ANALYSIS OF GAMMA-LINOLENIC AND OTHER FATTY ACIDS IN THE EDIBLE PART OF BORAGE (Borago officinalis L.).

M. del Roa, A. de Harob a C.I.D.A. Apdo. 4240. 14.080 CORDOBA b Inst. Agricultura Sostenible C.S.I.C. Apdo. 4084. 14080 CORDOBA.

ABSTRACT

Borage (Borago officinalis .! is currentl" used as a #egetable in t$e %ort$ o& Spain. '$e edible parts o& t$e plant are t$e basal lea& petioles( lea#es and ste). '$e ob*ecti#e o& t$is stud" $as been to deter)ine t$e oil( ga))a+linolenic acid and ot$er &att" acids content in t$e tissues o& borage. '$e lea#es ,ere t$e )ost i)portant source o& lipid ,it$ - . dr" ,eig$t( ,$ile basal lea& petioles and ste) ,ere 1.8 . and 0.-2 . respecti#el". Al&a+linolenic acid and / A are &or)ed b" desaturation o& linoleic acid b" delta+0 and delta+11+desaturases( respecti#el"( but t$e acti#it" o& t$e delta+11+desaturase predo)inates. Al&a+linolenic ,as t$e )a*or constituent ,it$ about 23. in tissues p$otos"ntetics and 24. in ste) and basal lea& petioles. Basal lea#es and )ain ste) lea#es s$o,ed t$e $ig$est content in / A 11.8 . and 11.1. respecti#el". In contrast to t$e seeds( all t$e tissues anal"4ed $ad stearidonic acid (1854 n+2!. '$e )a*or &att" acids ,ere5 pal)itic( al&a+linolenic and stearidonic acids in lea#es( and pal)itic( linoleic and al&a+linolenic acids in ste) and basal lea& petioles. In#erse relation ,as &ound bet,een

oleic acid and / A and bet,een linoleic #s. al&a+linolenic and stearidonic acids in basal lea#es. In )ain ste) lea#es and basal lea& petioles ,as also &ound in#erse relation bet,een al&a+linolenic #s. oleic and linoleic acids. . INTROD CTION

/a))a+linolenic acid (/ A( C518( n+0(-(12( cis! is one essential &att" acid precursor o& indispensable co)pounds in t$e bod" suc$ as prostaglandins (6/7 1! and leu8otrienes

( '!( o&ten 8no,n as eicosanoids. 9an" &actors $a#e been s$o,n to inter&ere t$e biocon#ertion o& linoleic to / A li8e ageing( stress( diabetes( $ig$ alco$ol inta8e( $ig$ c$olesterol le#els( #iral in&ections and nutritional de&iciencies. Since t$ere is so)e e#idence t$at 0+desaturase acti#it" is i)paired in eac$ o& t$ese situations( ,it$ an e:ogenous suppl" o& / A t$ese )a" be a)eliorated ;1<. Seeds o& se#eral plants suc$ as e#ening pri)rose (Oenothera biennis! and t$e co))on borage (Borago officinalis! and t$e &ruits o& Ribes spp.( $a#e been &ound to contain substantial proportions o& GLA as a constituent o& t$eir lipids ;2( 2( 4 <. =$ite &lo,ered borage is currentl" used as a #egetable in t$e %ort$ o& Spain( but no attention $as been paid till no, to stud" as source o& / A. '$e parts o& t$e plant t$at are pre&erentiall" eaten are t$e basal lea& petioles (be&ore t$e plant is co)pletel" de#eloped and be&ore &lo,ering! lea#es and ste). ipids containing GLA are unusual constituents o& tissues in $ig$er plants( ,$ic$ are generall" ric$ in al&a+linolenic acid (all+cis+-(12(11+octadecatrienoic acid!. Apparentl" a plant t$at s"nt$esi4es substantial a)ounts o& / A acid in t$e seeds( s$ould &or) t$is acid in t$e lea#es. >o,e#er t$e lea#es o& t$e e#ening pri)rose plants contain al&a+linolenic as )a*or constituent o& p$otos"nt$etic tissues but not / A ;1<. '$e ob*ecti#e o& t$e present stud" ,as to deter)ine t$e oil and &att" acids content in t$e edible portion o& t$e borage ,it$ e)p$asis on / A acid.

MATERIAL

Seeds o& 12 accessions o& ,$ite &lo,ered borage ( B. officinalis

.! ,ere pro#ided b"

'$e Center o& Agricultural Researc$ o& Rio*a . Borage plants ,ere gro,n in a 10 $. p$otoperiod at 11?C and an 8 $. dar8 at 10?C. Collections o& t$e basal lea& petioles and basal lea#es ,ere )ade be&ore t$e plants ,ere co)pletel" de#eloped and be&ore &lo,ering. '$e ste) and )ain ste) lea#es ,ere collected a&ter &lo,ering.

METHOD

L!"!d E#$ra%$!o& a&d A&al'(!(. Oil content ,as deter)ined according to t$e Association o& O&&icial Anal"tical C$e)ists. (AOAC( 1-84( 9et$od 3002! ;0<. '$e &att" acid co)position o& oil ,as deter)ined b" / C (/as i@uid C$ro)atograp$"! o& t$e corresponding )et$"l esters( using as @uanti&ier )et$"l$eptadecanoic acid as an internal standard . Digestion( trans)et$"lation o& lipids &ro) lea#es( petioles and ste) and e:traction o& &att" acid )et$"l esters in one step ,as reali4ed according to /arcAs and 9anc$a ;3<. '$e sa)ples ,ere $eated at 80?C ,it$ a reagent containing )et$anol5 $eptane5 toluene5 2+2+di)eto:"propane5 >2SO4 (235205205152!. A&ter 120 )in. t,o p$ases ,ere &or)ed( t$e upper one containing t$e &att" acid )et$"l esters read" &or / C anal"sis. '$e anal"sis ,ere per&or)ed using a 6er8in 7l)er Autos"ste) /as C$ro)atograp$ e@uipped ,it$ &la)e ioni4ation detector and split in*ector. '$e c$ro)atograp$ is e@uipped ,it$ a capillar" colu)n (21) : 0.21 ))( i.d. 0.21 ) &il)! ,it$ acidi&ied polietilengl"col as t$e stationar" p$ase. O#en te)perature ,as progra))ed &ro) 1-0?C to 210?C at a rate o& 2?CB)in. '$e te)perature o& t$e detector and in*ector ,ere 231?C and 200?C respecti#el". %itrogen ,as used as carrier gas.

RES LTS

'$e data gi#en in 'able I s$o, t$e oil and &att" acids content in t$e portion edible o& borage in co)parison ,it$ t$e content o& seeds ;8<. ea#es s$o,ed t$e $ig$est content o& oil ,it$ a )ean #alue o& -. dr" ,eig$t. Basal lea& petioles and ste) $ad a )ean #alue o& 1.8. and 0.-2. respecti#el". '$e )a*or &att" acid in all t$e tissues. ,as al&a+ linolenic acid . Catt" acids co)position ,as #er" si)ilar in basal lea#es and )ain ste) lea#es o& borage plant. '$e lea& lipids contained as t$e )a*or constituents( about 11. pal)itic acid( 23. al&a+linolenic and 13. steridonic acids. %e#ert$eless( basal lea& petioles and ste) s$o,ed as t$e )a*or &att" acids( about 20. pal)itic acid( 1-. linoleic acid and 24. al&a+linoleic acid.

'$e / A ,as present in all t$e tissues anal"4ed. '$e basal lea& petioles and ste) s$o,ed t$e $ig$est content o& / A ,it$ a )ean #alue o& 11.8 and 11.1 respecti#el". In contrast to t$e borage seeds( stearidonic acid ,as s"nt$esi4ed in substantial a)ounts in all t$e tissues e#aluated( ,it$ t$e $ig$est le#els( 13.04 and 18.32 in basal and )ain ste) lea#es. S$ort+c$ain &att" acids suc$ as lauric and )iristic acids and traces o& C20+ C24 &att" acids ,ere &ound in t$e tissues e#aluated. In#erse relation ,as &ound bet,een oleic acid and / A and bet,een linoleic #s. al&a+ linolenic and stearidonic acids in basal lea#es. In )ain ste) lea#es and basal lea& petioles ,as also &ound in#erse relation bet,een al&a+linolenic #s. oleic and linoleic acids.

DISC SSION

In contrast to t$e borage seeds( t$e lipids o& basal lea& petioles( lea#es and ste) o& plant contained t,o n+2 acids al&a+linolenic and stearidonic acids. Al&a+linolenic acid and / A ,ere detected in all t$e tissues o& borage( ,$ic$ indicates t$at bot$ delta+11 and delta+0 desaturases are acti#e( but t$e e&&ect o& t$e delta+11 desaturases predo)inates. '$ere&ore al&a+linolenic is t$e )a*or octadecatrienoic acid in t$e tissues o& borage. '$ese results support t$at al&a+linolenic acid is c$anneled al)ost e:clusi#el" to t$e p$osp$olipids and gl"colipids( ,$ic$ are t$e )a*or constituents o& t$e cellular )e)branes. =$ereas / A is esteri&ied )ainl" in t$e storage lipids( i.e triac"lglicerols o& borage seeds( suc$ as occur ,it$ e#ening pri)rose plant ;-<. %e, studies are being reali4ed in our group o& in#estigation &or deter)ining t$e proportions o& t$ese lipids in t$e tissues o& borage. '$e in#erse relation &ound bet,een / A and oleic acid in basal lea#es suggests t$at / A is &or)ed to desaturation o& oleic acid.( ,$ic$ corroborates t$e $"pot$etical bios"nt$etic pat$,a"s to &att" acids in borage seeds t$at proposes /unstone(;10< and corroborates results obtained b" C$err" et al. ;11< and =$ip8e" et al. ;12<( in ,$ic$ t$e se@uence o& alternating desaturations &ro) oleic acid to / A is signi&icant. >o,e#er( ot$ers patterns o& desaturations )ust be considered &or e:plain t$e n+2 acid s"nt$esis.

Considering t$e in#erse relation &ound in basal lea#es( bet,een linoleic acid #s. al&a+ linoleic and stearidonic acids( as ,ell as in )ain ste) lea#es and basal lea& petioles bet,een al&a linoleic acid #s. oleic acid and linoleic acid t$e results are co)patible ,it$ t$e &ollo,ing se@uential desaturations5 oleic acid linoleic acid ga))a+linolenic acid al&a+linolenic acid (1852( n+2!. stearidonic acid (1854( n+4!. Additional ,or8s are being reali4ed in order to e:plain t$e probable bios"nt$etic pat$,a"s to &att" acids in borage.

AC)NO*LEDGMENTS

'$e aut$ors ,is$ to e:tend t$eir appreciation to Dr. Dicente Do)ingue4 (Centre o& Agricultural Researc$ o& Rio*a! &or pro#iding t$e accessions o& borage. REFERENCES

1 2 2 4 1 0 3 8 8 10 11

D. C. >orrobin. 6lant ipid Res. Dol. 21( %o. 2( (1--2! 102. B. E. C. >udson. E. A). Oil C$e). Soc. 01 (1-84! 140. >. 'raitler( >. =inter( F. Ric$li and G. Ingenblee8. ipids. 1- (1-84! -22. R. B. =ol&( R. Hlei)an and R. 7. 7ngland. E. A). Oil C$e). Soc. 00 (1-82! 1818. H. D. 9u8$er*ee and I. Hie,itt. E. Agric. Cood C$e).( Dol. 21( (1-83! 100-. AOAC . O&&icial 9et$ods o& Anal"sis( =illia)s( Arlington( 1-84. R. /arcAs and 9. 9anc$a. Anal"tical Bioc$e)istr"( Dol. 211 ( (1--2! 12-. 9. del RIo( E.9.CernJnde4 and A. de >aro. /rasas " aceites. 44 (1--2! 121. /. ( otti and 9. C.( Kuartacci. Agroc$i)ica. 24 . (1--0! 242. C. D. /unstone. 6lant ipid Res. Dol. 21( %o. 2( (1--2! 141. E. >. C$err"( . Bis$op( 6. 9.( >asega,a and >. R.( e&&ler. Ibid. 24 .(1-81!. 223 A.( =$ip8e"( E. 7. Si)on and E. Eanic8. EAOCS. 01. (1-88! -3-.

TABLE I Fa$$' A%!d Co+"o(!$!o& a&d o!l %o&$e&$ o, $-e ed!.le "or$!o& o, .ora/e (da$a are /!0e& a( "ea1 area "er%e&$a/e(). o!l %o&$e&$ a +a!& ($e+ lea, .a(al lea, .a(al lea, "e$!ole( ($e+ (eed 9 9 2.8 5.92 :6.: 2345 . :.;6 5.938.36 :.:2 5-8.52 ------------------2645 2745 2742 2845 2842

28

5.28 26.:5 2.7 3.9< :.57 5-5.;9 23.63-28.58 5.:2-6.<3 2.<6-<.7: 2.2:-<.7; 2 5-8.; 2<.;: 23.67-28.29 2.2: 5-2.83 2.7: 5-:.86 2.62 5.<-3.56 ----------:.5; :.36 3.57-6.69 2.52-7.2< :.<9 5-;.2< 2.8; 5-6.38 :.87 3.6-9.3 :.;8 5-8.66 3.3: 5-<.37 28.6: 9.9-:5.3

9. ;.7;-

9. ;.56-

5.89 35.9 5-3.:6 29.59-32.92 2.:; 32.22 5-3.2: 2;.:9-37.<8 25.88 8.<-28.;

2; 2:.26

32 27-9;

---------- ---------

:8 37.;

a O!l %o&$e&$ !( /!0e& a( "er%e&$a/e( o, dr' =e!/-$. . Fa$$' a%!d( are de(!/&a$ed .' &>+.er o, %ar.o& a$o+(4&>+.er o, do>.le .o&d(. % T-e &-7 (or o+e/a-7) a&d $-e &-: (or o+e/a-:) de,!&e $-e "o(!$!o& o, $-e ,!r($ do>.le .o&d !& $-e +ole%>le ($ar$!&/ ,ro+ $-e %ar.o& a$o+ a$ $-e +e$-'l o, $-e %-a!&. dI&%l>d!&/ 3545, 3542, 3345 a&d 3342. eI&%l>d!&/ 3542 a&d 3342.