TIU CHUN VIT NAM

TCVN 3705-90
THY SN
PHNG PHP XC NH HM LNG NIT TNG S V PROTEIN TH
Aquatic products
Method for determination of total nitrogen and protein contents
Tiu chun ny thay th TCVN 3705-81, qui nh phng php xc nh hm lng nit tng s
v protein th i vi cc nguyn liu, bn thnh phm v sn phm thy sn.
1. XC NH HM LNG NIT TNG S
1.1. Ly mu
Ly mu v chun b mu th theo TCVN 5276-90.
1.2. Nguyn tc chung
V c ha mu th bng axit sunfuric m c, nit c trong mu th chuyn thnh amon
sunfat. Dng kim c y amoniac ra khi amon sunfat trong my ct m, to thnh amon
hydroxyt, ri nh lng bng axit.
1.3. Dng c v ha cht
- My ct m;
- Bnh kendan, dung tch 100, 250ml;
- Bnh nh mc dung tch 100ml;
- ng ong dung tch 10, 100ml;
- Buret 25ml;
- Pipet 10, 20, 50ml;
- Bnh nn, dung tch 250ml;
- Chn cn;
- Phu thy tinh;
- Tha nha;
- Bp in;
- Giy lc khng tro;
- Giy o pH;
- Cn phn tch, chnh xc 0,001 g;
- Axit sunfuric (H2SO4) m c v dung dch 0,1N;
- Natri hydroxyt (NaOH), dung dch 33% v dung dch 0,1N;
- Hn hp xc tc: ng sunfat (CuSO4) + Kali sunfat (K2SO4), t l 1/10 (theo khi lng);
- Ch th hn hp: 200mg metyl v 100mg xanh metylen ha tan trong 200ml etanol (C 2H5OH)
96%;
- Phenolphtalein, dung dch 1% trong etanol 60%.
1.4. Tin hnh th

Cn chnh xc 0,3 0,5g mu th vo mt mu giy lc khng tro, cun li, cho vo bnh kendan
sao cho mu th khng dnh vo c bnh, cho tip 1g hn hp xc tc v 10ml axit sunfuric m
c.
- Nu mu th l cc loi mm c: cho mt t mu th vo chn cn cng vi mt tha nha ly
khong 0,5g mu th vo mu giy lc khng tro, cun li, cho vo bnh kendan v c ha.
Tha nha li t vo bnh cn, cn khi lng cn li c M 2. Hiu s gia hai ln cn l khi
lng mu th ly phn tch.
- Nu mu th l nc mm: Dng pipet ly chnh xc 10ml nc mm lc cho vo bnh nh
mc dung tch 200ml, thm nc ct n vch mc, lc u. Ht chnh xc 20ml dch pha long,
cho vo bnh kendan, thm cht xc tc v 3 5ml axit sunfuric c vo v c ha.
Dng phu nh y bnh kendan, t bnh nghin 40 trn bp in trong t ht. un 15 -20
pht sao cho cht lng trong bnh khng si phng, khng bn ln c bnh (c th qua m ri
mi un). Sau t bnh thp gn bp hn cho ti khi dch v c ha trong bnh trong sut
hoc trong xanh (khng c c mu vng nht) mt trong bnh hon ton trong sch. Ngng
un, ngui.
Trong qu trnh un, nu thy mu khng trng, ngng un, ngui, cho thm khong 0,5g
cht xc tc vo ri tip tc un. Nu thy mu cn en m cn, th ly ra ngui, cho thm
khong 3ml axit sunfuric m c vo v tip tc un cho ti khi dung dch t yu cu nh trn.
Ly chnh xc mt lng axit sunfuric 0,1N khng ln hn 25ml (ty theo tng loi mu th) v 5
git ch th hn hp vo bnh nn dung tch 250ml, t bnh vo di ng sinh hn ca my ct
m sao cho u ng sinh hn ngp hn vo dung dch.
Cho cn thn dch v c ha vo bnh ct, trng bnh kendan nhiu ln bng nc ct cho
n khi nc trng ht phn ng axit (th bng giy o pH). Cho tip vo bnh ct 5 git
phenolphtalein 1% v dung dch natrihydroxyt 33% cho n khi dung dch trong bnh chuyn
thnh mu hng, cho tip vo mt t dung dch kim, trng nc ct cho sch kim phu ri
kha my li. Cui cng cho mt lp nc ct cao 1,5 2cm trn phu kim tra kn ca
my. (Ghi ton b lng nc ct dng bit lng nc ct cho vo khi chun mu
trng).
Cho nc lnh chy qua ng sinh hn v bt u chng ct lin tc trong 40 pht k t khi dung
dch trong bnh bt u si. H bnh hng ng sinh hn ln khi mt nc, dng bnh tia ra
u ng sinh hn, tip tc chng ct mt vi pht na. Sau hng nc chng chy ra u
ng sinh hn, th bng giy o pH thy khng c phn ng kim l c.
Dng natri hydroxyt 0,1N chun lng axit d trong bnh hng cho n khi dung dch trong
bnh chuyn t mu tm sang xanh l m.
Tin hnh xc nh mu trng vi tt c lng ha cht v nc ct v cc bc th nghim nh
trn, khng c mu th.
1.5. Tnh kt qu
Hm lng nit tng s (X7) tnh bng phn trm theo cng thc:
X7 =

(V1 V2 ) 0,0014 100

m

Trong :
V1 Th tch dung dch natri hydroxyt 0,1N tiu tn khi chun mu trng, tnh bng ml;
V2 Th tch dung dch natri hydroxyt 0,1N tiu tn khi chun mu th, tnh bng ml;
m khi lng mu th, tnh bng g;
0,0014 S g nit tng ng vi 1ml dung dch natri hydroxyt 0,1N;
100 H s tnh ra phn trm.

Ch thch:
i vi nc mm, mu th c pha long 20 ln, ly 20ml dch pha long xc nh.
Hm lng nit tng s (X7) c tnh bng g/l theo cng thc:
X7 =

(V1 V2 ) 0,0014 20 1000

2,8(V1 V2 )
10

Trong :
20 pha long ca nc mm;
10 Th tch nc mm pha long ly xc nh, tnh bng ml;
1000 H s tnh ra g/l;
Cc k hiu khc nh ghi trn.
2. PHNG PHP TNH HM LNG PRTEIN TH
Hm lng nit trung bnh trong phn t protein ca sn phm thy sn l 16%. V vy hm
lng protein th trong mu th bng hm lng nit tng s nhn vi h s 6,25.
Hm lng protein th (X8) tnh bng phn trm theo cng thc:
X8 = X7 . 6,25
Trong :
X7 Hm lng nit tng s, tnh bng phn trm;
6,25 H s chuyn nit tng s ra protein th (100:16 = 6,25).