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GVGD: TS. Lê Đình Vũ SVTH: BÁO CÁO ĐỒ ÁN CHUYÊN NGÀNH XÁC ĐỊNH HÀM LƯỢNG VITAMIN C TRONG VIÊN C SỦI BẰNG PHƯƠNG PHÁP SẮC KÝ LỎNG HIỆU NĂNG CAO HPLC/DAD TRƯỜNG ĐẠI HỌC CÔNG NGHIỆP TP HCM KHOA CÔNG NGHỆ HÓA HỌC Phạm Bảo Quý Lớp: ĐHPT7 MSSV: 11050991 1

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xác định vitamin C bằng pp HPLC

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GVGD: TS. L nh VSVTH: GVGD: TS. L nh VSVTH: BO CO N CHUYN NGNHBO CO N CHUYN NGNHXC NH HM LNG VT!MN C T"ONG VN C S# B$NG %H&NG %H% S'C () L*NG H+U N,NG C!O H%LC-D!DXC NH HM LNG VT!MN C T"ONG VN C S# B$NG %H&NG %H% S'C () L*NG H+U N,NG C!O H%LC-D!DT".NG / H0C C1NG NGH+% T% HCM(HO! C1NG NGH+ H2! H0CT".NG / H0C C1NG NGH+% T% HCM(HO! C1NG NGH+ H2! H0C%h34 B56 789%h34 B56 789L:;:H%T@@=MSSV:==>?>@@=1NA DUNGNA DUNG12 Tng quan l thuyt23Kt qu v tho lunThc nghim xc nh hm lng vitamint!ong vi"n#$i %&ng '()*+,+2=. TBnC D8En F9 Gh8HIG=. TBnC D8En F9 Gh8HIG=.=. TBnC D8En JK JLGE4Ln CMnh nChNE: -itamin . c/n c g0i l aci1 a#co!%ic. l m2taci1 h/a tan t!ong n3c t!ong ch4t 1inh 15ng c t6m th4yt!ongm2t#7lo8ithc9h:m;T!ongcho8t2ngnhm2t ch4t ch7ng oxy h>a. gi?9 %o v cc t %o @hAi nhBng thit h8i gCy !a %Di cc g7c t 1o;NC8OnCPQ:-itaminc>nhiEut!ongcclo8i!auqut hm lng cao t!ong !au xanh. Gc %it l %Hng ci xanh. ti"u. @hoai tCy. ci %!u##Il. !au ci. c chua. xoong cam. qut. chanh; T!ong cHng nghi9. cc #n 9h:m chJa vitaminch$ yu c tng h9 %&ng 9ha h0c i tK +Lgluco#I; 34TRnh QhSG JTG F9: Kh7i lng 9hCn tMN OPQ.OR g*mol > 18ngN %2t mu t!Sng n vng nh8t T@hanU. @hHng mVi. v chua Whit 2 n>ng chyN OXRo T9hCn h$yU Kh nYng h/a tan t!ong n3cN cao Kh> tan t!ong !u. @hHng tan t!ong cc 1ung mHi hBu c< ,ci1 a#co!%ic c> @h nYng h49 thu nh #ng tM ngo8i. Z-max [ \]^ nm. t8i 9' [ \. Z-max [ \Q] nm t8i 9' [ Q;^ ;TRnh QhSG hUE hVQ:Tnh acid+o hiu Jng li"n h9 c$a nh>m ca!%oxyl. n"n nguy"n tM hy1!o c$a nh>m hy1!oxyl D v t!F #7 R t!D n"n !4t linh 2ng. lm cho vitaminc> tFnh aci1 m8nh;__' '_''\_''__N> th= nh lng aci1 a#co!%ic %&ng 9ha%&ngccht!ung h/a cc g7c t 1o #n #inh !a tK cc 9hn Jng chuy=n h>a. nha > %o v c tFnh ton von c$a mng t %o;'pt!!4tl3ngi?9h49 thu#Stc$aca t! \ 1i 1ng c h49 thu #St h>a t! \. c/n #St h>a t! R mu7n c h49 thu 9hi c chuy=n thnh h>a t! \. m qu t!6nhchuy=nnycmnc>#x?ctcc$avitaminn"nnuthiu vitamin#q1igCythiumu1othiu#St.1ovy@hiu7ngvi"n #St cmn u7ng @rm v3i vitamin ;Mobile phase (Pha ng)Stationary phase (Pha tnh)=.a. TBnC D8En JK \b Gh8TG ZcQ \9 F]nC hLd8 nenC QE6(hfL nLd4:eSc@lAnghiunYngcaolm2t9h 9ha 2ng l ch4t lAng v 9ha tsnh chJa t!ong c2t l ch4t !Sn t c 9hCn chia 13i 18ng ti=u 9hCn hoGc m2t ch4t lAng 9h$ l"n m2t ch4t mang!Sn.haym2tch4tmangtc%ini%&ngli"n@tho h0c v3i cc nh>m chJc hBu c< ;-gut!6nhtch1avotFnhch4th>ah0c.vtlvh>alc$a cc ch4t;-+a t!"n \ qu t!6nhN'49 9hcuii h49 9hc-vy !a li"n tcc giBa \ 9haN (ha tsnhN ch4t !Sn hoGc lAng. giB ch4t 9hCn tFch(ha2ngNch4tlAngTOch4thoGchpnh9nhiEuch4tU.h/atan v 1i chuy=n ch4t 9hCn tFchL eSc @ lAng chia thnh \ nh>meSc @ lAng 9 #u4t thang T#Sc @ c i=nUeSc @ lAng 9 #u4t cao TeSc @ lAng hiu nYng caoN '()U%hgn F63L:+a vo %n ch4t c$a qu t!6nh #Sc @. '() chia thnhN%hgn F63L:+a vo %n ch4t c$a qu t!6nh #Sc @. '() chia thnhNHPLCS( Chh; Qi; L6n S( ;hgn jPS( ;hE Gh_`nC S( ;hE Y56 S\ GkE6 YBL L6n Khi n7i v3i mu 1/ T1ItIcto!U. '() cho 9hw9N-knh tFnhN 1a vo thai gian lu-knh lngN 1a vo chiEu cao hoGc 1in tFch 9Ia@ClU T/O MY H%LCClU T/O MY H%LC(1) Bnh ng dung mi - 'in t8i my '() thangc> x^ ang 1ungmHivomu%t l8i t!ong 1ung mHi 9ha 2ng;Wu nh t!ong qu t!6nh 9hCn tFch m 1ung mHi 9ha 2ng c/n #>t cc %0t @hF th6 m2t #7 hin tng #au Cy #q #y !aNL Tll9ha2ngc$accang1ungmHil4y@hHng?ng#qlm cho thai gian lu c$a (Ia@ thay i ;L T!ong t!ang h9 %0t qu nhiEu %2 @hM @hF @hHng th= lo8i t!K ht cth6c>th= (um9#q @hHng h?tc1ung mHi @hi > 9 #u4t @hHng l"n v my #Sc @ #q ngKng ho8t 2ng ;H+THsNGtU(HuN%SUlT HOrCTsC0DvNG%H!ANGw CvNG0LH+THsNGCUNGCl% DUNG M1: LC %UM% (3) Bm cao p (h !h"ng cung c#p dung mi)L | 9 #u4t t7i a ^O\ |a! TOat [ x;X}|a!U; T7c 2 1/ng x;OLX;XXX ml*9h?t ;($) B ph%n !i&m m'u ( in(ec!ion )a*)e)k=am~uvoc2t9hCntFchthIo9h9 l ] LOxxl ;>x\ cch l4y m~u vo t!ong c2t N |&ng ti"m m~u th$ cHng Tti"m %&ng tayU v ti"m m~u t 2ng T,uto#am9lIU ;B&M MxU B$NG T!Y B&M MxU Tq ANG m!UTO S!M%Lno) T T_)Z{n _-nWUN +Wu k kZ K'W W'T k T T'T T'N ) W v , gZ TW' T' 'T. u{ T ('W T' - T |_ -]; 2t tch TolumIU- Ct tch c kch c khc nhau- Chiu di: 10 25cm- n knh: 2 5mm(+) ,! !ch 2t chJa 9ha tsnh c coi l t!i tim c$a h th7ng #Sc @ lAng hiu nYng cao;L2t9hatsnhthHngthanglm%&ngthw9@hHng!`.chiEu1ic2t @hong Ox LRxcm .ang @Fnh t!ong OLOxmm .h8t ch4t nhbi c5 [ ]LOx m;Tngoi !a c/n c> m2t #7 t!ang h9 Gc %it vE @Fch th3c v @Fch c5 h8t;;;;UL-3ich4tnhbic2tc5[O;}L]mc>th=1Vngc2tngSnTRLOx cm U v nhA Tang @Fnh t!ong OL^;Q mmU lo8i c2t ny c> hiu nYng tch cao;L h4t nhbi c2t tVy thIo l0ai c2tv @i=u #Sc @L ThHng thang ch4t nhbi c2t l eilicagIl T9ha thunUhoGcleilicagIltceilanh>ahoGcc%aom2tl39mAng hBuca. @h7i 9h. (2) 3 !h"ng ghi nh%n )4 5 *6 !n hiu (in!e17ace8 so7!9a1e8:,); D! ?u@A. NGUYN T'C HO/T ANG MY H%LCL {~u ch4t lAng c a vo %ubng % c % @h nYng h49 thu ch4t 9hCn tFch g0i l 9ha tsnh. m2t 9ha 1i chuy=n qua 9ha tsnh g0i l 9ha 2ng; +o cc c4u tM ch4t 9hCn tFchc>[email protected]?ng1ichuy=nv3it7c2@hc nhau v tch !a @hAi nhau; L c qu t!6nh tch t!ong #Sc @ lAngN gu t!6nh quan t!0ng nh4t t!ong 9h ch?ng c chuy=n thnh tFn hiu in; TFn hiu ny c @huych 8i!bichuy=n#ang%2ghi;ctFnhiucxMlD>!bichuy=n #ang %2 9hn in v lu @t qu;LT!"n#Scbnhnc.#qc>cctFnhiuJngv3iccc4utMc tch g0i l 9Ia@; Thai gian lu c$a 9Ia@ l 8i lng Gc t!ng Tnh tFnhU cho ch4t cmn tch; /n 1in tFch 9Ia@ l th3c o nh lng cho ch4t cmn 9hCn tFch;{Thai gian ch4t 9hCn tFch % 9ha tsnh lu giB Tthai gian luU quyt nh %DiN |n ch4t c$a 9ha tsnh. c4u t!?c v tFnh ch4t c$a ch4t 9hCn tFch; |n ch4t v thnh 9hmn c$a 9ha 2ng 1Vng = !Ma gii ch4t 9hCn tFch !a @hAi c2t #Sc @ T9ha tsnhU; L uhi l8i ton %2 qu t!6nh tch #Sc @ c$a hpn h9 ch4t 9hCn tFch #Sc @ b gbm nhiEu 9Ia@;L kGc i=m c$a 9Ia@ 9hCn tFch N c 9Ia@ c> th= tch !ai nhau hon ton h9 nhau m2t 9hmn h9 nhau hon tonL eSc @ b 9hn nh qu t!6nh tch #Sc @ t!ong c2t t7t hay @hHng t7t; Tch t7tN hpn h9 c> %ao nhi"u ch4t c> %4y nhi"u 9Ia@ !i"ng %it @hHng ch9 nhau h4t no % lu giB m8nh #q c !Ma gii !a #au cVng. ch4t lu giB @wm #q !a t!3c Tu tKng ch4t cmn nghi"n cJu v cc iEu @in 9hCn tFch = ch0n cc 1ItIcto! @hc nhau;NG DNG C#! H%LCNG DNG C#! H%LC-h$ yu xc nh cc h9 ch4t hBu c< @h> %ay h th= 9hCn tFch bng thai nhiEu h9 ch4t-KhHng cmn lm %ay h 1ItIcto! +,+ D %3c #>ng \Q] nm;a. ThQ nChLd4 |fQ YMnh h4 F_WnC JLGE4Ln C Gk6nC JLn C ZXL jnC ;h_nC ;hf; H%LC-D!Da. ThQ nChLd4 |fQ YMnh h4 F_WnC JLGE4Ln C Gk6nC JLn C ZXL jnC ;h_nC ;hf; H%LC-D!DY8 Q8 Qh8nCT!ongqut!6nh9hCntFch.ch`#M1cngccthu7cthMlo8itinh @hit 9hCn tFch v n3c #M 1cng 9hi l n3c c4t hai lmn @hM ion hoGc lo8i O c$a T-W ^}]O Te_ RQXQUHUE QhSG(EFL }LhH}k6 ;h6Z;hEG^w TK'\(_^U XX;x y!Q^G6nLGkLF^w mF63L }nC Qh6 H%LCow T'RWU XX;x y%hE YynC Qh6 H%LC'/a tan OR;Q g @ali 1ihy1!o 9ho#9hatI t!ong Xxx ml n3c ng t!ong c7c c> mA; )0c qua %2 l0c c5 lp x;^] m T1ung 1ch ,U; Oxx ml acItonit!ilI c l0c qua %2 l0c x;^] m T1ung 1ch |UKhM @hF 1ung 1ch t!3c @hi #M 1cng. nu cmn;ChSG Qh8n ,xit a#co!%ic. TQ'}_QU XX;P yD8nC }MQh CPQ +ung 1ch g7c axit a#co!%icw TQ'}_QU O mg*ml TOxxx 99mU '/a tan m2t lng axit a#co!%ic t c cCn chFnh xc n x.O mg; hu:n % 1ung 1ch m3i t!ong ngy #M 1cngLSH 48 J | F9 Z jyT!3c @hi o cmn l0c 1ung 1ch qua mng l0c x;^] mCn chFnh xc n x;O mg @hong x;] g m~u vi"n #$i vitamin#au > h/a tan hon ton vo n3c. cho vo %6nh nh mJc ]x ml #au > nh mJc n v8ch. h?t O ml 1ung 1ch tK %6nh nh mJc ny cho vo %nh nh mJc Oxx ml #au > nh mJc %&ng n3c t3i v8ch. ta c 1ung 1ch m~u thM; ThLIG jMw }nC Q c 1cng cc thHng thang c$a 9h/ng thF nghim v cc thit % cc th= nh #auN|2 l0c 1ung mHi x;^] m|2 l0c m~u x;^] m' th7ng #Sc @ lAng hiu nYng caoN ubm c> % t!ong m~u; ko lmn lt cc 1ung 1ch chu:n c> nbng 2 Ox 99m. \x 99m. ^x 99m #au > 9ha 1ung 1ch chu:n c> nbng 2 3c lng t!ong @hong Ox ^x 99m !bi tin hnh o v3i cVng iEu @in nh o cc 1ung 1ch chu:n. hm lng vitamint!ong m~u #q c xc nh %&ng 9h a> >DLdn GRQh ;^E\m4!UZo}X];X^PR \xPO;]]x]^ Q]QQ;X}PRxKt qu o 1ty chu:n vitamin eSc @ b m~u cmn xc nhKh7i lng m~u cCnTgU+in tFch 9Ia@Tm,Z#UWbng 2 tFnh c T99mU %&ng 9h Qh GL8 Gk6nC hUE QhSG Q j5n\;Q; vc nh hm lng Wal t!ong m~u n3c mu7i nghro\;P; (hCn tFch hm lng Wa\e_R t!ong n3c mu7i nghro\;}; vc nh hm lng 'l t!ong m~u aci1 'l\;X; vc nh hm lng lo t 1o t!ong m~u aci1 'l \;Ox; vc nh hm lng Wa_' t!ong m~u #n 9h:m cu7i Wa_' giao hng5a.=. XfQ YMnh h4 F_WnC NEaO SLOa { M6}8F SLFLQEG Gk6nC Ghnh ;h4 NEGkL SLFLQEG m ThXH GLnh F]nCoa.=. XfQ YMnh h4 F_WnC NEaO SLOa { M6}8F SLFLQEG Gk6nC Ghnh ;h4 NEGkL SLFLQEG m ThXH GLnh F]nCoTi"u chu:n ny 9 1cng cho Wat!i eilicat 18ng lAng #n xu4t tK x?t v ct th8ch anh. 1ung = #n xu4t x 9h/ng. @Im giGt. %2t giGt tng h9 v cc mcc Fch @hc; Wguy"n tSc xc nh +a t!"n 9hn Jng chu:n 2 axitL %afo = xc nhy Wa\_. chu:n 2 ngc = xc nh y ei_\ vtFnh c mo1ul #ilicat c> t!ong m~u Wat!i eilicat thIo y Wa\_. y ei_\; (h{> 4L 48 Y FVQa{z CLVG %%Ch8n Yy jnCHCF>w= NTh4 awz CLVG MOCh8n Yy GLI; jnCHCF>w= NDnC Qh8nDnC Qh8n78H Gknh |fQ YMnh h4 F_WnC NEOH J NEaCOz YOnC Gh`L Gk6nC 48 48PL GLnh QhI13CnC Gh[Q GRnh G6fn |fQ YMnh h4 F_WnC NEOH J NEaCOz YOnC Gh`L Gk6nC 48 48PL GLnh QhICnC Gh[Q GRnh G6fn |fQ YMnh h4 F_WnC NEOH J NEaCOz YOnC Gh`L Gk6nC 48 48PL GLnh QhI Tg*lU Wa_' [L -ON Th= tFch 1ung 1ch chu:n 'l chu:n v3i ch` th (( TmlU L -\N Th= tFch 1ung 1ch chu:n 'l chu:n v3i ch` th {_ TmlU L -N Th= tFch 1ung 1ch m~u TmlU L N Wbng 2 a ch4t -icaco;1%(h8m vi 9 1cng k7i v3i m~u n3c mu7i c49 in gii Wguy"n tSc xc nh+a t!"n 9hn Jng chu:n 2 oxi h>a @hM 9hmn 1 = xc nh ch` ti"u ny. th"mvo1ung1chm~um2tlng1chFnhxcmu7i{o!ht!ongmHi t!angaci1'\e_^;Wal_R#q9hnJngv3ich4t@hMITW'^U\e_^ t!ong mHi t!ang aci1. D iEu @in n>ng; )ng ITW'^U\e_^ 1 #q c xc nh %&ng 1ung 1ch K{n_^ chu:n; ki=m t4F 48PL M6kh =>4F HaSO 8n YIn C z 4L n_:Q QSG? 4L MnSOLcQ YK8 nC8yL (MnO >wa? NHOnC nh3G jKnz>ZDnCQh8n78H Gknh |fQ YMnh h4 F_WnC NECFOz Gk6nC 48 n_:Q 48PL QS; YLdn CL5L78H Gknh |fQ YMnh h4 F_WnC NECFOz Gk6nC 48 n_:Q 48PL QS; YLdn CL5L21CnC Gh[Q GRnh G6fn |fQ YMnh h4 F_WnC NECFOz Gk6nC 48 n_:Q 48PL QS; YLdn CL5LCnC Gh[Q GRnh G6fn |fQ YMnh h4 F_WnC NECFOz Gk6nC 48 n_:Q 48PL QS; YLdn CL5L Tg*)U Wal_R [L -ON ) th= tFch 1ung 1ch K{n_^ chu:n 1Vng lm cho m~u thc TmlUjL -\N ) th tFch 1ung 1ch K{n_^ chu:n 1Vng lm cho m~u t!Sng TmlUjL -N ) th= tFch 1ung 1ch m~u TmlUjL N ) nbng 2 2lG9lai t2lG9l8i t 4L={a CLVG%%Qh8nYyjnC!CNOz>w>? NHG ? 4LNI8 QU 48 hOnC.Th4 GnC CLVG HaSOYIn \hL4SG 48NI8 \hnC QU 48 hOnC.Th4 GnC CLVG NEOH YIn \hL QU 48 hOnC kOL GLI; GQGh4GnCCLVGHaSOYIn\hL 4SG 48(IG GXEY] C3QhDnCQh8na{z CLVG(aCkaO78H Gknh |fQ YMnh h4 F_WnC NECF Gk6nC 48 n_:Q 48PL nCh678H Gknh |fQ YMnh h4 F_WnC NECF Gk6nC 48 n_:Q 48PL nCh630CnC Gh[Q GRnh G6fn |fQ YMnh h4 F_WnC NECF Gk6nC 48 n_:Q 48PL nCh6CnC Gh[Q GRnh G6fn |fQ YMnh h4 F_WnC NECF Gk6nC 48 n_:Q 48PL nCh6Tg*lU Wal [ L O N Wbng 2 = 4L }} a >w= NNEaSaOz >w>=>@ Nz CLVG Qh GhM hO GLnh jyGCh8n Yy T34 }nC Ch8n GLI;DnCF3L78H Gknh |fQ YMnh h4 F_WnC NEaSOz Gk6nC n_:Q 48PL nCh678H Gknh |fQ YMnh h4 F_WnC NEaSOz Gk6nC n_:Q 48PL nCh634CnC Gh[Q GRnh G6fn |fQ YMnh h4 F_WnC NEaSOz Gk6nC n_:Q 48PL nCh6CnC Gh[Q GRnh G6fn |fQ YMnh h4 F_WnC NEaSOz Gk6nC n_:Q 48PL nCh6Tg*lU Wa\e_R [ - ON ) nbng 2 a ch4t -icaco;3!(h8m vi 9 1cngk7i v3i m~u th9 \xxT t!ong nh my -,_Wguy"n tSc xc nh+at!"n9hnJngchu:n2aci1%afa ch4t -icaco; Tl\ ^x 99mULT4tcccm~u c>2til9 t