so 46 - ( E l Vo l wm etri c G rad u ate d An al yti cal fl as ks, of pi pe tte s , capaci ti e s of 0, capaci ti e s 20 an d an d 00 ml ml U se I SO as 5558 Setti n g l th e to th e m ark up th e by m e an s to of a 30 to m ark th e B H T) D i l u te to th e 0, l pg /m l mg, a 00 ml s o l ve n t a pi pe tte th e or : 250 m e th yl u n d e can o ate ( ) ( , to wi th th e n e are s t 0, l or m ark Tran s fe r a 20 ml of fl as k ml th e m eth yl (5 2) of vo l u m e tri c D i l u te th i s to S o l u ti o n , fl as k (5 2) an d s o l u ti o n s mg, ) wi th mg s o l ve n t an ti o xi d an t ( 30 vo l u m e tri c e xactl y i n to th e a 00 s o l ve n t 00 mg ml of an ti o xi d an t vo l u m e tri c ( ) fl as k Tran s fe r ml (5 2) of pre - co l u m n an al ys i s (5 ) an d by an d m ean s d i l u te of to a pi pe tte th e m ark (5 3) , wi th to th e a ml vo l u m e tri c th i s fl as k s o l ve n t OC I n to of n e are s t i n to th e (B H A s h al l be re m o ve d afte r a s e ri e s co n d i ti o n e d o ve rn i g h t at of fi ve ml vo l u m etri c fl asks (5 21 , i n tro d u ce , by e ach m e an s d ay th e S tan d ard We i g h , apparatu s (5 2) or S tan d ard d e vi ce Te m pe ratu re s l e e ve as fo l l o ws (4 4) wi th S o l u ti o n , Th e i n te rn al We i g h , I n j e cti o n - S tan d ard p re p are d u n d e can o ate P ro ce d u re I n te rn al S o l u ti o n , d i l u te m i xtu re s bal an ce D etecti o n See S tan d ard th e of a pi pe tte (5 3) , 0, 0, - - 0, - an d ,2 ml of th e te s t an ti o xi d an t S o l u ti o n Ad d , by m e an s of a pi pe tte , to e ach fl as k, te m pe ratu re ml of wi th N OTE - Ve ri fy passi n g , th e fro m th e ti m e proper to d i ti o n ti m e , a fat or of oi l th e of sl e e ve kn own or pre - co l u m n co m po si ti o n Th e s e an d co l u m n Te m pe ratu re N OTE - Fl o w rate e s tabl i s h e d by u n d er of th e i s o th e rm al carri e r g as co n d i ti o n s : Opti m u m : 60 OC val u e to fi ve - wi th fi rs t th e u se, carri e r i n tern al per by m e th yl d i l u te to th e m ark co n tai n , re s pe cti ve l y, 2, 5, 8, an d pg m i l l i l i tre m e an s of a bl an k u n d ecan oate If test, th ere i s, th at u se th ere i s no m e th yl i n te rferen ce as th e m yri state Stan d ard be th e fi l l ed co l u m n s fo r 24 h at 2 OC D e te rm i n ati o n pl o tti n g th e of cal i b rati o n th e cal i brati o n co e ffi ci e n t an d g rap h fl o wi n g I n j e ct e ach cal cu l ate an d O p e rato r co n d i ti o n g as s o l u ti o n s Ve ri fy, th e B e fo re (7 2 ) s o l ve n t an ti o xi d an t wi th - S tan d ard by of O ve n th e i n te rn al th rou g h ch rom atog raph th e S o l u ti o n th e (7 2 2) pro po rti o n al i ty i n to th e ch ro m ato g rap h K co e ffi ci e n t fro m th e an d fo rm u l a D e te cto r - Te m pe ratu re : 250 OC K =- A, X- As - Fl o w rate of au xi l i ary g as e s ms ma : wh e re h yd ro g e n : appro xi m ate l y 20 m l /m i n Aa r or O xyg e n : acco rd i n g to th e m an u factu re r’ s is th e are a of th e pe ak co rre s po n d i n g to th e an ti - o xi d an t; i n s tru c- ti o n s As is th e are a of th e pe ak co rre s po n d i n g to th e i n te rn al S tan d ard ; C al i brati o n m, U s e th e of i n te rn al a kn o wn n ot i n te rfe re wi th co rre cte d u si n g co m p are d wi th co m po u n d cal i brati o n co m po u n d , m e as u rem e n ts is in th e g ram s , of an ti o xi d an t in th e S tan d ard S o l u ti o n ; M e th o d th e m e th o d , pe ak o th e r pe aks , th e p e aks of th e i r th e in wh i ch i s ad d e d of re s pe cti ve m e as u re m e n t a kn o wn co rre s p o n d i n g th e to th e th e of th e i s th e m ass , in g ram s , of i n te rn al S tan d ard ad d e d d oes an d th e co n s ti tu e n ts , co e ff i ci e n ts , p e ak m, q u an ti ty wh i ch sam pl e vari o u s cal i brati o n of to are kn o wn If re q u i red , p e aks pl o t a g rap h co rre s po n d i n g co rre s p o n d i n g co n ce n trati o n s abs ci s s ae to of to th e h avi n g th e th e rati o s an ti o xi d an t i n te rn al an ti o xi d an t S tan d ard in th e to of th e as th e are a are a of o rd i n ate s s o l u ti o n s of th e an d i n j e cte d th e p e ak th e as