304 FLORIDA STATE HORTICULTURAL SOCIETY, 1970

Acknowledgments of compounds for the reduction of acidity in citrus fruit.

Proc. Fla. State Hort. Soc. 81: 1-6.
2. Pieninger, A. P., and W. F. Newhall. 1968. Deriva
We would like to thank Messrs. Glen Cop- tives of (_|_)limonene. Effect of chain length in n-alkyl-
quaternary ammonium derivatives on plant growth retard-
pock, Steve Ropicki, and Jack Lightner for ant activity. J. Agr. Food Chem., 16: 523-524.
3. Reese, R. L., and G. E. Brown. 1969. Legal ma
their help with the yield studies and Mr. Jim
turity of 'Temple' oranges as influenced by lead arsenate
Vice for the juicing operations. sprays. HortScience 4: 96-97.
4. Reitz, H. J. 1949. A study of certain factors affect
ing the acidity of Florida grapefruit following arsenic
sprays. Ph.D. Dissertation, Ohio State University.
LITERATURE CITED

1. Attaway, J. A., and B. S. Buslig. 1968. Screening

BIOCHEMICAL CHANGES IN GRAPEFRUIT AND 'MURCOTT'

CITRUS FRUITS AS RELATED TO STORAGE TEMPERATURE1
N. Vakis, J. Soule, R. H. Biggs storage. Glucose decreased at 50 °F and 60°F
and fructose increased at 50 and 60 °F. Changes
Department of Fruit Crops
in total reducing sugars were small. Internal
University of Florida
ethylene content of grapefruit was related to
Gainesville
fruit deterioration and respiration rates. Ethy
AND
lene treatments during storage of 'Marsh' grape
W. Grierson
fruit resulted in higher cellulase activity and
Florida Citrus Experiment Station internal ethylene content.
Lake Alfred
Introduction

Abstract
Citrus fruits do not undergo rapid chemical
Respiratory activity of 'Marsh' grapefruit at or physical changes ("ripening") after harvest
70 °F was highest following storage at 70 °F and (1,3,6,7,8,9,10). Their storage life, as with other
lowest subsequent to storage at 55 °F. Interme fruits, depends among other factors on the
diate respiratory rates were obtained after holding conditions, especially temperature.
storage at 34 and 40° F. Respiration of 'Mur- Knowledge of the biochemical changes during
cotts'2 at 70 °F was highest after storage at storage is essential if research to extend storage
34 °F and lower after cold storage, differences life and improve keeping quality is to be ef
among respiration rates after 40, 55 and 70°F fective. Various workers have reported in this
storage being small. Respiratory rates of 'Marsh* field (3,5,6,7,8,9,10,16,19,21,22), but more in
grapefruit peel were increased by wounding and formation is needed relating to local conditions
were higher than for intact fruit and pulp. and cultivars not included in earlier studies.

Cellulase and polygalacturonase activity in

grapefruit peel increased during storage while Materials and Methods
pectinmethylesterace activity decreased. Adeno-
'Marsh' grapefruit from several harvests
sine-triphosphate (ATP) levels in the peel in
and 'Murcotts' from a single harvest were ob
creased during the first 2 weeks at 40 °F and
tained from the Citrus Experiment Station, Lake
then declined sharply. Differences in ATP levels
Alfred. Ethylene-treated 'Marsh' grapefruit
among storage temperatures were negligible at
were provided by the USDA, MORD, Horticul
the end of 5 weeks storage. Total sugars and
tural Field Station, Orlando.
sucrose in grapefruit peel decreased during
Oxygen uptake at 70 °F subsequent to storage
was measured with an automatic sampling sys
lFlorida Agricultural Experiment Stations Journal Series
No. 3742. Grateful acknowledgement is made to the TJSDA,
tem (5) every 4 hours. Five replications, each
MORD, Horticultural Field Station, Orlando, for providing of up to 1 kg, were used for both grapefruit and
fruit from their experiments, and to Mrs. Sue J. Wells and
Mr. Sidney C-T Chen for technical assistance. 'Murcotts'. CO2 evolution at 70°F by grapefruit
2'Murcott* is a presumed tangor (tangerine x orange
was measured by infra-red analysis (23). Re-
hybrid).
 VAKIS, ET AL: GRAPEFRUIT - MURCOTT - STORAGE 305

suits were expressed as ml O2 or mg CO9 per to prior storage temperatures but did not exhibit
kg/hr.
visual symptoms of chilling injury (Fig. 4).
Enzyme activity, adenosine triphosphate 'Murcotts' responded differently than did
(ATP), adenosine diphosphate(ADP), an in
ternal ethylene were determined by methods
previously reported (21). Assays for cellulase
used methods previously described (13). Sam
ples of both cellulase extract and grapefruit
peel were frozen and held until assays were
made.
55 F
Grapefruit peel was extracted twice with
alcohol for sugar determinations and diluted 20
to 30-fold (on a fresh weight basis). Aliquots
i
were taken for colorimetric determinations of
total reducing sugar and apparent fructose. Su 3 4 6 7
DAYS
crose, glucose, and fructose contents were cal 1258-67-1
culated from these determinations (19).
Fig. 2.60, uptake of 'Marsh* grapefruit at 70 P after 4
weeks storage at 4 temperatures.

Results and Discussion

70 F
Respiration rates of 'Marsh' grapefruit at
70°F were higher after 2 or 4 weeks storage
at 70 °F than those of fruit stored at 34, 40 and
55°F (Figs. 1 and 2). Fruit stored at 40°F for
2 weeks had a higher respiration rate than that
stored at 34°F, but this difference disappeared
after 4 weeks storage (Figs. 1, 2 and 3).
Increased respiration rates following storage
at 70°F reflect the faster rate of deterioration
of the fruit probably stimulated by higher in
ternal ethylene content, as has been reported by
others (3, 23). High respiratory activity sub
sequent to storage at 40°F indicates chilling
injury that typically occurs in grapefruit (1, 3,
4). More mature 'Marsh' grapefruit from a later WEEKS STORAGE
harvest showed initial increases in respiratory
activity after 2 weeks storage inversely related Fig. 3. Effect of 2 and 4 weeks storage at various tem-
5S5illptal" at 70 p of 'Mareh> srapefniit
i 34 F
2 WEEKS STORAGE \*—-* 40 F 14-
~~\ 55 F
» 70 F 70 F
INITIAL
12

6
10

4-
I 8
2-

0
3 4 6
2 4
DAYS WEEKS
1258-67-1 1252-31-I

Fig. 1. 0, uptake of 'Marsh* grapefruit at 70 F after 2 4- InjtiaI rates of COO evolution of fully mature
weeks storage at 4 temperatures.
£UU!IIsubse<inent to t f
306 FLORIDA STATE HORTICULTURAL SOCIETY, 1970
grapefruit to holding temperatures. Prior stor declined rapidly during the first 48 hours as the
age at 34 °F caused increased respiration on time from wounding decreased (Fig. 7). Thus,
removal to 70 °F. There were essentially no incipient or apparent chilling injury to the peel
differences in respiration of 'Murcotts' stored at accounts for the increase in respiration of grape
40, 55 and 70°F (Figs. 3, 5, 6). Fruit stored fruit previously stored at 34 to 40°F. The in
at 70°F developed off flavors after 4 weeks; crease in respiration of 'Murcotts' previously
but there were no apparent differences in internal stored at 34° F indicates possible incipient chill
quality of fruit stored at 34, 40 and 55 °F. ing injury at that temperature.
The response to holding temperatures by Cellulase activity in 'Marsh' grapefruit peel
grapefruit and 'Murcotts' may be attributed to increased during storage and was highest follow
their degree of susceptibility to chilling injury ing storage at 70°F (Table 1). Increases in
and to their composition. The peel constitutes activity in fruit stored at other temperatures
approximately 20 to 25% of the fresh weight were not consistently related to temperature or
of the grapefruit but is a much lower proportion duration of storage. Ethylene in the storage at
in 'Murcotts*. Grapefruit peel tissue has a higher mosphere enhanced cellulase activity (Table 2).
respiration rate than does the pulp, and it re Ethylene treatments did not prevent low temp
sponds more readily to stress, e.g., wounding or erature pitting. Cellulase has been shown to be
chilling (Figs. 7, 8). Respiration rates of the necessary for the cellular breakdown involved
peel were high upon removal from the fruit, but in fruit abscission (13), and may contribute to
the changes associated with postharvest senes
34F cence. However, the results did not indicate a
2 WEEKS STORAGE A-™
55 F
""• 70 F
18
16
||4
fl2
*IO
PEEL
DECAY
o
STARTED
3 4 5 6 20 24 48
12 16
DAYS HOURS I258-8M
1258-76-1
Fig. 7. Oxygen uptake at 70 F immediately after wound-
Fig. 5. Effect of 2 weeks storage at various tempera
tures on oxygen uptake at 70 F. of 'Murcotf tangors. ing of 'Marsh' grapefruit tissues.
4 WEEKS STORAGE 12
x a- 10-
uJ = 6
UJ
d PEEL
40F
1 "■*
CM

I4 o 2
-•a WHOLE FRUIT
3 4
1258-81-1
DAYS
1258-76-1
Fig. 8. 0o uptake at 70 F of component tissues of
Fig. 6. Effect of 4 weeks storage at various tempera •Marsh' grapefruit. (See Fig. 7 for respiration during the
tures on oxygen uptake at 70 F, of 'Murcott' tangors. period immediately following disection).
 VAKIS, ET AL: GRAPEFRUIT - MURCOTT - STORAGE 309

Table 9. Internal ethylene, as Table 10. Internal ethylene, as ppm, in non-waxed and
waxed 'Marsh' grapefruit subsequent to storage
ppm, in 'Marsh1 grape for 4 weeks at 40 and 50 F in artificially main
fruit and 'Murcotts' tained levels of ethylene in the storage atmosphere.

subsequent to storage C2H4 in Non-waxed fruit Waxed fruit

at 34, 40, 55 and 70 F storage Picked Mar. ' 70 Picked May '70 Picked Mar. '70
for periods of 2 and 4 atmosphere 40 F 50 F 40 F 50 F 40 F 50 F

weeks. 0 .022 .001 .036 .070 .040 .075

1 .021 .102 .023 .066 .024 .069

Storage weeks in storage
temperature 0 10 .026 .063 .065 .130 .030 .143

200 .028 .157 .069 .100 .041 .219

'Marsh*harvested Jan., 1970 500 .021 .042

34 F .008* .008 .058 1258-79-1

1258-90-1

40 F .008 .009 .024

LITERATURE CITED

55 F .008 .017 .021 1. Biale, J. B. 1961. The post-harvest physiology and

chemistry of the orange fruit. In W. B. Sinclair, ed. The
Orange: Its biochemistry and physiology, pp. 96-130. Univ.
70 F .008 .033 .350 Calif. Press, Berkeley. 475. pp.
2. Biale, J. B., R. E. Young and A. J. Olmstead. 1954.
Fruit respiration and ethylene production. Plant Physiol.
'Marsh'harvested March, 1970 29:168-174.
3. Eaks, I. L. 1970. Respiratory response, ethylene
production, and response to ethylene of citrus fruits during
34 F .027 .045 .023 ontogeny. Plant Physiol. 45:334-338.
4. Eaks, I. L. 1965. Effects of chilling injury on the
respiration of oranges and lemons. Proc. Amer. Soc. Hort.
40 F .027 .059 .023 Sci. 87:181-186.
5. Green, G. F., E. M. Ahmed and R. A. Dennison.
1969. An automatic sampling system for respiratory gases
55 F .027 .045 .048 and respiratory response of irradiated citrus fruits J.
Food Sci. 34:627-629.
6. Harding, P. L., B. A. Friedman, M. B. Sunday, J.
70 F .027 .078 .154 Kaufman and H. W. Hruschka. 1952. The effect of pre-
storage treatments and storage temperatures on the keeping
quality of Florida grapefruit at Orlando, Fla. and New York
'Murcott' harvested March, 1970 City, N. Y., U. S. Dept. Agr. H. T. & S. Rep. No. 285.
28 pp.
7. Harding, P. L., J. Soule and M. B. Sunday. 1958.
34 F .018 .060 .179 Storage studies on 'Marsh' grapefruit, 1955-56 season. Citrus
Ind. 39 (2) :8-10, 12-14, 35-36.
8. Harding, P. L., J. Soule and M. B. Sunday. 1957.
40 F .018 .103 .176 Storage studies on 'Marsh' grapefruit, 1955-56 season. I.
Effect of nitrogen and potash fertilization on keeping qual
ity. II. Effect of different temperature combinations on
55 F .018 .081 .102 keeping quality. U.S. Dept. Agr. AMS-202. 16 pp.
9. Harvey, E. M. and G. L. Rygg. 1936. Field and stor
age studies on changes in the composition of 'Marsh' grape
70 F .018 .121 .392 fruit in California. J. Agr. Res. 52:747-787.
10. Jones, H. B., W. R. Buford and A. L. Ryall. 1951.
Storage tests on Florida and Texas grapefruit U. S. Dept.
Agr. H. T. & S. Rep. No. 237. 17 pp.
*Average for 7 fruit, 5 of 11. McDonnell, L. R., E. F. Jansen and H. Lineweaver.
1945. The properties of orange pectinesterase. Arch. Bioch
which values were zero. 6 :383-401.
12. Miller, E. V. and H. A. Schomer. 1939. Physiologi
cal studies of lemons in storage. J. Agr. Res. 59:601-607.
1252-31-1 13. Pollard, J. E. 1969. Enzymological studies of abscis
1258-67-1 sion in 'Valencia' oranges (Citrus sinensis, L.) and cala-
mondin (Citrus madurensis, Lour). Ph.D. Dissertation, Univ.
1258-76-1 Fla., Gainesville, 101 pp.
14. Pratt, D. E. and J. J. Powers. 1953. The thermal
effect on internal ethylene levels after subsequent destruction of pectic enzymes in grapefruit juice. Food Res.
18:152-161.
equilibration at 70°F (Table 10). Residual 15. Rouse, A. H. and C. D. Atkins. 1955. Pectinesterase
and pectin in commercial citrus as determined by methods
ethylene levels were higher subsequent to storage used at the Citrus Experiment Station. Fla. Agr. Exp. Sta.
Bull. 570. 19 pp.
at 50 °F than at 40 °F. They were increased by
16. Rygg, G. L. and E. M. Harvey. 1938. Behavior of
waxing prior to storage and appeared to be pectic substances and naringin in grapefruit in the field
and in storage. Plant Physiol. 13:571-576.
related to the degree of senescence of the fruit. 17. Stahl, A. L. and A. F. Camp. 1936. Cold storage
studies of Florida citrus fruits. I. Effects of temperature
Increased internal ethylene levels in citrus fruits and maturity on the changes in composition and keeping
are associated with fruit deterioration and in quality of oranges and grapefruit in cold storage. Fla. Agr.
Exp. Sta. Bull. 303. 67 pp.
creased respiration, and may exert secondary 18. Steward, J. McD. and G. Guinn. 1969. Chilling in
jury and changes in adenosine triphosphate of cotton seedl
effects on the rate of fruit senescence. ings. Plant Physiol. 44:605-608.
310 FLORIDA STATE HORTICULTURAL
19. Ting, S. V. 1956. Fruit juice assay. Rapid color-
metric method for simultaneous determination of total re
ducing sugars and fructose in citrus juices. Agr. and Food
Ghem. 4:263-266.
20. Trout, S. A., F. E. Huelin and G. B. Tindale. 1960.
The respiration of 'Washington' navel and 'Valencia'
oranges. Aust. Comm. Sci. and Ind. Res. Dir. Tech. Paper
*21. *Vakis, N. I., W. Grierson and J. Soule. 1970. Chill
ing injury in tropical and subtropical fruits. III. The role
EFFECT OF TEMPERATURE ON CENTRIFUGAL SEPARATION OF
COLD-PRESSED ORANGE OIL
Robert E. Berry and Pedro Casals1
U. S. Fruit and Vegetable Products Laboratory2
Winter Haven
Abstract
Centrifugal separation of cold-pressed orange
oil from aqueous media is greatly influenced by
densities and viscosities of the two principal
(oil and water) phases. Both densities and
viscosities are likewise affected by temperature.
Differentials between densities of oil and water
were calculated for different temperatures and
increased as temperature increased. Viscosities
of both phases decreased with increasing tem
perature. Based upon measured differentials in
density and changes in viscosity, predictions were
made for temperature at which optimum separa
tion of oil could be achieved from an aqueous
phase upon centrifugation. Optimum tempera
ture for separating oil was 125°F.
Introduction
Cold-pressed orange oil is an important by
product of the citrus industry and is usually
obtained by centrifugation of an aqueous emul
sion from pressed peel. This yields two streams,
one of which is predominantly orange oil and the
other an aqueous centrifuge effluent which may
contain up to 1% residual oil. In studies being
carried out at our laboratory on methods for con
centrating the solids (mostly sugars) in these
centrifuge effluents using the reverse osmosis
lForeign Research Associate sponsored by Ministry of
Education and Science, Barcelona, Spain.
2 One of the laboratories of the Southern Utilization
Research and Development Division, Agricultural Research
Service, U. S. Department of Agriculture.
References to specific products of commercial manufac
ture are for illustration only and do not constitute endorse
ment by the U. S. Department of Agriculture.
SOCIETY, 1970
of COn in suppressing chilling injury of grapefruit and
avocados. Proc. Amer. Soc. Hort. Sci. (Trop. Region).
Submitted.
22. Vines, H. M., W. Grierson and G. J. Edwards. 1968.
Respiration, internal atmosphere, and ethylene evolution of
citrus fruit. Proc. Amer. Soc. Hort. Sci. 92:227-234.
23. Vines, H. M., G. J. Edwards and W. Grierson. 1965.
Citrus fruit respiration. Proc. Fla. State Hort. Soc. 78:
198-202.
technique, these residual amounts of oil inter-
ferrd with concentration methods. Consequently,
treatments were sought by which the residual
oil content in aqueous effluents might be reduced.
Cursory studies had indicated the possibility of
a relationship between temperature of centri
fugation and efficiency of oil separation.
Centrifugal separation of oil from water
would be expected to be greatly influenced by
the relative densities of the two phases. This
is shown by the Stokes Law equation for the
theoretical sedimentation of a solid through a
liquid in a centrifugal force field (Perry 1963).
Although an oil-water system, such as referred
to here, is different from a solid-liquid system,
nevertheless, the general rate of centrifugal sep
aration could probably be expected to follow the
Stokes Law relationship. Rate of separation of
oil would therefore be expected to be influenced
by any change in viscosity in the aqueous phase.
Density and viscosity of both phases would be
expected to vary with temperature.
Several studies have been made of the specific
gravity (density) of cold-pressed orange oil. As
early as 1932, Poore reported specific gravities
of California cold-pressed orange oil at 25 and
15.6°C and these were verified in reports by
Kesterson and McDuff (1948) and Kesterson and
Hendrickson (1953). In the latter two studies
the specific gravities of orange oils made by dif
ferent processes and at different times of the
year were compared. All measurements were
made at 25°C, however, and no attempts were
made to determine changes in specific gravity
with changes in temperature. Kesterson in 1949
reported no significant changes in specific grav
ity of cold-pressed orange oil obtained under con
ditions resulting in different oil yields. In 1953,
Valiente et al. reported on physical characteris
tics of cold-pressed orange oil from several dif-