Professional Documents
Culture Documents
=
32
Trong :
m: khi lng thc phm cn ban u (g), hoc th tch ca thc phm o
ban u
m
1
: trng lng ca becher cha lipid (g)
m
2
: trng lng ca becher
33
BI S 8: XC NH HM LNG PROTEIN TRONG SA THEO
PHNG PHP KT TA
8.1. L THUYT
- Cc phng php xc nh protein
- Tnh cht ca protein trong sa
8.2. DNG C V THIT B
- My ly tm
- 4 ng ly tm chu nhit
- 1 ni cch thy
- a thy tinh
- ng nh git
- Cn phn tch
- Bnh ht m
- T sy
8.3. HA CHT
- Acid trichlor acetic 50 %
- Acid trichlor acetic 1%
8.4. THC HNH
8.4.1. Chun b mu
- Tng t nh bi xc nh cht bo theo phng php Adam Rose Gottlieb.
- Ly phn chit lng xung di y bnh lng.
8.4.2. Phng php xc nh
- Cho phn lng vo ng ly tm sy kh, cn sn.
- t ng ly tm ln ni cch thy cho bay ht hi ether, cn v ammoniac (khong
30 35 pht).
- Cho tng git acid trichlor acetic 50 % vo ng ly tm cho n khi kt ta. Ch ,
nh tng git theo thnh ng v quan st ch tip xc gia 2 dung dch.
- Dng a thy tinh khuy u.
- t ln ni cch thy un si khong 30 pht protein kt ta ht.
- Quay ly tm, protein s lng xung di y ng. B kt ta pha trn.
- Ra ta nh trn thm 2 ln na, cui cng cht kit nc.
34
- Cho ta v ng ly tm vo t sy nhit 100 105
0
C n khi lng khng i
(khong 4 h).
- Ly ra cho vo bnh ht m, ngui v cn.
8.4.3. Tnh ton kt qu
Hm lng cht bo c trong 100 g hoc 100 ml thc phm l:
1 2
( ) 100 m m x
X
m
=
Trong :
m: khi lng thc phm cn ban u (g), hoc th tch ca thc phm o
ban u
m
1
: trng lng ca ng ly tm cha m (g)
m
2
: trng lng ca ng ly tm
35
Bi s 9: XC NH M NGUYN LIU
9.1. Khi nim m nguyn liu
Nguyn liu m, c th coi nh hn hp c hc gm cht kh tuyt i v nc
t do:
m = mo + w (1)
Trong : m - Khi lng chung ca nguyn liu m
mo - Khi lng cht kh tuyt i (khng c m)
w - Khi lng nc cha trong nguyn liu.
+ m tng i () ca nguyn liu m: L t s gia khi lng nc trn
khi lng chung (m) ca nguyn liu m, tnh bng phn trm:
o
o
o
o
o
w m
w
m
w
100 100
+
= = (2)
+ m tuyt i (
o
) ca nguyn liu m: l t s gia nc (w) v khi
lng cht kh tuyt i (m
o
) ca nguyn liu m, %:
o
o
o
o
m
w
100 =
Mun quan st qu trnh sy bng ng cong ly mt cch r rng (to thnh
cc im un cc giai on sy) ngi ta thng s dng m tuyt i, cn m
tng i biu th trng thi m ca nguyn liu.
V vy trong sn xut, xc nh thnh phn m hay m ca nguyn liu
ngi ta thng xc nh v tnh ton m tng i () bng % theo biu thc (2)
trn.
9.2. Xc nh m tng i
9.2.1. Phng php sy khi lng khng i
9.2.1.1. i vi cc loi nguyn liu c m khng qu 18%: qu, c, ht,
vt liu ri v bt (vt liu rn)
Dng c th nghim
- Cc thy tinh hay hp kim loi ng mu
- My nghin hoc ci nghin, a thy tinh
- T sy, bnh ht m
- Cn phn tch
36
Cch tin hnh
X l nguyn liu
- i vi qu, c ti c th tho hoc bm nh, nghin nh i vi qu, c
kh
- i vi cc loi ht, cc loi bnh quy, vt liu dn c kch thc ln
em nghin nh
- i vi vt liu ri nh ng ct, bt ngt, cc loi bt kch thc nh
khng cn x l
- X l nguyn liu thc hin nhanh, x l xong cho vo hp kn
Dng cn phn tch cn chnh xc 5 g mu c x l cho vo cc sch, kh
bit trc khi lng
Dng a thy tinh san u lng mu trong hp ng mu m bc hi
nhanh v u
a mu vo t sy nhit 105
0
C n khi khi lng khng i. Sau khi
sy cho mu vo bnh ht m lm ngui v cn. Khi kt qu gia 2 ln cn cui
cng c sai s + 0,5% coi nh khi lng khng i.
Tnh kt qu: Nu gi
- G
1
: Khi lng hp v nguyn liu trc khi sy (g)
- G
0
: Khi lng hp khng (g)
- G
2
: Khi lng hp v mu sau khi sy (g)
- G: Khi lng mu cn xc nh (g)
G = G
1
G
0
m tng i hay m tng i c xc nh theo biu thc sau:
100
2 1
x
G
G G
=
9.2.1.2. i vi cc loi nguyn liu c m ln hn 18 %: bt nho, bt
st, ng non, du thc vt
i vi nguyn liu ny nn trn u vi mu ban u trn 0,5kg, sau ly mu
th nghim trn 20g.
Dng cc hp ng mu c th tch ln hn hp dng ng vt liu rn, a
thy tinh ngn dng trn mu, c th yn mu trong hp sau khi sy. Trc khi
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cho mu vao hp, hp v a thy tinh cng c ra sch, sy kh, lm ngu v cn
bit trc khi lng.
Sy nhit 105
0
C cho n khi m mu t 18%, nghin nh, cn 5g, sy
130
0
C trong 40 pht ri lm tip tc nh phn 4.1. Cng thc tnh:
W = 100 G.g (%)
Trong : G: Khi lng 20g nguyn liu sau khi sy s b nhit
105
0
C n m di 18% (khong 30 pht)
g: Khi lng 5 g nguyn liu (ly t G) sau khi sy 105
0
C n
khi lng khng i
9.2.1.3. Xc inh m nguyn liu dung dch c
- Nguyn tc: Cho nc bc hi ht, ta thu c lng m trong dung dch
- Dng c:
Cc ng mu
a thy tinh, mung
Ni un cch thy
T sy
Bnh ht m
Cn phn tch
- Tin hnh: Dng dng c thch hp ly 10 gam mu cho vo cc cng a thy
tinh bit trc khi lng. t cc c mu v a thy tinh vo ni nc cch thy
c cn nc trong cc. Tip theo ly cc ra cng a thy tinh cho vo t sy
105
0
C n khi lng khng i. Ch trong qu trnh c cn ni un cch thy
cng nh trong qu trnh sy dng a thy tinh khuy rt ngn thi gian.
- Tnh kt qu:
100
2 1
x
G
G G
=
Trong : G1: Khi lng mu + hp + a thy tinh trc khi sy
G2: Khi lng mu cn li + hp + a thy tinh sau khi sy kh
n khi lng khng i.
G: Khi lng mu (ban u)
9.2.1.4. Xc nh m nguyn liu dng dung dch ha tan
38
i vi loi dung dch ny thng hm lng m ln hn rt nhiu so vi lng
cht kh trong dung dch. V vy, ngi ta thng xc nh hm lng cht kh suy ra
hm lng m ca dung dch.
Cc phng php xc nh kh hay xc nh thnh phn cc cht ha tan hin
nay thng dng cc phng php sau:
- Phng php sy kh
- Phng php t trng
- Phng php quang hc
- Phng php ha hc
9.2.2. Xc nh m bng phng php o dn in
- Nguyn tc
+ Cht kh tuyt i ca mi loi nguyn liu c mt gi tr dn in nht
nh.
+ dn in tng theo t l thun vi m ca nguyn vt liu.
+ Da trn cc nguyn tc trn ngi ta c th o dn in ca nguyn vt
liu ri tnh ra m ca vt liu.
- Dng c:
+ My nghin hoc ci nghin
+ My xc nh m "Freuchtron"
+ My xc nh m Gralner II PM - 300
- Cch tin hnh
+ i vi loi my Freuchtron c th xc nh m ca nhiu vt liu khc
nhau: cc loi ng cc, ca cao, ch, c ph, thuc l si, bng, len, si v.v. s dng
loi my ny, nguyn vt liu phi c nghin nh, kch thc bt nghin ln nht
bng 1mm. Cn bng, len, si phi c ct ngn t 2 - 3mm. Cho nguyn vt liu
vo y o (khong 2 - 3gam), bt cng tc, my o lm vic s ch trn thang o
dn in ca mu bng "ohm", t gi tr ny tra theo bng lp sn, ta s tnh c
m ca nguyn vt liu theo %. Loi my ny c chnh xc cao, sai s 0,5%.
+ i vi loi my Grainer II PM - 300 thng dng cho cc loi ng
cc: la m, thc, go v.v. Nguyn vt liu dng cho loi my ny c th nguyn
ht, nhng tt nht l nghin nh. Cho nguyn vt liu cn o m vo y o, bt
39
cng tc, my o lm vic s cho ngay gi tr m ca nguyn vt liu tnh bng %,
v khi chuyn t "ohm" ra %, ngi ta nhn h s chuyn (h s ny ph thuc vo
cht kh tuyt i ca mi loi vt liu). Do my ny ch o vi mt s nguyn vt
liu c thnh phn ha hc gn ging nhau, c bit l hm lng tinh bt.
* u im: ca loi my ny dng pin, gn, nh, tin li cho vic s dng trn cc
cnh ng trng ng cc, kho tng bo qun, sn phi, trm sy... xc nh m
ca nguyn vt liu kp thi phc v cho cng ngh thu hoch.
* Nhc im: Ch xc nh m c i vi mt s ng cc c tnh cht gn ging
nhau. chnh xc thp, sai s 2 - 3%.
9.2.3. Phng php kt hp
- Nguyn tc: i vi mt s nguyn vt liu nh rau, qu thng c m ban
u rt cao t 60 - 90%. Nu ch dng phng php nhit (sy n khi lng khng
i) th thi gian sy di, nn ngi ta c th s dng phng php nhit kt hp
phng php o in tr bng cch: giai on u dng phng php nhit sy n
m cn li trong vt liu < 30%, sau tip tc dng phng php o in tr, tt
nht nn dng loi my o Freuchtron v n cho chnh xc kh cao.
- Cch tin hnh: xem cch tin hnh ca phng php nhit v phng php
o in tr rt ra qu trnh tin hnh thch hp.
40
Bi s 10: PHNG PHP PHT HIN NHANH HN THE
10.1. Nguyn l
Hn the c phn ng kim vi phenoltalein cho dung dch mu hng. Nu cho
dung dch ny tc dng vi glycerin trung tnh, dung dch s chuyn thnh acid, s mt
mu hng (khng mu) do to thnh acid glycero boric.
10.2. Ha cht v dng c
a- Ha cht
- Ch th mu phenoltalein 1% trong cn
- Glycerin trung tnh
b- Dng c
- Cc 100 ml
- Giy lc
- ng nghim
- Phu lc
- a thy tinh
- Ko
10.3. Tin hnh
- Cn chnh xc 1g thc phm cho vo bnh kjedahl c di, cho vo 10ml H
2
SO
4
m c v 5g cht xc tc. Cho vo thm khong 50ml nc ct
- t bnh Kjeldahl ln bp in trong t ht, nghing bnh v un t t cho n
khi bc hi v hnh thnh khi trng SO
2
, khi bt tan un si cho n khi dung dch
mu trong sut hoc c mu xanh l ca CuSO
4
. ngui.
- nh mc dung dch c v c ha thnh 250 ml vi nc ct. Cho c dung
dch trn vo bnh cu ca my ct m, trng bnh bng nc ct v cho vo bnh cu.
- Nh vo vi git thuc th tashiro, trung ha bng NaOH 50% cho n khi
dung dch chuyn sang mu xanh l cy th dng (khong 20ml).
- Lp bnh cu vo my ct m.
- Chun b bnh hng: cho vo bnh tam gic 50 ml acid boric v 5 git thuc th
Tashiro
- Bt my ct m, khong 15 pht. Khi dung dch trong bnh hng chuyn sang
mu xanh un tip 15 pht th dng.
41
- Dng H
2
SO
4
0,1N chun (mu xanh l cy chuyn tr li mu tm nht).
Ghi li s ml H
2
SO
4
0,1N dng.
10.4. Kt qu
- Nu c mu hng xut hin, nh tip 2-3 git glycerin trung tnh, mu hng s
mt i thnh khng mu: kt lun sn phm c hn the.
- Nu khng c mu hng xut hin: kt lun sn phm khng c hn the.
10.5. Sinh vin t chn b mu th:
- Thc hin th nghim vi hai mu thc phm khc nhau.
- Sinh vin t mang mu n PTN phn tch.
42
TI LIU THAM KHO
1. Trn Linh Thc, 2002. Phng php phn tch vi sinh vt trong nc, thc phm
v m phm. Nh xut bn Gio Dc, 230 trang
2. B Mn Cng ngh Thc phm, 2004. Gio trnh thc hnh Kim nghim thc
phm 1, 2. Trng i Hc Cng Nghip Tp.HCM.
3. Quy trnh kim nghim Salmonella tht. Tiu chun Vit Nam, 1998.
43
PH LC: THNH PHN MT S MI TRNG S DNG TRONG
PHN TCH CC CH TIU VI SINH
4. Mi trng Violet Red Bile Agar (VRB)
- Cao nm men 3 g
- Peptone hoc gelysate 7 g
- NaCl 5 g
- Mui mt 1,5 g
- Lactose 10 g
- Neutral red 0,03 g
- Crystal violet 0,002 g
- Agar 15 g
- Nc ct 1000 ml
5. Mi trng Brilliant Green Bile Lactose broth (BGBL)
- Pepton 10 g
- Lactose 10 g
- Mt b 20 g
- Brilliant green 0,0133 g
- Nc ct 1000 ml
6. Mi trng Baird Parker
- Trypton 10 g
- Cao tht 5 g
- Cao nm men 1 g
- Sodium pyruvate 10 g
- Glycine 12 g
- Lithium chloride.6H
2
O 5 g
- Agar 20 g
7. Mi trng Trypticase Soya Agar (TSA)
- Trypticase peptone 15 g
- Phytone Peptone 5 g
- NaCl 5 g
- Agar 15 g
44
- Nc ct 1000 ml
8. Mi trng Eosine Methylene Blue Agar (EMB)
9. Mi trng MR VP
- Buffered Pepton Water Powder 7 g
- Glucose 5 g
- K
2
HPO
4
5 g
- Nc ct 1000 ml
10. Mi trng Simmons Citrate Agar (SCA)
- Sodium citrate 2 g
- NaCl 5 g
- K
2
HPO
4
1 g
- NH
4
H
2
PO
4
1 g
- MgSO
4
0,2 g
- Bromothymol Blue 0,08 g
- Agar 15 g
- Nc ct 1000 ml
11. Mi trng Xylose Lysine Deoxycholate Agar (XLD)
- Cao nm men 3 g
- L Lysine 5 g
- Xylose 3,75 g
- Lactose 7,5 g
- Sucrose 7,5 g
- Sodium deoxycholate 2,5 g
- Ferric ammonium citrate 0,8 g
- Sodium thiosulfate 6,8 g
- NaCl 5 g
- Agar 15 g
- Phenol red 0,08 g
- Nc ct 1000 ml
12. Mi trng Triple Sugar Iron Agar (TSI)
- Cao tht 3 g
45
- Cao nm men 3 g
- Peptone 15 g
- Proteose peptone 5 g
- Glucose 1 g
- Lactose 10 g
- Sucrose 10 g
- FeSO
4
0,2 g
- NaCl 5 g
- Na
2
S
2
O
3
0,3 g
- Phenol red 0,024 g
- Agar 12 g
- Nc ct 1000 ml
13. Mi trng Urea broth
- Urea 20 g
- Cao nm men 0,1 g
- Na
2
HPO
4
9,5 g
- K
2
HPO
4
9,1 g
- Phenol red 0,01 g
- Nc ct 1000 ml
14. Mi trng Lysine DeCarboxylase (LDC)
- Dextrose 1 g
- KH
2
PO
4
0,5 g
- Nc ct 100 ml
- L Lysine HCl 0,5 g
15. Mi trng Trypton
- L tryptophan 1 g
- NaCl 1 g
- K
2
HPO
4
3,13 g
- KH
2
PO
4
0,27 g
- Nc ct 200 ml
16. Mi trng Rappaport Vassiliadis (RV)
46
Mi trng c bn
- Tryptone 5 g
- NaCl 8 g
- KH
2
PO
4
1,6 g
- Nc ct 1000 ml
Dung dch MgCl2
- MgCl
2
.6H
2
O 400 g
- Nc ct 100 ml
Dung dch Malachite green oxalate
- Malachite green oxalate 0,4 g
- Nc ct 1000 ml
Mi trng hon chnh
- Mi trng c bn 1000 ml
- Dung dch MgCl
2
100 ml
- Dung dch Malachite green oxalate 10 ml
- Tng th tch 1110 ml