ANALYTICA CHIMICA AC-I-A 455

STEAM DISTILtATION METHODS FOR I)ETERMINATION OF AMIMONIUM,

NITRATE AND NITRITE*

Numerous colorimctric and distillation methods of determining ammonium,

nitrate, and nitrite have been clescrihed I-7, but most of these methods arc not suitable
for analysis of biological materials. The calorimetric methods are subject to intcr-
iercnce by various substances which occur in extracts of I~iological materials, and are
;tot readily applicable to turbid or colored extracts. The chief disadvantages of the
distillation methods are that they arc generally tf2dicJUS and tirnc-consuming, rccluire
a, relatively large amount of nitrogen for accurate analysis, and arc subject to inter
fercncc by amides and other alkali-labile organic nitrogen compouncls.
For studies in 0ur laboratory of nitrogen transformations in soils, it w;~$ neccs-
sary to have metl!ocls of clctcrmining ammonium, nitrate, and nitrite in soil extracts
that wcrc rapid, specific, iltld precise and would permit accurate dctcrmination of
0.1-z mg of thcsc forms of nitrogen. Additional rcquiremcnts wc’rc that the methods
should hc readily applical~lc to turbid or colored extracts, and should permit nitrogen
isotope-ratio analysis of ammonium, nitrate, and nitrite in tracer studies of the fate of
IaN-enriched compounds in soils. The methods to be described have been found to
meet thcsc requirements. In these methods, the form of nitrogen under analysis is
convertecl to. and determined as, ammonium, which is readily oxidized to nitrogen
g<as for mass spectrometer assay of nitrogen-15 n.T1~e ammonia is scparatcd by steam
distillation, collected in boric acid-indicator solution, and determined by titration
with standarcl acid. Magnesium oxide is used for distillation of ammonia and finely
divided Dcvarda alloy is used for reduction of nitrate and nitrite to ammonium.
The methods arc based on the findings (a) that interfcrcncc by glutamine ancl other
alkali-labile organic nitrogen compounds in alkali-distillation methods of determining
ammonium ion can be eliminated by performing the distillation with steam using a
small amount of magnesium oxide and a very short p-id of distillation and (6) that
both the amount of magnesium oxide and the time required for quantitative reduction
of nitrate and nitrite to ammonium by steam distillation with magnesium oxitle and
Devarda alloy can be reduced by use of ball-rnillcd alloy. The method for dctcrmina-
tion of nitrate in the presence of nitrite depends upon the fact that sulfamic acid
decomposes nitrite rapidly and quantitatively to nitrogen at room temperature, but
does not react with ammonium or nitrate or interfere with their determination by
steam distillation with magnesium oxide and Devardx alloy.
-__
+ Journal Paper No. J-489G of the lowa Agricultural nnd Hotnc Economics Expcrimcnt Station,
Ames, Iowa. Project No. 1450.

Arrul. Clriw. A&n, 32 (rgG5) 485-495

486 J. M. RREMXER, D. R. KEfSSBY

.Sleum ~istit~utio~l u~~u~utl~s. The apparatus used (Fig. I) is designed so that

flasks fitted with standard-taper (x9/38) grouncl-glass joints can be used as distillation
chambers. The stc:tm required for distillation is generated by heating distilled \vatcr
in :L 5-l flask that contains pumice or glass beads (to promote smooth boiling) rend

* E 19 S?x?PPER

about 3 ml nf conccntratcd sulfuric acid (to trap any ammoniumin the distilled water).
l3cforc USC, the apparatus shauld bc stetrmcd out for about x0 min to remove traces
of zmzmonirt, and the rntc of stealrn gcner;\tion should be adjusted so that 7-8 ml of
distillate arc collected per minute. ‘The flow of cold water through the condenser of
the apparatus shoufd be such that the temycrature of the &&ilk& obtained by using
this rntc of C~isti~~~Ltion does not exceed 22*. The desired rate of distillrrtion is readily
obtained if tha steam generator f&k is heated by an electric heating tnantle and the
power supply to the mantle is controlled by a variable transformer. The trap at the
base of the water j,?ckct on the conclcnscr is to prevent water condensing on the external
surface of the condenser from entering the flask used to collect the distillate.
~~~st~~t~t~~~~ flnsik The flrtsks used are roo-ml Kjelclahl flasks fitted with a
side-rtrm, st~~nc~arcl-tal3~rground-glass joints , and glass hooks, as shown in Fig. 2.
STEAM DISTILLATION FOR NH%, NOo- ANI) Non,- 487

The neck of the side-arm is fitted with a standard-taper polyethylene stopper. The
flask dimensions should be sucl1 that, when the flasks are connected to tl1e distillation
apparatus, the distance between the tip of the steam inlet tube and the bottom of the
flask is approximately 4 mm.

Reagents
Mag?tesium oxide. Heat heavy magnesium oxide (U.S.Y.) in an electric muffle
furnace at 600-700~ for z 11.Cool the product in a desiccator containing KOH pellets
and store it in a tightly stoppered bottle.
Boric acid-intlicator solution. Dissolve 20 g of reugcnt-gracle boric acicl in about
700 ml of hot water and transfer tl1ecoolecl solution to a x-l volumetric flaskcontaining
zoo ml of g5”/0 ethanol and zo ml of mixed indicator solution prepared by dissolving
0,330 6 of brornocresol green and 0.~65 E: of methyl red in 500 ml of 95% ethanol.
After mixing the contents of the flask, add cn. 0.05 N sodium hydroxide cautiously
until a color change from pink to pale green is just dctcctable when I ml of the solution
is trcatcd with I: ml of WiLtCX. Then dilute tlic solution to volume with water and mix
it tl1orouglily.
Deztarda rrlluy. I3all-mill reagent-grade alloy (Fisher Scientific Co., Chicrrgo,
Illinois; J. T, Baker Chemical Co., Phillipsburg, New Jersey) until the product will
puss a roe-mesh screen ant1 at least 75 “/o of it will ljass a 300~rncsli scrccu. Store tlic
finely clividcd alloy in a ti&tly stoppcrcrl bottle.
S~r;tfunzic acid. Purify reagent-grade sulfamic acid by crystallization from 11ot
watcrB.10, and clissctlsc 2 g of tt1e purificcl reagent in xoo ml of water. Store the solution
in a refrigerator.
Sm’Juric acid. o.oog N stantlard.
Standard (arurnoGuuc -t_ ktrut$-N sotrrtimt. IXssolvc 0.236 6’ of ammonium
sulfate and 0.361 6 of potassium nitrate in water, dilute the solution to 2 1 in a volu-
metric flask, and mix tl1oroughly. If pure clry reagents are used, this solution contains
25 ~(6 of ammonium-N and 25 ,q of nitrate-N per ml. Store in a refrigerator.
.%mduvd (ant!lro#cizrr,t -t_ &vale + nitritc,J-N solutimr. Dissolve 0,23G 6’ of
ammonium sulfate, 0.246 g of sodium nitrite, and 0.361 6 of potassium nitrate in
water, clilutc the solutiorl to 2 1 in a volumetric flask, and mix tl1oroughly. If put-c
dry rcugcnts are usccl, tl1is solution contains 25 ,I_L~ of ammouiurn-N, 25 pg of nitrite-N,
and 25 ~66’of nitrate-N per ml. Store in a refrigerator.

Pvocedwcs in the jhrcsertce of &rite

Awzmo?tiwz-N. Add 5 ml of boric acid-indicator solution to a so-ml IMenmeyer
flask marked to indicate a volume of 30 ml and place the flask under tl1c condenser
of tl1e steam distillation apparatus so tl1at the end of the condcnscr is about 4 cm
above the surface of the boric acid. Pipet an aliquot of sample containing up to 2 mg
of inorganic-N into a distillation flask, add water until the total volume is cr2.20 ml,
and add 0.2 g of magnesium oxide tllrou& a dry powder funnel wit11 a long stem
reaching into the bulb of the fl,ask. Attach the flask by springs as shown in Fig. I
and commence distillation by closing the stopcock on the steam by-pass tube of the
distillation apparatus. When the distillate reaches the go-ml mark on the receiver
flask, stop the distillation by opening tile stopcock on the steam by-pass tube, rinse
the end of the condenser, and determine ammonium -N in the distillate by titration with

A~lul. Clrirtl. /l&u, 32 (1gG5) 485-495

488 J. M. URE~~NER, D. I<. KEENEY

0.005 N sulfuric acid from a s-ml microhuret graduated at o.oI-ml intervals. ‘fhe color
change at the end-point is from green to a permanent, faint pink. ’
(Nitrate -t_ nitrite)-N. After removal of ammonium-N from the sample as
described in the previous section, remove the stopper from the side-arm of the flask,
add 0.2 g of I~cvardn alloy through a dry powder funnel reaching into the flask about
x cm below the base of the ground joint on the side-arm, and immediately replace
the stopper in the neck of the side-arm. Then determine the ammonium-N liberated
by steam distillation as described in the previous section.
(rl,)izlilolliz/llL -I- nitvatc + nitrite)-N, Pollow the procedure described for detcr-
mination of ammonium-N, but add 0.2 g of Dcvarda alloy to the distillation flask
immediately after addition of magnesium oxide and before connection of the flask
to the distilIation apparatus.
(A~ttmo~ru’sr~t + nitrate)-N. 1~0110~ the pXJCcdUrc described for determination
of (ammonium + nitrate + nitrite)-N, but, before addition of magnesium oxide and
Dcvarcla alloy, treat the sample in the distillation flask wit11 I ml of sulfamic acid
solution, and swirl the flask for a few seconds to destroy nitrite.
Nitmte-N. I~ollr.~,v the procedure clescribcd for ctctcrmination of (nitrate +
nitrite)-N, but perform the analysis on a sample that has been treated with sulfamic
acid to destroy nitrite as in the procedure for determinatian of (ammonium+nitrate)-N.

Procedwes in absence of ,riitrite

Aw~nco~tiwrc-l\r. L;ollow the procedure dcscribcd for determination of ammonium-
N in the presence of nitrite
Nitrate-N. l~ollow the procedure described for determination of (nitrate -I-
nitrite)-N in the prescncc of nitrite.
(Arn~r~o~t~i~~~-I- nitrate)-hr. 1~0110~ the proceclurc described for determination
of (atnmonium -I- nitrate $- nitrite)-N in the presence of nitrite.

‘I’he steam distillation apparatus was designed so that it can bc rapidly

cIisassembled ancl washed bctwccn distillations to prevent cross contamination of
samples when it is usecl for clistillati~tl of ammonia in 1bN analysis. It is very conve-
nient fur the methods described, but a similar type of apparatus in which the parts
are conncctcd by rubber or plastic tubing instead of ground-glass joints can be used
if 1fiN analysis of the ammonia Iibcratcd by distillation is not required. It is not neccs-
sary (or clesirable) that the grouncl-glass joints of the distillation apparatus bc greased,
but the stopcock an the steam by-pass tube sl~oulcl be lubricated. The Teflon (DuPont
fluorocarbon resin) clry-film stopcock lubricant sold under tlte trade name of Fluoro-
Glide is satisfactory.

The tnagncsium oxide employecl is ignited before use to remove the magnesium
carbonate usually yresent in the magncsiutn oxide supplied commercially, and it is
stored in an air-tight container to protect it from atmospheric carbon dioxide.
Magnesium oxide containing carbonate could liberate carbon dioxide during the distilla-
STEAM I)ISTILLATION FOR NH3, NOa- AxD NOz- 4%

tion (particularly when the sample under analysis is acidic), and carbon dioxide inter-
feres with the titration of ammonia. When standard hydrochloric or sulfuric acid is
used for collection of ammonia, it is customary to boil the distillate for a few minutes
to remove carbon dioxide before titration with alkali, This is not permissible when
boric acid is used for collection of ammonia, because ammonia would be lostll*l~,
The use of heavy magnesium oxide is recommended, because the light variety tends
to creep up the walls of the distillation flask and enter the spray trap of the distillation
apparatus.
Finely divided Dcvarda alloy can be readily prepared from the alloy supplied
commercially by ball-nlillin~ it for several hours using a ceramic vial and ceramic
balls.
It is not necessary to use accurately wci.ghed amounts of magnesium oxide or
Devarda alloy, and calibrated glass spoons* 3 can bc used for addition of thcsc reagents.
f&agent-grade sulfamic acid sometimes contains an appreciable cluantity of
~~rnrn~~niu~nand usually requires purification. The method of purification reconlmenc~ed
is simple and effective, and only traces of ammonium were dctectcd in sulfamic acid
purified by this method, dried thoroughly and stored for z years in an air-tight bottle.
Aqueous solutions of sulfamic acid arc not completely stable, because sulfamic acid is
slowly hydrolyzed to form ammonium bistllfatc. However, this hydrolysis is extremely
slow at 50, and only traces of ~~nlnl~~~iulll were detected in sulfamic acid solutions
prepared as rccommtnded and stored for 6 months in a refrigerator. The 20/Osulfamic
acid solution used for destruction of nitrite dots not liberate measurable amounts of
ammonium under the conditions empltrycd.
The sharpness of the end-point obtainccl in titration with 0.005 N sulfuric acid
to determine ammonia in distillates collected in boric acid-indicator solution depends
upon the source of the indicators, and it may be necessary to alter the rccommcndcd
proportions of bromocrcsol green and methyl red to obtain a satisfactory end-point.
The sharpness of the end-point also depends upon the quality of the boric acidIs*.
The 5 ml of 2% boric acid solution used can effectively absorb about 5 mg of ammo-
nium-N14.15. It is rccomtnended that the o.oog IV sulfuric acid be standardized against
tris~i~y~lroxymctl~y~)~~minornet~ta~~e ~z-~~n~ino-~-(i~yc~roxymctl~yl)-r,~-propn-nediolJ,
and that titmtitions be performed using a variable-speed magnetic stirrer and a teflon
or glass-coated bar,

Frocedrr Y&s
The Dcvarda mcthodla has been widely adopted for nitrate analysis of ferti-
lizers’, but its use for ~~et~rrn~nation of nitrate in soils and other biological materials
has been limited because of the interference by organic nitrc,scn compounds which
decompose with formation of ammonia when distilled with sodium hydroxide.
Attempts have been made to modify the Devarda procedure for determination of
nitrate in solutions containing alkali-labile organic nitrogen compounds, and several
modifications involving distillation with magnesium oxide instead of sodium hydrox-
ide have been describccll~-20, However, fike the original Devarda procedure, these
methods are tedious and time-consuming and are subject to interference by amides
and other organic nitrogen compounds.
The procedures described developecl from the finding that interference by
alkali-labile organic nitrogen compounds in distillation methods for ammonia can be
490 J. >I. IiREM?*‘ElC, I). R. KI!EN;EY

eliminated by performing a short steam distillation with a small amount of magnesium

oxide, This is illustrated by l’ablc I, which sl~ows the amounts of ammonia-N liberated
by steam distillation of zo-nil aliquots of solutions containing IO ,ug of nitrogen per ml
in the form of glucosamine or galactosamine with cliffcrcnt amounts of magnesium
oxide and sodium hydroxiclc. These two compounds were selected because they arc
exceptionally labile towards alkaline reagents and cause serious interference in most
distillation methods. The data in Table I show that both compounds are extensively

hlg0 0. I 0 0 0.5
BlgO 0.z 0 o.fi 0.C)
M&g3 1.0 ‘ 1.0
N:10f-1 0. I Ho.5 8::: Hz:;
NaO i-1 1.0 8cL.I 88.5 go.2

Gnlnctoairminc hlgC) 0.1 0 a I ..)

XlgO 0.2 0 0.8 2.5
hly0 1.0 I.8 2.g 6.3
NdJH 0. I as.‘{ 7.3‘0 74.6
N.?C)I-I I.0 74.2 76.5 HO.,&
._- ..-_ .__..--_ ..- _.._. __._.._._I.._. . . __._ .. . .. . . . . _ . _ . . - .. - -.-..

n 12;it.c01 tlistilhtion, cu. 7.5 Inl/min; Ii, period uxotl in tlwthcdf4 dwcrilwd (~34. 3.3 min).

clccoml~osccl with formation of ammonia when steam-clistillccl with sodium hyclroxiile,

but do not yield ammonia when steam-cIisti1lccf with o. r-o.2 g of magnesium oxide for
z min, and release very little atnmonia w11cn steam-distilled with 0.2 g of magnesium
oxide for 3.3 min. ‘I’csts si~owccl that. clumtitativc recovery of ammonium was readily
obtainecl by stcanl distillatirm of 20 ml of ammonium sulfate solution containing
2 mg of ammonium-N with 0.x or 0.2 g of mngncsium oxide for 2 min; this indicated
thnt it might be possihlc to detcrminc nitrate and nitrite without significant intcr-
ferencc by :~lk~~li-la1~ile organic nitrogen compounds bv steam distillation in the
presence of Dcvnrda alloy and 0.1-0.2 g of mqpsium oxide, if the period of distilla-
tion required for quantitative recluction of nitrate and nitrite clicl not exccccl 3.3 min.
It was found that cluantitntivc rccluction of nitrate or nitrite could not be achieved
within this prior1 when reagent-grade Dcvarcla alloy or the c soo-mesh fraction of
this alloy was used, but was readily obtained if the reagent-grade alloy was ball-milled
before use. The effect of the particle size is illustrated by ‘fi~blt.! II, whiclt also shows
the effects of varying the amounts of magnesium oxide and Devarcla alloy on the
recuvcrics obtained in analysis of zo-ml aliquots of potassium nitrate solution con-
taining IOO pg of nitrate-N per ml. Similar results were obtainccl using zo-ml alicluots
of sodium nitrite solution containing IOO p,g of nitrite-N par ml. It was found that
3.3 min were required for quantitative rcco&ry of nitrate and nitrite using 0.2 g of
magnesium oxide and 0.2 g of ball-milled Dcvarcla alloy as recommended in the proce-
dures described. The amount of magnesium oxicle used has a marked effect on the

A?,&. Chiru. Acln, 32 (1005) 485-,tg5

STEAM DlSTlLLATION FOR NHs, NOs- ASD NOz- 491

rate of reduction of nitrate and nitrite by reagent-grade alloy, and quantitative reduc-
tion can be obtained within 3.3 min if 1.0 g of magnesium oxide and x.0 g of reagent-
grade alloy are used. However, distillation with x.0 g of magnesium dxide can lead
to significant interference by alkali-labile organic nitrogen compounds (Table I).
Quantitative rccovcry of nitrate and nitrite can be achieved within 3.3 min with only
0.1 g of magnesium oxide if the amount of ball-milled alloy is increased to 0.5 g
(Table II). Use of 0.2 g of magnesium oxide is recommended because tests using zo-ml
aliquots of soil and plant extracts showed that methods involving distillation with
less than 0.2 g of magnesium oxide for 3.3 min gave low recoveries of nitrate and nitri-
te, even when x.0 Q of ball-milled alloy was used.

LSI’PILCTS 01’ AMOUSTS OF SlAGNlSSlUhl OS11315 AND DISVAl<l3A ALLOY AND OF I’AHTICLIS SIZfr: OF ALLOY
ON RISCOVRRY OF’ NITRATR
r._. “---- ---- -.. _ .--. -.. .- . --- . .._._ ..- - .- - -. - _... ,..-.- -... .._ .- . _-. ._-- .._ -.__._..- I_._.____-._-__--
A t,roro1t fg)
--- .--- .-----. _I...__--_---- Purl iclc si:c! Ittxorwry of lr
ilvulc (“/“) a
.~zu~msrtr~rr I.hvnrdn rq u!foy” Pcriorl 01 distilln~ivn /trtiu)c
oxide dl0y a It 5 IO
..-._. ._ _. .-.-__.____- _-.. ..- ..__. .__._ __.__ -._- .-..-._ __.. _._. ..__.___.....___________
0.1 0.2 kc; (i I2 2L 39
0.2 0 .1 I<c; 23 40 57 80
0.5 0.2 1<c; 43 09 or 100
X.0 0.2 IN; 6.3 9.) IO0 IO0
0.2 0.5 1% 24 49 04 100
1.0 0.5 IG 5fi 97 x00 100
0.2 I.0 Lx; 46 79 !lG 100
1 .o I.0 INi 92 100 100 100

0.2 0.2 IO .I00 1IlCSII” 7 If 20 50

0.2 0.2 I oo- r 50 Itlc!HII” 23 39 ‘J3 97
0.2 0.2 150-300 1ncsiVJ 4s 7r 88 zoo
0.2 0.2 < 300 IX\CYlI” 68 92 IO0 100

0.1 0.2 IN 69 87 100 100

0.2 0.2 in-I 91 100 100 IO0

I.0 0.2 In1 92 100 100 100

0.1 0.5 Ini 91 IO0 100 100

0.2 I.0 In4 92 100 100 IO0
-- 1. .” .._...- --..--.-, . .. I. .I. _-.- __.... - ._.... _.. .- __.____.... _. . . ..I.. - ___. .-.._. _._. . .____..__ .__, ^._. _______ _ .._.
l iG.xovcrice bctwccn 99.7 and lC)O.30/u Rrc reportcri ioo”j$. ilR

$1RCG, reagent-Kratlc alloy (rooy{, < to mesh, 29’& < IOO ttwsh, 220/ < x50 mesh, 7% < 300
mcwh); 1331, bail-t~~illctf alloy ( roocyu ( LOO ~~11. 91X ( rgc3 tllcsll, 7694 ( 300 11~11).
c Rate of distillation. m. 7.5 rnl/nlin; 12, period used in rncthotls tlcscribctl (cu. 3.3 min).
d Material obtainctl by siwing rcngcnt-gmtlc alloy.

No difficulty due to foaming was experienced in analysis of soil extracts by the

steam distillation methods described, fi)ut.cxtensive frothing does occur when some
plant extracts arc steam-distilled with magnesium oxide and Devarda alloy. This
problem can be alleviated by addition of a small amount of mineral oil or silicone
spray, but this can lead to low or erratic results in determination of nitrate or nitrite.
It is recommended, therefore, that use of tlwse anti-foam reagents be avoided, and
that larger flasks (x50- to zso-ml capacity) be used for distillation when foaming is
a problem.
Controls should be performed in each series of analyses to allow for ammonium-

Asal. Clrirn. Actn, 32 (1965) 485-495

492 J, BI. RREMNEH, D. R. KEBNEY

N &rived from tlrc reagents used. Also, the methods should he checked at intervals
by analyzing IO-ml aliquots of the stanclard (ammonium -+ nitrate)-N and (ammonium
f- nitrate + nitrite)-N solutions. 130th soiutiuns are stable for several monthsif stored
in a r~fri~erat~)r.

‘3’11~ methods give quantitative results with zo-ml aliquots of solutions can-
taining up to 100 pg l,cr ml of nitrogen in the form of ammonium, nitrate, or nitrite.
Their :Lccuracy anti precision arc: illustrated by Table XII, which gives the results of
6 an:rlyscs by each rncthctcl of zo-ml aliquots of a solution containing x0 jog per ml of
amrne~nium-N [Ias (NI-Z&SOL~], nitrate-N (as T<N%), and nitrite-N (as NaNOs).

TA1’31.,13 I I I

._*_. _. _...._..
_.. ,_. - . ..‘ . . -..-----. .-_.._.,_
,...._. 1 _. .._. .-. ._ . .., * . . ._ .- . . . . . . . .

~l~~l(fJCi rtrr N Ilclrctivi?

._. . ._.. .____.__.. .._.,_,” I .. .. ._ . 1.1. . --

Atnmmnium-N
Nitrnte-N
(~x~~~llr~~i~~!~l -I- nitrutc)-N
(Nitrntc -I- rtitritcf-N
(Amttwniun~ -I- !litritto +- tritritcj-N
____. “.. .,.___-... -.-. . ..-.._._
-- - _...e...“---
88Avcmgc! of G ~letct*Itlit~ntionH.

The specificity of the mctltods is illustrated by ‘Crrblc IV, wllich shows that
only 2 of GCJorganic nitrogen compounds tcstcd yicldctl ammonia under the conditions
of tlic procedures tlcscribcd (glucosnminc and galactosamine in methods I, 2, and 4).
‘The methods wcrc applied to ztn amount of compound cc,ntnining zoo pg of nitrogen
clissalvccl or suspcnclcd in 20 ml of water, If gluctsaminrJ or galactosaminc are prcscnt
in the sample under analysis, it~t~rfcr~ll~~ by these compounds in ~l~t~rnlination of
:tmmonium, nitrate, and nitrite cikn be eliminated by use of methods I, 3, and 5 if
the period of clistillatian in nrctltocl I is rcriuccd to 2 min.

l’ablt: V shows the: results obtaincct in studies of ttlc effect of various cztians
and nnions on clctcrmination of nitrate by the method rccommcndcd furd&ermination
of (;tmmonium +- nitrate + nitrite)-N. Thr tolernncc limit recorded for each ion
represents the concentration at which the ion did not interfere with determination of
nitrate in 20 ml of potassium nitretc solution containing 2 mg of nitrate-N. Similar
rcsutts were obtttined in studies of the effect of thcsc ions on the determination of
nitrite in 20 ml of sorlium nitrite solution containing z mg of nitrite-N by the method
for clotcrmination of (ammonium + nitrate -t_ nitrite)-N. None of the ions listed in
Table V interfered with the method for c~~t~rnlillation of amrno!liu~, even when the
concentrations of these ions greatly cxcccded the tolerance limits fur cletcrmination
of nitrate. For example, this method wits not affect& when the concentration of
srEAnf DISTILLATION FOR NHa, NOa- AND NOa- 493

AMOUNTS Ott AMMONIA-N LIBRKATED FnoM VAKIOUS ORGANXC NITROGEN COMPOUNDS USDBR
COE;DITXONS OV MF,TIIOL)S LlESCHIUl'.D
-_-.-____-_I_._ ---.I_ ----
.4wtwroniu-N fibcrufed (%, of tofd N)
Illcflrod=
I a 3 4 5
- ^_..-- __--._._ _ - . ._ .._..__.^._I
-*.-.-. --.._.--.-- ____... .,--.- --_--_-
Amino acids (29) b 0 0 0 0 0
Hcxosntnincs:
CXucosaminc 1 I 0 3 0
Chlactoanmlnc I 3 o 5 0
Purincs (5)c 0 0 0 0 0
IJyrimidirm3 (4)” 0 0 0 0 0
Amides fc,)* 0 0 0 0 it
Xlisecifaneous (2# 0 0 0 a 0
-_- ..-_ -._.. I._._...” -__._._ “.. I_. -._-__-L”-_ .__. - ._.- . ...-..._-_I.
I^_ . .
* Mcthotlv: I, ammonium-N; 2, nitrntc-N; 3, (nmmonium + nitrate)-hi; +I# (nitrate f nitrite)-N;
5, (ornmonium -+ nitrate + nitrite)-Xi.
b Asptrtic acid. glutnmic wkl, glycinc, duninc, valino, norwlinc, leucinc. isdcucinc, norlcucino,
err-;unino-rr-l~utyric acid, phcnylnlnninc, tyrwinc, trvptophan, cystcic acid, mctiiioninc, swine,
tltrconinc. phcnylscrino, t@roxylysine, ilr@inu. fy&lc, histidinc. omithint. citrulfitrc, prolinc,
hytfrcJxyprolinc, fl-ataninc, )I-untintt-,t-r)ut~ric a-Ad, gIucos:rrninic acid,
C Admine, guanino. xnnthinc, ttgwxmti3Jno, uric acid.
~3‘f’hyminc, uracil, cytoaine, illiOX;~n.
0 Urcn, biurot, rLYl>;trilgillc, glutnminc, nicotinnmidc, filycinatnitlc.
f Crcatinc, cruntinino, Ily~lillltc~in, nlhntoiri, bctainc, choline, ~t.hiIt~C~li~tlliIl~, tlicvnnrli:amitlc,
nicotinic ;~cirl, gltlcosirminic :rcitl. chitin, h7-acct~lglucoH:rnlillc. hippric ilCit1. sarcot;it~u, p-hcto-
globulin, g:cl;ttin. CitScill, gliwiin, glycflglycinc. glycinol, :~lilninot. C~-i~ll~i~~O~~i~C~~OI, nucleic ilCid
( ptsq.

- _.__.. _-. . _... ,. . -..._.. I.._._ . .. . . ..I_ ._.,..., .., . ,.- ..-.----.” ._.‘. . . . . _.... _._“_” r....l.__.. _..._
lU?f sattrcrc nlolffriry lot1 Sl?tWCt* il folwiiy
prrtniffrd p&rwiiffPd
-__I-_- - . . ._.I._ *- . . . . .-.- I ” -... .- ..-_ _- I.__.. --_I.--__-..--. .. . . I .- _.__ _.__.. _. . . . ._ . ” . ..“_.._ ._ .*_-_._

Ii+ KC1 .$.O” %nzc ZnS0.I * 71 IzO’* IQ”3

Na+ Ndl +o* f*.($ 1. 1~CSO.i’ 7 1420” 10-3
Li+ I,i&T~ . II& 3.@ Ai”+ :\l?(S(k))n * IHf I&” IO-‘8

CaP+ c:x32 * 2 l-I& 10-1 &cl c FC#30&$’ IO”‘&

E3n s+ IjilCf2 * ZIf&> 10-i Cl-. NnCi ‘$A9
hlgzc MgCkz * bH!&3 Io-a 5042” T.&&Q * Hz0 3.0’

Co?.+ CoCl3 * Ctf-f&lr 10-16 l’O,r3- NaatiPOI lo-.’

Mns+ MnSC)., . Ii&l~ 10-J SiOp NilpSiOa + ~I-120 ro-6
cua+ CUSVI 10-2 133(_)7” - Nn4S07 * IOH& IO-in
------1--..- I_._-_-_-I.I-__.. ..- -._----..-.--_l_
- _--- -.---,_- ._____-- . .. *.*-__---------
* Highest molclri ty tcstctl,
h Used ncutralizcrl solution.

silicntc or phosphate wna 0.1 Ad, whcrcns both of these anions interfered at much lower
concentrations with dctcrminntion of nitrate or nitrite (‘Table 1’). The effect of @OS-
phate on reduction of nitrate and nitrite by Dcvnrda alloy has apparently not been
noted previously, but I3a~nsos AND KAXHBISI~~? and T>I-COUINEAU AXI> GOUNY$@
found that Devarda methods involving prolonged distillation with strong alkali
were affected by silica dissolved by this alkali from the glass I<jelclahi flrruks used.
 J. 11. BRE~ISEI<, D. R. KEENEY
494

The methods have been used mainly for analysis of extracts obtained hy shak-
ing soil samples with water or neutral salt solutions (ti.g., M K&50,1, 2 dl N&I).
‘I’lGr applical3ility to extracts of other biological materials has not hen studied in
clehil, but they appear satishctory for analysis of plant extracts. ‘This is illustrated by
‘rablc IfI, which ShOWS that the methods gave quantitative recovery of ammoni~lm,
nitrate, and nitrite adclcd to soil and plant extracts. ‘l’he cxtr:tcts used were prepared
by shaking 10 g of Soil or 0.5 6’ of plant leaf mnterial with zoo ml of M potassium
chloride, and zo-ml diclUc)tS of extract were CLnalyzcd before zlnd after addition of
5 ml of ;Lsolution containing 20 pg per ml of ammonium-N [as (NH.&SO,,), nitrate-N
(as KNO:)), and nitrite-N (as NaNOz).

I’~r01l
Alf;1lf;1 ‘)C).fJ loo..! 100.0
SCl~l>Cilll I Cm. I 39.H W*‘J
13romojirass IOO.0 w.!, X00.0
Corn 100. I 100. J 9% 7
Cm. 91).8 zoo.0 100. f
- ._C..._ .._. -.- .-.. -_.__- _._. -__ __.___ __.___-__ -... . ..___. . .. ._.. . .. - ______

Steam clistill:ttictn mctlmds of determining amnlonium, nitrate, and nitrite in

the prcscncc of alkali-labile Cwgilllic nitrogen compounds arc described. ‘lhy involve
the use of magnesium oxide for distillation of ammonium, ball-milled lhvarcla alloy
for reduction of nitrate and nitrite to ammonium, and sulfamic acid for destruction of
nitrite, ‘The mctliocls are lXpiC1, accurate, and precise, and thy permit nitrogen iso-
tope-ratio analysis of ammonium, nitrate, and nitrite in tracer studies using IfiN-
enriched ct~mpourtcls. They give quantitntivc recovery of ammonium, nitrate and
uitritc aclclcd to soil and plant extracts, and appear suitable for analysis of biological
matcrinls.

Lcs autcurs cl&rivent des mbthocles de distillation B la vapcur pour le dosage

dc l’ammot.lium, dcs nitrates et des nitrites, en prdsence de compos& organiqucs

A?rrbt.Chirtr. Acta, 32 (rc$~g) 485-495

STEAM T)lSTII,I.ATION FOR NHn, Non- ASD NO?- 495

aaotb. On utilise l’oxyde de magnCsium pour le dosage de l’ammonium, l’alliage

Devarda pour la rCduction des nitrates et dcs nitrites en ammonium, et I’acidc sulfa-
mique pour la destruction dcs nitrites. Ces m&hoc& sont rqides, exactes et prCcises
et ellcs pcrmettcnt dcs analyses lors d’&udes de traceurs, utilisant dcs compos&
enricliis en IOX. Elk3 donnent dcs r&ultats quantitatifs en ammonium, nitrate et
nitrite, additions-&s h des extraits de sots et de plantes et semblcnt applicables B
I’analysc de substances biologiques.

2:uS~\~lhI~sI’r\SSUS(;

Es werclcn Dam~)fdestillationsrnct!l~~den bcschriehen zur Bcstimmung von

Ammonium, Nitrat und Nitrit in Gegenwart von aikaIiumbestiitl<tigcn organischcn
Stickstoffvcrbindungen. Sic verwenclcn Mn~ncsiumoxyd ftir die Destillation des
Ammoniums, Devnrdaschc ~,cgicrung fiir die Rcciuktion von Nitrat und Nitrit zu
Ammonium und Aminosulfonsiiure ftir die Zcrstiirung clcs Nitrits. Die Methodcn
sind schncll, genau und rcprocluzicrbar und sin<\ verwendbar bci der Analyst dcs
Stickstoffisot~~~~ex~vcrll~~ltnisscs van Ammonium, Nitrat und Nitrit bci Tracer-
Untcrsuchungcn mit l&W angcrcichertcn Verbindungt3t. Das xu 13odcn- uncl Pflanzen-
extr;&tcn gcgebcnc Ammonium, Nitrat uncl Nitrit wurde quantitativ wicclcrgewon-
nen. Die Mctlrodcn schcincn fiir clic Analyse von biologischem Material gccignct xu scin.

ii I<IXI< 15X;cES