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Study of the application of the microscope to fat and oil analysis.

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Study of the application of the microscope to fat and oil analysis.

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Content S T U D Y O P T H E APPLICATIO N Of T il M IC B O SQ Q P E
T O fAf A N D OIL ANALYSIS
A Ties is
Presented to
t i e fa c ility o f t ie Department of d em ist ry
U niversity of Southern C alifornia
In fsrfcial fu lfillm e n t
of t i e Requirements for t i e Degree
Master of Science
Ly
V irg il G * Mellenbacher
la y 1936
UM I Number: EP41480
All rights reserved
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D f S M r t a t k s f ) R a b T f l t f i n g
UMI EP41480
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This thesisy written under the direction of the
candidate's Faculty Committee and approved by
all its memberSy has been presented to and ac
cepted by the Council on Graduate Study and
Research in partial fulfillment of the require• *
ments for the degree of
M S T E R . 5 P .S C I M .G S
\etary
Dean
11^
Date * . i . §!
9AP m m sq i§m $s
OHAPflE P A G E
I . HfH03)U0(DI0H . . ...................... , . * 1
X X * OEBIISfRY A N D AHAIYSIH Of fAfS AID OIHS. . . - 3
Composition . . . . . . . . . . . . . . 3
Aooepted Methods of Analysis . . . . . . . 4
Advantages of Microscopic Id en tifica tio n . * , 6
I I I t EI3T0RI0AL . . . . . ...................... . .. . . . . . f 7
IT. MSPlRIMMfAIi ..................... 11
R osenthale^s Method 11
React ions with Potassium Hydroxide -
without Heat 11
Reactions with Potassium Hydroxide -
with Heat ................................. 16
Reactions with Phenylhydrazine . . . . . . 16
Reactions with Bromine . . . . . . . . . . . HI
Patty Acids ............................. HI
f r e e z in g ............................. . 30
Y., COIOI.ESIOI . . . . . . . . . . . . . . . . . . . . m
B IB L IO G R A PH Y ................................................................ 35
Ill ST O F T A B L E S
T A B L E P A G E
I* O ils and Fats Used in Microscopic Study . IS
II# Reactions with Potassium Hydroxide -
without Heat . . . .......................... 14
III* Reactions with Potassium Hydroxide -
w ith Heat 17
I?* Beaotions with Phenylhydrazine . . . ............................... 19
Y* Beaotions with Bromine • • • • • » » • » • * * • « SS
JuISf OF FIGURES
FIG U RE P A G E
feaseod O il w ith Potassium Hydroxide . . . 15
2 Corn O il with Potassium Hydroxide 15
3 B e r illa O il with Potassium Hydroxide . . . . . . . 18
4 Olive O il with Phenylhydrazin© ..................... . . . 18
8 Sardine G il with Phenylhydrazine i . . . • • • , • 80
6 Walnut Oil with Phenylhydrazine # • • • • • • • • 80
f Whale O il, Patty Aoids . . . . . . . . . . . . . 83
8 Domestic Cottonseed O il, fa tty Acids . . . . . . . 23
9 We. 1 Oriental Cottonseed O il, fa tty Acids . . . . 24
10 Io< 8 O riental Cottonseed Oil* fa tty Acids . . . . 84
11 Palm O il, fa tty Aoids ................................ . . . . . 25
18 H om e O il, fa t t y Aoids • • • • • • • • « • • • • • 25
13 Bahassu O il, fa tty Aoids • .... . . . . . • ,. . • . . 26
14 Olive O il, fa tty Aoids • 26
15 Hayok O il, fa tty Aoids 87
16 Bard, fa tty Aoids . . . . . . . . . . . . . . . . 27
17 Beef f a t, fa tty Acids . . . . . . . . . . . . . . 28
18 Sardine O il, fa tty A c id s ................................. , . . . . 28
19 Coconut O il, fa tty A o id s .................... . .. . . .. 29
20 Palm O il, C r y s ta lliz e d ....................... ..................................29
21 Bahassu O n , C rystallized . . . . . . 51
22 Gooonut O il, C rystallized . . . . . . . . . . . . 31
C H A P T E R I
IB 3 J H 0 I K J Q TI O B
Within recent years, the microscope has become an
important to o l in nearly every laboratory* I ts uses are
^pwiifold and i t s purposes many# Organic and inorganic
a n a ly sis, wood, sugar, leath er, food s tu ffs , m inerals, and
numerous other substances have yielded th eir secrets to the
m icroseopist, Probably, the two most important reasons for
th is development are the greater speed and accuracy attain ab le
by microscopic methods as compared with the ordinary macro
methods.
R etailed explanations o f the various microscopes and
th eir operations are given in so many catalogues and standard
reference works that no attempt w i l l be made here to describe
them.
Although the microscope has been applied to many
f ie ld s of s c ie n tific endeavor, there are some which have, and
s t i l l do, p r o fit l i t t l e from any application o f th is valuable
instrument. Among these are the an alysis and id e n tific a tio n
of animal and vegetable fa ts and o ils*
Everyone, s c ie n tis t and layman a lik e , is at le a st
somewhat fam iliar with that cla ss o f substances known as fa ts
and o ils# fh eir place in the human diet i s an important one,
and one for which there i s l i t t l e substitute* Besides the
large amounts consumed as food, great q u an tities are processed
£
annually which enter into candle making, soap manufacture and
various other uses*
fhe accepted methods o f an alysis o f fata are time
consuming and, for many other reasons as w e ll, can stand
improvement. It was with an appreciation of what might he
gained that th is in vestigation into ways and means of id e n ti
fying fa t and o i l substances w ith the microscope was begun*
In th is research the effo rt has been made to in v estig a te
the few previously suggested procedures, carrying these out
with types of o ils which have not heretofore been studied#
A few changes have been recommended in th ese methods which
are b elieved to be improvements* further than th is , the
methods have been augmented so as to include te s t s foy a
greater variety of o ils* fhe ultim ate attainment of any
such study as th is would be to be able to define reactions
and methods which would y ie ld d efin ite r e su lts id en tifyin g
each and every individual f a t or o il* An a ltern a tiv e to th is
would be to estab lish procedures defining certain groups or
types, again subdividing these groups with further character
is t ic t e s t s , and eventually arriving at a p o sitiv e id e n tific a
tio n of any fa t or o il sample# It i s toward th is goal that
the in v estig a tio n herein discussed has been directed#
OHiPEER 11
O H E M IS S R Y M 2 AHA1YSIS O F FA ES A H D OILS
O QHmositiQn. Some w riters, in th eir terminology,
r e s tr ic t o ils to those fa tty sub stances, whiqhiare -liquid at
normal or room temperatures, and c la s s ify fa ts as those so lid
under the same conditions* Inasmuch as th is d iffe r e n tia tio n
is very in d e fin ite , chemically as w e ll as otherwise, no such
r e s tr ic tio n in nomenclature is resorted to i$ th is paper, and
the terms, fa ts and o i l s , are used in discrim in ately.
In general, the fa ts and o ils are composed largely of
triglycerid es* Yhe tr ig ly cerid es are combinations o f fa tty
acids with glycerin e, wherein the three hydroxyl groups (OlQg
in the la tte r are replaced by fa tty acid members* I t i s
known that the most common tr ig ly ce rid e s occurring in natural
fa ts and o ils are o f a mixed nature containing two or three
d ifferen t fa tty acid groups in conciliation with the glyceryl
|5gJIg] r a d ic le , She exact combinations or proportions in
these combinations are not known* Other substances normally
occurring in fa ts and o i l s , though in minor q u a n tities, are
hydroxy a cid s, higher alcohols such as the s te r o ls , and other
m iscellaneous unsap o n ifia b le m atter. In the subsequent con
sideration of microscopic slid e reactions these la tte r sub
stances must not be neglected because they are d e fin ite ly
reactive and may w e ll form the b asis fo r certain ch ara cteristic
4
forms obtained* Y ariations in composition and cbapaeteris
t ic s are w ell known phenomena exem plified by the common fa ts
v .
and o i l s , These are considered to be complex mixtures of a
higher order with varying proportions of the con stitu en ts
discussed above* (This high degree o f heterogeneity causes
much d iffic u lty in the chemical or physical a n a ly sis of these
substances, whether q u alitative or quantitative*
Accepted Methods of A nalysis. §h® accepted and
O ffic ia l methods of a n alysis of o ils and fa ts , though some
what scattered in the literatu re* and a lso s lig h tly lacking
in standardization, are fa ir ly completely described in the
1 %
A .0*0*3* Manual and the O ffic ia l Methods o f the A.O.A.C.
I t should be understood and appreciated at the outset th at
the accurate analysis o f fa ts for the exact determination o f
a l l o f the p o ssib le individual con stitu en ts i s such as to
be w ell nigh an im p o ssib ility . The methods available are
en tirely too elaborate and lengthy to be p ra ctica l for com
m ercial u se. Of course, the high degree of complexity o f th e
fa t mixtures Ahemselves, and the clo se sim ila rity between the
various con stitu en ts, esp ecia lly the glycerid es, are the bages
1
O ffic ia l Methods of the American O il Chemist*s
S ociety, le fa x Manual, T S ’ ew Orleans: American O ii &hemist1 s
S ociety, Eev* 1935)*
2 O ffic ia l and Tentative Methods of Analysis of the
A^ociation^of^^ffo^X^^Tc^t^aX^5s^3S>fc'^®J lir i6^ °^ >D ^ ♦
A ssociation of O ffic ia l A gricultural Chemists, Bnd ed*, 1934) •
5
for th is d iffic u lty .
Fats and o ils in general are id en tified by th eir
p hysical constants as w ell as by a few sp e c ific reaction s
such as q u a lita tiv e color t e s t s . She former includes such
measurements as iodine absorption value, saponification
3
number, refractive index, t it e r , and a few oth ers. Golor
4
te s t s include the Halphen t e s t for cottonseed oil* the
5
Besson te s t for kapok oil* the Baudouin te s t fo r sesame
6
o i l , and others, these color t e s t s are performed by add-
ing a sp e c ific reagent to the sample and noting the color
obtained, which estab lish es or denies the presence of the
p articular o i l in question. Golor te s ts are not always
r e lia b le being influenced by certain steps in processing
as w ell as the nature of the o i l s , and for these reasons
7
should be performed and interpreted with discretion*
3
A .0 .0 .9« Manual^ 0£* o i t . . pp. 29-37*
4
Ib id ., p* 36.
' ® T. 0. Mehlenbacher, 1 1 Color te s ts for Kapok Qfl%
O il and Soap ( Chicago, 111.* G ille tte Publishing Go.*)
I h S r e s s .
^ O ffic ia l Methods of the A.G.A.G. 2nd ed ., 1924,
p. 296.
^ Mehl enbac her, 0£ . c i t •
Other analyses which apply to fa ts and o ils include
the separation of the saturated and unsaturated fa tty acids
with id e n tific a tio n of the individual members, measurement
9
of color, free fa tty acid content, and unsaponifiable matter.
determination of the solub le and in solu b le fa tty acid s,
and various other t e s t s for sp e e ifie purposes*
d istin c t and outstanding advantages should be gained through
the development of microscopic methods and technique for fa t
and o il id e n tific a tio n . F ir st, the time required would in
many eases be m aterially lessen ed * and sin ce time is the most
expensive reagent in laboratory use, any saving thereof is
advantageous. Second, good supplementary evidence could
be obtained to confirm in terp retation s based upon resu lts
obtained by the ordinary methods of a n a ly sis. Then too,
there are certain o ils whose id en tity cannot be d e fin ite ly
established by the Imown methods, but which might be proved
by microscopic te sts*
a
1 0
o f Mioroscopie Id e ^ tlfie a ti on. T w o
Gr« S. Jamieson, Vegetable O ils and. Fats (lew York:
Chemical Catalogue Go., 1982)p* v
9
A.O.C.S. Manual, op. c i t . , p. 30.
Benn lim ited , 1936) pp« li'4~158T
10
0* B» Elsdon, Edible O ils and Fats (London: Ernest
osrapwsr m
HISTORICAL
for years, stain in g methods have “ been used by b io
chemists and p h ysiologists for the purpose of id en tifyin g
1 1
the glycerides* However, there is l i t t l e b a sis for
b elievin g that stain in g would have any ap plication in fa t
d iffe r e n tia tio n , and, in fa c t, Green reports l i t t l e success
12
in th is direction*
the potassium soap needles formed by the addition o f
aqueous potassium hydroxide to the substance under examination
followed by subsequent id e n tific a tio n of these needles by
means o f the microscope, have been used by some to prove the
presence of fa ts in body tis s u e .
Methods for the detection of carnauba wax as an adul-
13
terant in beeswax have been described by Watson, and
14
Baughman and Keenan*
Bolton has described microscopic methods for the exam-
15
ination o f lard and b eef fat# These te s ts are conducted by
^ 0. E* McOlung, Handbook of M icroscopical Technique
(lew York: Saul 1* H oeb erIS W r'p T 'i^ S . * ' 1
IS
L. W . Green, "Chemical Microscopy of fa ts and Waxes"
O il and Soap. 11, 31, (1034).
1 %
Watson. American Bee Journal. March (1930).
14
Baughman and Keenan. American Bee Journal. 714
376-8, (1931)
E, R. Bolton, O ils, fa ts and fa tty food s, (P hila
delphia; P* B lak iston ^ Son & d o 2nd ed ., 1928) pp. 33-34*
6
c r y sta lliz in g portions of the fa t from ether at room tempera
tu res, and then examining the c r y sta lliz e d substance with
the microscope. Typical and ch a ra cteristic specimens are
obtained. The lim ita tio n s of th is method having been accepted,
variation s of the same are used to a considerable extent in
in d u stria l lab oratories. Bolton h3tso discusses the cry sta ls
obtained from shea nut o i l , palm o i l , hydrogenated f a t s f and
coconut and palm kernal stea r in e s.
Yan Alphen studied th e action o f phenylhydrazine, and
16
hydrazine on fats* Ore an reported on the use of phenyl-
hydrazine as a reagent for the microscopic id e n tific a tio n of
17
fa ts and o i l s 4 Sreen'S method consisted o f in tim ately
mixing a drop of the reagent with a drop of the sample on
® o
the s lid e and heating to 130 - 150 0.
Some workers have used the microscope for the detection
1®
o f certain substances in e sse n tia l o ils .
The microscope has been used in the examination of
drying o ils to obtain information r e la tiv e to p orosity, hard-
19
n ess, and resistan ce to water.
16 ,
Y. Alphen, Reo. Trav. Ohim. . 44, 1064, (1925);
Qhem , Abst. , 19, 1235, (1926)"'cite d by E llis , Hydrogenation
o f Organic Substances, (lew York: B. Yan Hostrand do.,' Srd'
M i j g i.-M.. ■ wwwJt r-' • j l w .......
eoL«, 1 9£u, p# o % i, .
17
L « W « Sreene, Boo. c i t .
18 Ibid, c itin g B u ll, S o i, Pharmacol. 27, 210 (1920).
19
Circular Humber 110, Paint Manufacturer's A ssocia
tio n of U.S. , lovember, l9&(5.
9
Bosenthaler, using a lcoh olic sodium and potassium
hydroxide, examined the soap cry sta ls obtained by mixing two
20
mgs* of the o i l with one drop of the reagent on the slid e*
He claimed to have obtained cry sta ls with peanut and o liv e
o i ls , and ch aracteristic forms with rape, castor and poppy
seed o ils*
Maclennon, using polarized lig h t, studied the cry sta l
21
forms of commercial soaps*
Hartwich and Uhlmarm used a concentrated solution o f
y
aqueous potassium hydroxide containing twenty per cent
ammonium hydroxide for id en tifyin g individual fa ts*
Elsdon has described lin k s method for the q u a lita tiv e
23
detection o f coconut o il* This method is rather am elaborate
process of c r y sta lliz a tio n to obtain cry sta ls ch aracteristic
for coconut o il* Trimen has reported that th is same method
might be used to d iffe r e n tia te between buffalo and cow butter
24
fa t, and also between sheep and goat butter fat*
20
Green, L oc*0it. c itin g Hosenthaler, Schweig, Apoth-
2 t g ., 58, 545-9, 55£-Tr~578-83, (1920)*
Maclennon, £* goo* Qhem * Ind. , 42, 393-4015, (1923)*
22
Sartwieh and Uhlmann, Arehiv* Pharm*, 240, 470,
(1902); 241, 111, (1903)*
23
llsd o n , Edible O ils and f a t s , 1926, p* 331 c itin g
Hinks*
24 Ibid, c itin g Trimen, (A nalyst, 38, 246, (1913)*
10
fit# American A ssociation of A gricultural Chemists
describe a microscopic procedure fo r the examination o f
25
butter f a t . 3?h#y recommend d irect observation of the fa t
after spreading out- on the s lid e with a covey gla ss SS|"Well
as examination of the cry sta ls obtained by c r y sta lliz in g from
other.
I»ewkowitsch discusses the microscopic appearance of
26
a few f a ts and o ils , and gives methods for the examination
o f the unsaponifiable matter for the presence o f ch o lestero l
m
and p hytosterol,
fh@ foregoing references include very nearly every-*
thing that has been published to date on the application o f
microscopy to f a t s , fhe l i s t is not imposing in i t s length,
furthermore, recent years have seen the use o f o ils o f kinds
theretofore almost unknown in commercial p ractice. A ll of
th is would seem to indicate th at further study should be made
Of thp a p p lic a b ility o f the older methods to the present day
used o i l s , and also attempts should be made to elaborate upon
these so as to make the scheme su itab le for the id e n tific a tio n
of further and additional species*
85 O ffic ia l Usthods o f the A.O.A.O,, 2nd ed ., 1924,
p. 277♦
26
y. lewkowitsch, technology and Chemical Analysis
o f O ils, fa ts and Waxes» (London': Macmillan & Bo,1 , l t d .. 2nd
¥ 1 ., 1921} V ol. I , pp.* 552-3,
27
Ibid, pp. 598-9.
/ /
CHAPTER IY
W fg P T ffi TMy < M T ,
The o il sample© which were used in these experiments
were of a known purity and represented an ordinary commercial
production grade. Although, the group of samples worked with
" b y no means represented a l l of th e known typ es, those o ils
most commonly used were included* A l i s t o f the o ils and
fa ts used in the experiments i s given in fa b le I*
I t was found desirable to use a dark f ie ld in the ex-
amination of a l l o f the fa t and o i l cry sta ls obtained# This
afforded much greater d istin ctiv en ess in ou tlin e and form*
I t w ill be noted in the tab les th at follow that some
attempt has been made to divide the o ils into groups depending
upon the ^ ch a ra cteristicp a ttern s, forms, and cry sta ls obtained
with a certain reagent*
R p yen th a ler1 a Method* A lcoholic sodium and potassium
hydroxide were applied to a l l of the o i l samples lis te d in
fa b le I* according to the procedure described by Rosenthaler.
The r e su lts obtained were far from satisfactory* H o d efin ite
patterns were obtained which were s u ffic ie n tly ch aracteristic
t© permit being used for d iffer en tia tio n o f the samples*
Reactions with Potassium Hydroride - without Heat, fhe
reagent was a saturated solu tion of potassium hydroxide in
12
T A B L E I
OILS AIL EA TS USM ) H M IG B G SO O P IO S T U B S '
S A M E L ® S O U H O E
28
Olive
Kapok
Seyf Beam
Bio© Bran (Eome)
P e r ilia
Oom (Maize)
Palm
Peanut
Walnut
lompgesd
Mustardgeed
Sesame
Cottonseed
Teaseed (feanut)
Bahassu
Coconut
Linseed
Bap©seed
Eyuit of the tr e e , m m eurosaea.
Seed of the tr e e ,
Seed of the legume, Glycine
Seed o f the p lant, Oryze e s tiv a .
Seed of the P e r llla ocymofdes.
Seed o f the p lan t, le a mays (Indian corn),
Eleahy portion of the fr u it of the palm,
Elaeia gulmeensfa.
Seed of the legume, Araohls hypogaea.
Kernel of the tr e e , Juglans regia*
Seed of the Cannabis gatlva.
Seed of the Slnapig alb a*
Seed o f the Sesamum indicum*
Seed of the plant
Seed o f several sp ecies of tea p la n tsf
Kernel o f the nut of the palm, Orblgnya#
Kernel of the nuts o f the palm. Cocoa maolfera
Seed of the fla x p la n t.
Seed of the Bgaaiioa ra p ,.
28
$• S. Jamieson, Vegetable O ils and E ats, (1932)
• 31— 285*
IOTE: Lard, beef f a t , whale o i l and sardine o i l were
Included in some of the experiments,
15
n-butyl alcoh ol. A drop of the sample was placed on a clean
s lid e , and about fiv e m m . from th is was placed a drop o f the
reagent, fhe s lid e was t ilt e d ca refu lly so as to cause the
reagent drop to flow on to the o i l drop. Xsanedlately a fte r
the two had touched, a cover g la ss was applied, ih e slid e
was then allowed to set at room temperature u n til cry sta ls
appeared a t the junction of th e two liquids* fh sse were
then examined with the microscope using a dark fie ld sub-
stage element. Several d istin c tiv e and d ifferen t forms and
cry sta ls were obtained. Continued rep etitio n of the t e s t s
seemed to in dicate that th e re su lts obtained were sp e c ific and
ch aracteristic fo r each of the o i l samples. A grouping o f the
o i l and fa t samples according to their reactions in th is t e s t
i s given in fab le IX.
Some o f the resu lts obtained with potassium hydroxide
in n-butyl alcohol without the ap plication of heat were yery
d efin ite* Among the more ch aracteristic were the cr y sta ls
with teaseed and o liv e o i ls , the fa ilu r e to produce any
patterns under the condition o f the t e s t , and the fibrous
forms obtained with a l l o f the marine o ils te ste d , feaseed
O il was read ily distinguished from o liv e o i l by the rate of
formation of the c r y sta ls f as w ell as by rep etitio n o f the
te s t with the application of h eat. (See fig u re l t page Id .)
3.4
u a m i i
im O I IO I S WISH POTASSIUM HYDROXIDE
WITHOUT H l i f
Group 1 Group 8 Group 3 Group 4 Group & Gfoup 6
Teaseed Palm Hape Seed Pish
Sesame
Peanut
O live Mustard * Sardine la p e l H om e
lempseed Whale lin seed
Walnut Oom
l e r i l l a
Soya Bean
Explanation o f Groups;
1. large cr y sta ls, concentric n eed les, three to twelve needles
per cr y sta l.
5 . Small spherical globular forms*
3* H o crystals or ch a ra cteristic forms.
4 . Irregular fibrous shapes, no regular pattern*
3. Small concentric n eed les, many needles per crystal*
6. Like group 1, but very sm all, used a t le a s t 200 magnifica
tions*
15
F ig u re 1
2?easeed O il with.
P o ta ssiu m Hydroxide,
w ith o u t H eat, 80 X.
F ig u re 2
Corn O il with.
P o tassiu m H ydroxide,
w ith H eat, 80 X.
16
witii PctM'tttoft fearexid e ~ w ith iss$^ f
reagent and general procedure were the same as that described
for th e use of potassium hydroxide in the cold w ith one ex~
* oeption. A fter placing a drop of the sample in th e center*
the f lid e was heated by passing over the flame of a micro
burner u n til the g la ss f e l t hot to the hands w hile being held
at the extreme ends# The reagent was applied t® th e s lid e
and a fte r bringing th is and the sample drop together, i t was
immediately covered with a cover g la ss. I t was necessary
to act rapidly in th is case to prevent evaporation o f the
a lco h o l, the ch a ra cteristic forms were developed w ithin a
very short tim e, the r e su lts obtained with th is method are
tabulated in fa b le III# (See fig u re £, page 15, and figu re
5 , page 18.)
Esaetions w ith Phenylhy^Lraz in e » The preferred procedure
j |ir hrihgiug about the reaction was b est affpctpd by placing
abouh one ml* of the sample ,in a th ree by tbroe~eighths te s t
tube and adding to th is one fourth ml* of phenylhydrazine#
The t e s t tube and contents were them heated in an o i l bath
to S00°C* With a sm all p ip e tte, a drop of the substance in
the tube was transferred to a slid e and covered with a cover
g la s s , the cry sta ls appeared w ithin a short time and could
be hept for some tim e. The re su lts obtained by the reaction
of phenylhydrazine with the fa t and o i l samples are given in
Table 17. (8ee fig u re 4, page 18, and fig u res 5 and 6, page SO.)
w I I I
X B t M X Q M WOT PQ fASSItJM HIDBO ZIDE
W OT
Group l Group E Group f Group 4
■ y - n K i p r i n r ■ i i m . B w t # w r a . w ■ i j m i i j ." W h
feaseed Palm
Rape Seed Sesame
Mustard n Hempseed
Oottonseed Whale
Sardine
Walnut
Group $ Group 6
sassaniB^eaea^^
P e r illa
B orne
Soya Bean
Kapok
Peanut
Coconut Olive
Babassu
Explanation of G rom pa:
i
1, large oonoentrio needles#
2 . Small bunchy tu fts w ith many oonoentrio needles protruding*
3* Branching forms, m ossy-like covering en tire f ie ld almost
at once•
4 . Branching m ossy-like forms, not as rapid growing or as
large as Group 3.
5* Resembling bunches of knotted thread*
6* Resembling groups 1 and 4 , but more lik e the latter*
18
F ig u re 4
O live O il w ith
P h e n y lh y d ra z in e , 80 X.
F ig u re 3
P e r i l l a O il w ith
P o tassiu m H ydroxide,
w i t h H eat, 80 X.
19
(M B IE IV
E m c u o s s w ish mwnwzM & zzn%
ia sssr i^ .- rsr. , « ■ , r - ............:u; taU'-iaagass'V
Group l Group 2 Group 3 Group 4
Mustard Kapok Whale feaseed
Linseed O live Sardine Walnut
Rape Seed Corn B a rilla
Cottonseed Palm
Sesame Coconut
Kempsead Babassu
Peanut
Some
Soya Bean
Explanation o f Groups:
1; Ip c r y sta ls with, phenylhydrazine •
2* Small groups of concentric needles* circu lar in o u tlin e •
5* Resembling Group 2* hut much larger*
4* Resembling tu fts of cotton with irregular protruding
needle s f
20
ty> -
* M
* * ' ♦ *
T ‘
»
« * «
> • *
^
\*
F ig u re 5
Sardine O il
w ith P h e n y lh y d raz in e ,
80 X.
F ig u re 6
W alnut O il w ith
Phenylhydraz in e , 80 X.
21
Reactions with Bromine* About one ml* samples of the
fa ts and o ils were dissolved in a mixture of chloroform and
aoetic acid (1 -3 ), in a t e s t tube. A fter solu tion , bromine
was added u n til an excess was reached as evidenced by the
color* The tubes were allowed to set at room temperature
u n til cry sta ls appeared, These were then removed by an open
end glass tube, placed on a gla ss s lid e , and covered with a
cover g la s s , The resu lts obtained by bromination are shown
in Table V*
Fatty A cids. The fa tty acids were prepared by
saponifying with potassium hydroxide in g lycerol, and then
lib era tin g the acids from the soaps by adding an excess of
d ilu te sulphuric acid . These were then washed with d is t ille d
water u n til the washings were neutral to litmus paper*
S lid es of the fa tty acids were prepared by placing
a drop of the substance on a glass s lid e , warming s lig h tly
and covering with a cover g la ss. For the reason that the
so lid ify in g points o f a great portion of the fa tty acids are
above normal room temperatures, these w ill c r y sta lliz e without
farther a tten tio n . S o lid ific a tio n o f the acids which have
tit e r s below room temperature must be brought about by using
a cold sta g e. Photographs of fa tty acid patterns are shown
on pages 25-29.
zz
T A B L E V
R E A C T IO N S W IT H B H O M IH E
©roup 1 ©roup Z ©roup 3 ©roup 4 ©roup 5
Coconut f is h Linseed © lire Kapok
Bah&ssu Whale P eri 11a Palm
Corn Sardine Walnut
lap® Seed *B©annt
feaseed Mustard
Cottonseed
Sesame
Hempseed
H orns
Soya Bean
Explanation of ©roups:
l f W o cry sta ls with, bromine*
Z f Thick mass of pin point forms*
3* Small circu lar c r y sta llin e forms*
4 . lesem bling small hunches of fat* irregular ou tlin e and
shape*
Bf ©roups composed of threadlike file r s *
* Unoh larger in size than Obtained with other o ils in the
same group*
23
F ig u re 7
Whale O il, F a t t y
A c id s, 80 X.
Domestic C otton seed
O il, F a t t y A cids,
80 X.
24
No. 1 O r i e n t a l
C ottonseed O il,
P a t t y A c id s, 80 X
No. 2 O r i e n t a l
C o tton seed O il,
P a t t y A c id s, 80 X.
25
F ig u re 11
Palm O il,
F a t t y A c id s, 80 X
F ig u r e 12
Kome O il, F a t t y
A c id s, 80 X.
26
F ig u re 13
B atassu O il,
F a t t y A c id s ,
80 X.
27
F ig u re 15
E&pok O il,
F a t t y A c id s ,
80 Z.
I
“ *
F ig u r e 16
l a r d , F a t t y A cid s,
80 X.
28
K ' < $
i ? y j . \ ^
• > ? ^ j ' t l y
Z & & & 1
f i g u r e 18
S a rd in e O il,
F a t t y A c id s,
80 X.
F ig u re 17
Beef F a t,
F a t ty A c id s,
80 X.
29
F ig u re 19
Coconut O il,
F a t t y A c id s,
80 X.
W ' £
t
f t
• i
F ig u re 20
Palm O il,
C rystallized,
80 X.
Freezing. A dsop of the liqriidflBt or o i l sample
was plaeed on a slid© and ©oyer g la ss applied at omee* She
S lid e was then ©hilled below the glyoeride m elting point
and fin a lly examined with the microscope• (See Figures 30.t
21 and 22, pages 29 and 31) •
31
F ig u r e 22
Coconut O il,
C r y s t a l l i z e d
80 X .
CH AM SB. T
O G H G IU SIO H
Maeh more experimentation i s necessary "before d e fin ite
methods can be recommended covering a ll phases of fa t and o i l
microscopy* Other o ils mast be included with those already
studied, and farther ch aracteristic reactions should be
sought* I t most be remembered that before any reaction or
te s t can be said to be ab solu tely d ls t in t t it e any o i l ,
i t should be tried with samples of that o i l from a l l p ossib le
sources so that a ll of the variation s in nature and character-
. . * * ■ '
i s t i c s w ill have been accounted for*
F am iliarity and experience with the chemistry of the
fa ts and o ils* general chemical microscopy, and more s p e c ifi
c a lly with the appearance of fa t and o i l crystals are
necessary before proficiency in judgment can be attained*
H o great attempt has been made to study mixtures o f
d ifferen t o ils but a s u ffic ie n t amount o f work has been done
to prove that some o ils are e a sily id e n tifie d in m ixtures,
w hile others o ffer d iffic u lty * I t has also been indicated
th a t, whatever the mixture, concentration is an important
fa c to r ,
fhere Is l i t t l e lik elih ood that microscopic procedures
cap ever be su bstituted for the regular chemical methods in
the q u a lita tiv e an alysis of the fats* However, there Is no
denying the fact that anyone experienced with the microscopy
33
of til® fa ts can gain very u sefu l information and confirmatory
evidence by means of th is method of examination* A f m re
action s and procedures have been described which w ill quite
d e fin ite ly prove the id e n tity of certain o ils , although group
reactions are apparently more d istin c tiv e and r e lia b le than
sp e c ific t e s t s for individual o ils#
She task yet remaining before a complete and r e lia b le
scheme o f analysis can be recommended is immense# Only a
portion of the fa tty substances known to mankind have been
included in th is study, but, insofar as i t haS; gone, the
r e su lts obtained have been very encouraging.
As m an learns and progresses he str iv e s to lessen h is
manual burden. It is altogeth er f it t in g and proper th at he
should do th is* For th is purpose he invents labor saving
machines and devises new, shorter, and more p recise methods*
I t i s to th is end that the in v estig a tio n herein discussed has
been dirooted#
B IB L IO G R A PH Y
35
Baughman and Keenan, American Bee Journal. 71, 376-8, (1931) *
Bolton, E# !♦ , O ils, fa ta and Fatty Jtoodsu (Philadelphia;
P. BlakistonTs Son A d o,, 2nd ed*, 1928)«
Qhsmob & Mason, Handbook o f Chemical Microscopy, Y ols. I and
I I , (Hew j o r ^ T i m 3 ^ 7 1 ^ 1 1 *
U l i s , 0 ., Hydrogenation of Organic Substances (How York:
D. Yan Hostraud d o .,“3rd e&., 1950) •
His don, G. D .* Edible O ils and Fats (london: Ernest Bonn
lim ited , f lw r r r *
Green, 1* W., "Chemical Microscopy of Fats and Waxes ” , O il
and Soan^ 11, 31 flfM j •
Hanausekand Winton, Microscopy o f f e o teio a l Products (lew
York: W 11 ey & Soms, i95Y) •
Hartwich and Uhlmann, Archir# Pharm., 240, 470, (1902); 241,
I I I , (1905)#
Jamieson, G. 8 ., Yeiretgbls O ils and Fats (Hew York: Chemical
Catalogue Co., 19521,
lewkowitsch, J#f legbnology and Chemical Analysis of O ils.
Fats^and^Waxes. (jondonV^IfacmllIan& Co., l t d , , 2nd e d .f
Maelennon, * [# Soc. Qhem . Ind. , 42, 393-4015?, (1923)#
MaOlung, 0# 1#* Handbook of Mlorosoopioal leoim igne (Hew York:
Paul B. Hoeber, 192977
Mehlehbaeher, v , c# "Color fe a ts for Kapok O il", O il and 8eap
(Chicago, 111,, G ille tte Publishing Co#,) In- Press#
O ffic ia l and fe n ta tir e Methods of A nalysis o f th e A ssociation
AgMculiu^T"CEemTat's. (WaiFin^on7T>7Q« ------
IssoclatT on or O ffloS -i AgricuTtura 1 Chemists, 2nd ed .,
1924)#
O ffic ia l Methods of the; American Oil Chemist1 a S ociety,
Lefax Hanual, (few Orleans; American O il Chemist’s
S ociety, Bev. 1935).
Paint Manufacturer^ A ssociation o f U*8#, Circular Humber 110,
Borember, 1920# V
Watson, American Bee Journal, March (1930)#

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Asset Metadata

Creator Mehlenbacher, V. C. (author)

Core Title Study of the application of the microscope to fat and oil analysis.

Degree Master of Science

Publisher University of Southern California (original), University of Southern California. Libraries (digital)

Tag chemistry, analytical,OAI-PMH Harvest

Language English

Contributor Digitized by ProQuest (provenance)

Permanent Link (DOI) https://doi.org/10.25549/usctheses-c17-786890

Unique identifier UC11348158

Identifier EP41480.pdf (filename),usctheses-c17-786890 (legacy record id)

Legacy Identifier EP41480.pdf

Dmrecord 786890

Document Type Thesis

Rights Mehlenbacher, V. C.

Type texts

Source University of Southern California (contributing entity), University of Southern California Dissertations and Theses (collection)

Access Conditions The author retains rights to his/her dissertation, thesis or other graduate work according to U.S. copyright law. Electronic access is being provided by the USC Libraries in agreement with the au...

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