A. S. SAMSON S. J.,’ R. N. KHAUND, C. M. CATERand K. F.

MATTIL
Food Protein R&D Center, Texas Engineering Experiment Station
Texas A&M University, College Station, TX 77843

EXTRACTABILITY O F COCONUT PROTEINS

SUMMAR Y-Studies were made on nitrogen (proteinl solubility of coconut meal in aqueous media Coconut meal was prepared from the fresh
over a range of pH’s. The point of minimum solubility was found at pH 3.9 and nitrogen solubility coconuts by procedures that would not be ex-
increased toward the acidic and basic sides. Cbconut proteins displayed a markedly different pected to denature the proteins. The nuts were
pH-nitrogen solubility profile in salt solutions, showing minimal solubility at acidic pH’s IpH 4 31, cracked and the coconut water discarded. The
meats were removed from the shell, washed
a sharp rise of solubility and a maximum at neutrality. Coconut meal prepared from fresh coconuts
with water and then sliced into smaller frag-
with and without testa removed, from parings and from a sample of sun-dried copra showed
ments using an Urschel mill (cutting head
comparable solubility characteristics. However, meal prepared from commercial desiccated coconut 2-J-030510B or 2-K-020060B). The shredded
showed low solubility, even under acid and alkaline conditions IpH 2 and 1OJ. The most efficient meats (which included the testa) were dried un-
solvent-to-meal ratio was found at 20: 1 fv:wJ for a single extraction, and a two-step 15: 1 process der vacuum at 40°C for 24 hr to a moisture
for multiple extractions. Extractions using salt solutions show that the efficiency of extraction is content of less than 2%. The dried meats were
greatly dependent upon pH; concentrations > 0.25M effect maximum extraction. Osborne classifi- further comminuted by means of the Urschel
cation studies of coconut meal indicate that 90% of the proteins would be classified as albumins mill, this time using a finer cutting head
and globulins. (2-K-020035C or 2-K-010030B), prior to defat-
ting by several extractions with hexane. Residu-
al solvent was removed with dry air.
INTRODUCTION have made use of meats dehydrated by Typical analyses of coconut meal prepared
the application of heat (Sison et al., in this fashion are given in Table 1. Moisture,
COCONUTS are an indigenous product 1968). Studies by Chandrasekaran and ash, oil and crude fiber analyses were made by
and a major article of commerce in many King (1967) suggest that enzymatic proc- standard AOCS (1969) procedures. Table 1 also
countries of the world which are suffering esses may be required for the efficient gives typical analyses of fresh coconut meats
from protein deficiency. A conservative and desiccated coconut meats involved in the
extraction of coconut protein (from a
estimate, probably low, of world coconut preparation of coconut meal. It is to be noted
control value of 50-60% to 85%); how- that the moisture content of the dried meats
production in the early 1960’s gives 5.6 ever, they do not indicate how their equilibrated to a level of 7-10% on standing at
million tons a year. This is equivalent to starting coconut flour has been prepared, ambient temperature and humidity.
more than 200,000 tons of crude protein, except that it was supplied by the Central
only a minor proportion of which is Extraction of protein
Food Technological Research Institute,
presently being utilized for human food Mysore, India. Extraction of coconut proteins from coco-
consumption (Orr and Adair, 1967; PAG, nut meal was carried out at room temperature,
1965; Teply, 1967). EXPERIMENTAL for at least 30 min, using a solvent:meal ratio of
2O:l (v:w), unless otherwise specified. The pre-
The major proportion of coconuts is Reparation of coconut meal determined pH of extraction was obtained by
converted to coconut oil and coconut The coconuts used were food-grade fresh co- addition of OSN NaOH or 0.5N HCI solution;
meal via copra. Commercial processes for conuts obtained from the wholesale market in the pH was rechecked and readjusted after 30
extracting coconut oil commonly utilize Houston, Texas. Indications are the coconuts min of stirring. Sample size was 2.Og and the
expellers at temperatures which denature came from Central America. As is common volume of solution was brought to 40 ml after
proteins. Furthermore, prevailing world with commercial coconuts, these were received the final pH adjustment. After centrifugation at
practice in handling coconuts for the dehusked. The meats (kernel) are enclosed in a 4300 x G (Sorvall RC2-B Refrigerated Centri-
production of copra are not conducive to hard shell (endocarp). Between the shell and fuge) for 26 min, the supernatani extracts were
further utilization of the coconut meal the kernel is a thin, brown seed coat (testa) filtered through Whatmafr Filter Paper No. 1 to
which adheres firmly to the kernel. In the man- remove flocculent materials. A 20 ml aliquot of
for human consumption.
ufacture of desiccated coconut, the testa is re- each extract was taken for nitrogen analysis.
These studies were undertaken as a moved by paring. Nitrogen was determined either by macro-Kjel-
necessary preliminary step to the study of
basic properties of coconut proteins in
connection with the larger objective of
finding methods for the better utilization Table l-Coconut meal: Proximate analyses
of coconuts as protein-food sources. To
insure that the present work was being Moisture
conducted on good quality coconut pro- and volatile Protein Crude
tein, care was taken to prepare coconut material Oil N (%N x 6.25) Ash fiber
meal by processes not expected to de- Sample % % % % % %
nature proteins. Fresh meats 47.0
Previous studies on coconut meal have Desiccated
commonly used meal prepared from com- meatsa 1.8 13.0
mercial coconut oil processing (Chelliah 1.0 71.1
and Baptist, 1969) or from sun-dried 1.5 69.3
copra (Butterworth and Fox, 1963), or Coconut meal Ab 9.9 1.1 3.6 22.2 5.2 7.5
Coconut meal Bb 1.2 4.8 3.8 23.9 3.9 5.9
Coconut meal Cb 5.1 0.4 3.8 23.8 4.4 7.6
’ Present address: Dept. of Chemistry, Aten- ‘These are analyses of different lots dehydrated in the freeze drier.
eo de Manila University, Loyola Heights, Que- bCoconut meals A, B and C are different samples, prepared from three lots of fresh coconuts
zon City, Philippines. obtained at different times.

Volume 36 (1971)-JOURNAL OF FOOD SCIENCE-725

726 -JOURNAL OF FOOD SCIENCE- Volume 36 (1971)

IOO-

so-
5
5
3 60-
5:
5 40-
0”
6
0 zo-

0’ I I I I I I 0-1
2 4 6 8 IO 12 2 4 6 8 lo 12 2 4 6 8 IO 12
PH PH OH

Fig. l-Protein solubility profiles of coconut Fig. Z-Protein solubility profiles of coconut Fig. d-protein solubility profiles of a desic-
meals prepared from three lots of fresh coco- meal in several different salt solutions at varied cated coconut and a specially prepared copra.
nuts obtained at different times (see Table 1). pH’s adjusted by the addition of 0.5N NaOH or
0.5N HCI.

dahl (AOAC, 1965; AOCS, 1969) or micro- over a range of pH’s. The data shown in cipitation at pH 3.9. Sison et al. (1968)
Kjeldahl methods (Aminco, 1959; Miller and Figure 1 are for extracted meals prepared reported the best pH for protein precip-
Houghton, 1945). In most cases, the data from three different lots of coconuts. The itation in a citrate-phosphate buffer at pH
shown represent duplicate analyses, using two solubility profile for coconut meal C 3.4-3.5. Chelliah and Baptist (1969)
samples of meal.
represents the average of four replica- precipitate coconut protein at pH 4.5.1
RESULTS & DISCUSSION tions; the 95% confidence limits for a Nitrogen solubility increased with in-
single mean is f 2.95%. creasing acidity and alkalinity. The inflec-
pH-Nitrogen solubility profiles The point of least solubility was fur- tion in the curves near pH 8 suggests that
Studies were made on nitrogen (calcu- ther refined by extractions at pH’s 3.0, there may be two main protein fractions:
lated as protein using the customary 6.25 3.3, 3.6, 3.9, 4.2, 4.5, 4.8 and 5.0. one soluble at pH 8-8.5 and the other at
factor) solubility or extractability of Minimum solubility was obtained at pH pH 10.5-l 1.
three coconut meals in aqueous media 3.9. [Peters (1960) found maximum pre- The solubility profile of coconut meal
is strikingly different in salt solutions
(Fig. 2). In 1M NaCl solution, there is
rather low solubility on the acid side, up
to pH 4. This is followed by a steep rise
Table Z-Classification of the proteins of coconut meal according to in solubility to over 90% at pH 6 and
solubilitva more alkaline conditions. Solubility in
1 M CaCl, displays a similar low solubility
Fractions Extraction conditions % of Total N at acidic pH’s, followed by high solubility
Albumin CO, -free water, pH 6.7 30.6 near pH 5-6. An unexplained decrease in
Globulin 1M NaCl solution, pH 7.0 61.9 solubility at more alkaline pH’s is also
Prolamine 70% aqueous ethanol 1.1 observed.
Glutelin 0.2% NaOH solution 4.7 Similarly, coconut proteins show low
Insoluble residue -- -- 1.8 solubility at lower pH’s for 0.25M
Nonprotein nitrogen 12% sodium tungstate to Naa SOa and 0.25M dibasic sodium phos-
urecipitate oroteins 0.1 phate. The pH-solubility profile rises
steeply beginning at pH 4 and reaches a
aCkborne classification. maximum of almost total solubility at pH
8. A decrease of solubility at more
alkaline conditions follows.
The unique solubility characteristics of
Table 3-Pared coconut meal and coconut parings: Proximate analyses coconut proteins in salt solution have
been used to prepare an interesting isolate
Moisture
by extraction in 1M NaCl at pH 7
and volatile Protein Crude followed by precipitation at pH 2. The
material Oil N (%N x 6.25) Ash fiber
Sample preparation and characteristics of this and
% % % % % %
other isolates prepared by extractions at
Fresh pared pH 2, 8, and 10.5 are the subject of
meats 47 another manuscript now in preparation.
Fresh testa 35 The classical Osborne classification of
Desiccated proteins (Lund and Sandstrom, 1943), as
pared meats 1.1 73.5 applied to coconut meal, confirms the
Desiccated solubility data obtained in aqueous and
testa 2.2 54.7 salt media. Table 2 indicates that over
Pared meal 10.6 5.2 3.6 22.5 4.2 5.6 90% of the proteins in coconut meal
Defatted would be classified as albumins and glob-
testa (meal) 8.9 3.6 3.2 20.0 2.9 12.1 ulins. The sodium tungstate test indicates
 EXTRACTABILITY OF COCONUTPROTEltVS-727

Efficiency of protein recovery log. After centrifugation and gravity fil-

Protein solubility or extractability is tration, the supernatant volume was
not the only factor governing efficiency measured. Under these conditions, three
of protein extraction. Other variables to five times the weight of coconut meal
such as length of time of extraction (i.e., 30-5Og) was absorbed by the meal,
meal:solvent ratio, temperature of extrac- and was not recovered as part of the
tion and the effect of added salts are also separable solution. The amount not sep-
very important. arable under practical conditions would
Table 4 gives data obtained by varying vary depending upon the efficiency of the
the meal:solvent ratios, while extracting separation techniques used. The data in
at pH 8 in aqueous solution and pH 7 in Table 4 indicate that a solvent:meal ratio
,- Gas’, 1 M NaCl (conditions chosen as represent- of 2O:l (v:w) would be the most effi-
/ I
0
02 04 06 08 IO ative for aqueous and salt extraction). cient, for single extraction.
MOLAF3ITY OF SALT SOLUTION The sample size for these extractions was Table 5 gives efficiency of protein

Fig. 4-Solubility of the proteins of coconut

meal in different concentrations of several salts.
(Data next to solubility points indicate result-
ingpH’s.1 Table 4 Effect of solvent:meal ratio on extraction

AtpH8 At pH 7, 1M NaCl
Solvent : meal
ratio (ml:n) % N recovereda % N extracteda % N recovereda % N extracteda
5:l 11.9 60.9 17.1 95.1
that there is very little nonprotein nitro- 7.5:1 25.8 62.3 38.3 101.0
gen in coconut meal. 10 :l 32.2 60.1 53.6 97.4
The solubility curves (Fig. 1 and 2) 15 :l 40.8 61.2 63.3 94.0
were made on coconut meal which in- 20 :l 47.1 63.9 75.0 97.4
cluded the testa. In addition, some coco- 25 :l 46.6 59.2 77.9 91.7
nut meal was prepared with the testa 30 :l 49.4 62.4 83.6 100.3
removed by manual paring. The parings =The percent nitrogen extracted (or soluble) is based on the total nitrogen present in the
were also dried and defatted and the coconut meal sample and the total volume of solvent used for extraction. The percent nitrogen
solubility profile of the proteins deter- recovered refers to that nitrogen in the separable supernatant extract, excluding the amount of
mined. Table 3 summarizes some proper- extract (solvent) absorbed by the meal. The difference between the percent nitrogen extracted
(dissolved) and recovered represents the nitrogen in solution in the absorbed solvent. Percent
ties of coconut meal without the testa, nitrogen recovered is the more useful data for process development purposes.
and also of the coconut parings. The
pared coconut meal gave a slightly higher
protein analysis than the parings (22.5%
protein compared to 20.0%; 22.8% pro-
tein compared to 17.7% in another deter- Table 5-Effect of successive extractions with different solvent:meal
mination). The solubility curves of pared ratios
coconut meal and of the parings are Solvent:meal AtpH2 AtpH8
comparable to those of coconut meal ratio (ml:& Extraction %N recovereda %N recovereda
with the parings. 1011 First 45.8 33.7
Solubility profiles of the proteins in Second 17.7 12.5
meals prepared from commercial coconut Third 7.1 8.2
products have been obtained. Figure 3 TOTAL 70.6 54.4
gives the solubility profile for coconut 15:l First 65.5 40.7
meal obtained by defatting commercial Second 17.9 13.3
desiccated coconut and sun-dried copra TOTAL 83.4 54.0
prepared under a “heat tent.” The meal (Third) 5.9 9.5
from sun-dried copra gave a pH-solubility 2O:l First 61.3 43.6
profile comparable to the laboratory- Second 14.1 14.8
prepared coconut meal. That from desic- TOTAL 75.4 58.4
cated coconut, however, displayed rather aBased on actual recovery of extract.
low solubility at both acidic and basic
pH’s. The very low solubility (26%, com-
pared to about 7 0 % in the laboratory-
prepared meal) at pH 8 is also striking.
Table 7-Effect of temperature on extrac-
The behavior of commercial desiccated Table 6-Effect of time on extraction tiona
coconut is very puzzling, inasmuch as the
product is perfectly white. Preliminary Time AtpH8 At pH 7, 1M NaCl Temperature (“C) %N extractedb
work done at Texas A&M University (to (min) %N extracteda %N extracteda 4 49.0
be reported elsewhere) has indicated that 5 58.6 96.9 10 57.7
browning might be a useful indication of 15 59.6 99.5 15 61.5
incipient denaturation, as shown by loss 30 58.4 99.0 29 71.6
of solubility. This, however, does not 45 59.3 98.6 52 96.1
seem to be the case with commercial 60 60.5 103.0 75 92.3
desiccated coconut, which, though pure 90 65.4 104.0 ?Solvent: 0.2M phosphate buffer, pH 7.0.
white, has rather poor solubility. aBased on total solvent used. bBased on total solvent used.
I 728 -JOURNAL OF FOOD SCIENCE- Volume 36 (1971)

extraction and recovery with multiple Figure 4 summarizes the effect on 10th ed. Assoc. of Official Agricultural
Chemists, Washington, DC.
extractions, using 10: 1, 15:l and 20: 1 nitrogen solubility of several salts at AOCS. 1969. “Official and Tentative Meth-
ratios. From these data it appears that a various concentrations. The tests were ods.” 3rd ed. American Oil Chemists’ Soci-
two-step 15: 1 extraction may be most run using the salts without adjusting the ety, Chicago.
Butterworth. M.H. and Fox, H.C. 1963. The
efficient. Though using one step less than pH. In Figure 4, data next to the solubil- effect of heat treatment on the nutritive
a triple 1O:l extraction, the overall yield ity points indicate the resulting pH at value of coconut meal and the prediction of
nutritive value by chemical methods. Brit.
of the two-step 15: 1 is almost as high. each salt concentration. J. Nutr. 17: 445.
Compared to the two-step 2O:l extrac- Concentrations at 0.25M or higher of Chandrasekaran, A. and King, K.W. 1967.
Enzymatic modification of the extractabil-
tion, it uses almost a third less solvent, dibasic sodium phosphate or sodium sul- ity of protein from coconuts (Cocos nucif-
with only a small loss of efficiency of fite extract most of the available nitrogen era). J. Agr. Fd. Chem. 15: 305.
protein recovery. The most practical ex- in coconut meal. NaCl is most efficient at Chelliah, J. and Baptist, N.G. 1969. Extraction
of protein from expeller- and solvent-ex-
I traction conditions would be selected on concentrations of 0.75M or higher. CaCla tracted coconut meal by dilute acid, alkali,
I the basis of total process economics. and salt solutions. J. Sci. Fd. A&c. 20: 49.
solutions, at 0.5M or higher concentra-
Gheyasuddin. S., Cater, C.M. and Mattil. K.F.
Other factors affecting protein tions, effect over 80% extraction. Mono- 1970. Effect of several variables on the ex-
solubility basic sodium phosphate, however, reaches tractability of sunflower seed proteins.
J. Food Sci. 35:453.
a maximum extraction of a little less than Lund. A.P. and Sandstrom, W.M. 1943. The
In determining the effect of length of 60% of the available nitrogen, at concen- proteins of various tree seeds. J. Agr. Res.
time of extraction on nitrogen solubility, trations of 0.5M or higher. This differ- 66: 349.
it usually took at least 5 min to attain the Miller, L. and Houghton, J.A. 1945. The mi-
, ence in solubility could probably be cro-Kjeldahl determination of the nitrogen
I desired pH, and then 15 min for centrifu- content of amino acids and proteins. J. Biol.
explained in part by noting the resulting Chem. 159: 373.
gation. Thus, the “time” variable em- pH’s: both Naa SO3 and Naa HP04 attain
Orr. E. and Adair. D. 1967. “The Production of
ployed was artificially set as actual stir- Protein Foods and Concentrates from Oil-
pH’s on the basic side of neutral, in which
ring time after the desired pH had been seeds,” p. 11. Tropical Products Institute,
coconut protein is very soluble (as seen in London.
attained. Table 6 indicates that maximum PAG. 1965. FAO/WHO/UNICEF PAG News
Figure 2) while NaHzP04 results in pH’s
nitrogen solubility is attained in 15 min Bulletin No. 5, p. 33. New York.
between 4.6-5.5. Perhaps by coinci- Peters, F.E. 1960. Preparation of amino acid
or less.
dence, CaCla and NaCl effect pH’s at or composition of selected seed protein frac-
Table 7 summarizes the effect of near the highest points in their pH-solu- tions. p. 20. Ph.D. thesis, Purdue University,
temperature on nitrogen solubility. A bility profiles (see Fig. 2.). Lafayette, Ind.
Sison. B.C., Fernandez. E.B., Capulso, S.A., de
temperature increase to about 5O’C in- It should be noted that the very high la Fuente, F.S., Gonzales. A.L., Claudia.
I creases nitrogen solubility. The decrease solubility of coconut proteins in dibasic T.R. and Manalac, G.C. 1968. Coconut pro-
tein studies: Studies on the optimum condi-
observed at 75” may be due to partial sodium phosphate and sodium sulfite is in tions of extraction and precipitation of pro-
coagulation of protein at the higher tem- marked contrast to the rather poor solu- tein from coconut meat. NIST Technical
perature. A similar decrease in solubility Bulletin No. 2, National Institute of Science
bility of sunflower seed proteins under & Technology, Manila, Philippines.
at higher temperatures has also been similar conditions, in work done in the - -. L.J. 1967. Whole fresh coconut meat
TeDlv.
reported in studies on sunflower seed same laboratory and simple preparations from it as sources
(Gheyasuddin et al., of protein and other nutrients for young
proteins in the same laboratory (Gheya- 1970). This confirms the need to study children. FAOIWHOIUNICEF PAG Docu-
suddin et al., 1970). each plant seed protein system individ- ment 916, New’York.’
MS received 5126170; revised 4114171; accepted
The effect of various added salts on ually and makes generalizations impos- 4125171.
nitrogen solubility has also been studied. sible prior to actual experimental work. Portions of the data were obtained in re-
From Figure 2, giving the solubility search conducted by R.N. Khaund in partial
curves of coconut proteins under varying REFERENCES fulfillment of the Ph.D. degree.
pH’s for different salt solutions, it is This work was supported by the Agency for
Aminco. 1959. The determination of nitrogen International Development, Contract PIO/T
evident that these salt solutions, except by the Kjeldahl procedure including diges- 931-11-199-864, 73-3192010; the Agency’s
toward the acid end of the pH scale, are tion. distillation, and titration. Aminco Re- help is gratefully acknowledged.
print No. 104, American Instrument Co., The authors thank John M. Bulder, FAO.
rather effective solvents for coconut Inc.. Silver Spring, Md. Paramaribo, Surinam for samples of sun-dried
meal. AOAC. 1965. “Official Methods of Analysis.” copra.