The fatty acid composition of grapefruit seed oil

Item Type text; Dissertation-Reproduction (electronic)

Authors Teles, Francisco Franco Feitosa, 1941-

Publisher The University of Arizona.

Rights Copyright © is held by the author. Digital access to this material

is made possible by the University Libraries, University of Arizona.
Further transmission, reproduction or presentation (such as
public display or performance) of protected items is prohibited
except with permission of the author.

Download date 29/09/2019 16:59:55

Link to Item http://hdl.handle.net/10150/565233

 THE FATTY ACID COMPOSITION

OF GRAPEFRUIT SEED OIL

by

F ra n c is c o F ranco F e ito s a T e le s

A T h e s is Subm itted to th e F a c u lty of th e

COMMITTEE ON AGRICULTURAL BIOCHEMISTRY AND NUTRITION

In P a rtia l F ulfillm ent of th e R equirem ents

For th e D eg ree of

MASTER OF SCIENCE

In th e G rad u ate C o lleg e

THE UNIVERSITY. OF ARIZONA

19 7 1
 STATEMENT BY AUTHOR

T his t h e s i s h as b e e n su b m itted in p a rtia l fu lfillm en t of r e ­

q u irem en ts for an a d v a n c e d deg ree a t The U n iv e rsity of Arizona and is
d e p o s ite d in the U n iv e rsity Library to be m ade a v a ila b le to borrow ers
un der ru le s of th e Library.

Brief q u o ta tio n s from th is t h e s i s are a llo w a b le w ith o u t s p e c i a l

p e r m is s i o n , provided th a t a c c u r a te ack n o w led g m en t of s o u rc e is m a d e .
R e q u e sts for p e rm is sio n for e x te n d e d q u o ta tio n from or re p ro d u ctio n of
th is m a n u sc rip t in w hole or in part may be g ranted by th e h ead of the
m ajor d ep artm en t or th e D ean of th e G rad u ate C o lle g e w hen in his ju d g ­
m ent th e p ro p o sed u s e of th e m a te ria l is in th e i n t e r e s t s of s c h o la r ­
s h ip . In a ll o th er i n s ta n c e s , h o w e v e r , p e rm is sio n m u st be o b tain ed
from th e au th o r.

SIGNED:

APPROVAL BY THESIS DIRECTOR

T his t h e s i s h a s b een approved on th e d a te shown below:

L x a te
J . W arren Stull
"ofessor of D airy and Food S c ie n c e s
 ACKNOWLEDGMENTS
< '

The a u th o r w is h e s to e x p r e s s h is s in c e re a p p r e c ia tio n to th e

follow in g mem bers of The U n iv e rsity of A rizon a, T u c s o n , Arizona:

D r. J . W arren S t u l l , P ro fe s so r of D airy and Food S c i e n c e s , for

h is a d v ic e th ro u g h o u t t h is s tu d y . I am s in c e r e ly g r a te fu l for his p a tie n t

u n d e rs ta n d in g and a s s i s t a n c e e x te n d e d to me in th e p re p a ra tio n of th e

m a n u s c rip t. D r. G erald H. S to tt, H e a d , D epartm ent of D airy and Food

S c i e n c e s , D r. W illiam H . Brown, A s s o c ia te P ro fe s so r of D airy and Food

S c ie n c e s and D r. Thomas N . W e g n e r , A s s i s ta n t P r o fe s s o r , D ep artm en t

Of D airy and Food S c i e n c e s .

G ra titu d e is e x te n d e d t o D r. W illiam J. P i s t o r , for h is h elp

and e n c o u ra g in g w o r d s .

A p p reciatio n is a l s o e x te n d e d to Dr. W illiam G. M atlo ck , C am pus

co o rd in a to r of th e U n iv e r s ity of A r iz o n a /U n iv e rs ity of G e a r a C o n tr a c t.

D r. Ralph L. P r i c e , A s s i s ta n t P ro fe s s o r, is th a n k e d for th e

d o n a tio n of fruit s a m p le s and for h is h e lp during a p o rtio n of my s tu d y .

I w a n t to th a n k a ll my c o lle a g u e s w h o se h e lp e n a b le d me to

co m p lete t h i s s tu d y .

S p e c ia l re c o g n itio n is g iv en to my w i f e , E n e i d a , w h o s e

p a tie n c e and u n d e rs ta n d in g made t h is work p o s s i b l e .

 TABLE OF CONTENTS
< ■
Page

LIST OF TABLES . ........................................................ v

ABSTRACT ................... vi

INTRODUCTION . . . .. ............................. 1

LITERATURE REVIEW................................ 3

H i s t o r i c ' ................ .... . .................................... 3

B o t a n i c ................................... 4
G rap efru it Seed O il . . . . A '..................... 5
O il P ro c e s s in g . ........................................ 8
Im portance of H a rv e stin g Time ........................................ 9
S eed C o m p o sitio n . ........................ 11
F atty A cids .................... 13

MATERIALS AND M E T H O D S ................................ . . ' . .....................................15

S a m p l e s ............................................................................................ 15
Sample C o l l e c t i o n ........................................ 16
S eed P r e p a r a t i o n ........................................................ . 17
Oil E x t r a c t i o n ................................................................................ 17
M ethy l E s te rs P rep aratio n . ..................................... 17
G a s -L iq u id C hrom atography . . . . . . . ......................................... 18

RESULTS AND DISCUSSION . . . . . . . . . . . . . . . . . . . . . . 19

CONCLUSIONS ........................... 37

. LITERATURE C I T E D ...................................................... 38
 HST OF TABLES

T able Page

1. F atty Acid C o m p o sitio n of Burgher G rapefruit S eed O il

R elated to H a rv e st D ate . . . ................................................ 20

2. F atty Acid C o m p o s itio n of C h a n d le r G rapefru it S eed

Oil R elated to H a rv e st D a t e ............................................ 21

3. F atty Acid C o m p o sitio n of Yuma Yellow G rap efru it S eed

Oil R elated to H a rv e st D ate . ........................ 22

4. F a tty Acid C o m p o sitio n of Yuma Pink G rapefru it Seed

O il R elated to H a rv e s t D a t e .................................... 23

5 . F atty Acid C o m p o s itio n of U . of A. G rap efru it Seed

Oil R elated to H a rv e st D a t e ........................................ . . 24

6. A verage C o n te n t of S atu ra te d and U n s a tu ra te d F atty Acids

of A riz o n a 's G rapefruit S eed O i l s ................................ 28

7 . T otal Lipids , Petroleum Ether E x tract and I n d iv id u a l Air

D ried S eed W e ig h t of Burgher G r a p e f r u i t ......................... . 30

8. T o tal L ip id s , P etroleum E ther E x tract and I n d iv id u a l Air

D ried S eed W e ig h t of C h a n d le r G r a p e f r u i t ........................ 31

9. T o tal Lipids , Petroleum E ther E x tra c t and I n d iv id u a l Air

, D ried S eed W e ig h t of Yuma Yellow G r a p e f r u i t .................... 32

10. T o tal Lipids , Petroleum Ether E x t r a c t , and I n d iv id u a l

Air D ried Seed W e ig h t of Yuma Pink G r a p e f r u i t ................ 33

11. T o tal Lipids , Petroleum Ether E x tr a c t, and In d iv id u a l Air

D ried Seed W eight of U. of A. G r a p e f r u i t . 34

12. The Average C o n te n t of T otal E x tra c ta b le L ipid, Petroleum

Ether E x tract and T heir R atios . ........................................ . 35

v
 ABSTRACT

S eed s from five Arizona g r a p e f r u i t , C itru s p a r a d i s i (M acf.)

v a r i e tie s w ere a n a ly z e d by g a s liq u id chrom atography for f a tty a c id

c o m p o sitio n . Three M arico p a co u nty g ro v es (Burgher, U. of A. , and

C handler) and two in th e Yuma a re a (Yuma Pink and Yuma Yellow) w ere

u s e d as s o u rc e s of g ra p e fru it. Fruit w ere p ic k e d a t a p p ro x im a te ly

2 w eek in te rv a ls during th e 1969-70 growing s e a s o n .

Six fa tty a c i d s in clu d in g m y r is tic , p a lm itic , s t e a r i c , o l e i c ,

l i n o l e i c , and lin o le n ic w ere m e a s u re d . There w as s u c h a s im ila rity

b e tw e e n a ll s a m p le s t h a t n e ith e r v a rie ty no r h a r v e s tin g tim e could be

d i s tin g u is h e d on t h e b a s i s of s e e d fa tty a c id a n a l y s i s .

The A rizona g rap efru it s e e d o il a n a ly z e d in t h i s stu dy had an

av e ra g e of 65% u n s a tu r a te d and 35% s a tu r a te d f a tty a c i d s . The air

d ried s e e d s c o n ta in e d a p p ro x im a te ly 30% t o ta l e x tr a c t a b le lip id s of

w h ich 25% w a s petroleum e th e r s o lu b l e .

 I

INTRODUCTION

The a v e ra g e y ie ld of s e e d s from g r a p e f r u it, C itru s p a ra d is i

( M a c f . ) , is about 4 .7 per c e n t of w e t s e e d c o n ta in in g 55 to 60 per c e n t

m o i s t u r e , b a s e d on th e w e ig h t of th e w hole fruit (16). A ccording

to th e sam e au th o rity th e a ir d rie d s e e d s c o n ta in 28 to 35 per c e n t

o i l , a ty p ic a l a n a l y s is bein g g iv en a s 3 0 .3 per c e n t e th e r e x tr a c t from

s e e d c o n ta in in g 1 1 .9 per c e n t m o istu re . S eed s from tw o v a r i e tie s

grown in th e W e s t In d ie s w ere found to c o n ta in from 41 to 53 per c e n t

o il. In t h e s e s e e d s , o n e -fo u rth of th e w eig h t w as h u ll and th r e e - f o u r th s

w as k e rn e l (3).

The s e e d s of g rap efru it and o th er c itru s fru its are ric h in oil

and alth o u g h th e prop ortio n of s e e d to fru it is s m a l l , th e to n n a g e a v a i l ­

a b le a s a byproduct of th e c itru s fruit p r o c e s s in g in d u s try is c o n s id e r ­

a b le. M eth o d s for s e p a r a t i n g , drying and m illing th e s e e d have b e e n

d e v e lo p e d and com m ercial prod uction of c itru s s e e d o ils h a s become

large enough to be e x p r e s s e d in term s of t a n k - c a r s p er m onth (3).

There are no p u b lish e d re p o rts of the c h a r a c t e r i s t i c s of

Arizona g rap efru it s e e d o i l . W ith th e p o s s ib i l it y of in c r e a s e d in t e r e s t

in g rap efru it s e e d o i l , th e re a p p e a re d to be an a d v a n ta g e in stu d y in g th e

s e a s o n a l and v a r ie t a l d iffe re n c e s of A riz o n a 's s e e d o il p o t e n t i a l .

F u rth e rm o re , th e a d v e n t of improved g a s ch ro m ato g rap h ic p ro c e d u re s for

1 '
 2

th e q u a lita tiv e and q u a n tita tiv e d e te rm in a tio n s of fa tty a c i d s h as

g re a tly f a c i l i t a t e d a stu d y of t h is t y p e . In th is w o r k , th e s e a s o n a l and

v a r i e t a l d iff e r e n c e s in th e fa tty a c id c o m p o sitio n o f five A riz o n a 's

g rap efru it s e e d o ils w ere s tu d ie d .

/
 LITERATURE REVIEW

H is to ric

G rap efru it , C itru s p a r a d is i ( M a c f .) , is a n a tiv e of

M a la y s ia and P o l y n e s i a . It w as in tro d u c e d in to J a m a i c a , w here it

b ecam e of im p o rtance co m m ercially and w a s grown e x t e n s iv e ly (4).

G rap efru it may h ave come in to e x i s t e n c e a s a m utant or s e e d lin g in th e

W e s t I n d i e s , p erh ap s from s e e d in tro d u c e d by S h ad d o ck . C a p ta in

° S h a d d o c k , an E a s t In d ia n s e a c a p t a i n , is c r e d ite d w ith bringing s e e d of

th e s h a d d o c k (pummelo) to th e W e s t I n d i e s .

The e x a c t d e t a i l s of th e in tro d u c tio n of th e pom elo (grapefruit)

into F lorida are u n k n o w n . In all p r o b a b i li t y , it w as b rought to Florida

t o g e th e r w ith o th e r mem bers of the g en u s by the S p a n i a r d s , w h o , under

th e l e a d e r s h ip of Ponce de L e o n , fir s t la n d e d upon th e c o a s t of F lorida

in th e y e a r 1513. It is known th a t Don P h illip s , a S p a n is h n o b lem an ,

s e t t l e d in F lorida in ab ou t 1809. He brought g rap efru it s e e d s w ith him

and p la n te d th em . W ith th e tr e e s o b ta in e d from t h e s e s e e d s , and w ith

o th e f t r e e s , he s e t out a c itru s grove a t t h a t point and brought it in to

b e a rin g (10).

3
 Botanic

In a rev iew w h ich d i s c u s s e s th e c h e m ic a l c o m p o sitio n of a

n a tu ra l p la n t p r o d u c t , it is d e s ir a b le to id e n tify as a c c u r a te ly a s

p o s s ib l e th is p a r tic u la r fruit u nder d i s c u s s i o n . G rap efru it b e lo n g s to

the Rue F am ily — R u t a c e a e . The Rue fam ily in c lu d e s a b o u t 100 g e n era

and 900 s p e c i e s of h erb s , sh ru b s , and t r e e s , m ost of w h ic h req u ire

tro p ic a l or s u b tr o p ic a l c lim a te s but some of w h ich e x te n d in to te m p e ra te

r e g io n s . The d ev elo p m en t of oil g la n d s c o n ta in in g frag ran t e s s e n t i a l o il

in l e a v e s , fruits , or o th e r p arts of th e p la n ts is c h a r a c t e r i s t i c of th e

fam ily . Some of the members, of the fam ily are fam iliar o rn am en tal

p l a n t s , but th e m o st u s e fu l and w ell known members are th e c itru s fru it

t r e e s , w here g rap efru it is a s p e c i a l i z e d form or v a r ie ty (3) . Unfor­

t u n a t e l y , th ere is a la c k of ag reem en t among ta x o n o m is ts a b o u t th e

e x a c t b o ta n ic a l n o m en clatu re of c itru s s p e c i e s , and th e m a tte r is

fu rther c o m p lic a te d by th e e x i s te n c e of many m utan ts , hy brids , and

h o rtic u ltu ra l v a r i e t ie s (12).

It h as b e e n commonly a c c e p t e d , but not e s t a b l i s h e d , th a t

g r a p e f r u it , C itru s p a r a d is i (M a c f. ) , is a spo rt from the sh a d d o c k or

pum m elo. C itru s grand is (Linn.) O s b e c k , b e c a u s e of th e re s e m b la n c e of

th e tw o . It is p o s s i b le how ever t h a t th e tw o may not be r e la te d or th a t

th e g ra p e fru it may be a n a tu r a l c ro s s b e tw e e n s h a d d o c k and orange (8).

Although th e re is som e d is a g r e e m e n t, in 1938, H arold Hume (10)

referred to C itru s p arad is 1 (M acf.) (p a ra d is i of p a r a d is e or g a r d e n s ) .

 C o m m e rc ia lly , th e fruit i s known a s g r a p e fr u it. This a p p e lla tio n w as

g iv e n b e c a u s e th e fruit is so freq u en tly borne in g r a p e - l ik e c l u s t e r s

or from th re e or four to a d o z e n and a h a lf .

G rap efruit S eed Oil

One of th e e a r l i e s t re fe r e n c e s to g rap efru it s e e d o il a s a

com m ercial product w a s th a t of Ja m ie so n (11) who in 1932 rep o rted

both s o lv e n t e x tr a c te d and A nderson e x p e lle r o il s a m p le s fa tty a c id

c o n te n t, io dine num ber and o ther c h e m ic a l and p h y s ic a l c h a r a c t e r i s t i c s .

He s ta te d t h a t J a m ie s o n , Baughman and G e rtle r Oil and F at In d. ,

‘ 7, 181 (1930) . found th a t th e a ir d ried s e e d c o n ta in e d 30 per c e n t o il.

The e x p r e s s e d o il g av e th e follow ing c h a r a c t e r i s t i c s : s p e c i f i c g rav ity

at 25°C 0 .9 1 7 0 ; re fr a c tiv e in d e x 1 .4 7 0 0 ; iodine n o . (Hanus) 1 0 6 .3 ;

s a p o n if ic a tio n v a lu e 1 9 4 .1 ; a c id v a lu e 2 .5 ; a c e ty l v a l u e 7 .7 ; u n s a p o n i-

fia b le m a tte r 0.7% ; s a tu r a te d a c id s 28.60% ; u n s a tu r a te d a c i d s 68.50 % .

The o il c o n ta in e d th e fo llow ing p e r c e n ta g e s of fa tty a c i d s : o le ic 1 9 .6 6 ,

l i n o le ic 4 8 .8 4 ; p a lm itic 1 9 . 2 , s te a r i c 7 . 2 5 , and lig n o c e r ic 0 . 1 6 .

The s o lv e n t e x tr a c te d m eal w as ex am in ed w ith th e fo llo w in g

p e rc e n ta g e c o m p o sitio n rep o rted : m o istu re 9 .7 ; a s h 3 .8 ; fat 4 . 3 ,

p ro te in 2 1 .4 ; crude fib e r 2 2 .0 ; and n itro g e n - f re e e x tr a c t 3 8 .8 (11).

The b it t e r n e s s of the crude g ra p e fru it s e e d o il a p p e a re d

in i ti a l ly to p reclu d e its u s e in a n y th in g but s o a p m an u factu re s in c e a

f e a s i b l e m ethod for th e rem oval of th is t a s t e w as not a v a i l a b l e . By

 1940, h o w e v e r , N olle and von L o e seck (16) rep o rted p ro c e d u re s for the

com m ercial pro d u ctio n of g rap efru it s e e d o il in clu d in g e x p r e s s i o n ,

b l e a c h i n g , and w in terin g of th e o il in F lorida and p o in te d out further

th a t th e ex trem ely b itte r crude oil co uld be e a s i l y re fin e d to a bland

ta s t i n g pro du ct (5). In 1 9 3 8 -3 9 , th e p ro d u ctio n of c itru s s e e d o il in

Florida w as only 45 t o n s , and it w a s e s tim a te d t h a t a p p ro x im a te ly

2 ,0 0 0 to n s t o t a l cou ld h av e b e e n re c o v e re d (6).

A pproxim ately 40 to n s of g ra p e fru it crude o il w ere p roduced

an n u a lly in 1940 w hen a p o te n tia l of a lm o s t 2 ,0 0 0 to n s w a s s ti l l

t h e o r e tic a lly a v a i l a b l e . I n te r e s t in p ro d u ctio n of g rap efru it s e e d o il in

the s u b s e q u e n t 20 y e a r s a lm o st d is a p p e a r e d u n til r e c e n tly w hen p u b lic

. a tte n tio n w as c a lle d to th e p o s s ib le th e r a p e u tic u s e f u l n e s s of hav ing a

h ig h er proportion of u n s a tu r a te d fa ts in th e d ie t a s is fu rn is h e d by some

v e g e ta b le o ils (5). In 1954, M . J. W . K e ste rso n , c h e m is t at the

C itru s Experim ent S ta tio n , Lake A lfred, Florida d e c la r e d th a t one h a r ­

v e s ti n g s e a s o n can produce ab o u t 5 ,000 to n s of c itru s s e e d o il (9). In

th e l a s t few y e a rs , a n n u a l p ro d u ctio n h a s in c r e a s e d to ab o u t 1 ,50 0

to n s. On th e b a s i s of b o x es of s e e d y fru its p r o c e s s e d , it is e s tim a te d

th a t a th e o r e t ic a l p o te n tia l a s high as 1 4 ,7 2 0 to n s of s e e d oil could h a v e

b een rem oved in th e 19 61 -62 s e a s o n (6). According to th e U. S. D. A. Agri­

c u ltu ra l H and b oo k, 1968 (19), h o w e v e r, it h as b een e s tim a te d th a t if a l l th e

s e e d s from c i t r u s - p r o c e s s i n g p la n ts in Florida alo n e w ere u s e d for th e

e x tr a c tio n of o il around 4 or 5 th o u s a n d to n s of oil co u ld be o b ta in e d

a n n u a l l y . Only a s m a ll f ra c tio n of th e s e e d s are so u s e d , a s m ost of

them are d rie d w ith t h e w a s te c a n n e ry p e e l and rag to p ro d u ce dried

c itru s pulp for s to c k f e e d .

For o u r p u r p o s e , one s h o u ld n o tic e t h a t in F lo rid a , c itru s s e e d

o il is produced from a m ixture of p redo m inan tly g rap efru it and s m a lle r

q u a n titie s of orange s e e d s (6). Crude c itru s s e e d o ils are in g e n e ra l

s u it a b l e for th e u s u a l n o n e d ib le u s e s to w h ich s e m i- d r y in g o ils are p u t.

They in c lu d e s o a p m ak in g , s u lfo n a tio n for d e t e r g e n t s , and th e p r e p a ra ­

tio n of fa tty a c id d e r i v a t iv e s . G rap efru it s e e d o il h a s found some

com m ercial a p p lic a tio n in th e tre a tm e n t of t e x t i l e s and F eath er (19).

The fla v o r of a re fin e d c itru s s e e d o il r e s e m b le s g re a tly th a t .

of o liv e o i l , for w h ic h it some tim es h a s b een s u b s t i t u t e d . It is p ale

y e llo w in c o lo r, c o n s id e re d w h o le so m e and w e ll s u ite d for food and h a s

b een u s e d m o st s u c c e s s f u l l y a s a co o k in g oil and s a la d o i l . In ad ditio n,

c itru s s e e d o il c an be h y d ro g e n a te d to b ecom e a b u tte r s u b s t i t u t e or

cooking f a t . Som ewhat s im ila r ly , it c a n be bro m in ated to s e c u re a h ig h

s p e c if ic g rav ity o il (approxim ately 1 . 3 0 ) , w h ich may be u s e d by th e

b ev e ra g e in d u stry to a d j u s t the d e n s it y of flavoring o ils ad d ed to

beverages. Brominated c itru s s e e d o il is co m p etitiv e w ith other

b rom inated o ils , s u c h as a p ric o t k e rn e l s e e d o il and s e s a m e oil (6).

 O il P r o c e s s in g

C itru s s e e d s are o b ta in e d from c itru s p r o c e s s in g p l a n t s ,

w here larg e q u a n titie s of fru it h av e b e e n c o lle c te d an d u t i li z e d

prim arily for ju ic e or s e c tio n s p r o d u c tio n . The s e e d s are o b ta in e d

in tim a te ly m ixed w ith rag and pulp but s e p a r a t e from the p e e l . S e p a ra ­

tio n of s e e d s from the rag and pulp may be a c c o m p lis h e d in s e v e r a l

w a y s , but th e d iffic u lty of so do ing i s one of the c h ie f d e te r re n ts to

i n c r e a s e d p ro d u ctio n of s e e d o i l . S e p a ra tio n may be a c c o m p lis h e d

w ith a p a d d le - ty p e f in is h e r or s im ila r eq u ip m e n t. A nother p roced ure is

to lime th e m ixture of s e e d and w e t p u lp , a s in th e p re p a ra tio n o f drie d

c itru s f e e d , and dry in ro ta ry d r ie r s . The s e e d s can th e n be s e p a r a te d

from th e d rie d pulp by s c re e n in g and w innow ing (19).

D ried s e e d s from w h ich th e h u lls may or may not be rem oved

are p a s s e d through c ra c k in g r o l l s , and th e o il is e x tr a c te d in screw

e x p e lle r s of th e typ e commonly u s e d in p rep arin g c o tto n s e e d and tun g

o ils. T h e se m a c h in e s p roduce a p r e s s c a k e w ith an oil c o n te n t rang ing

from 14 to 16%. The oil c o n te n t of th e p r e s s cak e c a n be further

re d u c e d in t h e s e m a c h in e s but a t th e e x p e n s e of h e a t d am ag e to th e o i l .

M ore m odern m a c h in e s h av e b een d e v e lo p e d th a t w ill r e d u c e th e r e s id u a l

o il to 7 p e rc e n t w ith o u t h e a t d a m a g e , but t h e s e m a c h in e s are not y e t

(1968) in g e n e ra l u s e . S o lv en t e x tr a c tio n would r e d u c e th e r e s id u a l o il

e v e n f u r th e r , but so far a s is known t h i s h a s not b e e n a tte m p te d com ­

m e rc ia lly (19).
 The e x p r e s s e d o il is tra n s f e rr e d in to ta n k s w h ere it is allo w ed to

s e t t l e b rie fly and th e n pumped th ro u g h f ilte r s and in to s to ra g e t a n k s ,

w here it u n d erg o es a fin a l s e ttl i n g b efo re further p r o c e s s i n g . S ubse­

q uent tre a tm e n t of o il d e p e n d s on th e u s e for w h ich it is d e s ti n e d . For

e d ib le p u rp o s e s it is a lk a li r e fin e d . This tre a tm e n t rem o v es th e b itte r

p rin c ip le and free f a tty a c i d s . The l o s s e s in th is s te p u s u a ll y do not

e x c e e d 2%. The b la n d , n e u tra l o il r e s u l ti n g may th e n be h y d ro g e n a te d .

for u s e in s h o rte n in g p re p a ra tio n s o r, if n e c e s s a r y , may be w in te riz e d

if in te n d e d for a s a la d o il or liq u id s h o rte n in g . This l a t t e r tre a tm e n t is

= a c c o m p lis h e d by c h illin g u n til th e h ig h e r m eltin g g ly c e r id e s C ry s ta lliz e

and th e n rem oving them in a f ilte r p r e s s . Crude g ra p e fru it s e e d oil

w ill so lid ify around - 1 2 ° C to - 1 0 ° C (19).

Im po rtan ce of H a rv e stin g Time

In th e g re a t m ajo rity of p la n ts o il sto ra g e t a k e s p la c e in th e

s e e d in o i l - s t o r i n g - p l a n t - f r u i t s . The c o n d itio n s for th e fo rm atio n,

d ev elo p m en t and rip e n in g of the s e e d s are d eterm in ed by th e n atu re and

s tru c tu re of th e fruit t i s s u e s . Rapid s y n th e s i s of o il o c c u r s not only in

th e s e e d s of th e o i l - s t o r i n g p la n ts (su n flo w er, fla x , h em p , poppy, e t c . )

but in the s e e d s of th e g re a t m ajo rity of o th e r p la n ts (17 j .

Sam uel F r a s e r , in 1924 (4) p o in te d out th a t C a lifo rn ia is at a

d is a d v a n ta g e in com p eting w ith F lo rid a in th e e a s t e r n m a rk e ts on

a c c o u n t of th e g r e a te r e x p e n s e of r a il tra n s p o r ta tio n and th e tim e of y e a r

 10

at w h ich t h e ir crop m a t u r e s . C a lifo rn ia g rap efru it is c o n s id e re d a su m ­

mer fruit w h ile in F lorida it is a .w in te r and sprin g fru it. To m eet th is

s itu a tio n C a lifo rn ia is plan n in g to r a i s e m a te ria lly th e s ta n d a rd of its

pro du ct and to s u p p ly th e e a s t e r n m ark ets during l a t e sum m er and e a rly

f a l l w hen th e re is l i t tl e F lorida fru it. The M a rsh s e e d l e s s is th e v a r ie ty

w h ich h as b e e n c h o s e n by th e C a lifo rn ia in d u s tr y . This is a f l a t t e n e d ,

t h i n - s k i n n e d , j u i c y , s e e d l e s s v a r ie ty w h ich b e g in s to r ip e n in M ay (4).

The sam e a u th o rity s a id t h a t th e p ic k e rs sh o u ld h a r v e s t g r a p e ­

fru it for s i z e , b e c a u s e th e dem and in th e f a ll and e a rly w in te r is for

la rg e r s i z e s . M edium s i z e s are in l e s s dem and and s m a ll s i z e s are

at a d i s c o u n t . Toward th e l a t t e r end of th e s e a s o n c o n d itio n s are

rev ersed . The la rg e s i z e s are at a d i s c o u n t, many b u y ers p refer medium

s iz e and sm all fru it. The y ie ld and th e g ro w ers' incom e c a n be

in c r e a s e d by c a te r in g (4).

B e sid e s th is m arket problem th e r e are th e p h y s io lo g ic a l ones

co n c e rn e d w ith th e fru it i t s e l f su ch a s : "If ripe fru it is le ft on th e t r e e ,

th e s e e d s are apt to s p ro u t and th e fru its l o s e fla v o r, b u t in sto ra g e

th e a c id c o n te n t of th e fruit d e c r e a s e s and th e b itte r p rin c ip le b reak s

d o w n , w h ile th e s u g a r c o n te n t a p p e a rs to be u n a f fe c te d , so th a t the

fla v o r is im proved. Fruit h a r v e s te d in February or M a rc h may be h eld

for sh ip m en t in M ay to Ju ly . In F lo rid a , h a rv e s tin g p r o c e e d s from

N ovem ber to A ugust. Some fruit may be on t r e e s th ro u g h o u t the

y e a r (4). "
 . ' 11
A c o m p ariso n of th e p e rc e n ta g e of s e e d in th e d iffe re n t

v a r i e tie s of orange and g rap efru it during th e normal p r o c e s s in g period

w a s m ade by R. H e n d ric k so n and J. W . K e s t e r s o n , in 1965 (6). They

found th a t in the. l a te r s ta g e s of m a tu ra tio n th e p e r c e n ta g e of s e e d s by

w eight in c itru s is m arkedly in flu e n c e d by fruit m a tu r ity . Fruit w eigh t

i n c r e a s e s during t h i s p erio d a t a g r e a te r ra te th a n s e e d w e ig h t and

b rings about a c o n s ta n tly d e c r e a s i n g p e rc e n ta g e of s e e d w e ig h t in the

f r u i t . In many i n s t a n c e s th e re w a s a l s o a s m a lle r num ber of d e v e lo p e d

s e e d in th e fruit p ic k e d a t th e end of th e s e a s o n . Although th e re w ere

some i n c o n s i s t e n c i e s , th e s e e d s of a ll c itru s v a r i e tie s d e c r e a s e d in

m o istu re c o n te n t a s th e y m a t u r e . P e rc e n ta g e of o il in d rie d c itru s s e e d s

is v a r ia b le and d e p e n d s upon m a tu r ity . H ig h e s t o il c o n te n t u s u a lly

c o in c id e d w ith th e optimum m aturity n e e d e d for p r o c e s s in g (6).

Seed C o m p o sitio n

C itru s s e e d o ils are sem i drying o ils w h ich re s e m b le c o tto n ­

s e e d o il in th e n a tu re and d is tr ib u tio n of th e probable com ponent

g ly c e r id e s (12). It c a n be. d is t i n g u is h e d from the o t h e r s , h o w ev er, by

th e a b s e n c e or p r e s e n c e of c e r ta in fa tty a c id s or by th e p ro p o rtio n al

d iffe re n c e s in fa tty a c id c o n te n t (6).

A t w o - y e a r su rv ey of th e oil c o n te n t of th e d rie d s e e d s of

P in eap p le o ran g es and D u n can g rap efru it e s ta b li s h e d a v e ra g e s e a s o n a l

v a lu e s of 38% and 34%, r e s p e c t i v e l y . Although the oil c o n te n t of t h e s e

 12

s e e d s did not v ary w i d e l y , th e re w as a te n d e n c y for maximum oil

c o n te n t to o c c u r w hen th e fruit had r e a c h e d th e g e n e r a lly a c c e p te d

m a tu r ity . During th e 1965-66 c itru s s e a s o n , many pro blem s a s s o c i a t e d

w ith th e com m ercial p ro d u ctio n of c itru s s e e d o ils w ere i n v e s t i g a t e d .

|
It w a s show n t h a t (a) prolonged w et s e e d s to ra g e s e r i o u s ly i n c r e a s e d

the free fatty a c id c o n te n t of th e e x p e lle d o il , (b) th e w a te r c o n te n t of

c o lle c te d w e t s e e d s v a rie d from 5 6% to 71%, (c) im proved fin is h in g

te c h n iq u e s co uld lo w er drying c o s ts and d e c r e a s e th e fix ed c o lo r of th e

s e e d o i l , (d) d rain ed w a s t e liq u id from w e t s e e d s to ra g e b in s had a

b io lo g ic a l oxygen dem and of from 1 1 ,0 0 0 ppm to 2 0 ,0 0 0 ppm, and (e)

c o n s e c u tiv e t a n k - c a r lo ts of c itru s s e e d o il w ere m ost s im ila r.

I n v e s t i g a t i o n s of th e p ro te in c o n te n t of s e e d s and o th er

com ponents of d ried c itru s pulp h as s ta b l i s h e d a v e ra g e v a l u e s of 16% in

th e s e e d s , 9.4% in th e rag and pulp f ra c tio n , and 6.2% in th e f la v e d o -

a lb ed o p o rtio n . Removal of s e e d s from d rie d c itru s pulp d e c r e a s e d th e

p ro tein c o n te n t 8% to 15% w ith s e e d y v a r i e t i e s , but only 3% w ith the

l e s s s e e d y c itru s fruit (7).

The s e e d y v a r i e tie s w ere n o te d to have ap p ro x im a te ly th e sam e

num ber of s e e d s p er fruit w ith a tre n d tow ard a few er num ber of s e e d s a s

full or o v e r-m a tu rity w as re a c h e d (5). The range in dry s o li d s c o n te n t

of th e co m ponents of g rap efru it and s h a d d o c k w as: j u i c e 8 to 12%,

 13

a lb e d o 15 to 22%, fla v e d o 19 to 29%, rag and pulp 12 to 18%, s e e d s

30 to 68 p e r c e n t (8).

Seedy v a r i e t ie s of c itru s fruit s u c h a s s e e d lin g an d P in eap p le

o ra n g e s and D u ncan g ra p e fru it c o n ta in ab o u t 3.5% s e e d s , w h i c h in tu rn

c o n ta in from 55 to 60% m o istu re and ab o u t 15% o il. The a v e r a g e '

c o m p o sitio n of a ir - d r ie d c itru s s e e d s , co m piled from v a r io u s p u b lis h e d

d a t a , is a s fo llo w s: a s h 2 .4 8 to 3 .5 7%, crude fib e r 5 .4 1 to 1 7 .7 9 % ,

crude p ro te in 6 .9 4 to 1 5 .9 4 % , e th e r e x tr a c t 2 1 .8 8 to 3 4 .4 4 % , n o n -

.n itro g e n o u s s u b s t a n c e s 2 2 .2 1 to 4 4 .6 3 % , and w a te r 6 .8 2 to 1 1 .8 6

p e r c e n t (19). R. H e n d ric k so n and J. W . K esterso n (6) s t a t e d , h o w e v e r ,

th a t p e rc e n ta g e of o il in d rie d c itru s s e e d s is v a r ia b le and d e p e n d s

upon m a tu r ity . T y p ic a lly , th e d ried D u n can (grapefruit) s e e d s v a rie d

X

from 2 9 .2 p e r c e n t to 3 7 .3 p e rc e n t o i l.

F atty A cids

Early work by M a tla c k in 1929 (14) and in 1940 (15), had

show n t h a t th e p e e l and th e e n d o carp of o ra n g e s c o n ta in lip id m a te ria l

m ade up of o l e i c , l i n o l e i c , l i n o le n i c , p a lm itic , an d s t e a r i c a c id s a s

g ly c e r id e s and p robably a l s o s te r o ls (12). By a n a lo g y , th e s e e d s

s h o u ld c o n ta in a t l e a s t th e sam e q u a l i t a ti v e co m p o sitio n ,, and th is

s u p p o s itio n w as proved by D unn, H ild itc h and Rileg (2) who made an

in v e s tig a tio n in 1948, on sa m p le s from W e s t In d ian g r a p e fru its and

found th e follow ing co m p o sitio n :

 14

Variety F o s te r M a rs h

C o n s titu e n t

O il C o n te n t (%)£■ 43 41

S a tu ra te d a c id s : - p e r c e n t of t o ta l -
M y ris tic 0 .8 1 .2
P alm itic 2 8 .9 2 7 .5
S te a ric 2 .1 2 .9

U n s a tu ra te d a c id s :
O le ic 2 5 .1 2 1 .1
. L in o le ic 3 6 .6 3 9 .3
L in o len ic 5 .9 5 .9

(a) P ercen t by w eig h t of air d ried s e e d s

Later in 1961, H e n d ric k so n ,a n d K esterso n (5) d oing th e sam e

d e te rm in a tio n w ith g rap efru it from F lo rid a found th a t a ty p i c a l a n a l y s is .

of a F lo rid a g rap efru it s e e d oil co uld be e x p e c te d to h av e th e five fatty

a c i d s , p a l m i t i c , s t e a r i c , o l e i c , l i n o l e i c , and lin o le n ic in th e follow ing

p r o p o rtio n s : 3 5 , 3 . 5 , 21, 36, 4 .5 % , r e s p e c t i v e l y . T his is c o n s id e ra b ly

d ifferen t from th e e a r l ie r p u b lis h e d a n a l y s e s (11) w h e re in th e

o ld e r more d iff ic u lt e s te r f r a c tio n and l e a d s a lt proced ure had to be u s e d .

In 1965, th e sam e a u th o r itie s (6) s ta t e d th a t g ra p e fru it s e e d

o il is co m po sed of the follow ing f a tty a c i d s : p alm itic 34%, s te a r ic

3 .4 % , o le ic 22%, l in o le ic 37%, lin o le n ic 4 .6 % . This a n a l y s is draw s

a tte n tio n to th e point th a t c itru s s e e d o il h a s b e e n c l a s s i f i e d a s a

lin o le n ic a c id o il and by o th ers a s one t h a t is ric h in b o th o le ic and

lin o le ic a c i d s .
 MATERIALS AND METHODS

S am ples

Sam ples of g rap efru it from w h ich s e e d s w ere o b ta in e d for

a n a l y s is w ere c o l l e c t e d from th e fo llow in g s o u rc e s :

1. Burgher Grove (M aricopa C o u n ty , A rizon a): Burgher g r a p e ­

fru it, C itru s p a r a d is i ( M a c f .) c u ltiv a r Burgher, are t r a d i - '

t io n a lly h a r v e s te d very l a t e , som etim es a s la t e a s M ay or

June. B ec au se of t h e ir l a t e m atu rity th e y are v ery so u r and

b itte r during th e r e g u la r h a r v e s tin g s e a s o n (D ecem ber).

2. C h a n d le r H e ig h ts (Sposito) (M aricopa C o u n ty , Arizona):

C h a n d le r g ra p e fru it. C itru s p a r a d is i (M acf.) c u ltiv a r

C h a n d le r , is h a r v e s te d e a r l y , a s soon a s th e B rix -a cid

r a tio is l e g a l , or before (som etim es in N ovem ber). It is

u s u a ll y q u ite s w e e t if a llo w e d to m a tu r e .

3 . Yuma Yellow (Yuma C o u n t y , Arizona): C itru s p a ra d is i

(M acf.) c u ltiv a r Yuma Y ellow , w as grown at The U n iv e rsity

of Arizona Yuma E xperim en tal Farm on C itru s m acrop hy la

r o o ts to c k . H a rv e st is a c c o m p lis h e d th ro u g h o u t the w in te r .

T his is a M a rs h g ra p e fru it a s are a ll th e M a ric o p a C o un ty

g ra p e fru it a n a ly z e d in t h i s w o r k .

15
 16

4. Yuma Pink (Yuma C o u n ty , Arizona): C itru s p a r a d is i (M acf.)

c u ltiv a r Yuma P in k , a red or pink g ra p e fru it (not M a r s h ) ,

grown at The U n iv e rs ity of Arizona Yuma E x perim ental '

Farm on C itru s m acrophyla r o o ts to c k , i s h a r v e s te d th ro u g h ­

out th e w in te r u s u a lly a l i t t le e a r lie r th a n M arico p a C ounty

g ra p e fru it b e c a u s e of th e h ig h e r a v e ra g e en v iro n m e n ta l

te m p e ra tu re in Yuma. They a re a lit t l e s w e e te r and the

B rix -a c id r a tio is a l i t t l e h ig h e r th a n t h a t of Yuma Y ellow .

5. U n iv e rsity of Arizona (M aricopa C o u n ty , Arizona):

U. of A. g r a p e fr u it. C itru s p a r a d is i (M acf.) c u ltiv a r U . of

A . , is grown a t The U n iv e r s ity of Arizona E xp erim en tal

Farm in M e s a . It is u s u a lly medium in s w e e t n e s s and

a c id ity and is h a r v e s te d in la te D ecem ber or throu gh ou t

Jan u ary .

Sample C o lle c tio n

Fruit w ere p ic k e d a t ap p ro x im a te ly 2 w eek i n te r v a ls during th e

1969-70 growing s e a s o n . The th re e M a ric o p a County g ro v e s were

c h o s e n as e x h ib itin g v ery b itte r (Burgher— la te h a r v e s tin g ) , medium

(U. of A . — la t e h a r v e s tin g ) , and s w e e t (C h and ler— e a r ly h arv estin g )

c h a ra c te ristic s. In a d d itio n , two Yuma v a r i e t ie s w ere a l s o c h o s e n . Ten

g ra p e fru it w ere p ick ed a t random e a c h tim e , th e s e e d s w ere rem oved and

s to re d in p l a s t i c b ag s at - 5 ° C . for ab o u t s ix months prior to a n a l y s i s .

 Seed P repara tion

After r e a c h i n g room t e m p e r a t u r e , a p p r oxim a te ly t e n s e e d s from

e a c h "varie ty " w ere s e l e c t e d a t random and w a s h e d w i th d i s t i l l e d w a t e r

followe d by s p ra y i n g w ith 3 t o 5 ml. of a c e t o n e to a s s i s t in th e removal

of w a t e r . The seed's wer e air dr ie d ov ernight at room te m p e r a tu re and

t h e n w e ig h e d .

'Oil E x tr actio n

The oil w a s e x t r a c t e d by p la c in g th e s e e d s in a high s p e e d

mixing b l e n d e r , u s in g a 500 ml. j a r ,f o r 10 minutes w ith 100 ml. of

c h l o r o fo r m :m e th a n o l, 2:1 s o lv e n t mixture (18).

The e x t r a c t w a s f ilter ed th ro u g h f il ter p a p e r (S. and S.

No. 489, b l a c k band) a n d .d rie d un d er n itro gen atm o s p h e r e a t 5 0 ° C . and

w e ig h e d a s t o t a l li p id (TL)., The e x t r a c t e d lipid m a t e r i a l w a s w a s h e d

w ith four 5 ml. portions of petroleum e t h e r , dried un d er re d u c e d

p r e s s u r e at 4 5 ° C . and t h e n w e ig h e d to o b ta in the petro leum e th er

e x t r a c t e d m a te r ia l (PE).

M eth yl Ester s. Prep ar at ion

In p r e p a ra t io n for m e th y la tio n approxim a te ly 0 . 2 ml. of oil w a s

dr ie d under vacuum at 45 C . for 60 m i n u t e s . The f atty a c i d methyl

e s t e r s wer e prep ar ed by t r a n s m e t h y l a t i o n , u s in g sodium metho xide (13).

The methyl e s t e r s wer e e x t r a c t e d in h e x a n e (1), and s to r e d under

r e fr ig e ra ti o n until a n a l y z e d by g a s - l i q u i d - c h r o m a t o g r a p h y .
 18

G a s - L i q u i d Chromatography

A Per kin -E lm er M od el 800 g a s c h r o m a t o g r a p h , e q u ipped w it h

d u a l flame i o n i z a t i o n d e t e c t o r , two 6 f t. x 1 /8 in . s t a i n l e s s s t e e l

columns p a c k e d w ith 15% d ie th y l g l ycol s u c c i n a t e (DEGS) on a 60/80

m e s h Chro mosor b W , and a printing i n t e g r a to r , w a s u s e d . Argon at

15 p s i of p r e s s u r e w a s u s e d as t h e , c a r r i e r g a s . ■The column tem p er atu re

w a s 17 0 ° C . Sample s i z e w a s abou t 0 , 5 u l . The e s t e r s w ere id e n tifie d

by co m p ar iso n w it h s ta n d a r d compounds and ch ro mato grams (5). Peak

a r e a s w e r e m e a s u r e d by th e printing in t e g r a to r and e x p r e s s e d a s a

. p e r c e n t a g e of t o t a l f atty a c i d s .
 RESULTS AND DISCUSSION

Grap ef rui t s e e d o i ls of th e Arizona v a r i e t i e s a n a l y z e d in t h i s

s tu d y c o n ta in e d pred om inantly s ix f atty a c i d s (Tables 1, 2, 3, 4, an d

5). The s a t u r a t e d a c i d s wer e m y r i s t i c , pa lm itic and s t e a r i c , and the

u n s a t u r a t e d o n es were o l e i c , l i n o l e i c , and l i n o l e n i c .

The c o n te n t of m y r is tic a c i d r a nged from 0. 46% (Yuma Yellow)

to 0 .81% ( C h a n d le r ) . The p r e s e n c e of m y r i s t i c a c i d in gra pefr uit s e e d

oil w a s r eport ed by Dunn e t a l . (2) in 1948 who made an i n v e s t i g a t i o n

on s a m p le s from W e s t In dia n grapefruit and found r e s u l t s from 0.8%

(Foster grapefruit) to 1.2% ( M a r s h ) . This somew hat diff e r e n t r e s u l t is

p o s s i b l y e x p l a i n e d by th e f a c t that t h o s e autho rs u s e d th e old er e s t e r -

f ractio n and l e a d s a l t proce dure on the d e t e r m in a tio n (5).

The p alm iti c ac id v a l u e s found v a r ie d from 28 .3 3% (Chandler) to

32.37% (U. of A . ) . T h e se r e s u l t s a g r e e w it h t h o s e of Dunn et a l . (2)

who found from 27.5% to 28.9% in W e s t Indian grapefr uit s e e d oil and

by H en d r ic k so n and Kes ter son (5) who re p o rte d a range in pa lm i tic a c id

c o n te n t from 20 to 39%, w h en a n a l y z i n g F l o r i d a ' s g r a p e fr u it s e e d oi l.

The same a u th o r itie s (6) s t a t e d in 19 65 t h a t a t y p i c a l c o n t e n t was 34%

p a l m it ic a c i d in F lo r i d a ' s g r a p e f r u i t . The r e s u l t s d iff er so m ew hat from

t h a t found by J a m i e s o n , Baughman and Gentler, in 1930 (in 11) who

rep orte d 19.2% pa lm itic a c i d in gr apefr uit s e e d o il.

.
 20

TABLE 1 . F atty A cid C o m p o s it i o n o f Burgher G rapefruit S e e d O il

R e la te d to H a r v e s t D a t e

Harvest F atty Acids (%)

D ate M y r i s ti c Palmitic S te a r ic O leic L inoleic Lino len ic

09/08/69 0. 79 34.73 1.25 1 9 .97 40. 74 2.50

09/22/69 0.34 31.04 1. 85 22.64 39.68 4.42

10/04/69 0.29 32.03 0.87 24.04 40.16 2.58

10/18/69 1 .21 32.24 2.71 20.95 37.24 5.63

11/05/69 0.67 33 . 70 1.67 2 1.63 40.19 2.13.

12/13/69 0.50 34.32 1.48 20.53 40.93 2.47

01/19/70 0.38 31.36 2.19 2 2.21 41.00 2.84

02/02/70 0.40 28.08 3.80 1 9 .65 36.93 4.97

0 3 /0 2/7 0 0.85 2 8.7 5 4.02 2 0 .83 38.03 7.51

03/30/70 0.53 2 8.93 5.08 20.24 38.64 6.57

X 0.59 31.52 2.49 21.27 39.35 4.78

T o ta l Saturated: 34.5 0%

T o ta l U n s a tu r a te d : 6 5 . 40%

99.90%
 21

TABLE 2 . F a tty A cid C o m p o s it i o n o f C h a n d le r G rapefruit S e e d O il

R e la te d to H a r v e s t D a te

Harvest Fat ty Acids (%)

Date M y r i s ti c Palmitic S te a r ic O le i c Lin oleic Linolenic

09/08/69 0,52 32.39 3.56 1 9.36 39.13 5.02

09/22/69 0.44 2 9 .11 5.30 24.64 37.91 2.60

10/04/69 0.44 28.00 4.47 21.16 34.80 11.12

10/18/69 0.49 22.66 3.35 29.54 33.89 10.07

11/03/69 0.70 27.70 6.27 23.66 36.07 5.60

12/13/69 1.08 27.05 4.61 21.4 1 36.23 9.61

01/05/70 0.57 29.68 4.29 22.29 37.22 5.95

01/19/70 2.27 28.98 4.24 2 1 .19 3 7.90 5.42

02/16/70 0.78' 29.02 3.71 20.95 35.33 10.1 9

03/15/70 0.87 27.74 3.90 21.42 36.98 9.08

X 0.81 2 8.23 4.37 22.56 36.54 7.46 .

To tal Satura ted: 33.41%

T o ta l U n s a tu r a te d : 6 6 . 56%

99.97%
 2.2

T a b le 3 . F a tty A cid C o m p o s it i o n o f Yuma Y e llo w G ra pefruit S e e d O il

R e la te d to H a r v e s t D a t e

Harvest Fatty Acids (%)

D ate M y r i s ti c Palmitic S te aric O le ic Li noleic Lin olen lc

10/13/69 0.40 29. 65 5.05 24.1 1 37.62 3.17 .

11/10/69 0.37 29.03 5.92 2 2.51 38.38 • 3.79

11/24/69 0.13 32.08 . 2. 68 23.56 37.43 4.12

12/08/69 0.36 , 33.03 3.26 21.76 38.18 3.41

12/22/69 0.43 31.84 3.47 22.20 39.42 2.64

01/05/70 0.42 28.60 3.71 22. 19 37.23 7.85

01/19/70 0.54 29.92 2.70 2 2.60 36.73 7.51

02/02/70 . 0.41, 28.64 3.09 21.66 36.26 8.93

02/23/70 1.00 28. 60 3V.89 21.76 38.95 5.80

03/16/70 0.49 2 8.93 3.03 21.74 38.38 7.43

X 0.46 30.03 3.68 2 2.41 37.96 5.46

Total Saturated: 34.17%

T o ta l U n s a tu r a te d : 65.83%

99.90%
 23

T a b le 4 . F a tty A cid C o m p o s it i o n o f Yuma P ink Grapefruit. S e e d O il

R e la t e d t o H a r v e s t D a te

Harvest Fatty Acids (%)

D a te M yristic Palm itic S te ar ici O l e ic Li noleic Lin o len ic

10/13/69 0.45 27.79 5.03 .22.44 33.83 10.46

11/10/69 0.42 27.60 5.95 21.53 36.65 7.85

11/24/69 0.52 25.77 4.35 21.34 38.14 9.88

12/08/69 0.72 37.92 3.04 1 7 .85 35.13 5.34 ,

12/22/69 0.27 28.69 5 . 63 21.78 35.96 7 .63

01/05/70 0.49 26.54 3.40 22.08 40.13 7.36

01/19/70 0.70 22.70 4.58 25.72 35.02 1 1.08

02/02/70 1.79 26.24 3.51 22.73 3 8.40 7.33

02/23/70 1.65 32.35 5.02 19.05 36.09 5. 83

03/16/70 0.48 28.17 4.45 21.30 34.88 10.78

X 0.75 28.38 4.49 21.58 36.42 8.37

Total S aturated: 33.62%

T o ta l U n s a tu r a te d : 66.37%

99.99%
 24

T a b le 5 . F a tty A cid C o m p o s it i o n o f U . o f A, G rapefruit S e e d O il

R e la te d to H a r v e s t D a t e . ' .

Harvest Fatty Acids (%)

D ate M yristic P alm itic S t e a r ic O le ic Li noleic Lino len ic

09/08/69 0.89 2 9 .15 2.89 21.32 40.02 5.74

10/04/69 0.35 32.59 2.73 22.88 37. 88 3.57

11/03/69 0.57 35.30 2.19 2 2.01 38.21 1.7 1

11/15/69 0.56 32.51 2.57 2 3.09 37.92 3.35

01/05/70 0.31 32.69 2.43 2 1.70 39. 84 3.03

01/19/70 0.36 31.84 1. 87 20.73 39.62 5.58

02/02/70 0.43 31.35 2.61 21.90 39.67 4.05

02/16/70 0.51 31.24 3.43 22.38 37.99 4.46

03/02/70 0.48 32.95 2.64 20.58 39.61 3.87

03/30/70 0.63 34.06 2.03 20.60 37.58 5.10

X 0.51 32.37 2.54 21.72 38.83 4.05 .

Total S atura ted: 35.42%

T o ta l U n s a tu r a te d : 6 4 . 60%

100. 02%
 25

Ste aric a c id w a s d e t e c t e d in pr oportions from 2.49% (Burgher)

to 4.49% (Yuma P i n k ) . T h e s e r e s u l t s g e n e r a l l y agree w ith t h o s e p u b - h

l i s h e d by H e n d r i c k so n and Kes te rs on (5) (2.2 to 5%) and w i th t h o s e of

the sa me aut ho rs in 1965 (6) who r eport ed an a verage of 3 .4% s t e a r i c

a c i d in F l o r i d a ' s grapefr uit s e e d o i l . The r e s u l t s found by Dunn et a l .

(2) wer e s om ew hat lo w er (2.2 and 2.9%) in s t e a r i c a c i d in W e s t I n d ia n

grap ef ruit s e e d s / e s p e c i a l l y when compared with th e 7. 25% re port ed

by J a m ie s o n in 1932 (11).

The s e e d oil a n a l y z e d in t h i s work s how ed no d e t e c t a b l e

arachidic acid. This a c id w a s in c l u d e d in th e a n a l y s i s of Dunn et a l .

(2) in 1948, in W e s t In d ia n g r a p e f r u i t s , and in p e r c e n t a g e s of 0 .6 and

2 . 1 , r e s p e c t i v e l y , for F o s t e r and M a rs h grapefruit s e e d o i l .

In the u n s a t u r a t e d fatty a c i d g r o u p , the o l e i c a c i d c o n ten t

found in t h i s s tu d y compared c l o s e l y to t h a t re porte d in oth er w o r k . The

per c e n t o l e i c ac id in the v a r i e t i e s a n a l y z e d in t h is r e s e a r c h range d from

2 1 , 2 7 (Burgher) to 2 2 . 5 6 ( C h a n d le r ) . T h e s e v a l u e s c a n be r e l a t e d to a

1932 report by J am ie s o n (11) show ing 19.66% o l e ic a c i d , one by Dunn

e t a l . (2) giving 21.1% and 2 5 .1 % , and l e v e l s found by H e n d r i c k so n and

Kes terson in v a r io u s re p ort s (5, 6, 7, and 8) of 2 1 . 2 2 to 27%. The

r a t h e r c l o s e ag reem ent in th e v a l u e s found and reported for ole ic a c i d i s .

of i n t e r e s t a s the s e p a r a t i o n of th is c om ponent from s t e a r i c and l i n o l e i c

a c i d s is one m e a s u re of column per formance in the GLC a n a l y t i c a l

procedure (6).
 26

Values for l i n o l e i c a c id ran g ed from 36.42% (Yuma Pink) to

39.35% (Burgher). Comparing t h i s 'component w ith t h e oth er f atty a c i d s

a n a l y z e d in t h is w o r k , l i n o l e i c a c id a p p e a r s in the h i g h e s t c o n c e n tr a tio n .

T h e se v a l u e s confirm o n e s by Dunn e t a l . (2) who r e p o rt e d 3 6.6% and

3 9 . 3 % , and by H e n d r i c k s o n and K e s t e r s o n , who found 35%, 36 and 38%

in 1961 (5), and 37% in 1965 (6) working w it h Florida gra pef ruit

varieties. The h i g h e s t r e s u l t w a s found by J am ies o n et a l . in 1930 (in 11)

who r eport ed 48.8 4% l i n o l e i c a c id from t h e i r a n a l y s i s .

F i n a l l y , sm all am ounts of l i n o l e n i c a c id w ere d e t e c t e d . In

t h i s w o r k , it v a r ie d 4 . 05% (U. of A.) to 8.37% (Yuma Pink). In th e

li t e r a t u r e c i t e d , v a l u e s r eport ed w ere 0 . 16% (11), 3%, 4 . 5 % , 5.1 % (5),

4. 6% (6) and 5.9% (2). T h e s e v a r i a t i o n s are u n d e r s t a n d a b l e b e c a u s e of

th e r e l a t i v e l y s m a l l , broad p e a k r e c o r d e d in t h e GLC p r ocedure for

t h i s co m ponen t.

In th e Burgher grap ef ruit s e e d t h e r e w a s a t e n d e n c y for l i n o l e n i c

ac i d to i n c r e a s e in c o n c e n t r a t i o n in t h r e e a n a l y s e s of s a m p l e s from

February to M arc h ( 4 . 9 7 , 6 . 5 7 and 7 . 5 1 % , r e s p e c t i v e l y ) w ith r e s u l t s a ll

above t h e a v e r a g e p e r c e n t a g e ( 4 .7 8 ) . The same p henom ena oc cu rred

w ith C h a n d le r (10.19 and 9.08%) w h e r e i n the a v e r a g e p e r c e n t a g e of t h i s

a c id w a s 7 . 4 6 , and w ith Yuma Yellow ( 8 . 9 3 , 5 . 80 and 7 .43) , the

av e r a g e v a l u e bei ng 5 . 4 6 .

In th e same period (February and March) t h e r e w a s a tre nd for

palm itic a c i d c o n t e n t to be lower in Burgher (28.08 t o 28.93%) and Yuma

Yellow ( 28.60 to 38.93%) t h a n th e a v e r a g e , 3 1 . 5 2 and 3 0 .0 3 % ,

respectively. A s im i l a r r e l a t i o n s h i p is s e e n in the o l e i c ac id c o m p o s i ­

t io n during th e s a m e period (February and March) in th e tw o v a r i e t i e s

c i t e d , Burgher (19 .60% to 20 . 83%) and Yuma Yellow (21. 66% to 21. 76%)

w her e the r e s p e c t i v e a v e r a g e s for t h e s e d e t e r m in a ti o n s wer e 21.27% and

22 .4 1 % . This o b s e r v a t i o n w a s a l s o found in C h a n d le r g r a p e fr u it, w h e n

th e o l e i c a c id c o n t e n t v a r ie d from 20 .95% to 2 1 .4 2 % , in February and

M a r c h , r e s p e c t i v e l y , and its a v e r a g e p e r c e n t a g e w a s 2 2 . 5 6 . T he se are

only s l i g h t d i f f e r e n c e s , h o w e v e r , w hic h are not s tr i c t l y d i s t i n c t i v e for

h a r v e s t i n g tim e or v a r i e t y , in th e fruit ex am in ed in t h i s study. This a l s o

a g r e e s w ith H e n d r ic k so n and K es ter son (5) who found t h a t th e r e w a s a

rem ark able s im i la r ity b e tw e e n all s a m p l e s s u c h t h a t n e i t h e r v a r ie ty nor

m aturity cou ld be d i s t i n g u i s h e d on th e b a s i s of fatty a c i d a n a l y s i s alone.

Comparing t h e t o t a l s a t u r a t e d and u n s a t u r a t e d f a t ty a c i d s as

a v e r a g e s of all s a m p l e s (Table 6) th e r e is only a s li g h t v a r i a t i o n around

th e m e a n , (+1.19 and - 0 . 8 1 % in th e s a t u r a t e d and + 0 . 0 8 and - 1 . 1 5 % in

the u n s a t u r a t e d o n e s ) . This further c o r ro b o r a te s th e c o n t e n t i o n t h a t a ll

v a r i e t i e s of Arizona gr ap ef ruit a n a l y z e d s e e d oils h a v e f a t t y acid

co m p o sit io n ex trem ely a l i k e , al th o u g h C h a n d le r and Yuma Pink have

a s lig h tly highe r u n s a t u r a t e d ac id c o n t e n t (66.56 and 66.37%) ,

r e s p e c t i v e l y , w he n compar ed w i th Yuma Yellow (65.83 % ), Burgher

(65.40%) a n d U . of A. (64.60%). T h e s e r e s u l t s show t h a t A r iz o n a 's

grap ef ruit s e e d o il s , have an u n s a t u r a t e d f atty a c i d c o n t e n t

 28

Table 6. Average C o n t e n t of S atu ra te d and Urisa turat ed F atty Acids of

A r iz o n a 's Gra pefru it Seed Oils

Fatty Acids (%)

Variety S at ura te d Un s a t u r a t e d

Chandler 33.41 ' 66.56

Yuma Pink 33.62 66.37

Yuma Yellow 34.17 65.83

Burgher 34.50 65.40

U. of A. 35.42 64.60

X 34.22 65.75

Variation around th e mean (X):

S a tu ra te d +1.19% U n s a t u r a te d : +0.08%

-0.81% -1.15%
 29

ap proxim a te ly t w i c e t h a t of th e s a t u r a t e d a c i d s . One may c o n s i d e r it

as an oil of medium u n s a t u r a t i o n co n t a in in g (65%). u n s a t u r a t e d fatty

a c i d s as co mpa re d w it h a lo w er c o n t e n t (16%) in c o c o n u t oil and a

higher one (81%) of s e s a m e s e e d oil (11).

Since th e e x t r a c t i o n proce dure l e n t i t s e l f to o b s e r v a t i o n s on

oil co n te n t of t h e grap ef ruit s e e d , t h e s e d e t e r m i n a t i o n s are in clu d ed

in th e r e s u l t s ' ( T a b l e s 7, 8, 9, 10, and 11). There w a s so me r e l a t i o n .

b e t w e e n dr ie d s e e d w e ig h t and y i e l d of t o t a l l i p i d s . The t o t a l lipids

c o n t e n t v a r i e d from 1 4 . 8 0 to 37.90% in Burgher; 2 5 . 6 0 to 36.40% in

° C han dl er ; 11.59% to 40 . 02% in Yuma Yellow; 1 5 . 4 4 to 3 4 . 1 3 % , in

. Yuma P i n k , and 27.11 t o 32.62% in U. of A. g r a p e fr u i t. Thos e r e s u l t s

are not g r e a t l y dif fer en t t h a n t h o s e found in th e l i t e r a t u r e , where

H en d r ic k s o n and K e s t e rs o n (6) found 29.2% to 37.3% oil in F l o r i d a ' s

Du n can g r a p e f r u i t . The same a u t h o r it i e s r e f e r to a lipid c o n te n t of

35% in t h e same v a r i e t y s u p r a c i t e d in a l a t e r report (8) and t h e

U. S. D. A. (19), r e g i s t e r e d a lipid c o n t e n t of 21.84% to 39.44% oil

from ai r dried grap ef ruit s e e d s . Comparing t o t a l l ip id s and petroleum

e h t e r e x t r a c t t here w a s a r atio of 1 . 2 0 in th e Yuma Pink and C handle r

v a r i e t i e s , 1 . 1 5 and 1 . 1 8 for U. of A. and Yuma Yellow v a r i e t i e s ,

and 1 . 2 4 for Burgher, r e s p e c t i v e l y (Table 12).

There w as a high er lipid c o n t e n t in the v a r i e t i e s c o n s i d e r e d to

be e a r l y h a r v e s t i n g (Chandler and Yuma Pink) around D e c e m b e r and January.

 30

T a b le 7 . T o ta l L i p i d s , P etroleu m Ether E x tra ct and I n d i v id u a l Air D r ie d

S e e d W e i g h t o f Burgher G rapefruit

Dr ied See d . Total E x t r a c t a b l e . Petroleum Ether

Harve st W e ig h t Lipid Extract
Date : „ JgL_.. .......... (%)____ (%)

09/08/69 0.2000 14.80 10 . 0 0

09/22/69 0.1810 18.00 15.0 0

10/04/69 0.2321 20.40 17.00

10/18/69 , 0.1920 26.30 20.00

11/05/69 0.1425 26.60 22.00

12/13/69 0.1626 36.80 30.80

01/19/70 0.1479 37.40 30.90

02/02/70 0.1627 37.50 31.50

03/02/70 0.1624 33.80 27.50

03/30/70 0.1584 35.40 26.90.

X 0.17417 28.70 23.16

 31

T a b le 8. T o ta l L ip id s , P etroleu m Ether E xtract and I n d i v id u a l Air D r ie d

S e e d W e ig h t o f C h a n d le r G rapefruit

Dried Seed Total E x tr a c ta b le Petroleum Ether

Harvest W eight Lipid Extract
D ate ............... (g).............. ...................... (%)....... (%)

09/08/69 0.0949 25.60 21.30

09/22/69 0.1605 26.30 21.60

10/04/69 0.1801 29.20 24.50

10/18/69 0.1958 30.80 26.40

11/03/69 0.1735 34.80 30.00

12/13/69 0.1775 34.70 28.90

01/05/70 0.1716 36. 40 31.10

01/19/70 0.1985 34.40 2 9.10

02/16/70 0.1889 34.60 29.20

03/15/70 0.1765 . 33.00 23.70

X 0.17178 31.98 26.57

 32

T a b le 9 . T o ta l L ip i d s , P etroleu m Ether E x tract and I n d iv id u a l Air D ried

S e e d W e i g h t o f Yuma Y e llo w G ra p efru it

Dried Seed Total E x tr actab le Petroleum Ether

Harvest W e ig h t Lipid Extract
D ate .. ... (g) (%) (%)

10/13/69 0.1438 32.68 27.40

11/10/69 0.1542 34.44 29.28

11/24/69 0.1456 32.77 2 4 .23

12/08/69 0.1677 36.05 30.77

12/22/69 0.1564 40.02 34.23

01/05/70 0.1562 22.00 19.44

01/19/70 0.1872 23.34 20.68

02/02/70 0.1433 17.84 14.6 9

02/23/70 0.0 8 5 1 11.59 9.99

03/16/70 0.1648 15.92 13 .73

X 0.15143 26.46 22.44

T a b le 1 0 . T o ta l L ip i d s , P etroleu m Ether E x tr a c t, and I n d iv id u a l Air
D r ie d S e e d W e i g h t o f Yuma Pink. G rapefruit

Dried Seed Total Extracta ble Petroleum Ether

Harvest Weight. Lipid Extract .
D a te (g)............. ..._ _ ..._ ...(%) .............. . _____ (%)

10/13/69 0.2237 26.25 22.4 1

11/10/69 0.2030 28.79 24.48

11/24/69 0.1946 31.56 27.2 1

12/08/69 0.1524 33.42 26.55

12/22/69 0.2091 29.60 24.17

01/05/70 0.1236 30.81 25.82

01/19/70 0.1714 34.13 29.34

02/02/70 0.1233 29.84 24.98

02/23/70 0.1221 33.44 25.84

03/16/70 0.1514 15.44 11 . 8 4

X 0.1674 29.32 24.26

 34

T a b le 1 1 . T o ta l Lipids-, P etro leu m Ether E x tr a c t, and I n d iv id u a l Air

D r ie d S e e d W e i g h t o f U . o f A . G rapefruit

Dried Seed Total E x tr acta b le Petroleum Ether

Harvest W e ig h t Lipid Extract
D at e ...... (g) (%)
...................................... ...................... ... (%)

09/08/69 0.1143 27.33 21.07

10/04/69 0.1659 31.01 25 . 7 0

11/03/69 0.1770 29.78 25.55

11/15/69 0.2003 29.27 24.74

01/05/70 0.2012 29.56 24.1 1

01/19/70 0.1793 29.60 23.64

02/02/70 0.1911 27.11 21.63

02/16/70 0.1658 32.62 25.48

03/02/70 0.1862 29.28 21.78

03/30/70 0.1650 28.86 22.27

X 0.1744 29.44 25.59

 35

Ta ble 12. The Average C o n t e n t of To tal E xtractable Lip id, Petroleum

Ether Extrac t and Their Ratios

Total E x tr a c ta b l e Petroleum Ether .

Variety Lipid (%) (TL) Extract (%) (PE) TL/PE Ratio

Chandler 31.98 26.57 1.20

IT. of A. 29.44 25.59 1.15

Yuma Pink 29.32 24.26 1.20

Burgher 28. 70 23.16 1.24

Yuma Yellow 26.46 22.44 1.18

 36

The l a t e h a r v e s t i n g v a r i e t i e s , h o w e v e r , vary with t h e h i g h e s t lip id

c o n t e n t bein g from O c to b e r to November (IT. of A . ) , D e c e m b e r (Yuma

Y e l l o w ) , to January and February (Burgher). In t h e s e r e s u l t s the

maturity of fruits w a s only i n d ir e c tl y s tu d i e d but we ag r e e w it h 1962

U . S. D . A. information (19) t h a t not enough s a m p l e s of cit ru s li p id s

hav e b e e n a n a l y z e d to sh ow w h a t v a r i a t i o n s in co m p o s it io n may be

expected. H e n d r i c k so n and Kes ters on (6) s t a t e d t h a t t h e p e r c e n t a g e

of oil in drie d c i tru s s e e d s is v a r i a b l e and d e p e n d s upon fruit m a t u r i t y .

H i g h e s t oil c o n te n t u s u a l l y coincided, w it h th e optimum maturity n e e d e d

for fruit p r o c e s s i n g . T h e s e same a u t h o r s , in 1967 (8), s t a t e d t h a t a

2- y e a r s u rv ey of the oil co n te n t of t h e dried s e e d s of D u n c a n gra pe fru it

(Florida) had an average- s e a s o n a l v a l u e of 34%. Although th e oil

c o ntent of t h e s e s e e d s did not vary w i d e l y , t here w a s a t e n d e n c y for

maximum oil c o n t e n t to o c c u r when the fruit had r e a c h e d t h e g e n e r a l l y

a c c e p t e d m a t u r i t y . It w a s a l s o shown t h a t prolonged w e t s e e d s to r a g e

s e r i o u s l y i n c r e a s e d th e free f atty a c i d c o n t e n t of th e e x p e l l e d o il .
 CONCLUSIONS

1. Gra pe fru it s e e d o ils of th e Arizona v a r i e t i e s a n a l y s e d in

t h i s s tu d y c o n t a i n pr edom ina ntly s i x f a tty a c i d s . The s a t u r a t e d a c i d s

were m y r i s t i c , pa lm i tic and s t e a r i c , and th e u n s a t u r a t e d o n e s were

o l e i c , l i n o l e i c , and l i n o l e n i c .

2. There w a s a re m ark able s im ila r ity b e t w e e n a l l s a m p le s

s u c h t h a t n e i t h e r v a r i e t y nor h a r v e s t i n g time could be d i s t i n g u i s h e d

on th e b a s i s of f a tty a c id a n a l y s i s .

3. Gr ap efru it s e e d o i ls of th e v a r i e t i e s a n a l y z e d in th is

s tu d y had an a v e r a g e of 65% of u n s a t u r a t e d and 35% s a t u r a t e d fatty

acids.

4. Air dried grap ef ruit s e e d s c o n t a i n e d ap p r o x im a te ly 30%

t o t a l e x t r a c t a b l e lip id s of w h ic h 25% wer e petroleum e t h e r s o l u b l e .

37
 LITERATURE CITED

1. Brown, W . H. , J. W . Stull, and G. H. S t o t t , 1962. Fatty Acid

C o m p o s i tio n of M ilk . I . Effect of Roughage and D ie ta ry Fat
Journal of Da iry S c i e n c e , 46; 191.

2. D unn, H. C . , T. P. H i l d i t c h , and J. P. Rileg, 1948. The

C o m p o s it io n of Seed F at s of W e s t Indian C itr u s F r u it s . J.
Soc. Chem . In d. (London), 67: 199.

3. E c k e y , E. W . , 1954. Veg etable Fat s and O i l s . American

C h e m ic a l S o c i e t y , Mon ograph S e r i e s . Reinhold Publish ing
C o . , N. Y. , 548.

4. F r a s e r , S. , 19 24. American F r u i t s . Orange Judd Publish ing C o .

I n c . , N. Y. , 664.

5 . H e n d r i c k s o n , R. , and J. W . K e s t e r s o n , 1961. Gra pefru it Seed,

O il. Florida State Ag ricultu ral S o c i e ty , 74; 219 .

6. , 1965 . By -P ro duc ts of Florida C itrus . Agricultural

Experiment S t a t i o n s , U n i v e r s i t y of F l o r i d a , Bulletin 698: 44.

7. , 19 66. See d Oils From Florida C it r u s F r u i t s . Florida

Agricultural Experim ent S t a t i o n s , Annual Report: 239.

8. , 1967. N a r i n g i n , a Bitter Principle of G r apefr uit.

Agricultural Experiment S t a t i o n s , U n i v e r s it y of F l o r i d a ,
T e c h n i c a l Bulletin 511 - A .

9. H o r a n , J. , 1964. M arga rine a Base d 'H u i l l e Extra it e de Pepins

d'A grumes. C a r ib b e a n A g r i c u l t u r e , Puerto Rico, 3: 832.

10. Hume, H. H a r o l d , 1938. The C u l t i v a t i o n of C i t r u s F r u i t s . The

M a c M i l l a n Com pany , N. Y.

11. J a m i e s o n , G. S . , 1932. Veg eta ble F at s and O i l s . American

C h e m ic a l S o c i e t y , Monograph S e r i e s , The C h e m i c a l C a t a l o g
C o m pany, I n c. , N. Y.

38
 39

12. Kefford, J. F. , 1959. The C h e m i c a l C o n s t i t u e n t s of Citrus

F r u it s . Ad van ce s in Food R e s e a r c h , Academic P r e s s ,
N. Y. , 9: 286.

13. Luddy, F. E. , R. A. Barford, and R. W . R ie m e n s c h n e id e r , 1960.

D ir ect C o n v e r s io n of Lipid C ompon en ts to Their Fatty Acid
M eth yl E s t e r s . J. A. O. C . S . , 37: 447.

14. M a t l a c k , M. B . , 1929. A C h em ic al Study of th e Rind of •

C a lif o r n ia n O r a n g e s . J. Am. Pharm. A s s o c . , 18: 24.

15. _________ , 1940. The Fatty C o n s t i t u e n t s of C a li f o rn ia Valencia

Or ange Pulp. J. O r g . C he m . , 5: 504.

16. N o l t e , A. J. , and H. W . von Lo e se ck, 1940. Gr ap efru it Seed

O i l , M an u factu r e and P h y s i c a l P r o p e rt ie s . In d. and Eng.
C h e m . , 32: 144 .

17. P a n to v ic h , V. E. , 1955. The U s e of C ^ in S tu d ie s of the

P hysiology of O i l - S t o r i n g - P l a n t F r u it s . C o n f e r e n c e of the
Academy of S c i e n c e s of the USSR on the P e a c e f u l U s e s of
Atomic E n e r g y , (in En glish t ra n s l a t i o n ) : 155.

18. S tu l l, J. W . , F. M. W h it in g , W . H. Brown, Mary M il b r a t h , and

G. W . W a r e , 1968. C o m p ar iso n of Ex tr ac tion M ethods for
Analys is of DDT, DDE, and DDD in Alfalfa Hay. J. of Dairy
S ci. , Vol. 51: 1039.

19. U. S. D. A . , 1962. C h em is try and Techno lo gy of Citrus , Citrus

P r o d u c ts , and B y-P roducts . Agriculture Handbook No. 98:
18, 6 0 - 6 1 .
125

t 6 2 0 8--I