATION
13 Borate 7'((er &o"'tion
+dd ** mF of 0.1 N Sodium )ydro8ide ?4g Na5H6F@ to 500 mF 0.025( Sodium tetra1orate
?Na
2
9
4
5
'
@ solution ?5.0 g Na
2
9
4
5
'
or &.5 gm Na
2
9
4
5
'
.10 H
2
56F@ and dilute to 1 F.
3 So*i'$ h8*ro)i*e &o"'tion 0 N
,issol!e 240 grams of sodium )ydro8ide ?Na5H@ in 1 liter of ammonia free 7ater.
+3 So*i'$ thio&'"(ate &o"'tion
,issol!e 3.5 grams Sodium t)iosulfate penta)ydrate ?Na
2
S
2
5
3
.5 H
2
5@ in ammonia free 7ater and
dilute to 1 F. repare fres) 7ee:ly.
-3 So*i'$ h8*ro)i*e 1 N
,issol!e 40 grams of Sodium )ydro8ide ?Na5H@ in ammonia free 7ater and dilute to 1 F.
.3 S'"('ric aci* 1 N
#arefully pour 2* mF of concentrated Sulfuric acid ?H
2
S5
4
@ into 500 mF of ammonia free 7ater.
,ilute to 1 F.
03 Boric aci* a7&or7ing &o"'tion
,issol!e 20 g of 9oric acid ?H
3
95
3
@ in ammonia free 7ater and dilute to 1 F.
43 S'"('ric aci* a7&or7ing &o"'tion ;3;- N
,ilute 1.0 mF of concentrated Sulfuric acid ?H
2
S5
4
@ in 1 F of ammonia free 7ater.
REAGENTS FOR COLORI%ETRIC %ET,OD
13 Stoc# A$$oni'$ &o"'tion <1E;;; $g9L=
,issol!e 3.*1& grams of an)ydrous ammonium c)loride ?NH
4
#l@< dried at 100B# for 1 )our< in
ammonia free 7ater and dilute to 1 F.
3 Stan*ar* A$$oni'$ &o"'tion <1;; $g9L=
,ilute 10.0 mF of stoc: solution to 1 F 7it) ammonia free 7ater.
+3 Ne&&"er reagent
,issol!e 100 g of (ercury ?II@ iodide ?HgI
2
@ and '0 g of otassium iodide ?3I@ in a small amount of
ammonia free 7ater. #aution: (ercuric iodide is to8ic. +!oid ingestion. +dd t)is mi8ture slo7ly<
#)apter & 6 +ppendi8 9 6 age 1
7it) stirring< to a cool solution of 1"0 g of Sodium )ydro8ide ?Na5H@ dissol!ed in 500 mF of
ammonia free 7ater. ,ilute to 1 F. Store t)is solution in a ru11er stoppered yre8 1ottle in t)e
dar:. 2)is reagent s)ould remain sta1le for up to 1 year.
2)e reagent s)ould 1e c)ec:ed to ma:e sure it yields t)e c)aracteristic color 7it) 0.1 mg NH
3
-N6F
7it)in 10 minutes after addition and does not produce a precipitate 7it) small amounts of ammonia
7it)in 2 )ours.
REAGENTS FOR TITRI%ETRIC %ET,OD
13 In*icating Boric aci* &o"'tion
,issol!e 20 g 9oric acid ?H
3
95
3
@ in ammonia free 7ater< add 10 mF mi8ed indicator solution< and
dilute to 1 F. repare mont)ly.
3 %eth8" re*B%eth8"ene 7"'e $i)e* in*icator &o"'tion
,issol!e 200 mg of met)yl red indicator in 100 mF &5H et)yl or isopropyl alco)ol. ,issol!e 100 mg
of met)ylene 1lue in 50 mF &5H et)yl or isopropyl alco)ol. #om1ine t)e solutions. repare
mont)ly.
+3 S'"('ric aci* &tan*ar* titrant ;3; N
,ilute 20 mF of 1.0 N Sulfuric acid ?H
2
S5
4
@ to 1 F 7it) ammonia free 7ater. Standardi%e 1y
titrating against sodium car1onate incorporated into t)e indicating 1oric acid to simulate test
conditions.
Ca"c'"ation:
Normality D mg Na
2
#5
3
added di!ided 1y ?53 8 mF H
2
S5
4
added@
+t 0.0200 N< 1 mF D 2*0 ug6F
REAGENTS FOR SPECIFIC ION ELECTRODE %ET,OD
13 So*i'$ h8*ro)i*e 1; N
,issol!e 400 g of Sodium )ydro8ide ?Na5H@ in *00 mF of ammonia free 7ater. #ool and dilute to
1 F.
3 Stan*ar* a$$oni'$ &o"'tion
See #olorimetric section a1o!e.
REAGENTS FOR TOTAL 5@ELDA,L NITROGEN
13 56e"*ah" Dige&tion Reagent
,issol!e 134 g of otassium sulfate ?3
2
S5
4
@ in "50 mF of ammonia free 7ater and add 200 mF of
concentrated Sulfuric acid. +dd< 7it) s7irling< a solution of 2 g (ercuric o8ide ?Hg5@ in 25 mF "N
H
2
S5
4
. ,ilute t)e com1ined solution to 1 F 7it) ammonia free 7ater. ,o not refrigerate.
3 So*i'$ h8*ro)i*e B So*i'$ thio&'"(ate reagent
#)apter & 6 +ppendi8 9 6 age 2
,issol!e 500 gm of Sodium )ydro8ide ?Na5H@ and 25 g of Sodium t)iosulfate penta)ydrate
?Na
2
S
2
5
3
.5 H
2
5@ in ammonia free 7ater and dilute to 1 F.
+3 Borate 7'((er &o"'tion
See reliminary ,istillation reagents a1o!e.
-3 So*i'$ h8*ro)i*e 0 N
See reliminary ,istillation reagents a1o!e.
#)apter & 6 +ppendi8 9 6 age 3
APPENDIH C
Sa$p"e Bench Sheet&
COLORI%ETRIC %ET,OD
A$$onia Nitrogen Ana"8&i&
Sample collection ,ate: KKKKKKK 2ime: KKKKKKK 9y: KKKKKKK
Sample preser!edC yes no reser!ati!e: KKKKKKKKKKKK
+nalysis run ,ate: KKKKKKK 2ime: KKKKKKK 9y: KKKKKKK
Sa$p"e ID I T re&'"t Conc3 (ro$
c'r?e
Ca"c'"ate* Conc3
'&ing (or$'"a
2o determine t)e amount of ammonia nitrogen< use t)e follo7ing formula:
mg6F ammonia N D +6mF sample 8 96#
+ D micrograms of N from cali1ration cur!e
9 D Golume of distillate collected ?500 mF@
# D Golume of distillate used for color de!elopment
mF sample D mF of original sample placed in distillation flas:
#)apter & 6 +ppendi8 # 6 age 1
Titri$etric %etho*
A$$onia Nitrogen Ana"8&i&
Sample collection ,ate: KKKKKKK 2ime: KKKKKKK 9y: KKKKKKK
Sample preser!edC yes no reser!ati!e: KKKKKKKKKKKK
+nalysis run ,ate: KKKKKKK 2ime: KKKKKKK 9y: KKKKKKK
Sa$p"e
ID
/o"'$e o(
&a$p"e '&e*
Titrant (or
&a$p"e
Titrant (or
7"an#
Ca"c'"ate*
Conc3
2o determine t)e concentration of ammonia nitrogen< use t)e follo7ing formula:
mg6F +mmonia N D ??+ - 9@ 8 2*0@6mF sample
+ D !olume of sulfuric acid titrated for sample ?mF@
9 D !olume of sulfuric acid titrated for 1lan: ?mF@
#)apter & 6 +ppendi8 # 6 age 2
Ion Se"ecti?e E"ectro*e %etho*
A$$onia Nitrogen Ana"8&i&
Sample collection ,ate: KKKKKKK 2ime: KKKKKKK 9y: KKKKKKK
Sample preser!edC yes no reser!ati!e: KKKKKKKKKKKK
+nalysis run ,ate: KKKKKKK 2ime: KKKKKKK 9y: KKKKKKK
Sa$p"e ID $/ rea*ing Conc3 Fro$
c'r?e
Di"'tion
(actor
Correcte*
Conc3
#)apter & 6 +ppendi8 # 6 age 3
A$$onia Nitrogen A& N
Sample ,ate: +nalysis ,ate:
2ime: 2ime:
Initials: Initials:
#omposite or Gra1:
Sa$p"e In("'ent E(("'ent B"an# Stan*ar* Stan*ar*
T8pe 1
,ec)lorination
#)ec:
,istillation
pH #)ec:
,ilution ;actor
Instrument
/eading
;inal /esult
?mg6F@
-8pected
/esults
#alculations:
#)apter & 6 +ppendi8 # 6 age 4
APPENDIH D
Preparation o( a Ca"i7ration C'r?e
In se!eral of t)e analyses presented in t)is manual< t)ere is a need for t)e preparation of a standard
cali1ration cur!e. 2)e cur!e esta1lis)es t)e relations)ip 1et7een a1sor1ance or percent transmittance
and t)e concentration of t)e desired parameter. =sing t)is cur!e< samples can 1e tested 1y t)e
measurement of t)eir a1sor1ance or percent transmittance and t)e corresponding concentration can 1e
read off t)e grap).
/ules for preparing cali1ration cur!es.
1. repare all standards in duplicate or triplicate. =se t)e a!erage of a1sor1ance or transmittance for
preparing t)e cur!e.
2. rocess t)e standards in t)e same manner as t)e samples 7ill 1e analy%ed. If t)e samples 7ill
undergo a digestion< t)e standards must 1e digested as 7ell.
3. + distilled 7ater sample ?1lan:@ must 1e processed in t)e same manner as t)e standards. 2)is
1lan: 7ill 1e used as t)e reagent 1lan: for setting t)e spectrop)otometer to read %ero a1sor1ance
or 100H transmittance.
4. + minimum of one standard s)ould 1e processed daily to c)ec: t)e accuracy of t)e cali1ration
cur!e and t)e .uality of t)e reagents.
5. A)en ne7 reagents are prepared< t)e cali1ration cur!e s)ould 1e c)ec:ed 1y at least 3 standards.
". If t)e cur!e appears to differ from a straig)t line< t)e cur!e may 1e straig)tened to appro8imate a
straig)t line 1y plotting t)e data on semi-logarit)mic grap) paper.
'. lot data 7it) t)e a1sor1ance or percent transmittance readings on t)e !ertical a8is and
concentration on t)e )ori%ontal a8is.
*. Select grap) paper 7it) appropriate graduations. ,o not use graduations 7)ic) re.uire constant
estimation of readings.
#)apter & 6 +ppendi8 , 6 age 1

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