Ni dung mn hc

GiI THIU MN HC
Tn mn hc
Thi lng
Ging vin

Bui

: Thc Hnh Ha sinh TP

: 30 tit TH
: ThS. Phm Hng Hiu

Bi

Bui 1 Bi 1

nh tnh acid amin bng sc k giy

Bui 2 Bi 2
Bi 3

Xc nh protein th trong thc phm

Xc nh hm lng acid hu c trong
sa ti

Bi 8

Trang web
:
https://sites.google.com/a/foodtech.edu.vn/phamhong
hieu
Email
: hieuphamhong@yahoo.com

Bui 3 Bi 4
Bui 4 Bi 5
Bui 5 Bi 7

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

Gio trnh v ti liu tham kho

1.
2.
3.
4.
5.
6.
7.
8.
9.

Thc hnh Ha sinh TP

ThS. Phm Hng Hiu

nh lng vitamin C (chanh)

Kho st hot tnh enzym amylase
nh lng ng tng s ha tan, nh
lng ng kh (to, mn)
Xc nh mt s ch s du m (du n)
Thc hnh Ha sinh TP

Bo co thc hnh

Thc hnh Ha sinh thc phm H Cng nghip TP.HCM

Gio trnh Ha sinh thc phm. H Cng nghip TP.HCM
Hong Kim Anh, Ho hc thc phm, NXB KHKT, 2006
Phm Th Trn Chu, Trn Th Ang, Ha sinh hc, NXB
GD, 1997.
Nguyn Th Hin, V Th Th Ha sinh hc (nng nghip)
NXB Gio Dc 2000
Phm Quc Thng, Nguyn Th Qunh Anh, Ha sinh
hc, HBK H Ni, 1994.
ng Th Thanh Thu, Ha sinh ng dng, T sch H
KHTN, 1996.
L Ngc T v tp th tc gi, Ha hc thc phm, NXB KHKT
H Ni, 1994.
L Ngc T v tp th tc gi, Ha sinh cng
nghip, H&THCN, H Ni, 1997

ThS. Phm Hng Hiu

Ni Dung Bi

Trang ba (theo mu)

Mc lc
Cc bi th nghim:
Tn bi th nghim
Mc ch th nghim
Nguyn tc
Ha cht v dng c
Cch tin hnh (trnh by theo s khi)
Kt qu th nghim
Bn lun
3

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

Quy nh chung

Quy nh chung

Sinh vin phi i cc bui thc hnh, nu vng phi

i b vo bui khc (nhng c 1 bui i b b tr 1 vo
im bo co thc hnh)
Sinh vin i lm thc hnh phi ng gi (tr qu 15
pht s khng c vo lp, phi i b vo bui khc)
Sinh vin phi mc o blouse, gi gn trt t v sinh
trong qu trnh thc hnh, th nghim, khng c t
ra ngoi khi cha c s cho php ca ging
vin, khng tip khch trong phng th nghim.
Dng c th nghim c bn giao v kim tra, nu h
hng, mt mt phi c trch nhim bi thng.

Bi bo co thc hnh s np sau 1 tun khi kt thc

mn hc nhng s liu th ca tng bi phi c ging
vin k nhn trong tng bui thc hnh v np km vi
bi bo co tng kt.
Trong qu trnh thc hnh, ging vin c th kim tra v
l thuyt, cch tin hnh th nghim, nu sinh vin no
khng nm vng kin thc, cch tin hnh th nghim s
c v nh n bi lm b vo bui thc hnh sau
Sinh vin c 2 ct im, mt ct im bo co thc hnh
v mt ct im bi kim tra thng k (vn p t
xut)

ThS. Phm Hng Hiu

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

Thc hnh Ha sinh TP

Pha ha cht

100 g NaOH 30% t NaOH 96%

100 g NaOH 30% t NaOH 96%

250 ml NaOH 1% t NaOH 96%
250 ml CuSO4 1% t CuSO4.5H20 98%
250 ml HCl 1% t HCl 36%
250 ml NaOH 1N t NaOH 96%
250 ml NaOH 0,1N t NaOH 1N
250 ml cn 800 t cn 960

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

C%w/w = m ct x 100 / m dd
m ct = C% x m dd /100 = 30 x 100 / 100 = 30 g
Do NaOH c tinh khit 96%:
mct = m ct x 100 / 96 = 31,25 g
m H2O = m dd mct = 100 31,25 = 68,75 g
VH2O 69 ml
7

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

250 ml CuSO4 1% t CuSO4.5H20 98%

250 ml NaOH 1% t NaOH 96%

C%w/v (g/100ml) C%w/w (g/100g)

Tng t nh trn:
m ct = C% x Vdd /100 = 2,5 g

C%w/v = m ct x 100 / Vdd

m ct = C% x Vdd /100 = 1 x 250 /100 = 2,5 g

m = m x 100 / 98 2,551 g

Do NaOH c tinh khit 96%:

mct = m ct x 100 / 96 2,604 g

Trong 250 g CuSO4.5H2O c 160g CuSO4

m = m x 250 / 160 3,986 g

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

250 ml HCl 1% t HCl 36%

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

10

250 ml NaOH 1N t NaOH 96%

Tng t nh trn:
m ct = C% x Vdd /100 = 2,5 g

m = NV = 1.40.250/1000 = 10 g
m = m x 100 / 96 10,42 g

m = m x 100 / 36 6,944 g
dHCl36% = 1,18 g/ml
V = m / d 5,9 ml

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

11

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

12

250 ml cn 800 t cn 960

250 ml NaOH 0,1N t NaOH 1N

Cn 96:

N1V1 = N2V2

96

80
80

V1 = N2V2/N1 = 0,1 x 250 / 1 = 25 ml

H20:

16

Vcn 96 / VH20 = 80/16 Vcn 96 = (80/16) VH20

m Vcn 96 + VH20 250 ml
(80/16) VH20 + VH20 = 250 ml
VH20 = 250 x 16 /96 41,7 ml
Vcn 96 = 250 41,7 = 208,3 ml
Thc t:
Vcn 96 = 80 x 3 = 240 ml
VH20 = 16 x 3 = 48 ml
ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

13

ThS. Phm Hng Hiu

Bi 1: NH TNH ACID AMIN

Nguyn l:

10 cm
Bt ch

1,5 cm
aa1

aa2

ThS. Phm Hng Hiu

15

10ml mu
5 git pp
NaOH 30% (+2ml)
H2O

H2O

5
9
25ml H2SO4 0,1N
3 git MR
ThS. Phm Hng Hiu

ThS. Phm Hng Hiu

1.
2.
1
3.
4.
5.
6.
2
7.
8.
3 9.
*

16

g lactic/l

17

Nguyn l:
H+ + NaOH Na+ + H2O
phenolphtalein
(khng mu hng)

phu
bnh cu ng nc
bp in
phu
bnh cha nc ngng
cc thu tinh
ng sinh hn
erlen
bnh ct
ng ni cao su

6
Thc hnh Ha sinh TP

Thc hnh Ha sinh TP

Bi 3: XC NH HM LNG ACID
HU C TRONG SA TI

Tin hnh:

Nguyn l:
1.
V c ha mu: Mu c v c ha bng H2SO4 m c
nhit cao vi cht xc tc
Cc cht cha nit (NH4)2SO4
2.
Ct m:

Dng dung dch NaOH ui NH3 ra khi dung dch:

(NH4)2SO4 + 2NaOH Na2SO4 + 2NH3 +H2O

NH3 bay sang bnh hng c cha dung dch H2SO4 ( bit
trc nng , th tch:
NH3 + H2O NH4OH
2NH4OH + H2SO4 (NH4)2SO4 + H2O
3.
nh phn lng H2SO4 d bng dung dch NaOH, t ta xc
nh c lng H2SO4 phn ng v tnh c hm lng
nit tng c trong mu:
H2SO4 + 2NaOH Na2SO4 + 2H2O

hh aa

Thc hnh Ha sinh TP

Bi 2: XC NH PROTEIN TH
TRONG THC PHM

14

Bi 2: XC NH PROTEIN TH
TRONG THC PHM

BNG SC K GIY

Thc hnh Ha sinh TP

(V1 V2) 0,09 N 1000

V

Lu : mu pht hng

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

18

Bi 4: XC NH HOT TNH
ENZIM ALPHA AMYLAZA

Bi 4: XC NH HOT TNH
ENZIM ALPHA AMYLAZA

Nguyn l:
Hot tnh -amylase hin th kh nng ca
enzym amylase xc tc phn ng thy phn
tinh bt n dextrin trong 1 pht 50C v
c biu hin bng s n v ca enzym
trong 1g mu
Khi phn ng thy phn tinh bt xy ra, lng
tinh bt cn li cha c phn hy s to
phn ng mu vi iode v c so mu bng
my so mu quang hc bc sng 620nm

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

Cch tin hnh:

Cn 10g la ny mm nghin vi
30ml nc ct yn 15 pht
lc 20ml nc lc thm vo
80ml cn tuyt i (kt ta)
lng lc ra kt ta 3 4 ln
vi 5 ml cn tuyt i ha tan
phn kt ta = 20ml nc dung
dch amylase (dch chit)
19

ThS. Phm Hng Hiu

Bi 4: XC NH HOT TNH ENZIM

ALPHA AMYLAZA

ng nghim
ng th (t)

ng chun (0)

Nc ct (ml)

Dch chit enzym amylase (ml)

Dung dch m (ml)

Dung dch tinh bt 1% (ml)

0.5

0.5

Dung dch NaCl 3% (ml)

Dung dch HCl 1N (ml)

Nguyn l:
Phng php ny da trn c s phn
ng to mu gia ng kh vi thuc
th acid dinitrosalisylic (DNS)
Cng mu ca hn hp phn ng t
l thun vi nng ng kh. Da
theo th chun ca glucose tinh khit
vi thuc th acid dinitrosalisylic s tnh
c hm lng ng kh ca mu
nghin cu
Bc sng so mu: 530nm

em 50C trong 30 pht

Dung dch HCl 1N (ml)

Nc ct (ml)

5.5

5.5

Thuc th lugol (ml)

0.05

0.05

ThS. Phm Hng Hiu

1 git

Thc hnh Ha sinh TP

21

ThS. Phm Hng Hiu

Bi 5: NH LNG NG TNG V
NG KH

Chun glucose 500ppm

Dch xc nh
Dung dch DNS

0.5

0.5

0.5

0.5

0.5

ng nghim
C (ppm)
A

M1

M2

0.5

0.5

Nc ct

9.5

8.5

7.5

Thc hnh Ha sinh TP

6.5

5.5

7.5

22

7.5
23

M1

M2

50

100

150

200

Cx

Cx

A0 = 0

A1

A2

A3

A4

Ax1

Ax2

Ax = (Ax1 + Ax2)
T s liu thu c lp ng chun
y = ax (A = aC)
T tm Cx = Ax/a

ppm bd C x
C% bd C x

un cch thy si 10 ph (y np ng nghim), chuyn mu nu

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

Bi 5: NH LNG NG TNG V
NG KH

Cch tin hnh:

Cn 4 6g to/mn trch ly nhiu ln bng cn
c kh v 30ml nh mc 100ml pha
long 10 ln dch chit
ng nghim

20

Bi 5: NH LNG NG TNG V
NG KH

Cch tin hnh:

Dung dch ha cht

Thc hnh Ha sinh TP

Vdo Vdm 2 Vdm1

Vxd2 Vxd1 mbd

Vdo Vdm 2 Vdm1 100

Vxd2 Vxd1 mbd 106

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

24

V d
ng nghim

C (ppm)

M1

M2

50

100

150

200

Cx

Cx

0,805 1,372 2,052 2,634 1,006 1,012

th biu din hm lng ng kh

trong mu
3

y = 0.013x
R = 0.994

2.5

Ta c:
Ax = 0,013Cx
m Ax = 1,009
Cx = 77,6

2.634
2.052

2
1.5

Gi s m = 5,230 g
ppmbd = 74202
C%bd = 7,42%

1.372

0.805

BI 7: XC NH MT S CH
S DU M
Xc nh ch s acid
Ch s acid = s mg KOH cn thit trung ha ht
cc acid bo t do c trong 1 g cht bo:
RCOOH + KOH RCOOK + H2O
Trong du m, lng acid bo t do khng ng k
nhng s tng ln trong qu trnh bo qun hoc
giai on ny mm nh gi du m c/mi, qua
ch bin hay cha

0.5
0

0
0

50

ThS. Phm Hng Hiu

100

150

200

250

Thc hnh Ha sinh TP

25

BI 7: XC NH MT S CH
S DU M

+ 3KOH

CH2OH
CHOH
CH2OH

Thc hnh Ha sinh TP

26

Bi 8 : nh lng vitamin C
Nguyn tc:
-It tng i khng tan trong nc, nhng iu ny c
th ci thin bng cch pha trn it vi ioua v hnh
thnh triioua:
I2 + I- <--> I3-Triioua oxy ha vitamin C to acid dehydroascorbic:
C6H8O6 + I3- + H2O --> C6H6O6 + 3I- + 2H+
-Chng no m vitamin C cn hin din trong dung
dch, th triioua c chuyn thnh ion ioua rt nhanh
chng
-Tuy nhin, khi tt c vitamin C b oxy ha, th it v
triioua s hin din trong dung dch v phn ng vi
tinh bt to nn mt hn hp mu xanh en

Ch s x phng:
Ch s x phng l s mg KOH cn thit trung
ha ht cc acid t do v lin kt c trong 1 g cht
bo
Ch s x phng cng cao chng t du m cha
nhiu acid phn t lng thp v ngc li
RCOOH + KOH RCOOK + H2O
CH2OCOR1
CHOCOR2
CH2OCOR3

ThS. Phm Hng Hiu

R1COOK
R2COOK
R3COOK

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

27

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

29

ThS. Phm Hng Hiu

Thc hnh Ha sinh TP

28