TIU CHUN VIT NAM

TCVN 6653 : 2000

ISO 14238 : 1997
CHT LNG T - PHNG PHP SINH HC - XC NH QU TRNH KHONG HO NIT
V NITRT HO TRONG T V NH HNG CA HO CHT N CC QU TRNH NY
Soil quality - Biological methods - Determination of nitrogen mineralization and nitrification in soils and
the influence of chemicals on these processes
Li ni u
TCVN 6653 : 2000 hon ton tng ng vi ISO 14238 : 1997. TCVN 6653 : 2000 do Ban K
thut Tiu chun TCVN/TC 190
Cht lng t bin son, Tng cc Tiu chun o lng Cht lng ngh, B Khoa hc, Cng
ngh v Mi trng ban hnh.

CHT LNG T - PHNG PHP SINH HC - XC NH QU TRNH

KHONG HO NIT V NITRT HO TRONG T V NH HNG CA HO
CHT N CC QU TRNH NY
Soil quality - Biologial method- Determination of nitrogen mineralization and nitrification in soil and the
influence of chemicals on these processes
1 Phm vi p dng
Tiu chun ny qui nh cc trnh t cho phng th nghim o s v c ho v nitrit ho ca nit
do h vi thc vt t.
kho st c tnh cht c bn hoc t vn, tiu chun nu ln nhng qui trnh I cng nhm
nh gi t l v qui m v c ho nit trong t hoc cc t bit hoc cha bit cht lng.
kho st tnh c tim tng ca cc ho cht i vi s khong ho nit trong t, a ra mt
qui trnh n gin cho php nh gi nh hng ca tng ho cht v lm c s so snh tnh c
ca cc ho cht.
2 Tiu chun trch dn
TCVN 5960 : 1995 (ISO 10381-6 : 1993) Cht lng t - Ly mu - Phn 6 : Hng dn thu thp,
x l v bo qun t nh gi cc qu trnh hot ng ho kh ca vi sinh vt trong phng th
nghim ; TCVN 5979 : 1995 (ISO 10390 : 1994) Cht lng t - Xc nh pH ;
TCVN 6642 : 2000 (ISO 10694 : 1995) Cht lng t - Xc nh cacbon hu c v tng cacbon sau
khi t chy (phn tch c bn) ;
TCVN 6646 : 2000 (ISO 11260 : 1994) Cht lng t - Xc nh kh nng trao i cation thc t v
bo ho kim dng dung dch bari clorua ;
TCVN 6498 : 1999 (ISO 11261 : 1995) Cht lng t - Xc nh nit tng - Phng php Kjeldahl
ci bin ;
TCVN 6651 : 2000 (ISO 11274 : - 1) ) Cht lng t - Xc nh c tnh gi nc - Phng php
trong phng th nghim ;
TCVN 6653 : 2000 ISO 11277 - 1) ) Cht lng t - Xc nh thnh phn cp ht trong vt liu t
khong. Phng php ry v khi lng sau khi ly I mui tan, cht hu c v cacbonat ;
TCVN 6648 : 2000 (ISO 11465 :1993) Cht lng t - Xc nh hm lng cht kh v hm lng
nc theo khi lng c bn - Phng php khi lng.
3 nh ngha
Cc nh ngha sau y c p dng trong tiu chun ny:
3.1 Khong ho nit (khong ho N): s phn gii mt cht hu c c cha nit nh vi sinh thng
qua cc qu trnh amni ho v nitrt ho, thnh cc cht v c cui cng tng ng, c trng l
amni v nitrat

3.2 Amni ho : s phn gii nit hu c thnh amniac nh vi sinh.

3.3. Nitrt ho : s xy ho amni thnh nitrit nh vi sinh sau thnh nitrat.
3.4 Liu lng c ch (ID %) : lng ho cht a vo t c ch c hiu qu s khong ho N,
sau mt thi gian cho, tnh bng phn trm, so vi iu kin khng x l ho cht, v d ID 25,
ID50 cho bit v c ch 25 % hoc 50 % qu trnh khong ho nit tng ng .
4 Nguyn tc
T l hay mc khong ho N trong t hiu kh c xc nh bng cch o nng amni, nitrit
v nitrat c gii phng ra trong qu trnh khong ho nit cha trong cc cht hu c ca t,
hoc trong qu trnh khong ho ca mt hp cht nit hu c a vo t. nh hng ca ho cht
i vi qu trnh khong ho N c xc nh bng cch ci to t vi mt ngun nit hu c mi
phn hu v o phn trm c ch ca s to thnh sn phm trong cc mu c x l vi
nhng lng khc nhau ca mt ho cht so vi iu kin khng x l ho cht.
5 Vt liu
5.1 t
5.1.1 Chn t
5.1.1.1 Php th khong ho c bn
i vi php th c bn dng so snh nhng kh nng khong ho nit ca cc loi t khc
nhau, hoc so snh s khong ho nit mt loi t ly cc thi gian khc nhau trong nm,
phi m bo rng s la chn cc loi t ph hp vi mc ch xc nh.
5.1.1.2 Th tnh c: xc nh nh hng ca ho cht n qu trnh khong ho N, dng mt loi
t c hm lng khi lng cacbon hu c thp (t 0,5 % n 1,5 %) v st bit c kh nng
khong ho cht hu c cha nit c thm vo thnh sn phm cui cng (nitrat)
Ch thch - Loi t nh vy th hin mt tnh trng xu nht do s hp th l km v s c sn ca
ho cht i vi h vi thc vt l ti a. th hng ngy, cc loi t c pH nh hn 5 khng thch
hp v t l nitrit ho c th qu thp cho php nh gi y nh hng ca ho cht n qu
trnh ny.
5.1.2 Thu thp, x l v bo qun t
i vi tt c cc php th phi theo ng cc kin ngh trong tiu chun TCVN 5960 : 1995 (ISO
10381-6) v thu thp, x l v bo qun t.
Cc thng tin sau y phi c dn chng
- ngy thu thp;
- ngy lm th nghim;
- iu kin bo qun, k c nhit , m;
- thi gian bo qun.
5.1.3 Tnh cht c trng ca cc loi t
phc v cho vic din gii s liu v so snh, cc tnh cht sau y cn c xc nh :
a) Tnh cht vt l :
- thnh phn cp ht o theo ISO 11277;
- m theo TVVN 6648 : 2000 (ISO 11465); v
- c tnh gi nc theo TVVN 6651 : 2000 (ISO 11274) v/ hoc kh nng gi nc theo ph lc A.
b) Tnh cht ho hc :
- pH ca t theo TCVN 5979 :1995 (ISO 10390) hoc pH xc nh trong dung dch KCl hoc CaCl2;
- kh nng trao i cation (CEC) theo TVVN 6646 : 2000 (ISO 11260);
- hm lng cht hu c theo TVVN 6642 : 2000 (ISO 10694);
- hm lng nit tng s theo TVVN 6848 : 1999 (ISO 11261).

5.2. Thuc th v vt liu

5.2.1 Ct thch anh mn v sch, c ht t 0,1 mm n 0,5 mm
5.2.2 Kali clorua, C(KCl) = 1mol/l.
5.2.3 Cht nn nit c nng cho khong 100 mg N/kg t. V d :
- bt c linh lng vi t l C/N =16:1 ;
- bt sng ;
- bt c ngun nit hu c thch hp no khc.
Cng c th o s khong ho nit t cht hu c ca t. Trong trng hp ny khng cn phi
a vo t bng mt ngun nit hu c no.
Php kh ch quan tm n ni trt ho th amni (NH4)2SO4 l mt ngun nit thch hp.
5.3 Cht th
Cht th ch cn khi mc ch kho st liu cht c nh hng n khong ho N hay khng. Cn
dng nhng cht th tinh khit nht c trn th trng. Trong mt s trng hp, c th dng cc
ho cht hoc hn hp loi k thut hoc thng mi.
Ch thch - Nu cc ho cht hoc thnh phn theo cng thc c trn vi cht th, th nh hng
ca chng n khong ho N phi c tnh n y.
Trong cc th nghim chun vi cht th bit trc, cc d liu sau y (nu p dng c ), cn
a ra l :
- tn (IUPAC);
- cu trc;
- s dch v tm tt ho cht (CAS);
- khi lng phn t tng i;
- tinh khit;
- bn vng trong nc;
- ho tan trong dung mi hu c;
- p sut hi;
- h s phn b octanol/nc (POW);
- hng s phn li axt (pKa);
- h s hp th (Koc).
6 Thit b, dng c th
Cc thit b thng dng cho phng th nghim gm:
6.1 My lc c hc.
6.2 My ly tm hoc giy lc gp (khng c nitrat).
6.3 Dng c o nng amni, nitrat v nitrit trong cht chit t t.
7 Cch tin hnh
7.1 Cc th nghim tu chn
7.1.1 Php th khong ho c bn
so snh cc kh nng khong ho N ca cc loi t khc nhau, hoc so snh s khong ho
nit mt loi t ly thi gian khc nhau trong nm, phi m bo s thit k th nghim v cc
phn tch thc hin ph hp vi mc tiu ca th nghim.
7.1.2 Th tnh c

 xc nh nh hng ca ho cht n qu trnh khong ho nit, x l t gii hn cc hot

ng vi sinh t nht vi 5 nng ca cht th. thun tin, gii hn phn tch vic o cc lng
nitrat to thnh (bng miligam NO3/kg t kh) trong cc mu x l v kim tra sau 28 ngy
mu. S dng thit k th n gin ny, mi quan h liu lng p ng c th c thit lp. Trong
mt s trng hp, v d khi cc nng t c bit hoc c th d kin c bng cch loi
s b (v d t c cha cc cht tr dch hi), thng tin liu lng p ng c th khng cn thit v
mt mu khng x l v mt nng thch hp ca ho cht th l .
7.2 X l mu t
7.2.1 Th khong ho c bn
Chn cht nn trong danh mc lit k 5.2.3 hoc d vic la chn cui cng ca mt cht nn hu
c c trng s dng ph thuc vo mc ch ca php th. Trn tht cn thn v u vt liu hu
c chn vo t. Nu nh s khong ho ca nit do cht hu c ca t ang c kho st,
cn b sung mt cht nn nit.
7.2.2 Th tnh c
xc nh nh hng ca ho cht n s khong ho N, dng mt cht nn nit bt k 5.3.2.
Ch thch - Cc hp cht hu c vi t l C/N thp c l l thch hp nht, v t nit thot ra trong qu
trnh khong ho v b c nh do h vi thc vt ca t.
Trn k v u cht nn nit (5.2.3) vo t. Sau chia mu lm 6 mu nh c khi lng bng
nhau. Trn 5 trong s cc mu nh ny vi cc nng khc nhau ca cht th (3 ln lp li cho
mi liu lng).Trn mu nh cn li nhng khng thm ho cht th no (nu dng mt cht mang
th ch trn vo t). Mu nh khng c ho cht c dng lm i chng khng x l. Nu c th,
chn dy nng cho php cc gi tr ID25, hoc ID50 c c tnh.
p dng cht th c s dng mt cht mang thch hp, v d:
a) trong nc, ph thuc vo ho tan ca hp cht;
b) hoc trn mt cht rn, v d trn vi ct thch anh (5.2.1), hoc vi mt phn t ang kho st.
Vi nhiu loi ho cht hu c, t hoc ct s dng lm cht mang c th c ph ho cht th
bng cch ho tan trong mt dung mi. Trong cc trng hp nh vy, trc khi trn vi t th phi
lm bay hi dung mi.
7.3 t
kho st s khong ho N, cc loi t bng mt trong hai cch sau :
a) cc mu chung ca mi phng n thc,hoc kiu x l;
b) nh mt lot cc mu nh c th cho mi phng n hoc kiu x l.
Khi s dng mu chung , chun b cc lng ln t v ly cc mu (chng hn t 1g n 100
g) trong qu trnh th nghim tu theo s cn thit y, lng t chun c xc nh theo c mu
ly, s ln lp li s dng v thi gian ko di th nghim. Cn trn k t trong mu chung,
trc khi ly cc mu t. Vi mu chung, dn t thnh lp khng dy qu 3 cm to iu kin xy
d di chuyn, cng trn mu hng tun.
Khi cc phng n c thc hin nh l mt lot cc mu c th, ngi ta chia mi phng n
ra thnh mt lot cc mu bng nhau, v b bt cc mu ny i khi cn. Trong cc nghin cu
khong cch ly mu nhiu hn, chun b lng mu cho tt c s ln lp li v cc ln ly mu.
Ch thch - Vic chn nhit , hm lng nc trong t v cc iu kin chiu sng trong qu trnh
, ph thuc vo mc ch th nghim.
i vi cc php th xc nh nh hng ca ho cht n s khong ho N, duy tr nhit t
(200C 20C) v p sut nc l rng khong 0,02 MPa chnh xc n 5 % t (40 % 5 %) n ( 60
% 5 %) sc cha nc ti a trong bng ti.
Ch thch - Nhit (20 0C 20C) c chn lm nhit chun so snh, v v n cho nhng kt
qu tng i nhanh. C th s dng cc nhit ngoi khong nhit ny nu nh chng thch
hp hn (v d, bi v cc iu kin a phng v thiu thit b lm lnh).

Trong tt c cc th nghim, thng ng t phi thun li cho vic trao i t do cc kh. iu ny

gip ngn cn s pht trin cc khu t hiu kh c th l nguyn nhn lm mt nit qua qu trnh
kh nit. Gim ti thiu s mt nc ca t bng cch t trong cc thng y. Xc nh m
ca t theo inh k, b lng mt I bng nc kh ion ho.
Ch thch - C th phun nc kh ion ho ln b mt mu.
Khi cn phi so snh cc tim nng khong ho ca cc loi t khc nhau, phi gi cho tim nng
gi nc ca t tng t nhau cng nhiu cng tt. Trong mt s th nghim, c th t c iu
ny bng cch dn mu t thnh lp mng trong y bnh v thm nc ph trn b mt mu
khong 1 mm.
7.4 Ly mu t th
7.4.1 Th khong ho c bn
S mu v tn s ly mu s ph thuc vo mc ch ca cc th nghim nhng phi va c
th xy dng c mt ng cong liu tc dng.
7.4.2 Th tnh c
xc nh nh hng ca ho cht n khong ho N, ly mu t trc tip sau khi x l (xem 7.2)
ngy 0 v sau 28 ngy (xem 7.3). hu ht nhng mu t cn nguyn vn v vi sinh, c bn
nit hu c, khng x l th t l khong ho ti a t c trong vng 28 ngy. Nh vy ly t
28 ngy thng cho c tnh c hiu lc cc gi tr ID25 hoc ID50.
Ch thch - Nn lm li t nht 3 ln vi mi x l tnh ton thng k.
7.5 Chit t
Chit amni, nitrit, nitrat t cc mu t th (xem 7.4) bng cch lc t vi dung dch KCl (5.2.2) (5
ml KCl cho 1 g t kh), vi mc 150 vng/pht trong 60 pht. tch chit ti u khng c
t v dung dch KCl qu na bnh. Tch phn ht mn ca t khi cht chit bng cch lc hoc ly
tm bng my ly tm hoc giy lc (5.2). Bo qun dch chit khng c ht t (- 20 oC 5oC) trong
6 thng. Khi s dng cc phng php phn tch mi, phi m bo rng s gy nhiu ca dch
chit nm trong phm vi cho php.
Ch thch - Vt liu giu N c kh nng khong ho c th ng gp cho nhng lng amni, nitrit v
nitrat to thnh. Khi yu cu nhng nng cao ca nhng cht o th c tnh, th iu ny phi
c xem xt n.
7.6 Phn tch
Tin hnh phn tch nh lng amni - N, nitrat -N v nitrit - N trong dch chit. 2)
8 Biu th kt qu
8.1 Th khong ho c bn
Cho cc php th c bn, xem xt vic chun b ng cong vn tc khong ho, c s dng cc
gi tr ca ion nit c th. Vi mc ch ny, cho php cc cn bng nit, chuyn cc gi tr (NH 4+),
(NO2- ,), (NO3-) biu th bng mg/kg c xc nh bng php phn tch (7.6) ra amni - N(NH4 + - N);
nitrit -N(NO2- - N) v nitrat -N (NO3- - N).
Trong nhng trng hp khc, c bit l so snh vn tc khong ho ca cc loi t khc nhau,
biu th gi tr khong ho N ti mi khong thi gian ly mu nh l ch mt gi tr gi l " N min"
Dng cng thc sau y xc nh Nmin

trong :
(NH4+-NT) l nng amni thi im ly mu, tnh bng mgN/kg t kh;
(NO2--NT) l nng nitrit thi im ly mu, tnh bng mgN/kg t kh;
(NO3--NT) l nng nitrat thi im ly mu, tnh bng mgN/kg t kh;
(NH4+- NS) l nng amni thi im bt u , tnh bng mgN/kg t kh;
(NO2- - NS) l nng nitrit thi im bt u , tnh bng mgN/kg t kh;

(NO3- - NS) l nng nitrat thi im bt u , tnh bng mgN/kg t kh.

8.2 Th tnh c
xc nh nh hng ca cc nng khc nhau ca cht th n qu trnh khong ho nit, th
phi so snh cc lng nitrat ( ngha l s mg NO3 - /kg t kh) c tm thy cc mu c x l
sau 28 ngy vi lng tm thy mu i chng khng x l.
V cc cn bng N khng cn thit cho vic nh gi c tnh, nn khng cn chuyn i cc gi tr
s miligam NO3- trn 1 kg t kh thnh cc gi tr NO3 - - N trn 1 kg t kh.
Tnh ton cc gi tr c ch, bng phn trm ca gi tr mu i chng cho mi mc x l nh sau:
S c ch ca qu trnh khong ho

=100 -

s miligam NO3- / kg t x l
s miligam NO3- / kg t khng x l

x100

Sau tnh ton n gin ny cho mi nng ca ho cht th, th hoc l:

a) chun b ng cong liu lng p ng nh hnh 1. Trong v d ch ra hnh 1, cc gi tr ID 25 v
ID50 ca cht th gi thuyt l 1 mg ho cht/kg t kh v 10 mg ho cht /kg t kh, hoc l
b) dng php phn tch hi qui ca s ph thuc gia lng cht th v hiu ng c ch gy ra.
Ch thch - Ti thi im kt thc ca cc php th vi mt s ho cht, t x l c th cha
nhiu nitrat hn mu kim chng khng x l. iu ny thng thng l do khong ho ca N trong
cc t bo vi khun b cht bi ho cht. Cc nguyn nhn khc c th l s khong ho N, do ho
cht ang c th hoc s kch thch qu trnh khong ho ccbon hu c ca t.
9 Bo co kt qu
Bo co kt qu bao gm cc thng tin sau :
a) theo tiu chun ny;
b) cc c tnh ca t (xem 5.1.2 v 5.1.3);
c) m t cht th (nu s dng) (xem 5.3);
d) thu thp t, x l, , bao gm ngy thu thp, thi gian bo qun mu, ngy th, phng php x
l (nu c), s lng ho cht dng(nu c), iu kin (xem 7.1 n 7.6);
e) ngy ly mu, ngy v trnh t chit;
f) phng php v thit b phn tch, gii hn pht hin, hiu sut thu hi;
g) hnh v v/hoc bng kt qu;
h) nh gi kt qu v kt lun a ra (nu c).

Hnh 1 - V d v ng cong liu lng p ng th hin cc nng ca cht th c ch

qu trnh khong ho N thnh NO3

Ph lc A
(tham kho)
Phng php o kh nng gi nc ca t
A1 Phm vi p dng
Phng php ny p dng c c mt hnh nh v kh nng gi nc cho cc ng dng m gi
tr chnh xc khng quyt nh.
A2 Nguyn tc
Mt ng tr c y c l mt phn cha t c bt u, ngm vo nc v ro nc. Lng
nc t ly I c xc nh bng cch cn, sy kh n khi lng khng i 1050C v cn li.
A3 Thit b th
A.3.1 ng hnh tr, bit trc th tch, di khong 50 mm n 150 mm v ng knh t 50 mm n
100 mm, c l y.
A.3.2 Ni cch thu nhit phng.
A.3.3. Khay, c l thot nc, ng mt lp ct thch anh, mn v t cao khong 20 mm n 50
mm. A.3.4 T sy, c th duy tr nhit (1050C 20C).
A.3.5 Cn, c chnh xc 0,01 g.
A.4 Cch tin hnh
Ph y c l ca ng hnh tr (A.3.1) bng giy lc v cn ng cng vi giy lc. Cho t vo mt
phn ng hnh tr v y np ng li. Ngm ng ny vo ni cch thu trong 2h nhit phng ,lu
mc nc thp hn u ng. Sau , h thp ng di mc nc trong 1 h. Ly ng tr ra khi
nc v t ng ln khay cha ct (A.3.3), ro nc trong khong 2h n 24h tu theo loi t.
Cn ng cha t, ly ring t ra, sy kh n khi lng khng i 1050C v cn li.
A.5 Tnh ton
Kh nng gi nc ca t, WHC, tnh bng % theo cng thc:

trong : ms l khi lng cht rn bo ho nc + ng tr + giy lc, tnh bng gam;

mt l khi lng b ( khi lng ng tr + giy lc), tnh bng gam;
md l khi lng t kh, tnh bng gam.
A.6 Biu th kt qu
Biu th kh nng gi nc ca t, WHC, bng % khi lng t kh.

Ph lc B
(tham kho)
Ti liu tham kho
[1] ISO 11266 : 1994, Soil quality - Guidance laboratory testing for biodegradation of organic
chemicals in soil under aerobic condistions.
[2] Andersch, I. and Anderson, J.P.E (1991), Influence of pesticides on nitrogen transformation in soil,
Toxicological and Environmental Chmistry, 30, pp. 153 - 158. 1991.
[3] Bremmer, J.M. Nitrogen availability indexes. In C. A. Black : Methods of soil analysis - Part 2 :
Amer. Soc. Agon. Madison, WI 53711, USA, pp. 1324 - 1345, 1965.
[4] Herickson, A. and Selmer Olsen, A.R. Automatic methods for determining nitrate and nitrite in
water and soil extracts. Analyst. 95, pp. 514 - 518, 1970.
[5] Selmer Olsen, A.R. (1971). Determination of ammonium in soil extracts by automated indophenol
method. Analyst, 96, pp.565 - 568.
[6] Stanford, G. and Smith, S.J. Nitrogen mineralization potential of soil. Soil Sci. Soc. Am. Proc. 36,
pp 445 - 472, 1972.