You are on page 1of 40

B GIO DC V O TO

TRNG I HC CN TH
VIN NGHIN CU V PHT TRIN CNG NGH SINH HC

BO CO PHC TRNH
MN THC TP SINH HA
MSHP: CS115
NHM 1.E

CN B HNG DN

SINH VIN THC HIN

Th.S V Vn Song Ton

Nguyn Th Bch Lin B1203218


Nguyn B Trng B1203178
Thch Hoi Thng B1203167

Cn Th, Thng 3/2014

i
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

MC LC
Ni Dung

Trang

BI 1. CHUN B HA CHT V S DNG MICROPIPETTE


I.

CHUN B HA CHT ........................................................................................... 1


1. Mt s khi nim nng : 3 loi nng thng c s dng trong thc
nghim: .......................................................................................................................... 1
1.1 Nng phn trm: ................................................................................................. 1

II. Dng c ...................................................................................................................... 3


III.

Tin hnh chun b ha cht ................................................................................... 3

3.1.

Chun b ha cht ................................................................................................ 3

3.2.

Kt qu v tho lun. ........................................................................................... 9

IV.

Cch s dng micropipette: .................................................................................... 9

4.1.

Mt s nguyn tc khi s dng:........................................................................... 9

4.2.

Cch cm micropipette: ..................................................................................... 10

4.3.

Cch ly mu: .................................................................................................... 10

BI 2. pH V DUNG DCH M ................................................................................... 10


1. Mc tiu .................................................................................................................... 11
2. Mt s khi nim ...................................................................................................... 11
2.1.

pH ...................................................................................................................... 11

2.2

S phn ly ca cht in ly yu ......................................................................... 11

2.3

Phng trnh Henderson-Hasselbalch v dung dch m ................................. 11

2.4

Dung dch m ................................................................................................. 11

3. Cc bc pha dung dch m: .................................................................................. 13


3.1.

Thit b, dng c, ha cht ................................................................................ 13

3.2.

Chun b 50ml mi loi dung dch m sau: ..................................................... 13


ii

Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

3.3.

Cc bc c bn o pH: .................................................................................... 18

3.4.

Kt qu chun b dung dch m v nhn xt:................................................... 18

4. Bi tp 2: in vo ch trng: .................................................................................. 19
BI 3: CARBOHYDRATE: XC NH M , TRO V HM LNG X TH
20

(CF)

1. Mc tiu:................................................................................................................... 20
2. Khi qut: ................................................................................................................. 20
3. Thit b, dng c, ha cht, mu vt ........................................................................ 21
4. Phng php ............................................................................................................. 21
4.1.

Chun b:............................................................................................................ 21

4.2.

Xc nh m: ................................................................................................. 22

5. Xc nh hm lng tro: (3 ln lp li) .................................................................... 22


6. Xc nh hm lng x th Crube fiber (CF) ....................................................... 22
7. Kt qu th nghim v tho lun: ............................................................................. 23
8. Ph lc s liu .......................................................................................................... 23
BI 4: LIPID

24

1. Mc tiu: .................................................................................................................. 24
2. Khi qut: ................................................................................................................. 24
3. Vt liu v phng php .......................................................................................... 25
3.1.

Vt Liu: ............................................................................................................ 25

3.2.

Phng php: Xc nh hm lng lipid th bng my Soxhlet. ..................... 25

4. Chun b mu: mu bt bp...................................................................................... 26


5. Kt qu v tho lun ................................................................................................. 27
6. Ph lc s liu .......................................................................................................... 27
BI 5: NH LNG PROTEIN TNG (kjeldahl)

27

1. Mc tiu:................................................................................................................... 27
iii
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

2. Khi qut: ................................................................................................................. 27


3. Vt liu v phng php: ......................................................................................... 28
3.1.

Thit b, dng c, ha cht v mu:................................................................... 28

3.2.

Phng php: ..................................................................................................... 28

4. Tin hnh th nghim: (ngu nhin 3 ln lp li) ..................................................... 29


4.1.

V c ha mu: .................................................................................................. 29

4.2.

Chng ct m: .................................................................................................. 29

5. Kt qu v tho lun: ................................................................................................ 30


5.1.

Cng thc tnh: .................................................................................................. 30

5.2.

Kt qu th nghim: ........................................................................................... 30

5.3.

Gii thch: .......................................................................................................... 31

BI 6: CHUN AMONIAC

31

1. Mc tiu:................................................................................................................... 31
2. Gii thiu tng qut .................................................................................................. 31
3. Vt liu v phng php: ......................................................................................... 31
4. Tin hnh th nghim: (ngu nhin 3 ln lp li) ..................................................... 32
5. Kt qu ..................................................................................................................... 33
TNG KT MU THC TP SINH HA

33

1. S liu mu bt bp .................................................................................................. 33
2. Kt lun .................................................................................................................... 34

iv
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

BI 1. CHUN B HA CHT V S DNG MICROPIPETTE


Mc tiu.

I.

Vn dng cc khi nim nng chun b dung dch.

Nm vng cc quy tc s dng micropipette

CHUN B HA CHT

1. Mt s khi nim nng : 3 loi nng thng c s dng trong thc


nghim:
1.1 Nng phn trm:
Nng phn trm trng lng - trng lng (w/w): l s gam ca mt cht ha tan hon ton
trong 100 gam dung dch.

i vi cht rn khng ngm nc ta tnh theo cng thc sau:


C=

mct
100(%)
mdd

Vi : C l nng phn trm dung dch(%).


mct khi lng cht tan cn s dng(g).
mdd khi lng dung dch cn pha(g).
-

i vi cht rn ngm nc:


C=

mct * M
100(%)
m dd * M '

Vi: C l nng phn trm dung dch cn pha (%).


mct : khi lng cht tan cn s dng (g).
mdd :khi lng dung dch cn pha (g).
M :phn t gram khng ngm nc (g/mol).
M :phn t gram c ngm nc (g/mol).
-

i vi cht lng c nng ha tan ti a c tnh theo phn trm:


Cmdd = mct* C
Vi: Cmdd : nng dung dch cn pha (%)
1

Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

mct : trng lng cht tan cn c (g)


C : nng cht tan hin c (%)
-

i vi cht lng ta c th chuyn thnh th tch:


V=

m ct
d

Vi:

V: th tch cn ly (ml)
mct : trng lng cht tan (g)
d: t trng cht tan (g/ml)

a. Nng phn trm trng lng-th tch (w/v): l s gam ca mt cht ha


tan trong 100ml dung dch. Ta p dng cng thc:
Mct =

C % Vdd
100

b. Nng phn trm th tch -th tch (v/v): l s ml ca mt cht ha tan


trong 100 ml dung dch. Cng thc:
V1*C1 = V2*C2
Vi V1 : th tch dung dch cn ly pha (ml)
V2 : th tch dung dch cn pha (ml)
C1 : nng dung dch cn pha (%)
1.2 Nng mol: L s mol cht tan c trong 1 lt dung dch.
mct= M*MW*V
mct : trng lng cht tan cn ly (g)
Mw : phn t gam (g)
V : th tch dung dch cn pha (lt)
M: nng mol cn pha (M)
i vi cc cht tan ngm nc:
mct=

M MW V 100
C
2

Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

vi -mct : khi lng cht tan cn s dng


Mw : phn t gam (g)
V : th tch dung dch cn pha (lit)
M: nng mol cn pha (M)
C : nng dung dch cn pha (%)
1.3

Nng ng lng gam : l s ng lng gam, ca mt cht c trong 1 lt

dung dch hay s mili ng lng gam ca mt cht c trong mt ml dung dch.
ng lng gam (E) ca mt cht l phn phn t gam cht ng vi mt in
tch hot ng. in tch hot ng trong phn ng trao i tnh theo s in tch thc
hin tham gia kt hp vi ion khc, trong phn ng oxi ha kh th tnh theo s in t
cho hay nhn.
II.

Dng c
- Cn in t
-ng ong 10, 25, 50, 100, 1000ml
-Micropipet
-Bnh tam gic
-Cc nha
-C t cc loi
-Mung cn ha cht

III.

Tin hnh chun b ha cht

3.1.

Chun b ha cht

Tng th

Bi

Ha cht vn

Nng

tch/K.

dng th

lng

nghim

(final)

(final)

Tng lng
ha cht
(final)

(gram/ml)

(gram/ml/t)

Din gii (ngn


gn) phng
php chun b

3
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Carbonhydrate
Sy 600 C trong
Ht ht m

300g/nhm

3h (tri ht gel
mng 0,5 cm trn
khay inox)

2 bnh (dng

Bnh ht m

cho 6 nhm)

Bnh kh, sch ,


np bnh thoa
vaseline
Sy cc 1050C

Cc ( m)

3 ci

n khi lng
khng i (4-5h)
Quy trnh thc
hin trong t ht
v eo gng tay,

ta tin hnh nh
sau: Cho 100ml
nc ct vo cc
H2SO4

5%
(v/v)

150 ml

7,65 ml

loi 250ml. Sau


ong 7,65ml
H2SO4(98%) bng
ng ong 10ml
cho t t vo cc
v thm nc ct
vo n
150ml.

NaOH

5%
(w/v)

Quy trnh thc


150 ml

7,5 g

hin trong t ht
v eo gng tay,

4
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

ta tin hnh nh
sau: ong 80ml
H2O cho vo cc
nha loi 250ml,
cn 7,5gr NaOH
ri cho t t vo
cc nha, thm
H2O vo cho
150ml.
Sy giy lc
600C n khi
lng khng i.

Giy lc (15x

3 t

15cm2)/nhm

Sau em giy
vo bnh ht m
v em cn bng
cn phn tch.

Lipid
Chun b 4 chai
Diethy ether

(500ml/chai).

Diethy ether

trc tip vo bnh


cu chun b.
(Trong t ht).
Sy giy lc

Giy lc

600C n khi

(15x15cm2/

lng khng i.

nhm)

Sau em giy
vo bnh ht m
5

Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

v em cn bng
cn phn tch.

Ct 6 si ch c
Dy ch

1 cun

chiu di khong
30cm.

Nit tng

Se: 0,1g
Se:K2SO4:CuSO4

1:100:10

11,1g

K2SO4: 10g

Ha cht c
thy chun b sn.

CuSO4: 1g
Quy trnh thc
hin trong t ht

v eo gng tay,
ta tin hnh nh
sau: Rt H2SO4
H2SO4 chun
0,1N/ ng

0,5 N

1 lt

50 ml

0,1N c sn vo
bnh nh mc
loi 1 lt c
50ml nc ct,
sau thm nc
ct vo cho n
vch c bnh.

6
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Cn 80g NaOH,
cho t t vo cc
nha loi 250ml
c sn 100ml
NaOH

40%
(w/v)

200 ml

80 g

nc ct, khuy
cho tan, khi tan
hon ton cho
nc ct vo n
vch 200ml,
khuy u.
Do khi lng

Thuc th acid
boric

Methyl Red v

10 lt

Bromoresol green
qu nh nn ta
pha hn hp vi t

Acid bric

100 g

2,5 g

l ca 5 lt thuc
th dng chung
cho 6 nhm.

250 ml
(1) Ha tan 33mg
Methyl Red vi
Methyl Red

66g

1,65 mg

50ml ethanol vo
cc thy tinh
100ml bao giy

Bromoresol
green

bc.
99 g

2,475 mg

(2) Ha tan
49.5mg

7
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Bromoresol green
vi 50ml ethanol
vo cc thy tinh
100ml bao giy
bc.
Cho (1) v (2) vo
cc 250ml bao
giy bc, khuy
u.
Mi nhm ly
Ethanol

200 ml

5 ml

5ml hn hp trn
vo bnh cc
250ml bao giy
bc, sau cho
2,5gr acid boric
vo, khuy u.
Cho thm nc
ct n 250ml.
Cho vo bnh ti
mu dn nhn,
y kn.

6
NH3 v Formol

8
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Cho 100ml H2SO4


H2SO4 chun
0.1N

chun 0.1N (Bi


O,05 N

1000 ml

200 ml

5) vo cc 250ml.
Cho nc ct vo
n vch 200ml.
Cho 100ml NaOH

NaOH chun 0,1


N

chun 0,1N (Bi


0,05 N

1000 ml

200 ml

5) vo cc 250ml.
Cho nc ct vo
n vch 200ml.

Formol 37%

3.2.

Kt qu v tho lun.

Kt qu: Ha cht c chun b xong theo ng quy nh, ghi r:


-

Tn ha cht, cng thc ha hc.

Nng dung dch.

Ngy thng pha ha cht.

Tn nhm.

Tho lun: S dng cc cng thc tnh ng khi lng, th tch cc ha cht,
bo co vi cn b ging dy v trnh by vi c nhm. Trong qu trnh chun b
c sai s do cc yu t cn, o, ong, m.
IV. ................................................................................................................ Cch
s dng micropipette:
4.1.

Mt s nguyn tc khi s dng:

Chn micropipette vi th tch thch hp.

Phi bit cch s dng.


9

Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Sau khi s dng phi lm sch u micropipette v t vo gi mt cch cn

thn.
4.2.

Cch cm micropipette:

a.

Ci

mc:

cm

Pipet

nghing v mt pha hi cch


lng lo trn sng bn tay khi
khng s dng, nh vy s lm c
tay t b qu sc.
b.

Cm cht bng c bn tay.

Trnh cm pipet qu cht gim


mi c tay v c vai sau khi s dng
Hnh 1.1: Cch cm micropipette

pipet lu.

l phn ghi s th tch: c th nhn trong sut thi gian c th kim

c.

sot th tch ly ngay c khi ang s dng.


Nt bt ln: mt khong cch thun li trong khong vi ca ngn tay

d.

ci v c mt din tch b mt rng lm gim mi ngn tay ci.


4.3.

Cch ly mu:

Cm micropipette t th thng ng.

n ti nc dng th nht. Nhng u pipet tip khong 3 4 mm vo dung


dch.

Th pittng ra t t.

cho mu, n ti nc dng th nht.

Chuyn tt c phn cn li ca cht lng u pipet tip ra, n ti nc dng


th hai ( pht ht dung dch ra khi pipet tip).

Ch : khi ly mu v cho mu phi cn thn, khng cho mu qu nhanh


trnh to kh dung, to bt, khng cm pipet cht qu, v cm pipet lu c th
lm nng pipet, s lm thay i kch thc pittng, dn n sai s.

BI 2. pH V DUNG DCH M

10
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Mc tiu

1.

Bit vn dng pH k chun b dung dch m.


2.

Mt s khi nim

2.1.

pH

H2O OH- + H+
[OH-] x [H+] = 1x10-14

H2O + H2O OH- + H3O


250C, [H3O+] ca nc bng 1x10-7 M, hay pH = 7,0

pH= -log10[H+]
2.2

S phn ly ca cht in ly yu

Acid yu c cng thc HA


S phn ly ca acid trn c biu din bng phng trnh: HA

H+ +A-

Hng s phn ly:


2.3

Phng trnh Henderson-Hasselbalch v dung dch m

i vi mt acid yu bt k HA, mi quan h gia gi tr pKa, nng cc thnh


phn trng thi cn bng v pH ca dug dch c cho bi phng trnh:
;
2.4

Khi [A-] = [HA] th pH = pKa

Dung dch m

Mc ch: Chng li s thay i trong h thng sinh hc.


Khi nim: Dung dch m l hn hp ca acid yu (HAc/ NaH2PO4...) v nhng
base lin hp (NaAc hoc NH4Ac/ NaH2PO4) ca chng.
Phng php chun b mt s dung dch m: Chun b dung dch m
A.Acetate (Walpole, J. Chem. Soc,. 105, 2501,1914) Stock Solutions:
11
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

A: 0,2M HAc (11,05ml in 1000ml)


B: 0,2M NaAc (16,4g of C2H3O2Na.3H2O in 1000ml)
x ml A + y ml B diluted to a total of 100ml.
pH

3,6

3,8

4,0

4,2

4,4

4,6

4,8

5,0

5,2

5,4

5,6

46,3

44,0

41,0

36,8

30,5

25,5

20,0

14,8

10,5

8,8

4,8

3,7

6,0

9,0

13,2

19,5

24,5

30,0

35,2

39,5

41,2

45,2

Dung dch m phosphate (Sorensen, Biochem, z 21, 131, 1909, 22, 352, 1909) Stock
Solutions:
A: 0,2M NaH2PO4 (27,8g, in 1000ml)
B: 0,2M Na2HPO4 (53,6g Na2HPO4.7H2O or 71,7g Na2HPO4.12H2O in
1000ml)
x ml A + y ml B diluted to a total of 200ml.
pH

5,7

5,8

5,9

6,0

6,1

6,2

6,3

6,4

6,5

6,6

6,7

6,8

93,5 92,0 90,0 87,7 85,0 81,5 77,5 73,5 68,5 62,5 56,5 51,0

6,5

8,0

7,0

10,0 12,3 15,0 18,5 22,5 26,5 31,5 37,5 43,5 49,0

pH

6,9

7,1

7,2

7,3

7,4

7,5

7,6

7,7

7,8

7,9

8,0

45,0 39,0 33,0 28,0 23,0 19,0 16,0 13,0 10,0 8,5

7,0

5,3

55,0 61,0 67,0 72,0 77,0 81,0 84,0 87,0 90,5 91,5 93,0 94,7

12
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Dung dch m Maleate (Temple, J. AM. Soc.51:1754, 1929) Stock Solutions:


A: 0,2M solution acid Na-maleate (8g of NaOH + 23,2g of maleic acid or
19,6g of maleic anhydride in 1000ml).
B: 0,2M NaOH
50 ml A + x ml B, diluted to a total of 200ml:

pH

5,2

5,4

5,6

5,8

6,0

6,2

6,4

6,6

6,8

7,2

10,5

15,3

20,8

26,9

33,0

38,0

41,6

44,4

3. Cc bc pha dung dch m:


3.1.

Thit b, dng c, ha cht

Thit b: Cn in t, my khuy t, pH k
Dng c: Cc thy tinh cc loi, mung cn ha cht,
Ha cht: HAc, NaAc, Na2HPO4, NaH2PO4, sodium carbonate, NaHCO3.
3.2.

Chun b 50ml mi loi dung dch m sau:


3.2.1. Dung dch im A.Acetate pH 4,0 v 5,0:
-

ong 0,5525ml CH3COOH cho vo cc ong (bng ng ong) sn


20ml H2O ct, khuy u, sau thm nc ct vo cho 50ml th ta
c dung dch CH3COOH 0,2M.

Cn 0,7708g CH3COONH4 cho vo cc ong sn 20ml H2O (bng


ng ong), khuy u, sau thm nc ct vo cho 50ml th ta c
dung dch CH3COONH4 0,2M.

Chun b dung dch m A.Acetate pH 4,0:

Dng ng ong, ong 20,5ml dung dch CH3COOH 0,2M (va pha trn)
v dng 1 ng ong khc ong 4,5ml dung dch CH3COONH4 0,2M (va
pha trn), ri cho c hai dung dch ny vo cc 50ml, thm H2O vo cho
n vch 50ml, khuy u ta c dung dch cn pha.

Chun b dung dch m A.Acetate pH 5,0

13
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Dng ng ong ong 7,4ml dung dch CH3COOH 0,2M (pha trn) v
dng 1 ng ong khc ong 17,6ml dung dch CH3COONH4 0,2M (va
pha trn), ri cho c hai dung dch ny vo cc 50ml, thm H2O vo cho
n vch 50ml, khuy u ta c dung dch cn pha.
3.2.2. Chun b dung dch m A.Acetate pH 5,6:
-

ong 0,5525ml CH3COOH cho vo cc ong (bng ng ong)


sn 20ml H2O ct, khuy u, sau thm nc ct vo cho
50ml th ta c dung dch CH3COOH 0,2M.

Cn 0,7708g CH3COONH4 cho vo cc ong sn 20ml H2O


(bng ng ong), khuy u, sau thm nc ct vo cho 50ml
th ta c dung dch CH3COONH4 0,2M.

Dng ng ong 2,4ml dung dch CH3COOH 0,2M (pha trn) v


dng 1 ng ong khc ong 22,6ml dung dch CH3COONH4 0,2M
(va pha trn), ri cho c hai dung dch ny vo cc 50ml, thm
H2O vo cho n vch 50ml, khuy u ta c dung dch cn pha.

3.2.3. Chun b dung dch m phosphate pH 6,0


-

Cn 1,5601g NaH2PO4 cho vo cc ong (bng ng ong) sn


20ml H2O ct, khuy u, sau thm nc ct vo cho 50ml th
ta c dung dch NaH2PO4 0,2M.

Cn 0,71628g Na2HPO4.12H2O cho vo cc ong sn 5ml H2O


(bng ng ong), khuy u, sau thm nc ct vo cho 10ml
th ta c dung dch Na2HPO4.12H2O 0,2M.

Dng ng ong 21,925ml dung dch NaH2PO4 0,2M (pha trn) v


dng 1 ng ong khc ong 3,075ml dung dch Na2HPO4.12H2O
0,2M (va pha trn), ri cho c hai dung dch ny vo cc 50ml,
thm H2O vo cho n vch 50ml, khuy u ta c dung dch cn
pha.

3.2.4. Chun b dung dch m phosphate pH 7,0 v pH 7,5

14
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Cn 0,62404g NaH2PO4 cho vo cc ong (bng ng ong) sn 10


ml H2O ct, khuy u, sau thm nc ct vo cho 20ml th ta
c dung dch NaH2PO4 0.2M.

Cn 3,5814g Na2HPO4.12H2O cho vo cc ong sn 20ml H2O


(bng ng ong), khuy u, sau thm nc ct vo cho 50ml
th ta c dung dch Na2HPO4.12H2O 0,2M.

Chun b dung dch m Phosphate pH 7,0

Dng ng ong 9,75ml dung dch NaH2PO4 0,2M (pha trn) v dng 1 ng
ong khc ong 15,25ml dung dch Na2HPO4.12H2O 0,2M (va pha trn),
ri cho c hai dung dch ny vo cc 50ml, thm H2O vo cho n vch
50ml, khuy u ta c dung dch cn pha.

Chun b dung dch m Phosphate pH 7,5

Dng ng ong 4ml dung dch NaH2PO4 0,2M (pha trn) v dng 1 ng
ong khc ong 21ml dung dch Na2HPO4.12H2O 0,2M (va pha trn), ri
cho c hai dung dch ny vo cc 50ml, thm H2O vo cho n vch 50ml,
khuy u ta c dung dch cn pha.
3.2.5. Chun b dung dch m Maleate pH 6,0
- Cn 0,16 g NaOH v 0,464 g maleic acid ri cho vo cc c cha sn 10 ml
H2O ct, khuy u, sau thm nc ct cho 20ml, khi ta c dung
dch acid Na-maleate 0,2M.
- Dung dch cn dng th 2 l NaOH 0,2M cn 0,08g NaOH ri cho vo cc
c cha sn 5ml H2O ct, khuy u, sau thm nc ct cho 10ml, khi
ta c dung dch NaOH 0,2M.
- ong 12,5ml dung dch acid Na-maleate 0,2M bng ng ng cho vo cc
50ml, dng ng ong khc ong 6,725ml dung dch NaOH 0,2M cng cho
vo cc trn, thm H2O ct vo cho n vch 50ml th ta c dung dch cn
pha.

15
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Tng th

BI

Ha cht vn
dng th ngim

Nng

tch/K.

lng

(final)

(final)
(gram/ml)

Tng lng
ha cht
(final)
(gram/ml/t)

Din gii (ngn gn)


phng php chun
b

Dung dch m A.Acetate


ong 0,5525ml
CH3COOH cho vo
cc ong (bng
HAc
(CH3COOH)

ng ong) sn 20ml
0,2M

50ml

0,5525 ml

H2O ct, khuy u,


sau thm nc ct
vo cho 50ml th
ta c dung dch
CH3COOH 0,2M.
Cn 0,7708g
CH3COONH4 cho
vo cc ong sn

NaAc
(CH3COONH4)

20ml H2O (bng ng


0,2M

50ml

0,7708g

ong), khuy u, sau


thm nc ct vo
cho 50ml th ta
c dung dch
CH3COONH4 0,2M.

Dung dch m phosphate


Cn 3,1202g
NaH2PO4

0,2M

100ml

3,1202g

NaH2PO4 cho vo
cc ong (bng

16
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

ng ong) sn 80 ml
H2O ct, khuy u,
sau thm nc ct
vo cho 100ml th
ta c dung dch
NaH2PO4 0.2M.
Cn 7,1628g
Na2HPO4.12H2O cho
vo cc ong sn
80ml H2O (bng ng
Na2HPO4

0,2M

100ml

7,1628g

ong), khuy u, sau


thm nc ct vo
cho 100ml th ta
c dung dch
Na2HPO4.12H2O
0,2M.

Dung dch m Maleate


Cn 0,16g NaOH ri
cho vo cc c cha
sn 10 ml H2O ct,
NaOH

0,2M

20ml

0,16g

khuy u, sau
thm nc ct cho
20ml, khi ta c
dung dch NaOH
0,2M.
Cn ln lt 0,16 g

Maleic acid

200ml

4,64g

NaOH v 0,464 g
maleic acid ri cho

17
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

vo cc c cha sn
10 ml H2O ct, khuy
u, sau thm
nc ct cho
20ml, khi ta c
dung dch acid Namaleate 0,2M
Tin hnh o pH bng pH k bn:
Cc bc c bn o pH:

3.3.

o Chun b mt s dng c sau: cc nha 500ml, bnh tia, giy thm.


o u tin l ta bt my
o Ra in cc (t 2 3 ln bng nc), v lau kh bng giy thm.
o o pH chun (nn chn pH gn vi pH cn o)
o Ra in cc v lau kh
o o pH dung dch m
o Ra in cc v lau kh
o t in cc vo dung dch tr in cc nh lc u v tt my.
3.4.

Kt qu chun b dung dch m v nhn xt:

Kt qu:
Chun b dung dch m Maleate pH = 6,0, kt qu pH ca dung dch m
sau khi pha o c l 5,947. So vi pH yu cu l chnh lch 0,053 =>
cha t yu cu, sai s cn qu ln (sai s ti a cho php l 0,02).

Nhn xt:
Nguyn nhn sai s:
-

Do ghi nhn tr s pH khi my cha n nh.

Do cch pha cc ha cha chnh xc: cc qu trnh cn v ong th tch

cha chnh xc.


18
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Do nh hng t cc yu t bn ngoi: cc cht trong khng kh (CO2,

SO2,), nhit v m nh hng n tc bay hi.


Cch khc phc sai s:
-

Ch tr s trn pH k n nh ri mi ghi nhn.

Pha ha cht cn c chnh xc cao.

Gim bt s nh hng ca mi trng (thao tc nhanh v chnh xc, hn ch


di chuyn cc ha cht,)

4. Bi tp 2: in vo ch trng:
1/. 7.
2/. 5.
3/. pH= 8.
4/. Hydrophilic.
5/. Solvent.
6/. pH value
7/. Equilibrium constant (Kw)
8/. A- ; HA
9/. Hydrophopic.
10/. logarithmic
11/. exchange reaction
12/. Titration.
13/. hydren, ion, hydrophohic, interactions, vanderwaals force.
14/. base solution.
15/. Buffer.
16/. Donor.
17/. equilibrium.
18/. Acceptor.
19/.pH=pKa.
19
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

20/. Phosphate.
21/. Bicarbonate.
22/. Acidic solution.
23/. Nng ca acid acetic v mui acetat trong dung dch bng nhau v bng
1,738105
24/. pH l ch s o hot ng ca cc ion H+ trong dung dch v v vy n th
hin acid hay base ca mt dung dch, thang o pH l 14, v l thuyt pH=7 l
trung tnh, pH<7 l c tnh acid v pH>7 l c tnh base, pH c tnh bng cng
thc pH = log10[H+].
pKa l pH m ti c 50% phn t acid phn ly thnh H+ v base tip hp, v
50% phn t cn li khng phn ly. Tnh acid ca nhm phn ly cng cao th gi tr
pKa cng thp, => pKa = -log10[Ka].
S khc bit gia pH v pKa: pH biu th mnh hay yu v tnh acid hay base
ca mt dung dch; cn pKa th mc phn ly ca acid v da vo pKa ta c th
xc nh c acid no mnh - acid no l yu, cn da vo pH th kh phn bit
hn.
25/. acid acetic l acid yu hn acid formic. Acid yu c khuynh hng cho proton
d dng hn acid mnh.

BI 3: CARBOHYDRATE:
XC NH M , TRO V HM LNG X TH (CF)

1. Mc tiu:
Kho st hm lng x th trong bt bp.
2. Khi qut:
- Phng php nh lng hm lng cellulose da vo kh nng ha tan nhng hp
cht khng phi cellulose (hemicelluloses, lignin, pectin) s b ha tan bng dung dch
acid sulphuric (H2SO4) v sodium hydroxide (NaOH).
20
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

- Bt bp rt giu cht x, gip gim


cholesterol. Cht x a tan cng gip ngn
nga s gia tng nhanh chng lng ng
trong mu, sau ba n. V vy, y l thc
phm tuyt vi cho nhng ngi b bnh
tiu ng, khng insulin v h ng
huyt.
- Bt bp c cha cc cht chng
oxy ha anthocyanins, c th gip c th
chng li bnh ung th. Bt bp giu cht
Hnh 3.1: Bt bp

x, cc folate v magi nn c li cho tim.


Magi gip ci thin mu, oxy v tt cho tnh mch, ng mch.

- Polyphenols hot ng nh cht chng oxy ha trong mu, ngn nga cc gc t


do t oxy ha.
-Bt bp t calo v khng cha cht bo. Do c hm lng st phong ph nn bt
bp rt tt cho ph n mang thai, tr em v thanh thiu nin ang thi k tng trng.
-Mangan c trong bt bp gip sn xut nng lng v hot ng nh cht chng
oxy ha.
-Tr phong nhit, gii c, gii nhit, bt bp rt thch hp vi ngi thn yu h,
suy nhc khi cm nng, l mn n gii nhit rt tt trong ma h, gip li tiu.
3.

Thit b, dng c, ha cht, mu vt


-

Thit b, dng c: t sy, l nung, h thng phn tch x, my lc nc, cn


in t, cn phn tch, bnh ht m, ht sillicagel (sy 60-70oC trong khong
5 gi).

4.

Ha cht: H2SO4 1,25%, NaOH 1,25%, nc lc, giy lc, cc tro ha.

Mu: bt bp.

Phng php
4.1.

Chun b:

21
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Sy ht ht m 60-70oC trong khong 3-4 gi; vo bnh ht m (sy

trc mt ngy).
-

Sy giy lc (15 x 15 cm2) 60oC n khi lng khng i.

Sy cc 70oC n khi lng khng i (4-5 gi) hoc 105oC trong 2 gi.

4.2.
-

Xc nh m:
Cn 1 gram mu bt bp (M1) cho vo cc ( xc nh trng lng khng
i) ri sy 1050C n trng lng khng i (2-3 gi) xc nh trng
lng khng i ca mu (M2)

cc ng mu trong bnh ht m cho n khi mu tr v nhit phng,


sau em cn, ghi nhn kt qu.

Hm lng m = (M1-M2)x100/M1 (%)

M1: khi lng mu (bt bp)

M2: khi lng mu sau khi sy.

5. Xc nh hm lng tro: (3 ln lp li)


Cn 1gram mu bt bp cho vo cc ( xc nh trng lng khng i),
em tro ha 550oC trong 3 gi; sau ngui nhit phng, mang
mu i cn xc nh hm lng tro.
6. Xc nh hm lng x th Crube fiber (CF)
-Cn 1 gram mu bt bng cn phn
tch (giy du cn mu), chuyn
mu vo my phn tch x.
-Thm 150 ml H2SO4 1,25%.
-un si mu trong 30 pht.
-Lc b H2SO4 bng bm rt chn
khng.
-Ra 3 ln vi 30 ml nc kh ion
Hnh 3.2: My x ha

nng.
-un si mu trong 30 pht.

-Lc loi b NaOH bng bm rt chn khng.


-Ra 3 ln vi nc kh ion nng.
22
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

-Xc nh khi lng mu x sau khi ha tan cc hp cht non-cellulose (x th,


khong khng ha tan, cutin, sillica).

Lc mu sau khi phn tch x qua giy lc gp np (khi lng


khng i).

Tip tc sy ti mu n khi lng khng i 70oC trong 4-5 gi.

Lm ngui ti mu trong bnh ht m v nhit phng trc khi cn;


cn khi lng mu phn tch.

7. Kt qu th nghim v tho lun:


Hm lng m ca mu khong 7,21%, hm lng tro khong 17,23%.

Hm lng x th l 5,52%. Hm lng trn tng i cao. T c th


thy bt bp rt giu cc khong cht.
Gi tr cc ln lp li c s chnh lch ng thi chch lnh v khng

chnh xc ca th do mu ht m, mu ln tp cht, dnh vo cc vt ng


mu nh giy, b tht thot mu, thi gian sy mu cha lu (mu vn
cha n khi lng khng i).
So snh vi nhm 1.a (mu u nnh):
-

Hm lng m ca mu khong 4,85%, hm lng tro khong 4,05%,


hm lng x th 8,3%. Lng x th trong bt bp khng nhiu bng u
nnh tuy nhin gi tr dinh dng l tng i cao.

8. Ph lc s liu
Bng gi tr 3 ln lp li m trong 1 gram bt bp:
Khi
Cc lng mu
(g)

Khi lng cc
(g)

Khi lng
cc + cht x
(g)

Khi lng
m (g)

% m

1,0090

23,7132

24,6532

0,94

6,84%

1,0070

24,8621

25,8001

0,938

6,90%

1,0095

26,6304

27,5674

0,9371

7,90%

23
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Bng gi tr 3 ln lp li hm lng tro trong 1 gram bt bp:


Khi lng cc

Cc + Tro

(g)

(g)

1,0042

22,50

1,0097

1,0086

Cc

Mu

Tro (g)

% Tro

22,6722

0.1722

17.15%

19,83

19,9973

0,1673

16.57%

22,44

22,6213

0,1813

17.98%

Bng gi tr 3 ln lp li hm lng x trong 1 gram bt bp:


Giy lc + x

X th (g)

% x

1,8431

0,0356

3,56%

1,8275

1,8845

0,057

5,69%

1,8679

1,9382

0,0703

7,03%

Cc

Mu

Giy lc (g)

1,0011

1,8075

1,0025

1,0002

th (g)

BI 4: LIPID
1. Mc tiu:
-

Kho st hm lng lipid trong mu bt bp.

2. Khi qut:
-

Lipid thuc nhm hp cht hu c khng ng nht, khng tan trong nc, nhng
d tan trong dung mi hu c khng phn cc nh: diethyl ether, chloroform,
benzene trong alcol cht bo tan c mc .

Trong phn t lipid c t nht mt chc ester ca acid bo cao phn t vi alcol.

Khi thy phn lipid trong mi trng kim mnh nh NaOH (hay KOH) trong alcol
un nng cho x phng v glycerin.
24
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Trong c th sinh vt, lipid c phn gii nh enzyme thy phn lipase to
thnh acid bo tng ng v alcol

Lipid: Lipid trong ht bp ton phn t 4-5%, phn ln tp trung mm. Trong

cht bo ca ng c 50% l axit linoleic, 31% l axit oleic, 13% l axit panmitic v
3% l Stearic. - Gluxit: Gluxit trong ng khong 69% ch yu l tinh bt. ht ng
non c thm mt s ng n v ng kp.
3. Vt liu v phng php
3.1. Vt Liu:
-

Thit b, dng c: bp un, h thng soxhlet dng trch ly cht bo, giy
lc, ch cun.

Ha cht: diethylether

Mu vt: bt bp.

3.2. Phng php: Xc nh hm lng lipid th bng my Soxhlet.


Nguyn l hot ng ca my Soxhlet:
Cht cn chit trong gi trong giy lc (kch thc 3x5 cm) ti v tr (5)
c np vo bung chnh tr chit
(4) ca soxhlet. Tr chit soxhlet
c t ln trn ming bnh cu
(cha 2 lt diethyl ether) v ni vi
ng sinh hn nh hnh bn. Diethyl
ether khi c cung cp nhit s bay
hi ln trn, hi diethyl ether s theo
ng r (3) v bay ln ng sinh hn
(8) s ngng t tr xung thm vo
gi giy lc cha mu, mt s cht
bt u ha tan vo dung mi diethyl
ether. Khi dung mi tng n v tr
cao nht ca ng thot (6) v khi cao

Hnh 4.1: S h thng Soxhlet

hn v tr mc (6) do p sut dung mi trong tr chit cao hn ng thot nn


25
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

dung mi s vt qua co ng thot ti v tr (6) theo ng xung (7) v


bnh cu. Chu k lp li n khi chit bo xong (khong 12h).
Nguyn tc:
Da trn kh nng ha tan lipid ca dung mi hu c khng phn cc
(diethyl ether), dng dung mi hu c ly trch lipid ra khi nguyn liu
c gi nhuyn (bt bp). Trong qu trnh ly trch lipid, cc hp cht tan
trong cht bo nh vitamin (vitamin nhm A, D, E, K v Q), sc t cng
b tch ra khi nguyn liu ban u. Do c ln cc tp cht khc, nn thnh
phn ly trch c gi l lipid th.
Ta xc nh gin tip da trn s chnh lch khi lng nguyn liu
kh trc v sau khi chit lipid ra khi nguyn liu, t suy ra khi lng
lipid trong mu.
4. Chun b mu: mu bt bp.
Trch ly cht bo
- Cn 1g mu vo t giy lc 15x15 cm2 ( xc nh trng lng khng i). Sau
gp giy li c gi khong 3x5
cm2, dng ch ct 2 u ti.
-

Cho mu vo ng hnh tr ca h thng


soxhlet

- S dng 2 lt diethyether trch ly


lipid c trong mu trong thi gian
khong 12 gi bng h thng soxhlet.
- Thu mu, kh mu trong t ht (1530 pht).

Hnh 4.2: Gi 1gram mu


bt bp

- Sy mu n trng lng khng i.


- Hm lng cht bo: (M2 - M1) x100/M1 (%)
M1: Khi lng mu ban u.

M2: Khi lng mu sau khi sy 1050C

26
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

5. Kt qu v tho lun
Hm lng lipid: mu 2 l 11,9%; mu 3 l 16,41%. Hm lng lipid gia 2 mu
chnh lch ng k. Trung bnh t 3 mu lp li l 13,99 tng i cao.
Nguyn nhn c th do sai trong qu trnh thc hin th nghim. Thi gian cng di
th hm lng lipid cn li trong b cng t. Mu tht thot trong qu trnh thc nghim.
So snh vi nhm 1.a ( mu u nnh):
Hm lng lipid: 23,07% ln hn nhiu so vi lng lipid ca mu bt bp

l 13,99%. Do u nnh cha rt nhiu lipid hn cc loi ng cc khc.


6. Ph lc s liu
Bng ph lc hm lng lipid 3 ln lp li trong 1gram bt bp:
Giy lc +

Khi lng

Giy lc

mu (g)

(g)

1,0006

1,7681

2,6319

0,1368

13,67%

1,0032

1,8048

2,6886

0,1194

11,90%

1,0071

1,7871

2,6289

0,1653

16,41%

Mu

mu (g)
sy

Khi lng
lipid

% lipid

BI 5: NH LNG PROTEIN TNG (kjeldahl)


1.

Mc tiu:
Xc nh hm lng Protein trong mu bt bp.

2.
-

Khi qut:
Protein c cu to t nhiu nguyn t nh C, H, O, N, S, P trong s cc
nguyn t ny, nit chim t l tng i cao (khong 15-17%) v tng i n
nh trong thnh phn ca protein. V vy v nguyn tc, protein trong thc phm
27

Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

c nh lng bng cch xc nh nit tng s v kt qu nhn vi 6,25 (xem


nit tng s chim 16%).
-

Bt bp c trung bnh 10,6% protein, protein chnh ca ng l zein, mt loi


prolamin gn nh khng c ly sin v tryptophan. Nu n phi hp ng vi u
v cc thc n ng vt th gi tr protein ng s tng ln nhiu.

3.

Vt liu v phng php:


3.1.

Thit b, dng c, ha cht v mu:

Thit b, dng c: t sy 105oC, l v c ha mu, my phn tch m bn t


ng, ng phn tch m tng, bnh ht m, c ng mu xch nh trng lng
khng i.

Ha cht: Se:K2SO4:CuSO4 = 1:100:10, H2SO4 , H2SO4 0,1N, NaOH 40%, Acid


boric, methyl red, bromoresol green, ethanol, MgO.

Mu vt: Bt bp.
3.2.

Phng php:

Da theo nguyn l Kjeldahl


Khi un si mu cha N trong H2SO4 m c vi s cht xc tc th cc hp cht
hu c b oxy ha cn NH3 c gii phng ra lin kt vi H2SO4 to thnh
(NH4)2SO4.

Ta c th xc nh hm lng m ny khi cho chng tc dng vi sodium


hydroxide (NaOH), chng s c li cun bng hi nc v i qua ng sinh hn,
ngng t xung bnh tam gic cha dung dich acid boric c cha cht ch th, to
thnh tetraborat amon.

NH 4 2 SO4 2 NaOH Na 2 SO4 H 2 O 2 NH 3


2 NH 4OH 4 H 3 BO3 NH 4 2 B4O7 7 H 2O

28
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Sau chun dung dch ny bng dung dch chun H2SO4 0,1N, NH3 c gii
phng v xc nh c lng Nit theo cc phn ng sau:

NH 4 2 B4O7 H 2 SO4 5H 2O NH 4 2 SO4 4 H 3 BO3


4. Tin hnh th nghim: (ngu nhin 3 ln lp li)
V c ha mu:

4.1.
-

Cn 1gram bt bp cho vo ng
nghim ng mu (ng Kjeldahl)
ca my phn tch m.

Thm 5ml H2SO4 m c v mt


t

bt

xc

tc

(Se:K2SO4:CuSO4=1:100:10) vo
ng Kjeldahl.
-

Lp ng vo h thng l v c
ha mu, m my v tin hnh v
c ha mu.

Hnh 5.1: h thng c ha

Vic v c ho mu hon tt khi


thy dung dch trong ng v c ho mu trong sut khng mu hoc c mu
xanh l ca CuSO4.

ng nghim v ton b h thng ngui hon ton, sau chuyn sang h


thng my chng ct m.

4.2.
-

Chng ct m:
Dng 15ml thuc th Boric cho vo bnh tam gic 250ml; t vo v tr thu
kh NH3 bay ra ( u thu mu ngp trong dung dch acid Boric); chuyn mu
( v c ha) vo bnh cha mu ca h thng.

29
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Tip tc thm t t 30ml NaOH


40% (m/v) vo; tip tc chng
ct ui ht NH3 ra khi
bnh cha mu (khong 15 pht
t lc thuc th i mu).
nh phn NH3 bng dung dch

H2SO4 chun 0,1N.


5. Kt qu v tho lun:
5.1.

Hnh 5.2: Chng ct m

Cng thc tnh:


a.

Mu rn:

N 0,0014 * V H 2 SO4 * 100 / M %

b.

Mu lng:

N 0,0014 * V H 2 SO4 * 1000 / V mgN / ml

N: hm lng nit tng s.


V: th tch nguyn liu em phn tch.
m: khi lng mu em phn tch (g).
0,0014: s gam nit tng ng vi 1ml H2SO4 0,1N
Protein tng s = Hm lng Nit tng*6,25
6,25: h s c trng cho protein
5.2.

Kt qu th nghim:
Bng gi tr Nit tng trong 3 ln lp li 1 gam mu bt bp:
Hm lng nit

Mu

Khi lng mu (g)

H2SO4 chun (ml)

1,0006

4,9

0,69%

1,0019

7,8

1,09%

1,0008

2,5

0,35%

tng s (%)

30
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Nhn xt: Hm lng nit tng trung bnh l: 0,71%


Hm lng protein tng s = 0,71*6,25= 4,4375%
Vy hm lng protein tng s trong mu bt bp em phn tch l
4,4375%.
Kt qu thu c cc ln thc nghim c s chnh lch.
5.3.

Gii thch:

Mu b dnh thnh ng, thnh phu,v trng nc cha sch.

Lng m cn ng li trong h thng.

Trong nc vn c ngun m.

S dng nhiu nc trong trng ra mu.

Hp th cha ht sn phm.

Do chun sai.

Tho lun v kt lun: Rt kinh nghim v phng php tin hnh th nghim cho
ng quy cch hn ch sai s, hm lng m trong cc loi lng thc hay trong
cng mt loi c trng cc iu kin khc nhau v trong cc giai on sinh trng
khc nhau th c hm lng khc nhau, do kh c th cho mt kt qu th nghim
chnh xc.

BI 6: CHUN AMONIAC
1. Mc tiu:
Xc nh hm lng amoniac trong mu nc
mm c cm Hng i.
2. Gii thiu tng qut
Vt liu: mu nc mm c cm Hng i.

3. Vt liu v phng php:


- Dng c th nghim: ng ong, h thng
nh lng ammoniac, pipet, bnh tam
31
Chuyn ngnh Vi Sinh Vt K38

Hnh 6.1: Nc
mm Hng i
Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

gic 250 ml, my ct m, ng Kjendahl, ng ong, Pipette, Erlen, bnh nh


mc, Burette, Becher
- Ha cht: MgO, thuc th Acid Boric, NaOH 40% (w/v), H2SO4 chun 0,05N.
- Mu vt: nc mm c cm Hng i.
Phng php
Nguyn tc: y mui amon ra th t do (thng di dng mui NH4Cl)
bng cht kim mnh hn amoniac nh Mg(OH)2. Dng hi nc ko
ammoniac c gii phng ra th t do sang bnh hp th cha dung dch
acid Boric (H3BO3) vi cht ch th mu (hn hp bromoresol green v methy
red) to thnh mui
o amon tetraborat.

MgO + H2O Mg(OH)2


2 NH4Cl + Mg(OH)2 MgCl2 + 2 NH3 + H2O
2 NH3 + 4 H3BO3 (NH4)2B4O7 + 5 H2O
Sau nh phn Nit trong dung dch (NH4)2B4O7 bng dung dch acid mnh
H2SO4 0,05N (chun) n khi dung dch chuyn t mu xanh sang nu.
(NH4)2B4O7 + H2SO4 + 5 H2O (NH4)2SO4 + 4 H3BO3
4. Tin hnh th nghim: (ngu nhin 3 ln lp li)
Tin hnh chng ct:
-

Cho vo ng Kjendahl 50ml nc

ct, ht chnh xc 1ml mu nc mm c


cm Hng i cho vo ng Kjendahl.
Thm 3gram MgO. Lm kn h thng v
chng ct cho hi nc v NH3 to thnh
NH4OH vo bnh tam gic 250ml cha
20ml dung dch acid Boric- cht ch th
mu. Ch khng cho ng dn kh ngp
vo dung dch ch th v s ht ngc dung

Hnh 6.2: H thng chng ct.

32
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

dch tr li. Chng ct trong khong 15 pht t khi thuc th bt u i mu. Sau
hng khong 50ml dung dch c mu xanh nc bin.
Sau khi chng ct xong cc mu ta tin hnh mu th khng (thay lng

mu bng lng nc ct) loi t sai s.

5. Kt qu
Bng gi tr hm lng Nit amoniac:
Mu

H2SO4 0,05N chun mu (ml)

Nit amoniac (%)

2,7

1,82

3,2

2,17

4,9

3,36

m amoniac: =

0,7 xV V0 0,0007xV Vo

x1000 2,45( g / l )
m
M

V: s ml H2SO4 0,05N chun.


V0: s ml H2SO4 0,05N chun mu khng (nc) (V0=0).
0,007: s gram nit tng ng vi 1ml H2SO4 chun.
m: s gram hay ml mu tht.
Nhn xt: Hm lng ammoniac trong nc mm theo kt qu th nghim l
2,45(g/l). Theo s liu tham kho v hm lng ammoniac trong mu nc mm c cm
Hng i l 35. Qu thp so vi s liu mu.
Gii thch v kt lun: Nguyn nhn sai s c th do, ha cht pha cha chnh
xc, ra h thng khng sch, do chun sai, mu b bin cht, doanh nghip
cha cung cp ng ch s dinh dng ca mu.

TNG KT MU THC TP SINH HA


1. S liu mu bt bp

33
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

Trong qu trnh thc tp vi mu bt bp, kt qu hm lng cc cht thnh phn


c tng hp nh bn sau:

STT

Thnh phn

Hm lng trung bnh (%)

7,21

Cht x

5,52

Tro

17,23

Lipid

13,99

Protein

4,44

2. Kt lun
-

Trong cc thnh phn o c, x v lipid c hm lng cao.

Tng phn trm hm lng cc cht trn khng t 100% v mt s cht khc
khng c tin hnh o c v tht thot trong qu trnh thc hin th nghim.

Hm lng tro qu cao (17,23%) l do sai st trong qu trnh thc hin.

34
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

PH LC BNG
Ni dung

Trang

BI 1. CHUN B HA CHT V S DNG MICROPIPETTE

Bng chun b ha cht

BI 2. pH V DUNG DCH M

10

Bng pha dung dch m phosphate

12

Bng pha dung dch m Maleate

13

Bng pha dung dch m A.Acetate

16

BI 3: CARBOHYDRATE: XC NH M , TRO V HM LNG X TH


(CF)

20

Bng gi tr 3 ln lp li m trong 1 gram bt bp:

23

Bng gi tr 3 ln lp li hm lng tro trong 1 gram bt bp:

24

Bng gi tr 3 ln lp li hm lng x trong 1 gram bt bp:

24

BI 4: LIPID

24

Bng ph lc hm lng lipid 3 ln lp li trong 1gram bt bp:

27

BI 5: NH LNG PROTEIN TNG (kjeldahl)

27

Bng gi tr Nit tng trong 3 ln lp li 1 gam mu bt bp:

30

BI 6: CHUN AMONIAC

31
33

Bng gi tr hm lng Nit amoniac:


BNG TNG KT

33

PH LC HNH
Ni dung

Trang

BI 1. CHUN B HA CHT V S DNG MICROPIPETTE


Hnh 1.1: Cch cm micropipette:

1
10

v
Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

Trng HCT

Phc trnh thc tp Sinh Ha- 2014

BI 2. pH V DUNG DCH M

10

S phn ly ca cht in ly yu

11

BI 3: CARBOHYDRATE: XC NH M , TRO V HM LNG X TH


(CF)

20

Hnh 3.1: Bt bp

21

Hnh 3.2: My phn tch x

22

BI 4: LIPID

24

Hnh 4.1: S h thng soxhlet

26

Hnh 4.2: Gi 1gram bt bp

27

BI 5: NH LNG PROTEIN TNG (kjeldahl)

27

Hnh 5.1: H thng l v c ha mu

29

Hnh 5.2: Chng ct m

29

BI 6: CHUN AMONIAC

31

Hnh 6.1: Mu nc mm c cm Hng i.

31

Hnh 6.2: H thng chng ct

33

PH LC NGUN
-

Trong Bi bo co c tham kho ngun ti liu thc tp ca thy V Vn Song


Ton, bi bo co ca kha 38.

C s dng hnh nh v thng tin t cc trang web:


http://www.wtionline.org/file.php/168/Water_Reaction.png
http://upload.wikimedia.org/wikibooks/en/7/76/Chemical_Principles_Pic_5.1.png
(4:43PM ngy 19/03/2014)
http://www.nihe.org.vn/new-vn/dao-tao-ngan-han--tap-huan412312272/392/Cach-su-dung-cac-loai-Pipet-trong-phong-thi-nghiem.vhtm
http://vi.scribd.com/doc/154773860/Giao-Trinh-Thuc-Tap-Sinh-Hoa
(10:35AM ngy 20/03/2014)
vi

Chuyn ngnh Vi Sinh Vt K38

Vin NC&PT Cng ngh Sinh hc

You might also like