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BO CO THC HNH
Mn GVHD
: :
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
LI M U
Knh cho thy, y l bi bo co ghi li ton b cng vic v kin thc hc c trong mn Thit b phn tch thc phm ca Nhm 1 trong thi gian thc hnh qua. Nhm 1 cm n thy Nguyn Thanh Nam tn tnh ch dy chng em trong qu trnh hc tp v thc hnh, cn nhiu thiu st trong qu trnh hc cng nh trong qu trnh lm bo co nn mong thy b qua v cho chng em li nhn xt cng nh gp hon thin hn v mt kin thc trnh by trong bi bo co ny. Mt ln na xin cm n thy, chc thy nhiu sc khe v thm nhiu thnh cng trong cng vic ging dy.
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
Cn k thut
Cn phn tch
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
L nung
My UV-Vis
GVHD Nguyn Thanh Nam 4
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
My AAS
My HPLC
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
BI 2: Xc nh hm lng nitrit, nitrat trong mu rau bng phng php UV-Vis ------------2.1 Nguyn tc:
Trong mi trng acid (pH = 2) nitrit s b diazo ho acid sulphanilic, sau kt hp vi alpha naphthylamin cho hp cht naphathylamino azobenzen sulphonic c mu khng bn. Nitrat trong nc, sn phm tht v rau c cadimi kh thnh nitrit. nh gi nitrit trc v sau khi kh nitrat thnh nitrit th chng ta c th tnh ton hm lng nitrat bng php so snh gia nitrit trc v sau khi kh.
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
Dng c: - Bnh nh mc 50ml, 100ml, 250ml, 500ml, 1000ml - Cc thu tinh 50ml, 100ml, 200ml - ng ong 100ml - Chai l ng thuc th - Pipet 1ml, 2ml, 5ml, 10ml - Qu bp cao su - Bnh tia - Phu thu tinh Ho cht: - Dung dch nitrit chun 1000ppm - Dung dch sulphanilamin 0,2%. Cch pha: ho tan 2g sulphanilamin trong 800ml nc nng, lc, thm 100ml acid hydrochloric c, khuy k v pha long ti 1000ml vi nc ct (thuc th 1). - Dung dch N-1 naphthylethylen diamin dihydrochlorua 0,1%. Cch pha: ho tan 0,25g N-1 naphthylethylen diamin dihydrochlorua trong nc. Pha long ti 250ml vi nc ct. Bo qun trong l nu nt kn v gi trong t lnh (thuc th 2). - Dung dch acid HCL 44,5%. Cch pha: pha long 445ml acid hydrochloric c ti 1000ml vi nc (thuc th 3).
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
- Dung dch chun gc natri nitrit nng 10000mg/l. Cch pha: ho tan 1g natri nitrit trong nc v pha long ti 100ml (thuc th 4). - Dung dch lm vic natri nitrit nng 50mg/l. Cch pha: pha long 5ml dung dch gc ti 1000ml vi nc (thuc th 5). - Dung dch dihydrogen dinatri ethylene diamine tetra acetat dihydrat 1%. Cch pha: ho tan 20ml acid hydrochloric c vi 500ml nc ct, thm 10g dihydrogen dinatri ethylene diamine tetra acetat dihydrat v 55ml dung dch aminiac c. Pha long ti 1000ml vi nc. Chnh pH = 9,6 9,7 (thuc th 6). - Cadimi kim loi (thuc th 7). - Giy lc
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
Xy dng ng chun: Dung dch chun gc natri nitrit c nng 1000ppm nn pha long xung 10ppm. Ht 1ml dung dch chun gc natri nitrit c nng 1000ppm cho vo bnh nh mc 100ml v nh mc ti vch bng nc ct. Lc u v s dng dung dch ny xy dng ng chun nh sau. Ht ln lt 0; 1; 2; 3 v 5ml dung dch chun gc natri nitrit nng 10ppm cho vo 5 bnh nh mc 50ml, thm khong 25ml nc ct, 5ml thuc th 1 v 3ml thuc th 2 vo mi bnh. dung dch ny trong ch ti 5 pht. Tip theo thm 1ml thuc th 2 vo v lc u hn hp, vo ch ti 3 pht. Sau 3 pht ly ra v nh mc ti vch bng nc ct, nng nitrit ca cc dung dch ny s l 0; 0,2; 0,4; 0,6 v 1ppm, lc u hn hp v o hp thu bc sng 538nm. i vi mu nh lng nitrit: Ht 10ml dch lc t mu rau cho vo bnh nh mc 50ml. Thm vo khong 25ml nc ct, , 5ml thuc th 1 v 3ml thuc th 2 vo mi bnh. dung dch ny trong ch ti 5 pht. Tip theo thm 1ml thuc th 2 vo v lc u hn hp, vo ch ti 3 pht. Sau 3 pht ly ra v nh mc ti vch bng nc ct, lc u hn hp v o hp thu bc sng 538nm. Cn c vo ng chun dung dch o c c nng Cx (ppm).
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
Bm Dd chun (ml) Dch mu (ml) TT1 (ml) TT3 (ml) TT2 (ml) C (ppm) Abs
1 0
2 1
3 2
4 3
5 5
mu 0
0 5 3 1 0 0
0 5 3 1 0,2 0,133
0 5 3 1 0,4 0,278
0 5 3 1 0,6 0,449
0 5 3 1 1 0,670
10 0 3 1 Cx Tb 3 ln o=0,010
i vi mu nh lng nitrat: Ht 5ml mu sau khi lc v khong 1g cadimi t cho vo bnh nh mc 100ml, cho cadimi vo bnh nh mc bng phu nh v s dng mt lng nh nc ct trng phu. Thm 25ml dch lc, y nt v lc u. Pha long ti vch bng nc ct, trn tht k hn hp v yn khong 10 pht cho cadimi lng. Ht 10ml ca dung dch ny ( x l vi cadimi kh nitrat thnh nitrit) cho vo bnh nh mc 50ml v to mu nh xc nh nitrit trn.
GVHD Nguyn Thanh Nam 10
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
Bm Dd chun (ml) Dch mu (ml) TT1 (ml) TT3 (ml) TT2 (ml) C (ppm) Abs
1 0
2 1
3 2
4 3
5 5
mu 0 10 ( x
l vi cadimi)
5 3 1 0 0
5 3 1 0,2 0,133
5 3 1 0,4 0,278
5 3 1 0,6 0,449
5 3 1 1 0,670
0 3 1 Cx Tb 3 ln o=0,014
V phng trnh hi qui tuyn tnh: T kt qu Abs o c v phng trnh hi qui tuyn tnh th hin mi tng quan gia nng v hp thu (quy trnh phn tch nitrit v nitrat theo nguyn tc phi s dng ring ng chun nhng do hn ch v thi gian v dng c nn s dng chung mt ng chun).
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
Nitrit
0.8 0.7 0.6 0.5 Axis Title 0.4 0.3 0.2 0.1 0 0 1 2 3 Axis Title 4 5 6 y = 0.1365x + 0.0057 R = 0.9943
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
<=> => Hm lng nitrit trong mu tnh theo cng thc sau:
Xnitrit (mg/ml) =
= 0.016 (mg/L)
Vi Cx : nng ca mu (ppm) Vbm : th tch bnh nh mc (ml) Vmu : th tch mu ht (ml) K : h s pha long 1000: n v mg/L nn phi chia 1000 i t ml => L i vi mu nitrit tng, ta c kt qu Abs l 0,014 v phng trnh ng chun c dng vi R2 = 0.9943 t ta c th tnh c Cx ca mu bng cch th vo phng trnh ng chun: <=>
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
<=> => Hm lng nitrit tng trong mu sau khi kh nitrat tnh theo cng thc sau:
= 0.0305 (mg/L)
Vi Cx : nng ca mu (ppm) Vbm : th tch bnh nh mc (ml) Vmu : th tch mu ht (ml) K : h s pha long 1000: n v mg/l nn phi chia 1000 i t ml => L Hm lng nitrat = nitrit tng nitrit = 0.0305 0.016 = 0.0145 (mg/ml)
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
Dy chun nitrit
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
BI 3: nh lng Fe bng phng php UV-Vis (mu rau mung) ------------3.1 Nguyn tc:
Fe(II) trong dung dch, kt hp vi 1,10-phenaltroline thnh mt phc cht mu cam bn trong mi trng pH 3-9. Nu mun mu ny khng b nh hng ca thuc th d v c mt ca cc ion khc, phn ng cn tin hnh pH 3,5-4,5. Phc cht ny gm 3 phn t 1,10-phenaltroline kt hp vi 1 ion Fe(II). Phn ng c hiu cho Fe(II) nn phi chuyn ht Fe(III) v Fe(II) bng cch kh vi hydroquinone hay hydroxylamine clohydic. 2NH2OH + 4Fe3+ N2O + 4Fe2+ + 4H+ + H2O
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
- Bp in - L nung - Chn nung Dng c: - Bnh nh mc 50ml, 100ml, 250ml, 500ml, 1000ml - Cc thu tinh 50ml, 100ml, 200ml - ng ong 100ml - Chai l ng thuc th - Pipet 1ml, 2ml, 5ml, 10ml - Qu bp cao su - Bnh tia - Phu thu tinh Ho cht: - Dung dch hydroxylamine clohydric 1% - Dung dch m pH 4,0 - Dung dch 1,10-phenalthroline 0,1% - Dung dch HCl tinh khit - Dung dch HCl 20% - Giy lc
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
- Dung dch Fe2+ chun. Cch pha: pha long t dung dch Fe2+ 1000ppm xung 10ppm. Ht 1ml Fe2+ 1000pm cho vo bnh nh mc 100ml v nh mc ti vch.
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
5ml m pH 5, 1ml 1,10-phenontroline 0.1% vo tng bnh, lc nh v yn trong 5 pht, sau dng nc ct nh mc ti vch ta c nng tng ng mi bnh l 0; 0.2; 0.4; 0.6 v 0.8ppm. Mang cc bnh nh mc o hp thu = 510nm. Bnh nh mc Fe(II) 10ppm Mu Hydroxylamin 10% m pH 5 1,10phenantroline 0,1% Nc ct C(ppm) Lc nh, sau 5 pht dng nc ct nh mc ti vch 0 0,2 0,4 0,6 0,8 Cx 1 1 1 1 1 1 mu 0 10 1 (lc 1 pht) 5
1 0 0 1 5
2 1 0 1 5
3 2 0 1 5
4 3 0 1 5
5 4 0 1 5
Sau 15 pht em o = 510nm Khng Abs 0 0,039 0,056 0,110 0,152 pht hin
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
V phng trnh hi qui tuyn tnh: T kt qu Abs o c v phng trnh hi qui tuyn tnh th hin mi tng quan gia nng v hp thu.
Fe(II)
0.16 0.14 0.12 0.1 Axis Title 0.08 Fe(II) 0.06 0.04 0.02 0 0 -0.02 0.5 1 1.5 2 2.5 3 3.5 4 4.5 Axis Title Linear (Fe(II)) y = 0.0375x - 0.0036 R = 0.9785
Da vo Abs v phng trnh ng chun ta tnh c CFe(II) tuy nhin trong qu trnh o vi mu khng pht hin Fe(II) nn khng c kt qu c th. Nu c kt qu th tnh ton CFe(II) v sau tnh hm lng Fe(II) c trong mu.
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
Dy chun Fe(II)
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
BI 4: nh lng Fe bng phng php UV-Vis (mu sa) ------------4.1 Nguyn tc:
Fe(II) trong dung dch, kt hp vi 1,10-phenaltroline thnh mt phc cht mu cam bn trong mi trng pH 3-9. Nu mun mu ny khng b nh hng ca thuc th d v c mt ca cc ion khc, phn ng cn tin hnh pH 3,5-4,5. Phc cht ny gm 3 phn t 1,10-phenaltroline kt hp vi 1 ion Fe(II). Phn ng c hiu cho Fe(II) nn phi chuyn ht Fe(III) v Fe(II) bng cch kh vi hydroquinone hay hydroxylamine clohydic. 2NH2OH + 4Fe3+ N2O + 4Fe2+ + 4H+ + H2O
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
- Cn k thut - Bp in - L nung - Chn nung Dng c: - Bnh nh mc 50ml, 100ml, 250ml, 500ml, 1000ml - Cc thu tinh 50ml, 100ml, 200ml - ng ong 100ml - Chai l ng thuc th - Pipet 1ml, 2ml, 5ml, 10ml - Qu bp cao su - Bnh tia - Phu thu tinh Ho cht: - Dung dch hydroxylamine clohydric 1% - Dung dch m pH 4,0 - Dung dch 1,10-phenalthroline 0,1% - Dung dch HCl tinh khit - Dung dch HCl 20% - Giy lc
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
- Dung dch Fe2+ chun. Cch pha: pha long t dung dch Fe2+ 1000ppm xung 10ppm. Ht 1ml Fe2+ 1000pm cho vo bnh nh mc 100ml v nh mc ti vch.
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
yn trong 5 pht, sau dng nc ct nh mc ti vch ta c nng tng ng mi bnh l 0; 0.2; 0.4; 0.6 v 0.8ppm. Mang cc bnh nh mc o hp thu = 510nm. Bnh nh mc Fe(II) 10ppm Mu Hydroxylamin 10% m pH 5 1,10phenantroline 0,1% Nc ct C(ppm) Lc nh, sau 5 pht dng nc ct nh mc ti vch 0 0,2 0,4 0,6 0,8 Cx 1 1 1 1 1 1 mu 0 10 1 (lc 1 pht) 5
1 0 0 1 5
2 1 0 1 5
3 2 0 1 5
4 3 0 1 5
5 4 0 1 5
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
V phng trnh hi qui tuyn tnh: T kt qu Abs o c v phng trnh hi qui tuyn tnh th hin mi tng quan gia nng v hp thu.
0.14 0.12 0.1 0.08 0.06 0.04 0.029 0.02 0 0 0 0.2 0.4 0.6 0.8 1 0.061 0.087 Fe(II) Linear (Fe(II)) Linear (Fe(II))
Fe(II)
y = 0.153x - 0.001 R = 0.9975 0.124
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
<=> => Hm lng Fe(II) trong mu tnh theo cng thc sau:
XFe (mg/ml) =
= 0.0171 (mg/L)
Vi Cx : nng ca mu (ppm) Vbm : th tch bnh nh mc (ml) Vmu : th tch mu ht (ml) M : khi lng mu dng phn tch (g) K : h s pha long 1000: n v mg/l nn phi chia 1000 i t ml => L
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
BI 5: Xc nh hm lng Fe bng phng php AAS (mu sa) -----------------5.1 Nguyn tc:
Hm lng Fe trong mu c xc nh bng k thut quang ph hp thu nguyn t ngn la.
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
Nng 2ppm: - Ht 10ml (10ppm) => bnh nh mc 50ml => 2ppm Nng 1ppm: - Ht 5ml (10ppm) => bnh nh mc 50ml => 1ppm Nng 0.5ppm: - Ht 2.5ml (10ppm) => bnh nh mc 50ml => 0.5 ppm Nng 0.2ppm: - Ht 1ml (10ppm) => bnh nh mc 50ml => 0.2ppm Nng 0ppm: - Khng cn ht dung dch chun, nh mc ti vch bng nc ct. Sau khi pha xong dy chun c nng 0; 0.2; 0.5; 1; 2; 5ppm, chun b nc ct 2 ln v mang i o hp thu bng my AAS.
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
0 0 0
1 0.2 0.176
2 0.5
3 1
4 2
5 5
0 0.13
0.007 4.2
V phng trnh hi qui tuyn tnh: T kt qu Abs o c v phng trnh hi qui tuyn tnh th hin mi tng quan gia nng v hp thu.
Fe
0.25 y = 0.0395x + 0.0018 R = 0.9985
0.2
0.197
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
BI 6: Xc nh hm lng Acid benzoic bng phng php HPLC -----------------6.1 Nguyn tc:
Acid benzoic trong thc phm c chit v pha long bng nc ct, trong trng hp cn thit mu s c lm sch vi dung dch Carrezz. Acid Benzoic sau c xc nh bng phng php sc k lng hiu nng cao vi detector UV bc sng 226 nm.
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
- Bnh tia Ho cht: - Acetonitile: loi dng cho HPLC. - H3PO4 0.05%: loi dng cho HPLC - Methanol: loi dng cho HPLC. - Dung dch Acid benzoic chun 1000ppm. - Nc ct 1 ln, 2 ln.
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
- Ht 25ml (10ppm) => bnh nh mc 50ml => 5ppm Nng 2ppm: - Ht 10ml (10ppm) => bnh nh mc 50ml => 2ppm Nng 1ppm: - Ht 5ml (10ppm) => bnh nh mc 50ml => 1ppm Nng 0.5ppm: - Ht 2.5ml (10ppm) => bnh nh mc 50ml => 0.5 ppm Nng 0.2ppm: - Ht 1ml (10ppm) => bnh nh mc 50ml => 0.2ppm Nng 0ppm: - Khng cn ht dung dch chun, nh mc ti vch bng nc ct. Sau khi pha xong dy chun c nng 0; 0.2; 0.5; 1; 2; 5ppm, tim mu vo cc vial chun b sn chun b nc ct 2 ln v cc ha cht chuyn dng cho HPCL mang i phn tch = 226nm. iu kin chy my: - Ct: C18 (150mm 4,6mm 5m)
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
- Pha ng: H3PO4 0.05% v Acetonitile (60:40) - Detector (u d PDA) = 226nm cho Acid benzoic - Tc dng: 1ml/pht (tc dng cao th dung dch s i nhanh v c th lm h ct) - Nhit ct: ~30oC - Th tch tim mu: 20l. - Thi gian chy mu: 6 pht Bnh nh mc 50ml Cbenzoic (ppm) mAbs Din tch peak % din tch peak 0 43.4170 62.3405 80.1054 87.7340 94.7879 0 178.73540 421.15576 790.15509 1709.01038 4067.13184 0 0 0.2 5.069 0.5 5.016 1 4.976 2 4.924 5 4.862 0 1 2 3 4 5
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
V phng trnh hi qui tuyn tnh: T kt qu mAbs v din tch peak chy mu c, v phng trnh hi qui tuyn tnh th hin mi tng quan gia nng v din tch peak.
Acid benzoic
4500 4000 3500 3000 2500 2000 1500 1000 500 0 0 0 790.15509 421.15576 178.73540 1 2 3 4 5 6 1709.01038 Acid benzoic Linear (Acid benzoic) y = 814.66x + 13.102 R = 0.9995 4067.13184
Nhm 1
Trng i hc Cng nghip thc phm TP.HCM Khoa Cng ngh thc phm
a sang pha ng, pha ng chun b sn H3PO4 0.05% v Acetonitile t l 60:40 c lc qua ray 0.25m, sau qua b loi kh chn khng v cui cng c bm cng vi mu qua ct v bt u qu trnh ra gii. Kt thc qu trnh phn tch dng nc ct v dung mi ra ct.
Cc vial c tim mu
Nhm 1