HA PHN TCH THC PHM

Th.S. PHM KIM PHNG Chuyen nganh Hoa Phan Tch 45 TIT L THUYT

Ni dung hc

Cung cp nhng kin thc c bn trong phn tch nh tnh v nh lng. -Cung cp cc gii php xy dng phng php phn tch. -Cung cp k thut phn tch c in v hin i. -Cung cp nhng ng dng ca lnh vc ha hc phn tch trong nghin cu khoa hc v thc tin.
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CHNG TRNH GING DY

1. I CNG HA PHN TCH P DNG CHO PHN TCH THC PHM 2. CHUN AXT - BA Z 3. CHUN KT TA 4. CHUN OXY HA KH 5. CHUN PHC CHT 6. PHNG PHP O MAU 7. PHNG PHP PHN TCH QUANG PHO - K thut my hp thu nguyn t AAS - K thut my quang ph pht x Plasma - ICP 8. L THUYT K THUT SC K - K THUT SC K KH - GC - K THUT SC K LNG CAO P HPLC

PHAN 1

I/ I CNG HA PHN TCH

1/ NH NGHA TH NO L HA PHN TCH 2/ NH NGHA TH NO L HA PHN TCH THC PHM

3/ PHAN TCH NH TNH

4/ PHN TCH NH LNG 5/ NH GA PHNG PHP PHN TCH V S LIU PHN TCH

5.1/ O CHNH XAC, O UNG, O LAP LAI, TOC O PHAN TCH, O NHAY, PHNG PHAP AC TRNG , PHNG PHAP C CONG NHAN

6/ X LY THONG KE ( STATISTICAL ASSESSMENT OF QUAILITY OF DATA)

7/ CCH BIEU DIEN THANH PHAN DUNG DCH 7.1. chat chuan goc 7.2. cac nong o dung dch

9/ / DNG C O THU TINH CHNH XC V TNG I

1. NH NGHA TH NO L HA PHN TCH

Hoa phan tch l mt ngnh khoa hc chuyn nghin cu cc phng php phn tch nh tnh v nh lng mt cht hay nhiu cht , mt nguyn t hay nhiu nguyn t c trong sn phm ang nghin cu V d : Trong mt mu nc ung c b nhim d lng thuc tr sau hay khng ? Bc u tin chng ta phi nh tnh xem trong mu nc bao gm nhng cht g? Bc 2 : nh lng nhng cht c nh tnh Bc 3 : da trn cc mu chun tnh ton v cho ra kt qa cui cng

PHAN TCH THC PHAM

2/ Thc phm c p ng cc tiu chun ha hc v v sinh ? C b i thiu, h hng v bin thnh cht c hi hoc c cha nhng cht c do thi ra t bao b, ha cht cho thm vo Kim nghim phn tch thc phm bng phng php ha hc ngoi ra cn phn tch trng thi cm quan, vi sinh vt

2. TH NO L PHN TCH HA THC PHM

Thc phm l nhng thc n , nc ung l nhng cht dinh dng cn thit cho c th con ngi, vt nui .do vy p ng cc yu cu trn thc phm phi cn c kim nghim trc khi a ra th trng tiu th. Phn tch ha hc thc phm ( Analytical chemistry of Food) : nhm xc nh Thc phm c p ng cc tiu chun ha hc v phm cht v thnh phn dinh dng theo ng nh quy nh hoc c b gian di v gi mo hay khng?

oi tng cua Hoa phan tch thc pham la cac chat dinh dng nh am , beo, bot, ng co trong ca, tht sa, trng, nc uong
e nh lng cac chat dinh dng cac nguyen to hoa hoc hay nh danh cau truc thanh phan cua cac chat oi hoi phai co phng phap phan tch chnh xac va phu hp vi cac oi tng nghien cu.

PHAN TCH NH LNG

3. PHAN TCH NH TNH

3.1.Phan tch nh tnh : Nhm xc nh s hin din ca cc cu t ( ion, nguyn t hay nhm nguyn t ) trong mu phn tch ( thc phm, mu nc , t ) v ng thi nh gi s b hm lng ca chng :a lng, vi lng, vt nh vo cc thit b phn tch v cc phn ng ho hc c trng ln mu, kt ta i vi nguyn t cn xc nh 3.2. PHAN TCH NH LNG

Xc nh chnh xc hm lng ca nhng cu t trong mu. c th hin bng nhng gi tr sau: %, mg/kg,mg/L (ppm), g/kg, g/L (ppb), ng/kg, ng/L ( ppt)

cc phng php phn tch thch hp v x l s liu phn tch

4. Chn

Khi chn phng php s dng cho phn tch thc phm ph thuc vo cc yu cu sau : S lng, ch tiu yu cu phn tch thit b hin c ti phng th nghim t c nhng cn nhc khi s dng quy trnh phn tch. - Quy trnh phn tch phi tho mn cc iu kin sau:

- chnh xc, ng , lp li tt - Quy trnh phn tch c tnh chn lc cao ph hp vi yu cu cn phn tch - Thi gian phn tch nhanh v c kh nng phn tch ng thi nhiu nguyn t - Thit b s dng cho phn tch c nhy cao, cc tiu pht hin nh - Gi thnh phn tch khng cao - Thit b d s dng, d bo tr

- u tin s dng cc phng php phn tch c cng nhn bi cc t chc quc t sau: ISO : International Organisation for standardusation) AOAC : Association of Oficial Analytical Chemists , published in the AOAC book BSI : British Standards Institution TCVN : Tiu chun Vit Nam FAO : Food and Agriculture Organisation
T chc lng thc-nng nghip ca Lin Hip Quc

S la chon phng phap phan tch

Moi oi tng lai co nhieu chat, nhng chat nay co the la ham lng en phan tram (%) cho en ham lng nho nh mg/kg , mg/l ( 10-6 hoac nho hn g/kg, g/l (10-9) hoac nho hn na 10-12 picrogam ( ppt) .

 moi phep phan tch co nhng u va nhc iem rieng cua tng phng phap co ngha la co nhng phng phap thch hp cho phep xac nh ham lng ln, co phng phap phan tch cho phep xac nh ham lng nho v vay phong oan trc ham lng co trong mau e chon phng phap phan tch cho phu hp va giam s sai so trong qua trnh phan tch.

Vi ham lng ln va ban nh lng ngi ta thng dung cac phng phap hoa hoc co ien : Khoi lng, the tch, phan tch ban nh lng -Ham lng vi lng ( ppm ) va sieu vi lng ( ppb, ppt hoac nho hn ) phai dung cac thiet b phan tch hien ai e o nh AAS, ICP, ICP/MS, HPLC, GC, GC/MS,LC/MS, HRGC/HRMS .. Nhng thiet b nay thng s dung e o vet cac kim loai nang, cac d lng thuoc tr sau, vitamin, khng sinh trong nc, at, thuy san, thc pham

5. NH GI PHNG PHP PHN TCH

- chnh xc Precision Biu th qua cc kt qu phn tch do 01 ngi thc hin hoc do mt s ngi khc thc hin trong cng mt phng th nghim v s dng cng mt phng php phn tch trn cng mt thit b phn tch. - lp li Reproducibility Biu th qua cc kt qu phn tch ging nhau trn mt mu c chia ra lm nhiu ln phn tch do 01 ngi thc hin cng mt quy trnh phn tch v trn cng mt thit b phn tch. Mt mu lm nhiu ln c lap nhau khc vi mt mu o nhiu ln.

- o ung ( Accuracy):

Ket qua a ra gan ung vi so thc c bieu th qua gia tr trung bnh va gia tr thc va o lech chuan. o lech chuan cang nho th ket qua co c cang ung. Va cang ung hn na ngi phan tch phai tnh c o khong am bao cua phep o ( Uncertainty in measurement). lech chuan bang phng phap thong ke, tnh c nguon sai so ngau nhien va sai so he thong

o khong am bao cua phep o la phai tnh c o

Sai s ngu nhin (sai s khng xc nh) Sai s gy n lp li ca kt qu, sai s ngu nhin lun c trong qu trnh thc hin phn tch nh cn, o my gim sai s ngu nhin bng cch tng s ln phn tch ( n= 5, 7,8..)

la do cac nguyen nhan sau : Thiet b phan tch, dung cu s dung nh can , bnh nh mc, sai so do nha che tao, chng t hieu chuan, cac chat chuan tinh khiet ..., hoc phng php s dng c khuyt im nh cch c, chuyn i mu khng r Sai s h thng gy nh hng n ng ca php phn tch. Gim thiu sai s nay bng cch hiu chun thng xuyn cc thit b s dng

Sai so he thong ( sai s xc nh)

PHNG PHAP TNH X LY THONG KE 1/ Tnh gia tr trung bnh cua cac phep o Xtb = xi

n
2/ o lech chuan = ( Xi Xtb )2

n
-1 3/ Tnh o bien ong cua ham lng ( RSD) RSD = x100 Xtb Khoan tin cay (CI) - Confidence interval CI = Xtb tp x

H s bin thin hay ch s phn tn

= . 100 Xtb RSD 10% cc Xi t phn tn - tt RSD

10% < RSD < 20% cc Xi c th s dng c RSD > 20% cc Xi qu phn tn khng nn s dng

Trnh by kt qu KQ =

HLtb tp .

CHAP NHAN KET QUA

1/ Ket qua c chap nhan khi : o bien ong cua ham lng phai nho hn 5% oi vi phan tch co ham lng % RSD 5% 2/ Vi phan tch vet co ham lng: ppm,ppb co the chap nhan khi:

RSD 10 %

- Cc tiu pht hin ca u d thit b: LOD limit of detection LOD l hm lng ti thiu c pht hin bi thit b s dng phn tch. LOD khng ging nhau i vi tng cht v thit b phn tch. Nguyn tc: LOD ca mi loi u d c xc nh bng cch so snh trn cng mt thang o- chiu cao tn hiu - S (signal) ca cht cn phn tch vi chiu cao ca ng nn (n) noise

Thc hin mu trng o chiu cao ca ng nn (n) - Thc hin o mu c nng thp nht (Cmin ) o chiu cao ca tn hiu (S) sao cho 10 > T = S 3 n

LOD = 3Cmin LOD = Cmin T

LOD c th biu din theo nng hoc theo trng lng

LOQ : Gii hn nh lng ( limit of Quantitation) LOQ l gii hn ti thiu nh lng ca phng php phn tch. Nguyn tc: LOQ ca mi phng php v mi cht cn xc nh l khc nhau. Cch tnh LOQ : - Cho vo mu c trng lng hoc th tch m0 xc nh, cu t cn phn tch bit trc Cmin LOQ = 3Cmim . F T F : h s pha long hoc lm giu mu

- mu khng lm giu Vi = Uo LOQ = LOD - mu c lm giu ( ui bt dung mi, hoc c cn bt) Vi << U0 LOQ << LOD
Kt qu LOQ c th biu din theo nng hoc theo trng lng tuyt i.

YEU CAU PHONG TH NGHIEM THC HIEN QA/QC TRC KHI CHO PHAN TCH

oi vi mot phep phan tch nao nhat la oi vi cac phan tch vet trc khi phan tch phong th nghiem phai thc hien chng trnh QA/QC cho tng phep th.

QA : Quality assurance am bao chat lng la khi phan

tch cho mot ch tieu nao o th phai am bao rang co quy trnh phan tch co hieu lc cho ch tieu o, co chat chuan cua ch tieu o (chat chuan phai co giay chng nhan ISO cua ngi ban), ket qua phan tch phai co o tin cay cao. QA bao gom ca kiem tra chat lng (QC) va anh gia chat lng cua qua trnh phan tch (quality assessment)

tra bang cach them chuan vao mau, phan tch mau chuan (material reference - RM) hoc trn mu kim tra (CRM) t o tnh c hiu sut thu hi (recovery) cua phng phap. Quality assessment: nh gi cht lng Qua trnh phan tch c thc hien trong phong th nghiem thong qua cac anh gia noi bo va ngoai bo bi mot anh gia vien trng cua mot phong th nghiem khac hay noi cach khac anh gia phan tch bi cac lien phong th nghiem, kiem tra cheo gia cac phong th nghiem.

QC: Quality control La phng phap a c hieu lc kiem

CAC BC THC HIEN QA/QC

- Gii thieu nang lc cua phong th nghiem - Thc hien QC ve o ung va o chnh xac cua phep phan tch - Quy trnh lay mau

- Bao quan mau

- Phng phap phan tch

- Hieu chuan thiet b, hieu chuan phng phap theo nh ky - Thc hien QA,QC - Kiem tra cheo cac phong th nghiem ben ngoai va noi bo, anh gia ket qua - Bao cao ket qua

6. NH NGHA CHT CHUN GC

Cht gc C tnh n nh cao Bn vi mi trng ng lng ln Khi lng ng vi cng thc ha cht Phn ng nh lng
H2C2O4.2H2O ; C6H5COOH Na2B4O7.10H2O K2Cr2O7 ; NaCl KMnO4 ZnSO4.7H2O ; MgSO4.7H2O

NH NGHA CAC LOAI NONG O

NH NGHA :

Nong o la ai lng cua mot chat ( ion hoac phan t ) trong mot lng xac nh dung dch

1/ Nong o the tch: cua mot chat long la ty le the tch gia
chat long o va the tch cua dung moi V du : HNO3 1:3 co ngha la dung dch gom mot the tch HNO3 ac va 3 the tch nc HNO3 : HCl 1:3

2/ Nong o % khoi lng Cho biet so gam chat tan co trong 100g dung dch C% = So gam chat tan (a) x 100% So gam dung dch V du : Hoa tan a gam chat tan vao b gam dung moi th nong o % cua dung dch la : C% = a .100 a+b Trong hoa phan tch, nong o % c coi la gan ung V du : Muon co dung dch KNO3 1% , th can 1g KNO3 hoa tan vao 100ml H2O

3/ Nong o mol/l :
Cho biet so mol ( co the la ion hay phan t ) chat tan co trong 1lt dung dch, dung ch M hay mol/l CM = So mol chat tan (n) = n
The tch dung dch n : s mol cht tan V : the tch, M: phan t gam V (l)

n = m( chat tan) M

V du 1: Dung dch H2SO4 2M, la dung dch co cha 2 mol H2SO4 hay 2 x 98 = 196g H2SO4 trong 1 lt dung dch V du 2: Hoa tan 1,2g MgSO4 vao nc thanh 100ml dung dch ta c dung dch MgSO4 co nong o mol / L la: CM = 1,2g = 0,1M 120 . 0,1

ve mat hoa hoc tng ng vi 1mol Hydro hay 1mol Hydroxyt trong phan ng ma ta xet Trong cac phan ng hoa hoc, cac chat phan ng vi nhau vi cung so ng lng gam Lg khng phi l hng s nh s mol m ph thuc vo phn ng ha hc m cht tham gia CN = so ng lng gam chat tan (n) So lt dung dch (V) n = m( cht tan)

3/ Nong o ng lng gam ( lg) la so gam cua chat o

lg

4/ Nong o molan Cho biet so mol chat tan co trong 1kg dung moi

a/ Phan ng axt baz 1/ NaOH + HCl = NaCl + H2O 1 mol NaOH phn ng vi 1 ion H+ nen lgNaOH = MNaOH

Tng t 1 mol HCl tng ng vi 1 mol OH- nn : lg HCl = MHCl 2/ NaOH + H3PO4 th tuy theo phan ng co khac nhau ve lg : - NaOH + H3PO4 = NaH2PO4 + H2O lgNaOH = MNaOH , lgH3PO4 = M H3PO4

2 NaOH + H3PO4 = Na2HPO4 + 2 H2O lgNaOH = MNaOH , lgH3PO4 = MH3PO4 3 NaOH + H3PO4 = Na3PO4 + 3 H2O lgNaOH = MNaOH , lgH3PO4 = MH3PO4

Vay lg cua axt bang khoi lng mol cua axt o chia cho so ion H+ ma 1 mol cua axt o tham gia phan ng lg cua baz bang khoi lng mol cua baz chia cho so ion OHma 1 mol baz o a tham gia phan ng

1.5. Nong o ng lng trong phan ng ket tua

Al2(SO4)3 + 3 Pb(NO3)2 = 2Al(NO3)3 + 3PbSO4 Trong phan ng nay lg cua cac chat tham gia phan ng bang khoi lng mol cua chat o chia cho so ien tch cua 1 mol chat o tham gia phan ng lg (Al2(SO4)3 = M (Al2(SO4)3 hay lg Al = MAl 6 3 lg Pb(NO3)2 = M Pb(NO3)2 hay lgPb = Mpb
2 2

1.6. Nong o ng lng trong

phan ng tao phc

Trong phan ng tao phc phan ng xay ra phc tap cho nen e tnh lg cua cac chat tham gia phan ng tao phc ta phai quy c lg cua 1 chat roi t o tnh lg cua chat kia V du: Ag+ + 2CN Ag(CN)-2 Neu lgAg+ = MAg+ th lg CN = 2MCN V du: Hg2+ + 4I- = HgI42lg(Hg2+) = MHg2+ th lg (I-) = 4MI-

trong phan ng oxy hoa kh

1.7 Nong o ng lng

V 1 electron tng ng vi ion H+ nen lg cua chat oxy hoa hay chat kh bang khoi lng mol chia cho so electron ma 1 mol chat o cho hay nhan. 2KMnO4 + 10 FeSO4 + 8 H2SO4 = 2MnSO4 + K2SO4 + 5 Fe2(SO4)3 + 8 H2O lg KMnO4 = MKMnO4 hay lg Mn = MMn 5 5 lg FeSO4 = MFeSO4 hay lg Fe = MFe 1

2/ 3As2S3 + 28 HNO3 + 4H2O = 6H3AsO4 + 9 H2SO4 + 28NO As3+ - 2e As5+ S2- - 8e S6+ N5+ + 3e N2+ lg(As2S3) = MAs2S3 , lgHNO3 = MHNO3 2 3

Nong o ng lng gam chat tan co trong 1 lt dung dch ky hieu : N Theo nh ngha ve ng lng g trong cac phan ng hoa hoc cac chat phan ng vi nhau theo so lg nh nhau va cac chat tao thanh sau phan ng cung tng ng vi nhau ve so lg

V du: nA + mB pC + qD Mac du he so n, m, p, q cac chat phan ng khac nhau nh so lg cua chat A phan ng bang ung so lg cua chat B. Chat C va D tao thanh sau phan ng cung co so lg nh nhau

 CHUN (T)

chun (T) c biu din bng s gam (g) hay micro gam (g) cht tan c trong 1ml hay 1L dung dch V d : Dung dch NaCl c chun bng 0,1mg/ml c ngha l trong 1ml dung dich c cha 0,1mg NaCl - chun theo cht cn xc nh: TA/B c biu din bng s gam cht cn xc nh B tng ng vi 1 ml dung dch cht chun A

V d: Tnh chun ca dung dch KMnO4 0,02M theo Fe trong mi trng axt 5Fe + MnO4 + 8H+ 5Fe3+ + Mn2+ + 4H2O

Theo phan ng ta co : lg (MnO4) = MKMnO4

5 Do o nong o ng lng cua dung dch KMnO4 la : 0,02 x 5 = 0,1N. So mili ng lng gam KMnO4 co trong 1ml dung dch la 0,1 x 1= 0,1mlg. Theo phan ng lg Fe = 0,1mlg , vay so mg Fe tng ng vi 1ml dung dch KMnO4 la : 0,1 x 56 = 5,6mg Vay T KMnO4 / Fe = 0,0056g/ ml

PHA DUNG DCH T CHAT RAN 1/ Tnh toan lng can 2/ Hoa tan 3/ nh mc 4/ Chuyen vao chai cha 5/ Hieu chnh nong o Mcan = CM . V. M 10. P% V : the tch tnh P% : o tinh khiet cua hoa chat M : phan t gam CM : nong o mol V du : Tnh lng can NaOH co P = 96% e pha c 200ml dung dch NaOH 0,1M mcan = 0,1. 200. 40 = 0,83g NaOH 10 . 96

1/ Tnh toan the tch can dung 2/ Pha loang 3/ nh mc 4/ Chuyen vao chai cha 5/ Hieu chuan lai nong o neu can

Pha dung dch t chat long

CM = C% . 10. d , V can lay = Ccan pha . Vcan pha M CM V du : Tnh the tch H2SO4 98%, d= 1,84 can lay e pha 200ml dung dch H2SO4 co nong o 1M CM = 10. 98. 1,84 = 18,4 M 98 V can lay = 1. 200 = 10,87 ml 18,4

Moi lien he gia 3 loai nong o

CM = C% . 10. d M

, CN = C% .10. d

CN = M. CM , CM = so mol chat tan (n) Lg so kg dung moi M : khoi lng mol phan t chat tan : ng lng g chat tan d : Ty trong rieng cua chat tan CM = a gam chat tan. 1L dung dch M .V(L) M: khoi lng phan t chat tan, V : th tch dung dch

5. Nong o vi lng va sieu vi lng ppm ( mg/kg, mg/l) 10-6 ppb ( microgam / kg, microgam/l) 10-9 ppt ( ng/kg, ng/l) 10-12

6. DNG C O THU TINH CHNH XC V TNG I