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(2&$$R  s *@P0  0/B0 W L Vitamin A & 2 &^  6A?pXH  0޽h ? ̙33 800(  R  s *   0cB  p b - Carotene F 2"' &"^  6A?0`H  0޽h ? ̙33 RJ@(  R  s *@ ^  6A?p  S   @`   (Vitamin A and b - Carotene Determination@)(2#c#H  0޽h ? ̙33 aYP(    0lrB 3b-CAROTENE STANDARD ABSORBANCES AT 440 AND 620 nm J4(2%$0&R  s *0^  6A?pH  0޽h ? ̙33 H@`(  d  0|B Use absorbances at 440 nm and then convert this to absorbance at 620 nm and subtract from the absorbance at 620 nm to determine the absorbance at 620 due to Vitamin A.  2  R  s * ^  6A?`  0$B BCarotenoid Absorbance at 440nm Vitamin A Absorbance at 620 nm *C 2A& 9j  0܇Bp@ L Carotenoid (mg/ml) Vitamin A mg/cuvette (sample) nM 2 $ &'!&'&,' H  0޽h ? f3f3[  p (   R  s *P  0HB 07 PTHIAMIN DETERMINATION (2&^  6A?xm  0LB 0 zpyrimidine thiazole , 2&&   0B` AExcite thiochrome at 365 nm and measure the absorbance at 435 nm *B 2@&$ 1H  0޽h ? ̙33a  4(  4i 4 0B0`\  Thiamin Determination in Foods 5 g enriched flour 75 ml 0.1 N HCl Make volume to 100 ml Digest at 1000C for 30 min Centrifuge and filter : 2 &"n BJH 4 0޽h ? ̙33d  $(  $ $ 0B`  Oxidation of Thiamine Tube 1 Tube 2 5 ml Sample 5 ml Sample 2.5 g NaCl 2.5 g NaCl 3 ml K3FeCN6 3 ml NaOH 13 ml Isobutanol 13 ml Isobutanol Centrifuge Centrifuge Tube 3 Tube 4 5 ml Standard (thiamin) 5 ml Standard (thiamin) 2.5 g NaCl 2.5 g NaCl 3 ml K3FeCN6 3 ml NaOH 13 ml Isobutanol 13 ml Isobutanol Centrifuge Centrifuge Thiamin Standard Solution = 0.2 mg/ml Calculation of B1 (mg of B1/5 ml assay sample) = (Tube 1  Tube) / (Tube 3 - Tube 4 )0F2&"""""""""" "#"#."""J d RB $@ s *DP PH $ 0޽h ? ̙33t $0(  0 0 0pBPp0 bRIBOFLAVIN DETERMINATION ,(2&R 0 s * P0 0 0B  (6,7 Dimethyl-9-D-1-Ribitylisoalloxazine ) 2)$, ^ 0 6A?TP H 0 0޽h ? ̙33O ((  ( ( 0B@  7Fluorometric Method Autoclave in 0.1 N HCl for 30 min Adjust pH to isoelectric point to precipitate proteins Filter Determination: Test Tubes 10 ml sample, + 1 ml of riboflavin standard or 1 ml H2O + 1 ml acetic acid, + 0.5 ml KMnO4 (4%) + 0.5 ml 3% H2O2 -- KMnO4 color should disappear in 10 seconds, + 20 mg powdered Na2S2O4 Calculation mg Riboflavin/ml Final Sample Solution B0P2"f"""H  6R ( s *0P^ ( 6A ?"@?H ( 0޽h ? ̙33g 8(  8 8 0Bp ONIACIN ,(2&R 8 s *P ^ 8 6A ?v H 8 0޽h ? ̙33? ,(  , , 0BRJB___PPT9$ :1. Digestion an Hydrolysis 2. PPT Protein with (NH4)2SO4 3. Centrifuge and Filter 4. Rupture of Pyridine Ring with CNBR (Cyanogen Bromide) 5. Color Formation with Sulfanilic Acid 6. Determine the Absorption at 470 nm 7. Plot the Standard Curve of Niacin vs. Concentration 8. (Straight Line of Best Fit) v0P20P20P20P2  " ,"  @` , 0B@ ONiacin Determination(2&R , s *P0^ , 6A ?0pH , 0޽h ? ̙33 yq8?< (  <R < s *  < 6@ i Q " 4B  <@ # 7:z4B <@ # 7vz4B < # 7:z4B < # 7g4B < # 774B < # 7g4B < # 7| < 6 _ AN$4B < # 7q24B <@ # 7q4B < # 7]M4B < # 7@4B < # 7b4B < # 7l < 6 b AC$ < 6 bc BOH$ < 6  AO$ < 6r I '  AO$ < 6T : B  BOH$  < 6P7 , I ^ A-$  !< 66 0  AC$4B "< # 7 :4B #< # 7 04B $< # 7v) z w4B %< # 7+I q4B &<@ # 7? K 4B '< # 7?% l  (< 6,P ]  AN$4B )<@ # 7 l4B *<@ # 7DI 4B +< # 7 K l4B ,< # 7lK L 54B -< # 7  4B .< # 7l  54B /< # 7lK m4B 0<@ # 7@ &4B 1< # 7 0  2< 6j %  BBr$ 3< 6  BCN$4B 4< # 7 !  5< BCxDEF7<x#<< @` I 6< tBCxDEF<x#<< @` I4B 7< # 7    9< tBCxDEF<x$<< @`~S :< 6 `+ CNBr$ ;< 6ȕ{+  NREARRANGEMENT $  << 68J|  NRING IS OPENED$  =< 64= J REARRANGED$  >< 6܄! J DERIVATIVE$ ^  < 6A ?   < 0\B_ 71 2$  < 0E_  72 2$ ?< Bى@ cNiacine Determination$H < 0޽h ? ̙33 1) @(  @R @ s *`^ @ 6A?@  @ S lۉ 0P   cNIACINE STANDARD CURVE 0#'H @ 0޽h ? ̙33 [SL(  L L 0 EPp0 R VITAMIN C , (2 &R L s *`P  L 0%E` 0 Ascorbic Acid Dehydroascorbic Acid 81 2$.&$^ L 6A? 0 + L 0D+E` P kHSCH2CH2(SH)CH2OH 2,3-DIMETHYLPROPANOL Reducing agent or converting dehydroascorbic acid to ascorbic acid "l 2k"DH L 0޽h ? ̙33 D5(  D D 04E`   Titrimetric Method for Reduced Vitamin C 1. Extract with metaphosphoric acid (HPO3) in HOAC. 2. Titrate with 2, 6-dichloroindophenol (blue). 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%          r # ( H/ 0|DTimes New RomanPbbv 0b( 0DArialNew RomanPbbv 0b( 0" DWingdingsRomanPbbv 0b( 00DSymbolgsRomanPbbv 0b( 0@DNew YorkRomanPbbv 0b( 0 b .  @n?" dd@  @@`` X        @ 2$QL$CA[(Y6vr}C2$_5eP CrC2$_: oqC$C2$زkQC$C2$}H)xO(rC2$h9j+T@%HC2$̒ͅ/42ʊsZTmC2$ 1g=Xm,!C2$.r`}K<*,#C$C2$ 2ߡ%}Xb&C2$)_pPMqGxLD)C2$eD= 9+C2$7 QK+̥ocj%/C 0@8{ ʚ;w4ʚ;g4BdBdv 0bFppp@ <4!d!d` 0b<b<4dddd` 0b<b<4BdBd` 0b<b ? %          v  &  ` (    09 3b-CAROTENE STANDARD ABSORBANCES AT 440 AND 620 nm J4(2%$0&R  s *04B  # jJhi4B  # jJh4B  # jJtp4B   # jJ tQ|:   3 jJ5 > m p :   3 jJ  * :   3 jJ+ :   3 jJRO:2  3 jJ B ~ :2  3 jJM ;v :2  3 jJI R:2  3 jJ @ 4B  # jJ h 4B  # jJT hU 4B  # jJ t 4B  # jJJhK4B  # jJt4B  # jJ@ 4B  # jJ@KL4B  # jJL  <>6fw V  M A at 440 nm $   <Ŧ`  J and 620 nm$   6["1  I at 440 nm$   6# |  I at 620 nm$   !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~Root EntrydO)URPictures 2Current UserASummaryInformation(UPRPowerPoint Document(*DocumentSummaryInformation8manHbbv 0b( 0DArialNew RomanHbbv 0b( 0" DWingdingsRomanHbbv 0b( 00DSymbolgsRomanHbbv 0b( 0@DNew YorkRomanHbbv 0b( 0 b .  @n?" dd@  @@`` X       @ 2$QL$CA[(Y6vr}2$_5eP Cr2$_: oq2$^. !-&w Lm2$زkQ$2$}H)xO(r2$h9j+T@%H2$̒ͅ/42ʊsZTm2$ 1g=Xm,!2$.r`}K<*,#$2$ 2ߡ%}Xb&2$)_pPMqGxLD)2$eD= 9+2$7 QK+̥ocj%/ 0@8{uʚ;w4ʚ;g4BdBdv 0bppp@ <4!d!d` 0b4b<4dddd` 0b4b<4BdBd` 0b4b ? %          ru au# (  ՜.+,0    $ iOn-screen ShowThe Ohio State Universityt*| Times New RomanArial WingdingsSymbol New YorkSoaringPowerPoint PresentationPowerPoint Presentation)Vitamin A and b - Carotene DeterminationPowerPoint PresentationPowerPoint PresentationPowerPoint PresentationPowerPoint PresentationPowerPoint PresentationPowerPoint PresentationPowerPoint PresentationPowerPoint PresentationPowerPoint PresentationPowerPoint PresentationNIACINE STANDARD CURVE PowerPoint PresentationPowerPoint PresentationPowerPoint Presentation  Fonts UsedDesign Template Slide Titles  6(ph  AA$  6 H  AA$H  0޽h ? ̙33rCn # ( H/ 0|DTimes New RomanPbbv 0b( 0DArialNew RomanPbbv 0b( 0" DWingdingsRomanPbbv 0b( 00DSymbolgsRomanPbbv 0b( 0@DNew YorkRomanPbbv 0b( 0 b .  @n?" dd@  @@`` X$        @ 2$QL$CA[(Y6vr}C2$_5eP CrC2$_: oqC$C$C$C2$}H)xO(rC2$h9j+T@%HC2$̒ͅ/42ʊsZTmC2$ 1g=Xm,!C2$.r`}K<*,#C$C2$ 2ߡ%}Xb&C2$)_pPMqGxLD)C2$eD= 9+C2$7 QK+̥ocj%/C 0@8{ ʚ;w4ʚ;g4BdBdv 0bFppp@ <4!d!d` 0b<b<4dddd` 0b<b<4BdBd` 0b<b ? %          a  p#(  d  0D Use absorbances at 440 nm and then convert this to absorbance at 620 nm and subtract from the absorbance at 620 nm to determine the absorbance at 620 due to Vitamin A.  2    6p` 0&:B  3 jJ:B   3 jJ :B   3 jJ; < :B   3 jJG :B   3 jJpf :B   3 jJ+ G   < //  (  < M  (  <t 6  D620 $   0Ky@` KCarotenoid Absorbance at 440nm Vitamin A Absorbance at 620 nm *L 2J& Bj  0tPp@ L Carotenoid (mg/ml) Vitamin A mg/cuvette (sample) nM 2 $ &'!&'&,'   HpI  IAbsorbance at 440nm&  <|`  LAbsorbance at nm&  0kF p e  3x 2  0  7x 2  0 @  7x 2  0 @z  7x 2  08 @  7x 2  00 p *  7x 2  0  7x 2   0TP pJ 7x 2 ! 0) p  7x 2 " 0+   7x 2 # 0M `  7x 2H  0޽h ? f3f3r(S # ( H/ 0|DTimes New RomanPbbv 0b( 0DArialNew RomanPbbv 0b( 0" DWingdingsRomanPbbv 0b( 00DSymbolgsRomanPbbv 0b( 0@DNew YorkRomanPbbv 0b( 0 b .  @n?" dd@  @@`` X$        @ 2$QL$CA[(Y6vr}C2$_5eP CrC2$_: oqC$C$C$C2$}H)xO(rC2$h9j+T@%HC2$̒ͅ/42ʊsZTmC2$ 1g=Xm,!C2$.r`}K<*,#C$C2$ 2ߡ%}Xb&C2$)_pPMqGxLD)C2$eD= 9+C2$7 QK+̥ocj%/C 0@8{ ʚ;w4ʚ;g4BdBdv 0bFppp@ <4!d!d` 0b<b<4dddd` 0b<b<4BdBd` 0b<b ? %          r Ի## ( H/ 0|DTimes New RomanPbbv 0b( 0DArialNew RomanPbbv 0b( 0" DWingdingsRomanPbbv 0b( 00DSymbolgsRomanPbbv 0b( 0@DNew YorkRomanPbbv 0b( 0 b .  @n?" dd@  @@`` X$        @2$QL$CA[(Y6vr}C2$_5eP CrC2$_: oqC$C$C$C2$}H)xO(rC2$h9j+T@%HC2$̒ͅ/42ʊsZTmC2$ 1g=Xm,!C2$.r`}K<*,#C$C2$ 2ߡ%}Xb&C$C2$eD= 9+C2$7 QK+̥ocj%/C 0@8{ ʚ;w4ʚ;g4BdBdv 0bFppp@ <4!d!d` 0b<b<4dddd` 0b<b<4BdBd` 0b<b ? %          w ' @(  @R @ s *`4B  @ # VXY4B  @ # VX4B  @ # Ve2  @ s *IV H  > F2 @ S VY @2 @ C V )m F2 @ S VP @ <x,Fp  K A at 470 nm$  @ 6 F2 @ S VY @2 @ C V I F2 @ S Vp @ <x,f0p  K A at 470 nm$  @ 6 F2 @ S VY @2 @ C V I F2 @ S Vp @ <x,f0p  K A at 470 nm$  @ 6?@ABCDEFGHIJKLMNOPQRSTVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~