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BO CO TH NGHIM HA SINH

BI 3: NH LNG NITO BNG PHNG PHP KJILDAHL

Ngy TN: 19/1/2017


Nhm: S5-02
SVTH: Nguyn Th Hng An MSSV: 61202141
Phm Vn Trc Mi MSSV: 61402453

I. Nguyn tc
Qua 2 giai on
Giai on v c ha:5
Nit tng s l tt c cc dng nit c trong mu. Khi t nng mu cn phn tch
vi H2SO4 m c c mt ca cht xc tc, nit tt c cc dng m c trong mu u
bin thnh dng v c (NH4)2SO4 tan trong dung dch.
Giai on ct m:

Sau khi v c ha. Ta ui NH3 ra khi dung dch bng NaOH. Lng NH3 c
li cun bng my Parnas v c dn n mt erlen c cha mt lng tha H2SO4
bit chnh xc nng .
(NH4)2SO4 + NaOH = Na2SO4 + H2O + NH3
H2O + NH3 + H2SO4 = H2SO4 d + (NH4)2SO4
nh lng H2SO4 cn li bng NaOH, qua tnh c lng nit c trong mu
H2SO4 d + NaOH = Na2SO4 + H2O

II. Tin hnh


1. V c ha
Tin hnh trong t Hotte
Cho mu vo bnh Kjeldahl: ht 1ml nc mm hoc nc tng (ch sao cho
nguyn liu khng dnh vo thnh c bnh). Thm 10ml H2SO4 v 0,5g xc tc.
Cht xc tc c th dng 1 trong cc hn hp sau:
1/ K2SO4 : CuSO4 : Se (100: 10: 1)
2/ CuSO4 : K2SO4 (1:3)
3/ Se kim loi (0,05g)
Sau khi thm xc tc un nh hn hp, trnh si tro v ch un mnh khi hn hp
hon ton chuyn sang dch lng. Trong qu trnh un thnh thong lc nh, trng kho
lo sao cho khng cn mt vt en no ca mu nguyn liu cha b phn hy cn st li
trn thnh bnh. un cho ti khi dung dch trong bnh hon ton trng.Chuyn ton b
dung dch v c ha ha xong vo bnh nh mc 100 ml, thm nc ct cho n vch
nh mc. Ch phi lm ngui v lc u.
2.Ct m
2.1.Ra sch bnh Parnas-Wagner
- Cho nc vo bnh A khong na bnh, thm vi git methyl ; thm dung dch
H2SO4 2N cho n khi nc trong bnh ha , kha van (1).
- Cho nc qua ng sinh hn D, kha van (2) v (3).
- un cho A si u, cho vo C mt t nc ct, m van (2) cho nc chy nit
nit xung B (lu : khng bao gi cho nc chy xung ht m gi li mt t h
thng lun lun kn). Hi nc A s t t qua E, nh vy cho ti khi E c khong
5 ml.Tt bp A, hi nc trong my s ngui lm gim p sut A v B do nc
B s rt qua G. Khi nc rt ht thm nc vo C, m van (2) nc s b rt ht qua G.
Ta c th lm nh vy vi ba ln. Nu nc G y, m van (3) x ht. Xong kha van
(1),(2), (3).
- Ch : khi erlen E vo hoc ly ra khng c h thng b h, c bit l
cc v tr ni 4,5,6.
2.2.Ct m
- Ly vo erlen E 10ml dung dch H2SO4 0.1N thm ba git phenolphthalein , lp
vo h thng.
- Ht 10ml dung dch v c ha nit bnh nh mc cho chy t t vo phu C.
un si A, m van (2) cho dch chy t t v B. Trng phu vi nc ct (khng cho
nc ct C chy xung ht). ng van (2). Cho 10 ml NaOH m c vo C v m (2)
cho NaOH chy t t, trng phu vi mt t nc ct, ng van (2). Qu trnh ct kt thc
sau khong 25 pht. C th kim tra xem NH3 ct hon ton cha bng cch ly E ra,
a mu giy qu vo u ng xem c i mu khng nu khng coi nh NH3 c
ct hon ton.
- Trng vi bng nc ct v ly erlen E ra. Tt bp, ra h thng.
- nh phn lng H2SO4 0,1N tha bng dung dch chun NaOH 0,1N cho n
khi xut hin mu hng nht.
3. Tnh ton
3.1. Tnh h s hiu chun F ca dung dch NaOH
- Ly 10 ml H2SO4 0,1N chun cho vo erlen, nh phn bng NaOH 0,1N.
- Tnh nng thc t ca dung dch NaOH em nh phn.
- F l t s gia nng thc t v nng l thuyt ca NaOH.
3.2. Tnh kt qu :
Hm lng nit trong mu c tnh theo cng thc:

Trong :
X : hm lng nit tnh bng g/l
a : s ml H2SO4 em hp th NH3
b : s ml NaOH 0,1N tiu tn khi chun H2SO4 tha
V : s ml mu em v c ha
0,0014 : lng gam nit ng vi 1 ml H2SO4 0,1N
F : h s hiu chnh nng dd NaOH 0,1 N.
Tnh hm lng protein th thng qua hm lng nit tng
Nit trong vt liu sinh hc ch yu l nit protein ngoi ra cn c mt lng nh
nit trong cc thnh phn khc gi l nit phi protein.
Nit tng = Nit protein + Nit phi protein
Hm lng protein trong cc vt liu sinh hc c tnh bng cch nhn hm
lng nit protein vi mt h s chuyn i xc nh do hm lng nit c t l n nh
t 15 n 18 % protein, i a s protein l 16%.
Trong cc nguyn vt liu sinh hc do hm lng nit phi protein nh v vic
tchring rt phc tp nn theo quy c ngi ta tnh hm lng protein theo nit tng v
gi l protein th hay protein tng. Cng th tnh hm lng phn trm protein th c
trong i a s cc nguyn liu l:
Protein (%) = Nit (%) x 6,25
Ring ng cc v u c h s chuyn i l 5,7
Sa c h s chuyn i l 6,38

III. Kt qu tnh ton:


1. Tnh h s F:
Chun NaOH:
Th tch 3 ln chun : V1=7.4mL , V2=7.45 mL,V3=7.5 mL

1+2+3
Vtb=
3
= 7.4+7.45+7.5
3
= 7.45 mL

100.1
CN (NaOH)=
7.45
=0.134 mL

0.134
F= = 1.34 >1 F 1
0.1
2. Hm lng nito trong mu:

(.)0,00141001000 (105.6x1)x0,0014x100x1000
X= = = 6.16
10 10x10

Hm lng protein th l: % protein=% nito x 6.25 = 6.16 x 6.25=38.5 %

IV. Bn lun:
Hm lng nito trong mu l 6.16 : kh thp
Theo TCVN 5107:2003, nc mm c phn thnh bn hng l c bit, thng
hng, hng 1, hng 2 da theo m.
Cc ch tiu ha hc ca nc mm gm hm lng nit ton phn tnh bng g/l
(theo th t l 30-25-15-10), hm lng nit axit amin, tnh bng % so vi nit ton phn,
khng nh hn 55-50-40-35, hm lng nit amniac, tnh bng % so vi nit ton phn,
khng ln hn 20-25-30-35 v hm lng mui t 145-295g/l.
Vai tr ca ha cht:
CuSO4 : K2SO4 (1:3) : trong th nghim n c tc dng lm cho nng
lng hot ha ca nito gim xung. Trong giai on v c ha mu, s
c mt ca xc tc CuSO4 :K2SO4 thch hp chuyn ton b hn hp
cha N c trong mu thnh NH4+.
H2SO4: H2SO4 m c, t chy hn hp hu c( khi t nng mu
vi H2SO4 m c, cc hp cht hu c c trong mu b oxi ha thnh
SO3, SO3 phn ly thnh SO2 v oxi nguyn t oxi c thi ra s oxi ha
hydro v cacbon ca hp cht hu c c trong mu to thnh CO2 v
H2 O
Cn nito c chuyn thnh dng NH3 v kh NH3 ny s kt hp vi
H2SO4 d to thnh (NH4)2SO4 tan trong dung dch.
Trong giai on hp th: H2SO4 0.1N c vai tr hp th NH3 bay ra
trong qu trnh chng ct:
H2SO4+2NH3 (NH4)2SO4
NaOH m c trung ha acid H2SO4 giai on v c ha.
Cho nc vo bnh A khong na bnh, thm vi git methyl ; thm
dung dch H2SO4 2N cho n khi nc trong bnh ha : nhn bit
h thng c b h hay khng,nu b h, dung dch NaOH b tro ngc
vo bnh A th bnh A s mt mu . Cn m bo h thng kn qu
trnh ct on din ra chnh xc.

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