TIU CHUN VIT NAM

TCVN 6688-1:2001
ISO 8262 1 : 1987

SN PHM SA V THC PHM T SA XC NH HM LNG CHT BO BNG

PHNG PHP KHI LNG WEIBULL BERNTROP
(PHNG PHP CHUN)
PHN 1: THC PHM DNH CHO TR NH
MILK PRODUCTS AND MILK BASED FOODS DETERMINATION OF FAT CONTENT BY
THE WEIBULL BERNTROP GRAVIMETRIC METHOD (REFERENCE METHOD)
PART 1 : INFANT FOODS
Li ni u
TCVN 6688 - 1 : 2001 hon ton tng ng vi ISO 8262 1 : 1987;
TCVN 6688 1 : 2001 do Ban k thut tiu chun TCVN/TC/F12 Sa v sn phm sa bin
son, Tng cc Tiu chun o lng Cht lng ngh, B Khoa hc, Cng ngh v Mi
trng ban hnh.
Li gii thiu
Tiu chun ny quy nh phng php xc nh hm lng cht bo ca sa, sn phm sa v
thc phm t sa, nm trong s mt lot cc phng php chun c hi ha ti mc ti a.
Cc phng php ny da trn cc nguyn tc Rose Gottlieb (RG), hoc Weibull Berntrop
(WB) hoc Schmid Bondzynski Ratzlaff (SBR).
Tiu chun ny lin quan n cc loi thc phm t sa v cc loi thc phm khc dnh cho
tr nh cha tinh bt hoc dextrin, hoc rau, qu, tht ln hn 5% (tnh theo cht kh),
phng php ny da trn nguyn tc Weibull-Berntrop (WB) v:
a) Sn phm cha cc thnh phn trn vi hm lng ln s khng th chit ht c cht bo,
do hm lng cht bo chit c b thp i, nu quy trnh Rose-Gottlieb (RG) l khng thch
hp.
b) Nhn chung, sn phm c hm lng cacbon hidrat cao s lm tng cc hp cht c th chit
c bng ete khi thy phn bng axit, do cho gi tr hm lng cht bo qu cao dn n
vic p dng quy trnh Schmid-Bondzynski-Ratzlaff (SBR) l khng thch hp.
c) Quy trnh Weibull-Berntrop (WB) mc d cng s dng qu trnh thy phn bng axit nhng
khng nh hng ngc n cc hp cht c th chit bng ete, v phn thy phn bng axit
c lc v ra, cn kh trn phu lc khng cn cha cc hp cht c th chit c bng
xng nh.
d) mt s nc s dng phng php m t trn y xc nh hm lng cht bo v
c y ban Codex v cc phng php Phn tch v Ly mu khuyn co s dng.
Phng php Weibull u tin c p dng cho bnh m; sau c Berntrop pht trin, sa
i mt cch ng k nh trnh by trong tiu chun ny. Phng php ny hin nay c
p dng rng ri xc nh hm lng cht bo trong nhiu loi thc phm.

TCVN 6688 - 1 : 2001

SN PHM SA V THC PHM T SA XC NH HM LNG CHT BO BNG
PHNG PHP KHI LNG WEIBULL BERNTROP (PHNG PHP CHUN) - PHN 1:
THC PHM DNH CHO TR NH
Milk products and milk based foods Determination of fat content by the Weibull Berntrop
gravimetric method (Reference method) - Part 1 : Infant foods
1. Phm vi v lnh vc p dng
Tiu chun ny quy nh phng php chun xc nh hm lng cht bo trong thc phm
dnh cho tr nh khi khng xc nh c bng phng php Rose-Gottlieb, tc l cc loi thc
phm t sa v cc loi thc phm khc dnh cho tr nh cha khng qu 5% (m/m) (tnh theo
cht kh) tinh bt hoc dextrin, hoc rau, qu, tht
Ch thch Nn dng phng php thng nht da trn nguyn tc ca RG a ra trong TCVN
(1)
6687 : 2000 (ISO 8381) kim tra cc loi thc phm khc dnh cho tr nh. (Cc Malto
Dextrin khng cha cc dextrin c phn t lng cao thng c trong thc phm dnh cho tr
nh khng gy nhiu qu trnh chit RG k c khi tn ti vi phn trm cao).
Khi sn phm cha hm lng ng k cc axit bo t do hoc sn phm cha cc mng
khng tan c ht trong amoniac th cng c th s dng phng php ny.
2. Tiu chun vin dn
TCVN 6400 : 1998 (ISO 707) Sa v sn phm sa Phng php ly mu
3. nh ngha
Hm lng cht bo: Tt c cc cht xc nh c bng phng php quy nh trong tiu
chun ny
Hm lng cht bo c biu th bng phn trm khi lng
4. Nguyn tc
Thy phn phn mu th bng cch un si vi axit clohidric long, lc phn thy phn nng
qua giy lc t thu ly cht bo, chit cht bo bng n-hecxan hoc xng nh t giy lc
kh, loi b dung mi bng cch chng ct hoc cho bay hi v cn lng cht va chit c.
(iu ny c gi l nguyn tc Weibull-Berntrop).
5. Thuc th v nguyn liu
Ch s dng cc thuc th loi tinh khit phn tch v khi tin hnh xc nh theo phng php
ny khng c li lng cn ng k. Ch s dng nc ct hoc nc c tinh khit
tng ng.
5.1. Dung dch axit clohidric, cha khong 20% HCl (m/m), 20 xp x 1,10 g/ml.
Pha long 100 ml axit clohidric m c (20 = 1,18 g/ml) bng 100 ml nc v trn.
0
5.2. Dung mi chit, khng cha nc: n-hecxan hoc xng nh c im si khong t 30 C
0
n 60 C.
kim tra cht lng ca dung mi chit, chng ct 100 ml dung mi ny t bnh chit (6.4)
chun b theo 8.4. Dng mt bnh chit rng chun b theo cng phng thc kim tra khi
lng (xem 10.1). Dung mi chit khng c li lng cn vt qu 1,0 mg.
Thay hoc chng ct dung mi khi khng tha mn yu cu trn.
5.3. Giy lc gp np, loi trung bnh, tt nht l loi kh cht bo, c ng knh 150 mm.
Tin hnh cc php th mu trng kim tra cht lng ca giy lc nh quy nh trong 8.3,
s dng dung mi tha mn yu cu trong 5.2. Dng mt bnh chit rng (6.4) chun b nh
quy nh trong 8.4 kim tra khi lng (xem 10.1). Giy lc khng c li lng cn vt
qu 2,5 mg.
Thay cc giy lc khng tha mn iu kin trn.
5.4. Giy qu xanh
5.5. Diatomit (ty chn; xem 8.5.3).
5.6. Lactoza tinh khit (ty chn; xem 8.5.3).
5.7. Si bng, kh cht bo bng cch chit bng dung mi (5.2) trong 1,5 h v sy kh.
6. Thit b, dng c

1
TCVN 6687 : 2000 Thc phm t sa dng cho tr nh - Xc nh hm lng cht bo
Phng php khi lng ca Rose Gottlieb (phng php chun)
Cnh bo V php xc nh ny s dng cc dung mi bay hi d chy, nn thit b in
c dng phi tun th cc quy nh an ton trong vic s dng dung mi.
S dng cc thit b phng th nghim thng thng v c bit nh sau:
6.1. Cn phn tch.
6.2. B trn, lm ng nht mu th nghim, nu cn; th d: my xay thc phm hoc b
trn tc cao vi bnh trn dung tch 1 lt, c np y.
6.3. Thit b chit, c th chit lin tc hoc bn lin tc; th d: loi Soxhlet, gm mt bnh chit
(y phng, c ngn) dung tch 150 ml, mt b chit c si phng dung tch t 40 ml n 60 ml v
b sinh hn gn vi ng sy hoc nt bng.
6.4. Bnh chit, dung tch 150 ml, y phng, c ngn
6.5. ng chit, lm bng giy lc kh cht bo, thy tinh, nhm oxit hoc bng PTFE
(polytetrafluoroetylen), khng c mt lng cn ng k trong mu trng, hoc lm bng
xeluloza, c ng knh trong 22 mm v chiu di bin 80 mm s dng vi thit b chit (6.3).
6.6. Ni cch thy, c th duy tr cc khong nhit sau:
0 0
t 40 C n 60 C (xem 8.1.1);
0 0
t 30 C n 40 C (xem 8.1.2);
6.7. Dng c gia nhit, dng cho thit b chit, th d: ni cch thy, b ct hoc bp in kim
sot c nhit .
6.8. Cht tr si, khng cha cht bo: ht thy tinh, hoc cc mnh s kh v, khng xp
hoc silicon cacbua.
6.9. Bnh nn, dung tch 250 ml, c gn vi b sinh hn, tt nht l loi Liebig.
6.10. Dng c gia nhit, dng cho bnh nn gn vi b sinh hn, th d: li kim loi v u t
bng kh, bp in hoc b ct.
6.11. Phu lc, thch hp s dng vi giy lc gp np (5.3).
6.12. Cc c m, dung tch 100 ml v 250 ml.
6.13. Thit b chng ct, c th chit c dung mi nh t cc bnh nhit khng qu
0
100 C.
0
6.14. T sy, t nng bng in, c cc l thng gi m hon ton, c th duy tr 102 C
0
2 C trong sut khoang sy. T sy c gn vi mt nhit k thch hp.
6.15. ng ong, dung tch 50 ml, 100 ml v 250 ml.
6.16. Kp, lm bng kim loi, thch hp gi bnh hoc gi cc.
6.17. Cp, c u t gi giy lc v ng.
7. Ly mu
Xem TCVN 6400 : 1998 (ISO 707)
0
Bo qun tt c cc mu th nghim dng lng, dng snh hoc dng nho nhit t 2 C
0
n 4 C, k t khi ly mu cho n cc bt u quy trnh th. Trong trng hp mu ng trong
0
bao b kn, th bo qun nguyn nh th nhit di 20 C.
8. Cch tin hnh
8.1. Chun b mu th
8.1.1. Sn phm dng lng
Lc v o chiu hp cha. M np hp, rt t t sn phm sang hp cha th hai (c np y
kn) v trn bng cch chuyn qua chuyn li sn phm, trnh cht bo hoc bt k thnh
phn no khc trong mu st li trn thnh v y ca hp cha th nht. Nu sn phm vn
cn cha cc thnh phn khc dng tng mng hoc tng ming th ng ha chng trong b
trn thch hp (6.2). Cui cng, chuyn ht sn phm sang hp cha th hai. y np hp.
 0 0
Nu cn, hp cha ang y kn vo ni cch thy (6.6) nhit t 40 C n 60 C. C 15
pht li ly ra v lc mnh. Sau 2 h, ly hp cha ra, dng khn kh lau mt ngoi ca hp v
ngui n nhit phng. M hn np v trn k lng cha bng tha hoc dao trn. (Nu cht
bo tch hn, th khng th mu). Chuyn ht sn phm sang hp cha th hai. y np
hp.
8.1.2. Sn phm snh hoc nho
M np hp cha v trn k lng cha bn trong bng tha hoc dao trn. Nu c th, s dng
my quay trn t di ln trn theo cch sao cho cc lp trn cng vi cc phn cc gc
di ca hp cha trn c vi nhau. Trnh cht bo hoc bt k thnh phn no khc
trong mu st li trn thnh v y ca hp cha th nht. Nu sn phm vn cn cha cc
thnh phn dng tng mng hoc tng ming th chng phi c ng ha trong b trn
thch hp (6.2). Cui cng, chuyn ht sn phm sang hp cha th hai (c np y kn). y
np hp.
0 0
Nu cn, hp cha ang y kn vo ni cch thy (6.6) nhit t 30 C n 40 C. Ly
hp cha ra, dng khn lau kh mt ngoi hp v m np. Vt sch tt c sn phm pha trong
hp cha cho sang a rng c th trn c k v trn cho n khi thu c mu ng
nht. Chuyn ht sn phm sang hp cha th hai nh trn. y np hp.
8.1.3. Sn phm dng kh
Trn k bng cch quay v o chiu hp cha. Nu cn, chuyn mu th nghim sang mt hp
cha kn thch hp c dung tch rng thc hin thao tc ny.
Nu sn phm vn cn cha cc thnh phn dng tng mng hoc tng ming th ng ha
chng trong b trn thch hp (6.2).
8.2. Phn mu th
Trn mu th (8.1) bng cch khuy (i vi sn phm dng snh, dng nho hoc dng kh),
hoc bng cch o chiu hp cha nh nhng ba hoc bn ln (i vi sn phm dng lng)
v cn ngay trong bnh nn (6.9) hoc bng cch khc, khong t 3 g n 20 g mu th, chnh
xc n 1 mg, tng ng vi 3,0 g n 3,5g cht kh. Phn mu th khng cha qu 1,0 g cht
bo; tha mn yu cu ny, c th cn phi ly phn mu th nh hn.
Phn mu th c chuyn ht sang y bnh nn (6.9).
8.3. Th mu trng
Tin hnh th mu trng ng thi vi php xc nh, s dng cng mt quy trnh v cng loi
thuc th, nhng thay phn mu th pha long (xem 8.5.1) bng 25 ml nc (xem 10.2).
8.4. Chun b bnh chit
Sy bnh (6.4) c cha mt t cht tr si (6.8), lm si nh trong qu trnh chit v loi b
0 0
tip dung mi, trong t sy (6.14) nhit 102 C 2 C trong 1h.
ngui bnh (bo v khi bi) t nht 0,5 h n nhit phng cn.
Ch thch Nn t bnh trong bnh ht m trnh khng ngui hoc thi gian lm ngui
qu lu.
Dng kp (6.16) (c bit trnh thay i nhit ), t bnh ln cn v cn chnh xc n 0,1 mg.
8.5. Tin hnh xc nh
0
8.5.1. Thm nc nhit 30 C vo phn mu th (8.2) c c tng th tch 25 ml (
thu c dung dch axit clohidric 4 mol/l trong 8.5.2) v lc nh.
Ch thch Xem ch thch ca 8.5.3. khi c b sung lactoza.
8.5.2. Thm 50 ml dung dch axit clohidric (5.1) vo phn mu th pha long, trong khi thm
trng lun thnh bnh v trn nh bng cch xoay bnh. Ni bnh vi b sinh hn, un nng bnh
cho n khi lng cha trong bnh bt u si v si nh trong 30 pht, thnh thong xoay
bnh.
0
8.5.3. Ly 150 ml nc nng ( nhit t nht l 80 C), dng khong 75 ml trng pha trong
b sinh hn, tho bnh nn ra khi b ngng v cho nt 75 ml nc nng cn li vo bnh sao
trng c c v pha trong thnh bnh. Nu cn, cho thm 1g diatomit (5.5) hoc khong 100
2
cm giy lc kh cht bo c x nh (c bit l trng hp cha mt hm lng nh cc
cht rn khng cha cht bo) lc nhanh hn.
Ch thch lc nhanh hn, c th thm 1g lactoza tinh khit (5.6) vo phn mu th pha
long trong 8.5.1.
8.5.4. Lc ngay lng cha trong bnh, rt cht lng qua a thy tinh vo giy lc gp np (5.3)
c lm t k bng nc nng v t vo trong phu lc (6.11). Trng k bnh ba ln bng
nc nng, cho nc ra vo giy lc qua a thy tinh v cui cng ra giy lc t nht ba ln
bng nc nng cho n khi nc ra khng cn cha axit khi th bng giy qu (5.4). Khng
s dng qu 400 ml nc. cho giy lc ro hn nc.
8.5.5. Dng cp (6.17) ly giy lc ra khi phu v t vo ng chit (6.5) sao cho mp trn ca
giy thp hn ming ng t nht l 20 mm. t ng vo cc c m dung tch 100 ml (6.12).
8.5.6. Sy cc cng vi lng cha bn trong v bnh nn cng vi a thy tinh trong t sy
0 0
(6.14) kim sot nhit 102 C 2 C t 1h n 1,5 h sy kh k. Ly cc v bnh cng vi
a thy tinh ra khi t v ngui.
Ch thch Nn sy kh giy lc, v cht bo thng khng chit c ht. Trong trng hp
giy lc qu t v dng b chit lin tc th vi git hp cht tan c trong nc c th ln
vo cht chit, iu lm cho cht chit c mu ti v hm lng cht bo thu c cao.
8.5.7. Dng cp (6.17) gi ng chit, nt nh ng bng si bng kh cht bo (5.7) v t
vo b chit. Dng ng ong ly 100 ml n-hecxan hoc xng nh (5.2); s dng cc phn dung
mi trng u kp, pha trong cc, bnh nn v a thy tinh, thu ly nc trng cho vo bnh
chit chun b (xem 8.4). Cho nt phn dung mi cn li vo bnh chit trng lun pha
trong c bnh.
8.5.8. Ni bnh chit vi b chit ng ng chit, ni b chit vi b sinh hn v un nng bnh
khong 4 h sao cho ng chit v lng cha trong ng chit c t nht l 1000 ml dung mi
(20 ln ht bng siphng).
8.5.9. Ly bnh chit ra khi thit b chit, trng pha trong c bnh v nh ca b ngng bng
mt t dung mi. Sau cn thn chng ct ht dung mi ra khi bnh. Nu s dng ni cch
thy, th lau tht kh pha ngoi bnh.
0 0
8.5.10. Sy bnh chit trong t sy (6.14) c kim sot nhit 102 C 2 C trong 1h (
sao cho dung mi c th thot ra c). Ly bnh ra khi t v ngui (khng trong bnh ht
m, nhng trnh bi) n nhit phng cn (t nht l 30 pht) v cn chnh xc n 0,1 mg.
Khng lau bnh ngay trc lc cn. Dng kp t bnh ln cn (c bit trnh thay i nhit ).
8.5.11. Lp li cc thao tc m t trong 8.5.10 cho n khi chnh lch khi lng ca bnh gia
hai ln cn lin tip khng qu 1,0 mg. Ghi li khi lng ca bnh v cht chit l khi lng
nh nht thu c.
9. Biu th kt qu
9.1. Phng php tnh v cng thc
Hm lng cht bo, c biu th bng phn trm khi lng, theo cng thc sau:
(m1 m2 ) (m3 m4 )
x 100
m0

Trong
m0 l khi lng phn mu th (8.2), tnh bng gam;
m1 l khi lng bnh chit cng vi cht chit xc nh c trong 8.5.11, tnh bng gam;
m2 l khi lng bnh chit chun b (xem 8.4), tnh bng gam;
m3 l khi lng bnh chit s dng trong th mu trng (8.3) v cc cht chit xc nh c
trong 8.5.11, tnh bng gam;
m4 l khi lng bnh chit chun b (xem 8.4), dng trong th mu trng (8.3), tnh bng
gam;
Ghi kt qu chnh xc n 0,01%.
9.2. chnh xc
Ch thch Cc gi tr v lp li v ti lp c biu th mc xc sut 95% v thu c
t cc kt qu ca mt th nghim lin phng th nghim tin hnh theo TCVN 4550-88 (ISO
1)
5725)
9.2.1. lp li
Chnh lch gia hai kt qu th ring r thc hin trn mu ging ht nhau, do mt ngi phn
tch trong mt khong thi gian ngn, khng vt qu cc gi tr sau y:
- i vi sn phm c hm lng cht bo ln hn 5% (m/m): 0,2 g cht bo trn 100g sn
phm;
- i vi sn phm c hm lng cht bo nh hn hoc bng 5% (m/m); 0,1 g cht bo trn
100 g sn phm;
- i vi sn phm dng lng: 0,05 g cht bo trn 100 g sn phm.
9.2.2. ti lp
Chnh lch gia hai kt qu ring r v c lp do hai ngi phn tch trong cc phng th
nghim khc nhau, thc hin trn mu th ging ht nhau, khng vt qu cc gi tr sau y:
- i vi sn phm c hm lng cht bo ln hn 5% (m/m): 0,4 g cht bo trn 100 g sn
phm;
- i vi sn phm c hm lng cht bo nh hn hoc bng 5% (m/m): 0,2 g cht bo trn
100 g sn phm;
- i vi sn phm dng lng: 0,1 g cht bo trn 100 g sn phm.
10. Ch v cch tin hnh
10.1. Th mu trng kim tra dung mi v giy lc
Trong cc php th mu trng ny, s dng bnh kim tra khi lng khi c nhng thay i
v iu kin khng kh ca phng cn hoc nh hng nhit ca bnh thu nhn cht bo th
cng khng nh gi sai s c mt hay khng c mt cht khng bay hi c trong cht chit ca
thuc th. Bnh ny c th c s dng lm bnh i trng trong trng hp cn c hai a.
Mt khc, chnh lch biu kin (m 3 m4 trong cng thc ca 9.1) ca bnh kim tra phi c
xem xt khi kim tra khi lng bnh thu nhn cht bo s dng trong th mu trng. Do s
thay i v khi lng biu kin ca bnh thu nhn cht bo, c hiu chnh cho khi lng
biu kin ca bnh kim tra, khng c vt qu 0,5 mg.
Rt t khi dung mi cha cht bay hi b gi li nhiu trong cht bo. Nu nh c mt ca cc
cht , th tin hnh th mu trng s dng bnh cht bo vi khong 1 g butterfat khan. Nu
cn, chng ct dung mi trong s c mt ca 1 g butterfat khan trn 100 ml dung mi. Ch s
dng dung mi va mi chng ct.
10.2. Tin hnh th mu trng ng thi vi vic xc nh
Gi tr thu c trong php th mu trng tin hnh ng thi vi php xc nh a ra khi
lng biu kin ca cc cht chit c t phn mu th (m 1 m2) dng hiu chnh s c
mt ca cht khng bay hi trong thuc th v trong giy lc v cng dng hiu chnh mi
thay i v iu kin khng kh ca phng cn v s chnh lch nhit gia bnh thu nhn
cht bo v nhit phng cn hai ln cn (8.4 v 8.5.11).
Trong cc iu kin thch hp (ch s mu trng ca dung mi v giy lc thp, nhit phng
cn cn bng v thi gian lm ngui bnh thu nhn cht bo), th gi tr ny thng nh
hn 3 mg. Cng thng gp phi cc gi tr hi cao hn, ln n 5 mg. Sau khi chnh li gi tr
mu trng, cc kt qu s chnh xc. Khi thc hin hiu chnh vi gi tr ln hn 5 mg th phi
nu thc t ny trong bo co th nghim (xem iu 11).

1)
TCVN 4550-88 (ISO 5725 : 1986) chnh xc ca phng php th - Xc nh lp li v
ti lp i vi phng php th chun bng cc th nghim lin phng th nghim.
Nu gi tr thu c trong php th mu trng ny thng vt qu 3 mg, th phi kim tra li
dung mi v giy lc (nu ngay trc cha thc hin) v thay hoc lm sch li (xem 5.2 v
5.3).
11. Bo co th nghim
Bo co th nghim phi ch ra phng php s dng v kt qu thu c. Bo co cng
phi cp n tt c cc chi tit thao tc khng quy nh trong tiu chun ny, hoc ty la
chn, cng vi cc chi tit bt thng khc c th nh hng ti kt qu. Phi bo co gi tr
mu trng (m3 m4, xem 9.1), nu vt qu 5 mg.
Bo co kt qu cng bao gm mi thng tin cn thit v vic nhn bit y mu th.

TI LIU THAM KHO

Weibull, M., Z. angew. Chemie (1892) 450.
Weibull, M., Z. angew. Chemie (1894) 199.
Berntrop, J. C., Z. angew. Chemie (1902) 11.
Koning, J.C. v Mooy, W. C., W.C., Pharmaceutisch Weekblad 53 (1916) 50.
ISO 1443 Tht v sn phm tht Xc nh tng hm lng cht bo.
Schuller, P.L., Bo co nghin cu lin phng th nghim ca CX/MAS v vic xc nh cht bo
trong thc phm dnh cho tr nh (Phng php phn tch v ly mu ca Codex, CX/MAS
75/10, 1975).