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Potgieter, M., J . Am. Dietetic (27) Spitzer, R. R., and Phillips, P. H.? (33) West, .4.P.. and Cruz.

.4.P.. and Cruz. A. O.,


As~oc., 16, 898-904 (1940). J . ,lutrition, 30, 183-92 (1945). Phil+pine J . S a . , 52, 1-16 (1933).
Rather, J. B., J. A m . Chem. SOG.,39, (28) Sreenivasan, A., ivature, 143, 244-5 (34) Wolff, E. T., Aschenanalysen Von
2506-15 (1917). (1 939). Landwirthschaftlichen Produc-
Reddi, P. B. V., Murti, K. S., and (29) Subrahmanyan, \., Sreenivasan, ten, Bd. 1, p. 29; Bd. 2, p. 22.
Feuge, R. O., J . Am. Oil Chem- A., and Das Gupta, H. P., Indian Berlin, Wiegandt and Hempel.
ists Soc., 25, 206-1 1 (1 948). J.Agr. Sci., 8, 459-86 (1938). 1871.
(30) Tani, T., et al., R e p . Food Re- (35) Yampolsky, C., Wallerstein Lab.
Sherman, H. C., Chemistry of search Inst. Tokyo, No.3, 11-25 Commun., 7, 7-26 (1944).
Food and Sutrition, 6th ed., (1950).
Xew York, hfacmillan Co., 194 1. (31) Thompson, A. R., J . Agr. Research, RECEIVED for review August 12, 1953. Ac-
cepted September 28, 1953. .Wention of the
Simpson, I. A., Chow, A. Y . , and 3. 425-30 (1915). names of j r m s or trade products does not tmpiv
Soh, C. C., Bull. Inst. M e d . Re- (32) U. S . Agr. itarketing Service and that they are indorsed or recommended b j the
search, Federation Malaya, New Regulatory Announcement No. U. S. Dppartment of Agriculture over othfr
Series S o . 5 , 1-28 (1951). 147, rev. 1941. j r m s or similar products not mentioned.

Pilot Plant Production of Low-Methoxyl


Pectin from Citrus Peel
R. P. GRAHAM and A. D. SHEPHERD
Western Regional Research laboratory, Albany, Calif.

Low-methoxyl pectins are versatile materials which make possible the preparation of
many new food products and the preparation of old food products in new or easier ways.
Use of various modified pectins is restricted, however, by limited availability. Suitable
grades and types of low-methoxyl pectin can be prepared in good yield from citrus peel.
Care is required throughout the manufacturing operation to avoid excessive loss of pectin
quality. Procedures have been developed on a pilot plant scale. Three stages are
involved: preparation of peel for extraction, extraction of pectin and clarification of the
extract, and chemical modification and isolation of low-methoxyl pectin, All operations
are conducted in an aqueous phase, in contrast to present production methods. An
integrated, practical process for making low-methoxyl pectin from citrus peel has been
developed. Operating conditions, procedures, and equipment are discussed in a manner
designed to assist prospective users of the process.

L OJV-METHOXYL PECTIS is a com-


paratively recent development, al-
though pectin has been manufactured
to form uniform strong gels in the pres-
ence of bivalent cations over a wide p H
range. This property makes low-meth-
uct. Processing of the pectin extract in-
volves the controlled de-esterification of
the pectin, isolation of the lo\v-methoxyl
in the United States for more than 35 oxyl pectin useful for numerous applica- pectin by acid precipitation, detvatering
years (75). The many methods re- tions in which ordinary pectin cannot and neutralizing of the lo\v-methoxyl
ported in the literature for the prep- be used. Applications suggested for pectin, and drving and grinding to make
aration of low-methoxyl pectins have low-methoxyl pectin include jellied fruit the final product.
been based on laboratory scale experi- cocktail ( 7 4 , tomato aspic (79), low- Pectin is a long-chain molecule ivhich
ments. Little if any information has solids gels (3, 5, 79, 28), milk puddings is de-esterified and degraded (reduced
been published on production methods (79,37), candy centers (70), and coatings in chain length) by heat, acidity, al-
used by the several manufacturers who for various food materials (27). Nu- kalinity, and enzyme action. The qual-
have made low-methoxyl pectin on a merous patented applications are not ity-Le., the gelling ability-of a pectin
commercial scale. reviewed here. product is controlled largely by the chain
Pectin is largely the partial methyl This paper summarizes the develop- length of the molecule-the longer the
ester of anhydrogalacturonic acid and, ment of a process for making low-meth- chain, the higher the quality. Most of
as extracted from common source ma- oxyl pectin from citrus peel. The proc- the pectin in fresh citrus peel is fairly in-
terials, contains ordinarily from 9 to ess is divided into three parts: prepa- soluble in cold \%rater. Heat and acid
12% by weight of methoxyl groups. ration of the peel, extraction of pectin are used to render the pectin extractable,
When a substantial portion of the methyl from the peel and clarification of the ex- but these agents also degrade the pectin.
ester groups is removed by hydrolysis, tract, and processing of the extract to pre- The amount of degradation must be
the modified pectin attains the ability pare the dry, low-methoxyl pectin prod- limited, so that satisfactory quality is

V O L . I , NO. 1 6 , O C T O B E R 2 8 , 1 9 5 3
993
-7I
I
T , MBINATION DEESTERIFIC4TION
D ACID PRECIPITATION TANK
WATER
IN

5TCRASC

ST E
FILTER

' Y
WASTE
RECYCLE OF P R E V I O U S

VACUUM

P
EV4POR4TbTIM
COOLLR

Figure 1.
SLUDGE
CEN'IRIFUGE

TO W A 5 T E

L NEUTRALIZER

Flow sheet for manufacture of low-methoxyl pectinic acid from waste citrus peel
maintained in the lobs-methoxyl pectin
product. Further degradation occurs
citrus peel. As finally developed, the
process covers the entire production of
C40INLT
DRlCR
HAMMER
UlLL

materials makes the pectin extract easier


to filter, less colored? and substantially
during the processing of the extract to lo\\-methoxyl pectin from citrus peel, tasteless. Refining of the extract in this
low-methoxyl pectin. The process em- rather than its production from commer- manner is reflected in the quality of the
ployed, therefore. is a compromise, de- cial pectin as a starting material. final lo\v-methoxyl-pectin product.
signed to accomplish the necessary physi- The flow sheet of the process is shown Blanching and Lvashing of ','a- to 3,'16-
cal and chemical operations in a rea- in Figure 1. inch slices of peel are effected in a single
sonable manner \vith a minimum of deg- continuous unit (9). The unit shown
radation. Preparation of Citrus Peel in Figure 2, used in a field pilot plant,
The steps involved in making 13w- For Extraction comprises a wooden trough, 12 feet long
methoxyl pectin have been investigated and 12 inches wide, with a semicylindri-
to varying degrees, a t this laboratory and Orange, lemon, or grapefruit peel can cal bottom. The trough is fitted with an
other places. Pectin has been de-esteri- be used as the starting material in this interrupted-flight screw and a perforated
fied experimentally by ammonia (78, process. Peel as produced by any of metal ramp and drag for removing peel.
79), acid (2, 4, 23. 33), and enzymes the commercial juicing machines is suit- At the peel feed end of the trough, steam
(72, 22, 25). The products obtained able. The presence or absence of the is injected over a S-foot section to main-
differ somewhat in properties ( 7 7, 24, 29, flavedo is immaterial. tain a Lvater temperature of 194' F.
3 2 ) ; the product from the aqueous- Several types of cutter have Freshly cut peel is introduced into the
Cutting
phase, ammonia de-esterification of pec- been used successfully. The heated section to accomplish blanching,
tin is readily adapted to the applications most suitable type for a particular case and is conveyed by the screw through
outlined above. Lo\\-methoxyl pectin depends upon whether the peel is dried the trough. Cold water, added as a
may be precipitated from solution by the for storage or extracted in its wet state. spray above the drag, flows counter 10
addition of solvents, polyvalent cations, For making dried peel, a kraut cutter the peel and out a n overflow pipe. Va-
or acid (20, 27). The first two methods producing slices from '/8 to 3/16 inch thick lencia orange peel has been washed suc-
impose solvent handling and recovery is suitable. If the peel is to be extracted cessfully in this unit a t a rate of 400
problems on a process, and,'or need for immediately, a finer cut is preferable. pounds per hour \vith a retenticn time
concentrating the solution before isola- -4 suitably cut peel was obtained from a in the trough of 30 minutes. About
tion of the low-methoxyl pectin. Acid high-speed comminutor fitted with a 3 pounds of water per pound of peel was
precipitation offers the advantage of sim- screen perforated with round holes '/4 required to \\ash the peel adequatel)..
plicity; excess acid is easily removed by inch in diameter. Dicing-type cutters The blanching was sufficient to inacti-
washing the precipitated low-methoxyl \viere too low in capacity to be useful vate the pectin enzymes without seri-
pectin with water. in preparing either type of cut peel. ously reducing the quality of the pectin.
Studies on the de-esterification of an Blanching of raw peel The blancher-washer described above
aqueous solution of pectin with am- and immediately after has not been tested on finely cut peel
monia (78, 79) and the isolation of the Water Washing cutting is necessary such as that produced with a high-
low-methoxyl pectin with acid (20, 27) to minimize the action of naturally oc- speed comminutor. ,Minor changes in
Jvere a part of the background work curring enzymes upon the pectin. Wash- operating conditions would probably be
leading to the development of the proc- ing of the cut peel removes substantial necessary for finely cut peel, particu-
ess described in this paper. However, quantities of soluble solids such as sugars larly for reduction of blanching time
as various process steps were studied in and acids, and colloidal materials such to correspond to the smaller piece size.
the laboratory, it became apparent that as pigments, glycosides, and finely No serious problems are anticipated in
meaningful results could be obtained divided material produced during the using the continuous blancher-washer
only by starting with the extraction of cutting operation. Removal of these for handling finely cut peel.

994 AGRICULTURAL A N D F O O D CHEMISTRY


V O L 1, NO. 16, O C T O B E R 28. 1 9 5 3
995
Pressing of the washed peel in this project was directed toward the imposed on the permissible fineness of
Pressing
is desirable for two reasons. production of a suitable pectin extract the peel (and in consequence the yield
If the peel is to be dried, pressing reduces which could be used directly-i.e., of pectin obtained), or on the permis-
the evaporative load imposed on the without further concentration-in sub- sible concentration of pectin in the ex-
dryer. If extracted immediately, pressed sequent processing. Proper balance be- tract. The pectin in the extractor was
peel absorbs the acidulated water more tween the many relevant factors re- also exposed to an undesirable amount
readily than unpressed peel, thus hasten- quired careful and extensive study of of degradation because of the charac-
ing the extraction of the pectin. The extraction procedures in pilot plant teristics of the countercurrent method.
pilot plant has been operated, however, equipment. Until extracted from the peel, the pectin
on both pressed and unpressed fresh The unit used in these traveled upstream in &e%&%ctor while
peel. Extracting
studies (Figure 3) was being exposed to high temperature and
A roller press of simple construction, as constructed for countercurrent extrac- acidity. After extraction from the peel,
shown in Figure 2, is adequate for press- tion of the peel (9). Its general con- the pectin retraced its course whi!e
ing the peel. Two solid rubber belts, struction was similar to the blancher- traveling downstream with the extract
both 3 feet in width, are run one above washer, except that all portions contact- under the same unfavorable environ-
the other a t the same linear speed. ing the peel or extract were fabricated ment. Attempts were made to improve
Washed peel is fed between the belts, of stainless steel. The screw was 16 feet the countercurrent extraction of dried
and is pressed during passage through long and 16 inches in diameter with a peel by preliminary cold acid recon-
the 4-foot length of the press. At a ca- 4-inch pitch. The trough was steam- stitution or pickling of the peel. This
pacity of 400 pounds of washed peel per jacketed in three sections, u p to shaft- procedure improved the extraction in
hour, the press raises the solids content center level. Spray nozzles for the intro- general, but failed to permit prbduction
of the peel from about 7% to about 13%. duction of hot water were provided above of a n extract at the desired concentration
Solids content of the washed peel can be the perforated ramp and drag. Sulfuric of pectin. Thus, the countercurrent
raised somewhat higher in a bag press acid, pumped by a glass metering pump, extraction method was proved to be
without serious loss of pectin. How- was introduced into the water between inherently unsuited to the process re-
ever, the gain is not sufficient to justify the screw and ramp. At the opposite quirements, and was abandoned.
the additional time and labor involved. end, the sol was discharged over a weir In order to obtain pectin solutions of
Citrus peel for use as a source which controlled the tank level. Dried higher concentration, the theoretically
Drying
of pectin must be carefully peel was fed a t a point about 3 feet from more efficient countercurrent extraction
dried to avoid damage to the pectin. the sol discharge end by means of a vi- method had to be abandoned. T o avoid
Initial phases of the drying may be brating feeder. Fresh peel or reconsti- the difficulty of forcing the peel to travel
carried out a t 200' F., but the tempera- tuted dried peel was fed by hand a t the against the flow of the extract, parallel-
ture must be reduced to about 130' F. same point. flow extraction was used. The peel
when the average moisture content of the This apparatus was modified for paral- and extractant were fed in the desired
peel is reduced to about 30%. Drying lel-flow extraction. The rotation of the proportion a t one end of the extractor,
of the peel to a moisture content of less screw was reversed and bars were pro- and the mixture was gently agitated and
than 10 to 12y0 should be avoided. Peel vided between the flights, to increase heated as it flowed through the treater,
so dried can be stored for a year a t 70' agitation. A mechanically cleaned screen and was discharged from the extractor
F. without serious loss of pectin quality was placed a t the discharge end of the as a slurry from which the extract was
(30). extractor for separation of the extract continuously separated. This method
from the slurry by drainage. The pulp was much easier to control than the
Extraction of Pectin from Peel on the screen was washed by water sprays countercurrent method. With ground
And Clarification of Extract after substantially all of the extract had grapefruit peel, the extractor was oper-
drained, and: the wash water was re- ated a t 194' F., with a retention time for
Factors related to the best procedure turned to the extractor near the dis- the peel of 20 minutes, and a p H of about
for extracting pectin from citrus peel in- charge end. The feed point for peel was 1.8 to 1.9 in the extract. The dried
clude quality of pectin in the peel ini- adjusted to give desired retention time. peel, ground to, pass a 40-mesh screen in
tially, amount of pectin recovered, qual- In order to eliminate the variables asso- burrstone mill, was fed a t 40 pounds per
ity of pectin in the extract, and concen- ciated with raw material, a quantity of hour with water fed a t 180 gallons per
tration of pectin in the extract. The ex- grapefruit peel which had been dried hour to yield an extract with a pectin
traction of high-quality pectin from as a pectin source material was used in content of about 0.7y0. Variations in
citrus peel is more difficult than extrac- many of the pilot plant operations. Un- the feed rate were used to alter the pectin
tion of low-quality pectin. The amount fortunately, the pectin in this peel had content of the extract. IVith fresh
of pectin recovered during extraction of been degraded during the processing and Valencia orange peel, the retention time
citrus peel with hot dilute acid increases drying to a quality much lower than that was increased to 30 minutes, blanched
as the extraction temperature and acidity of the pectin in fresh grapefruit peel.
increase. The rate of degradation of As the handling characteristics of the
the pectin also increases with increasing acid-precipitated low-methoxyl pectin Table 1. Parallel-Flow Extraction of
temperature and acidity, thus affecting are influenced by the pectin quality, Pectin
the quality of the pectin obtained in the extraction of the dried grapefruit peel Exfroct Recovery
extract. required great care to minimize further Pectin of
The concentration of pectin in the ex- degradation of the pectin. Fresh and content, (VI, Pectin,
tract is highly important. Handling dried Valencia orange peel, prepared Run % dl./g. %
characteristics of the de-esterified pectin a t the laboratory and containing pectin From Ground Dried Grapefruit Peel Con-
obtained in subsequent processing re- of high quality, were also used in a taining 42% Pectin of (?) 4.1
quire that the extract contain 0.6 to limited number of experiments. 1 0 9 3 8 68
1% pectin, depending upon the quality Countercurrent extraction was in- 2 1 .o 3 6 60
of the pectin. Production of a more 3 1.2 3.8 62
vestigated first, and found to be usable
dilute extract is easier and concentration only when dilute extracts were pro- From Fresh Valencia Orange Peel Con-
of the dilute extract is technically feasible. duced. Because of mechanical problems taining 3.6% Pectin of (?) 9.0
However, the procedure is expensive. in moving the peel and extract in op- 4 0.7 6.7 50
A large portion of the extraction studies posite directions, a working limit was

996 AGRICULTURAL AND FOOD CHEMISTRY


and washed peel was fed a t 400 pounds Filtration was carried out in a con- trol was maintained aver the concentra-
per hour, and the water at 100 gallons ventional plate-and-frame filter press tion of bivalent ions in the extract. The
per hour. precoated with a medium-slow filter aid. procedures used for ion exchanging and
Typical data for several parallel-flow One half per cent of the same filter aid vacuum concentrating the dilute ex-
extractions are given in Table I. I t is was used in the extract. Filtration rates tracts, and for de-esterifying the modi-
difficult to compare these recoveries of 5 to 10 gallons per square foot per hour fied extract will be described in another
with those obtained in commercial prac- were obtained with solutions containing paper.
tice, because pectin contents of raw ma- 0.8 to 1% pectin by weight.
terials are not usually determined. T h e Processing of Pectin Extract to
methods for determining pectin content Optional Processmg T h e process as Prepare Dry Low-Methoxyl Pectin
described u p to
and intrinsic viscosity, ( a ) , of the pectin Of Pectin Extract
this point is the Processing of the pectin extract to make
are those used previously (30). simplest and most straightforward that dry, powdered, low-methoxyl pectin
Because parallel-flow extraction is has been used with success in the course involves de-esterification of the pectin
really batch extraction on a continuous of the pilot plant work. If, however, with aqueous ammonia a t reduced tem-
basis, the influence of variables on the particular circumstances should dictate perature, isolation of the low-methoxyl
extraction process may be readily studied the production of a dilute extract, con- pectin by acid precipitation, delvatering
on a laboratory scale. In Tables I1 and centration of the extract prior to further and neutralizing of the low-methoxyl
111, the influence of variables upon the processing would be desirable. T h e pectin, and drying and grinding to make
yield and quality of extracted pectin reason for this is that the reagent cost the final product.
is shown. These are results obtained for de-esterifying pectin and acid-precipi- Preceding the pilot plant studies, lab-
from small batch extractions, from which tating the low-methoxyl pectin is almost oratory studies were made on the de-
the extract was separated and cooled directly proportional to the volume of esterification of pectin with ammonia
rapidly. Results given may be used extract handled, regardless of the con- (19) and acid precipitation of the Iow-
directly to predict operation of the paral- centration of pectin in the extract. Di- methoxyl pectin (21). In most of these,
lel-flow extractor. lute pectin extracts have been concen- solutions prepared from dry commercial
It is important that the ex- trated successfully in vacuum on a pilot pectin were used, Consequently, results
'"ling tract be cooled to about room plant scale, provided that adequate con- of many of the laboratory studies \\-ere
temperature as rapidly as possible after
its separation from the extracted peel.
~~

I n this manner, degradation of the pec- Table II. Batch Extraction of Ground Dried Grapefruit Peel
tin is arrested. Pectin Content
of Sol a t 700% Exfraction Conditions The0r.O Extract
T h e cooler used was a forced-draft
' evaporative cooler, shown in Figure 4.
W t . Wafer/ Theoretical Temp., Time, % Pectin
W t . Peel Recovery, % OC. PH min. Recovery (d
I t comprised a distributor pan discharg-
ing on a series of staggered wooden slats 42 1. o 85 1.7 15 73 4.1
30 78 3.6
to break u p the liquid flow, and a col- 45 82 3.5
lector pan to receive the cooled extract. 60 85 3.2
Forced draft was provided by a blower. 42 1.o
The size of the unit, exclusive of the ex-
tract-collecting pan and the blower, was
40 90 3.1
24 X 30 X 24 inches. This unit was
capable of cooling extract from 190' to 42 1 .o
90' F. a t a rate of about 3 gallons per
minute.
42 1.o 90 2.15 20 3.7 69
The extract from the 30 3.6 69
Centrifuging
extractor contains solid 40 3.3 77
material which can be readily removed 42 1 .o 95 1 . 7 5 3.7 72
by centrifugation. Removal of this i~.
n 82 3.6
material is desirable to increase capacity 20 88 3.2
and rate during subsequent filtration. 30 91 2.9
The centrifuge used was a n overflow type 33.5 1.25 90 1.85 20 82 3.8
with a 17-inch, stainless-steel, solid 2.0 20 69 4.1
basket. T h e extract was fed directly 28 1.5 90 1 . 8 5 20 77 4.0
from the cooler to the centrifuge; 2 to 2.0 20 72 4.0
4y0 of the weight of extract was re- a Calculated assuming that all water added contains pectin at concentration measured in
moved as sludge by the centrifuging oper- sol and all is removed from peel. Actual recoveries will be about 80% of these figures.
ation. The sludge was discharged from
the basket after 150 to 200 gallons of ex- Table 111. Batch Extraction of Ground Fresh Valencia Orange Peel
tract had been processed. Pectin Content
The filtering of pectin ex- of sol at 700% Exfraction Conditions Theor.a Extroct
Filtering %
tracts a t ambient rather W t . Water/ Theoretical Temp., Time, Pectin,
than elevated temperature represents a W t . Peel Recovery, % OC. PH min. Recovery (7)
minor departure from commercial prac- 3 1 .0 80 1.9 10 40 9.7
tice. Extracts filtered a t elevated tem- 20 55 8.8
peratures decrease in clarity as they 30 64 8.7
cool to room temperature. Conse-
quently, extracts filtered at lower tem-
peratures are clearer than those filtered
a t higher temperatures. T h e improved
clarity is carried through to a solution a Calculated assuming that all water added contains pectin at concentration measured in
prepared from the low-methoxyl pectin sol and all is removed from peel. Actual recoveries will be about 80% of these figures.
product.

VOL. 1, NO. 16, O C T O B E R 20, 1 9 5 3 997


signed so that pectin extract fed a t the Otherwise, alkali (ammonia) Iost from
Table IV. Changes in Methoxyl top flowed in thin films, in and out, in the sample could not be distinguished
Content of Pectin with Time During proceeding toward the bottom. The from alkali consumed in the saponifica-
Pilot Plant De-esteriffcation vacuum was provided by a four-stage tion. Typical data showing the change
Elapsed 0bserved steam ejector capable of operating a t an in methoxyl content with time as
Run De-esferificotion Methoxyl absolute pressure of 4 mm. of mercury. determined by this method are given
No. Time, Min. Content, % Extract was cooled from room temper- in Table IV.
1790 120 5.9 ature to 50' F. at a rate of 150 to 200 With experience gained from observ-
165 5.0 gallons per hour in this unit. The tem- ing many samples of various methoxyl
210 4.2 perature to which the extracts were
240 3.8 contents, the end point of the de-esteri-
cooled varied from 43' to 58' F. in vari- fication was usually determined from
1793 120 6.0 ous experimental runs.
165 5.1 the physical handling characteristics of
210 4.2 De-esterifying The de-esterification a small grab sample precipitated with
240 3.6 oDeration was carried acid.
180- 60 8.3 out in a batch manner. Two stainless- The results of several de-esterification
125 6.2 steel cylindrical tanks of about 180- runs made in the pilot plant are shown
180 5.1 gallon capacity were used alternately in Table V. Commercial dried grape-
240 3.7 (Figure 6). The tanks were insulated fruit peel was used in all these runs
by an air jacket. A slow-speed. opposing except 1804 and 1807. Valencia orange
agitator was provided in each tank for peel prepared a t the laboratory was
not directly applicable to the pilot plant mixing in reagents and breaking gels. used in these runs. R u n 1804 was made
because impurities in the peel extracts, The tanks were covered to minimize loss on washed peel that had been pressed
not present in the solutions prepared of ammonia and were provided with a but not dried. R u n 1807 was made on
from commercial pectin, greatly affected 3-inch center drain for discharging gel. part of the same peel dried prior to
the de-esterification and acid-precipita- To conduct a de-esterification, about extraction. The intrinsic viscosity of
tion steps. Calcium, a n important im- 165 gallons of cooled extract were col- the pectin was considerably higher in
purity in the peel extracts, caused the lected in one of the de-esterification the extracts in runs 1804 and 1807, and
formation of strong gels during the alka- tanks. About 5 gallons of concentrated the intrinsic viscosity of the final low-
line de-esterification. Much of the proc- aqueous ammonia were metered into the methoxyl pectins was correspondingly
ess development in this phase of the work tank and mixed into the extract, making higher.
involved methods and equipment for the ammonia concentration about 0.75y6 Table V includes data from extracts
handling large quantities of strong gel, by weight in the extract. During treat- of varying pectin contents, run a t
in both the de-esterification and acid- ment with ammonia, methyl ester groups different temperatures and undoubtedly
precipitation steps. of the pectin are hydrolyzed to ionized containing different concentrations of
The following discussion assumes that carboxyl groups, and the pectin solu- cations because different extraction
the pectin extract has been produced a t tion slowly sets to a gel. Gel formation methods were used. Even with all
the proper concentration (0.6 to 1.0% by is the result of a reaction between the these variables. the time required for
weight) for subsequent processing. More carboxyl groups of the low-methoxyl de-esterification in most of the samples
dilute extracts may be adjusted to the pectin and calcium or other polyvalent was in the range of 3.5 to 4.5 hours.
desired concentration by vacuum evap- cations in the extract. The gel must be Almost all of the samples could have
oration. broken up periodically by agitation dur- been stopped in this time range and still
The sol is cooled to reduce ing the de-esterification to avoid forma- have had a methoxyl content within the
"'ling the amount of degradation tion of a solid mass in the tank. desired range of 3.0 to 3.5%. In a plant
occurring during de-esterification of the In some instances the methoxyl con- where extraction is made in a uniform
pectin. The temperature to which the tent of the pectin \vas followed during the manner to yield a sol of reasonably
sol is cooled represents a compromise be- de-esterification to determine the proper constant pectin content, and where the
tween decrease in the amount of deg- time for stopping the reaction. Meth- de-esterification is carried out under
radation and increase in the time re- oxy1 content was determined by saponi- fixed conditions of temperature and
quired for de-esterification. fication of a small sample drawn from ammonia concentration, control of the
Cooling was accomplished by vacuum the batch. Since saponification involves time for de-esterification would be
evaporation. The unit used for cooling measurement of alkali consumed, the relativelv easv.
was a cylindrical stainless-steel vacuum conventional method for determining
Increasing He-este
body (shown in Figure 5). The unit methoxyl content had to be modified
Rate (Optional Pruccr
was 30 inches in diameter and 6 feet to avoid loss of ammonia from the
high, and contained a series of baffles de- sample during the determination. tion of pectin under alkaline condi-

Table V. Pilot Plant De-esteriffcations


De-esterification Conditions Final Product
Pectin in Concn. NH40H. Me0 Ca
Sam- Exiracf after gol.1 con- pH o f con-
ple Method of Concn., (q), Evop., Temp., 100 gal. Init. Time, fent, (q), 1% tent,
No. Extraction % dl./g. % O c. exfrocf pH min. % dl./g. sol %
1753 Pickled and countercurrent 0.5 3.4 1.2 10 3.0 10.8 195 3.2 2.7 4.3 0.03
1767 Pickled and countercurrent 0.5 3.8 1.4 10.5 2.85 10.5 200 3.6 2.9 4.5 0.06
1772 Pickled and countercurrent 0.6 3.9 1.2 6 3.0 10.8 255 3.2 2.9 4.5 0.05
1790 Pickled and countercurrent 0.4 3.8 1.3 9 3.0 10.6 240 3.8 3.1 4.2 0.06
1793 Pickled only 1. o 3.6 N o evap. 10 3.0 10.7 240 3.5 3.0 4.3 0.04
1794 PickIed only 0.9 3.7 No evap. 9 3.0 10.6 240 3.4 3.0 4.4 0.04
1804 Countercurrent, no pickling 0.5 6.1 1.2 11 3.0 10.4 350 2.7 3.5 4.6 0.05
1807 Pickled and countercurrent 0.2 7.5 0.9 12 3.0 10.5 275 3.2 4.2 4.5 0.07
1827 Parallel flow 0.9 3.6 No evap. 9.5 3.0 10.6 210 3.4 2.8 4.4 0.04
1830 Parallel flow 1 .o 3.6 2.4 9.5 4.0 10.5 255 3.4 2.9 4.2 ..
1832 Parallel flow 1 .o 3.8 N o evap. 11.5 3.0 10.6 180 3.8 3.0 4.4 ..

998 AGRICULTURAL A N D FOOD CHEMISTRY


tent is reduced in the final low-methoxyl manner was extremely effective because
Table VI. Effect of Sodium Sulfate pectin product. However, the amount the wash water tended to displace rather
Concentration upon Time Required of lead that can be tolerated is so low than to dilute the acidulated water.
for De-esterification that this method does not reduce the After washing, reeling was continued
(Details in text) lead content sufficiently. until the solids content of the gel reached
Time Required fo
Introduction of lead must be avoided a b x t 8%.
Produce low-Mefhoxyl in so far as possible. S o lead-containing These operations of draining and
Concn. o f Pectin o f 3.5% M e O , materials should be used a t any point in washing may be carried out in separate
Sample NanSOh, N Min. the plant where lead might be intro- reels if desired.
1 0 250 duced into the product, and reagents The water-Lvashed gel was
2 0.04 195 Pressing
should be scrutinized with respect to their removed from the reel and
3 0.1 135 pressed in a rack-and-frame hydraulic
4 0.2 100 lead content. The reagent of major
concern is the sulfuric acid used for pre- press. Pressure was applied at a rate
cipitation. The use of .4CS grade sul- such that the maximum pressure of about
furic acid Mould assure a very low lead 100 pounds per square inch on the
tions is greatly influenced by the pres- content in the lo\\ methoxvl pectin, but "cheese" was reached in 5 to 10 minutes.
ence of cations, particularly bivalent the acid is expensive. .Although con- This pressure was maintained as long as
cations (76, 77). The cation content centrated technical sulfuric acid has a the drainage rate was reasonable, usu-
of pectin extracts is usually high enough high lead content, a satisfactory method ally about 15 minutes.
so that de-esterification proceeds a t a has been found for reducing its lead con- T h e pressed gel was
reasonable rate. With extracts which tent to an acceptable range by dilution Shredding shredded by passing it
I * 1

have been treated to replace bivalent and settling of the lead sulfate precipi- through a hammer mill equipped with
with monovalent cations, the de-esteri- tate. I t was found that the minimum knives in place of the customary ham-
fication rate of the pectin in the extract lead content per unit of sulfuric acid was mers. A screen with 3jg-inch round
is only about three fourths that of pectin reached a t about 55 Ireight sulfuric perforations was used.
in the natural extract. acid. The lead content of sulfuric acid The gel was recycled through the press-
Laboratory tests were made in which diluted to varying degrees from concen- ing and shredding steps until the solids
monovalent cations were added to a pec- trated technical acid is shown in Table content of the material reached the range
tin solution to increase the de-esterifica- VII. For comparison, ACS grade sul- of 30 to 35%. This usually required
tion rate. Only monovalent cations furic acid must contain not more than a total of three pressings. Repeated
were used in order to facilitate their re- 0.00017, heavy mrtals [as lead) or 1 pressings may be avoided. if purity of
moval during subsequent washing of the p.p.m. (7). For use in the pilot plant, the product is not important, by par-
loiv-methoxyl-pectin gel. The tests were concentrated technical sulfuric acid was tially drying the gel to a solids content of
made 011 a solution containing 1.5% by diluted with ice in glass vessels. The 30 to 35% after the first pressing. If
weight of commercial pectin \vhich had dilute acid was allo\ted to stand 18 to this method of raising the solids content
a very lo\t ash content. The de-esteri- 24 hours, and \\as then decanted from were used, a rotary or continuously
fications were conducted a t 45' to 46" the precipitated lead sulfate as needed mixing dryer would be preferred to main-
F. in thr presence of 30 ml. of 28% for acid-precipitation of the loxr-meth- tain a uniform moisture content in the
aqueous ammonia pyr liter of solution oxy1 pectin. product. Low-methoxyl pectin is very
and sodium sulfate at four concentration The acidified gel was dis- sensitive to degradation under these pH
levels. The decrease in de-esterification Draining conditions, and should the drying be
charqed from the tank into
time with increasing salt concentration a loose-cloth-lined reel irhere the acidu- uneven, the overdried and overheated
is sho\\.n in Table VI. lated water was drained (ser Figure 7). portion 12ould be liable to damage.
T h e de-esterification is The solids content reaches about 6% in .After the solids content \\as adjusted
Acid
Precipitating terminated by the acid this step. Details of construction and to the desired range, the gel was finally
precipitation
- . of the low- the action of this type of reel have been ground through a screen n i t h '4-inch
methoxyl pectin. described ( 7 ) . round perforations. The final grinding
This is accomplished by breaking the When the draining was prepared the gel for neutralization and
gel by thorough agitation and introduc- Washing permitted easier grinding of the final
completed, a sprav of water
ing acid slo\vly, so that it is incorpo- was played on the gel as it was reeled. dried product.
rated as uniformly as possible. Sufficient The washing was continued until the T o render the lo\\.-meth-
acid is added to neutralize the ammonia Neutralizing
pH of the wash water draining from the oxvl
, Ipectin stable for
and reduce the p H to 1.5 or slightly gel had risen to about 1.8. The rate storage and to increase its rate of solu-
lower. Upon acidification, the gel is a t which the \rash water was added was tion. the product must be partially
converted from the calcium form to the controlled so that time of application neutralized. Any excess sulfuric acid
hydrogen form without any apparent was about 0.5 hour. \\'ashing in this remaining in the gel as well as a portion
change in structurr. For completion of of the free carboxyl groups of the 1 0 1 ~ -
the transition. the batch was allowed methoxyl pectin must be neutralized.
to stand for 20 to 30 minutes following the Seutralization must be carried out
incorporation of the acid. Table VII. l e a d Content of Diluted carefully to avoid local excessively alka-
Considerations of cost and corrosion Technical Sulfuric Acid Removed line conditions. Sodium bicarbonate
indicate the use of sulfuric acid for the from Precipitated lead Sulfate has been found to be ideal for this pur-
precipitation step. Bxause of the nature Sulfuric l e a d Content pose ( 8 ) because of the relatively low
of low-methoxyl pectin, concentrated Acid Lead Content Colcd. to p H produced even a t high concentra-
technical sulfuric acid with a high lead Content o f Diluted Sulfuric Acid, tions. To have the reaction proceed
content cannot be used directly. Low- W f . / W t .% Acid, P.P.M. P.P.M. uniformly throughout the material. a
methoxyl pectin binds bivalent cations 100 127 127 high moisture content of the gel is desir-
very effectively. At low p H , of course, 75 2 96. 41)
70 1 .42 2.0
able. However, neutralization a t too
the hydrogen ion concentration is suffi- 60 0.31 0.9 high a moisture content causes some
cient to displace these ions largely from 50 0.44 0.9 solution of the pectin; the material be-
the low-methoxyl pectin. This is the 40 0.79 2.0 comes sticky and lumpy, and is diffi-
mechanism by which the calcium con- cult to dry. T h e optimum range for

VOL. 1 NO 16, O C T O B E R 2 0 , 1 9 5 3 999


neutralization is between 60 and 65% for general applicability. The specifica- the lead content of low-inethoxyl pec-
moisture (30 to 35y0 solids). After the tions for intrinsic viscosity and methoxyl tin. No difficulty should be experienced
sodium bicarbonate has been thoroughly content were established by H. S. Owens in maintaining a lead content under
mixed into the finely ground lory-meth- of this laboratory. Details of the analyt- 10 p.p.m., provided the necessary pre-
oxyl pectin, a holding period of several ical methods used are published else- cautions are taken. Lead content is
hours is required to allow complete where (26). determined by a spectrographic (73) or
equilibrium to be established, even to An entirely dry product is dithizone method ( 6 ) .
Moisture
the centers of the pieces. During the not desirable. Excessive The p H of a 1% solution of the
Content
drying step to follow, any local condi- drying causes degradation pH final low-methoxyl pectin should
tions of high p H would cause degra- and a completely dry product would be between 4.0 and 4.5. This is
dation and further de-esterification of the absorb moisture in an opened package. near the point of maximum stability,
low-methoxyl pectin. In consequence, the weight required for and the low-methoxyl pectin has a rea-
The amount of sodium bicarbonate a specific purpose would vary, depend- sonably high rate of solution in this
required for neutralization is about 18 ing upon the amouyt of water absorbed. range. A IT0 solution of the product is
to 20% of the dry weight of the 1 3 ~ - A product with a moisture content in the prepared on a moisture-free basis and
methoxyl pectin. The actual amount range of 10 to 15% is reasonably stable the p H is measured with a glass-elec-
necessary was determined by checking in this respect. The moisture content trode pH meter.
the p H of a 1% solution prepared from was determined by drying the product
the low-methoxyl pectin. This p H for 16 hours a t 70 C. in a vacuum oven. Acknowledgment
should be between 4 and 4.5. The ash content of this type
Ash The authors wish to acknowledge the
The batch to be neutralized was spread Content of low-methoxyl pectin is
out on a sheet of canvas and a portion high, because the product is assistance and cooperation of all con-
of the sodium bicarbonate was sprinkled partially neutralized for stability. The cerned with the prcject. Pilot plant
on the surface from a sieve. The batch ash content as determined by heating at equipment and pilot plant facilities dur-
was partially mixed to present new sur- 600 C. is in the range of 10 to 14Yc ing part of the work were made avail-
face and again sprinkled with sodium (moisture-free basis), largely because able by Libby, McNeil & Libby under
bicarbonate. When all of the bicarbon- of the carbonate which is produced when a .Memorandum of Understanding with
ate had been added in this manner, the sodium pectinate is ashed. An ash con- the Bureau of Agricultural and Indus-
batch was mixed thoroughly by rolling trial Chemistry.
tent in the range of 5 to 8% is obtained
and tumbling on the canvas and then when a correction is made for the car- Administrative direction of the pilot
placed in a covered vessel to equilibrate. bonate. The correction is made by plant phases of the prcject was under I\.
For convenience the material was usu- titrating the ash for alkalinity, calculat- D. Ramage, head of the Engineering and
ally held overnight before drying. A ing the alkalinity as carbonate, and Development Division, and A. H.
tumble barrel or similar piece of equip- correcting the ash by this amount. The Brown. J. H. Thompsm \vas project
ment Lvould be satisfactory for this mix- value so obtained is referred to as ash- leader in the pilot plant phase. Admin-
ing step. minus-car bonate. istrative direction of the laboratory
Drying was accomplished in The methoxyl content of phases of the project was under IV. D.
Drying thin layers on trays in a forced- Me*oxy1 the kw-methoxyl pectin Maclay, head of the Field Crops Utili-
draft oven operating a t 130 F. Start-
Ontent
Droduct should be in the zation Division, and H . S. Owens. R.
ing temperatures as high as 150 F. were range of 3.0 to 3.5% on a moisture-free, hi. McCready \+asproject leader in the
used, but were of short duration. The ash-minus-carbonate-free basis. The laboratory phase. The authors are also
product was rabbled a t least once during methoxyl content is determined by sa- indebted to other IVRRL personnel ivho
the drying period. Heating beyond the ponification. contributed to the project: S o r m a n
point when the sample contained 10 to The intrinsic viscosity of the Fishman and George H. Nee1 for assist-
Intrinsic ance in operating the pilot plant;
15% moisture was avoided because it loiv-methoxyl pectin prod-
causes unnecessary damage to the prod- Viscosity uct should be at least 3.0 Roberta Erlandsen, Jack Guggolz, Eliz-
uct. Drying required about 3 hours. dl. per gram on a moisture-free, ash- abeth McComb, and E. J. Eastmond for
The dried low-methoxyl minus-carbonate-free basis. The in- analytical \vork; and Aaron Smith and
Grinding pectin was ground in a trinsic viscosity is measured by deter- Robert Patterson for general assistance.
hammer mill in two stages. It was first mining the efflux time of a n 0.170 pectin
passed through a screen with ,a-inch sol containing sodium chloride and sodium Literature Cited
round perforations, and was finished hexametaphosphate a t p H 6. (1) AM. CHEM.SOC.,Reagent Chemi-
using a screen with 0.024-inch round This method of measuring quality has cals, Am. Chem. SOC. Speci-
perforations. Again care is required to the advantage over any jelly-strength fications, p. 369, 1950.
avoid overheating of the lo~-methoxyl measurement, in that no control is (2) Baker, G. L., and Goodwin, M.
pectin, \rith attendant product damage. necessary over the calcium content. W., Del. Agr. Ex t. Sta., Bull.
In fact, the influence of calcium is re- 234, Tech. No. 28 6 9 4 1 )
moved by the presence of the sodium (3) Zbid.,246, Tech. No.31 (1944).
Characterization of Low-Methoxyl (4) Baker, G. L., and Goodwin, M.
hexame taphosph a te. W.. U. S. Patent 2,233.574
Pectin Prepared by Ammonia The calcium content of the
I .

De-esterification in Aqueous Solution (194 I).


Calcium low-methoxyl pectin should (5) Baker, G. L., Pollari, V. E., and
Content
Working specifications for low-meth- not exceed 0.1% on a mois- Murray. W. G.. Fruit Products J.,
oxyl pectin are set forth by each manu- ture-free and ash-minus-carbonate-free 24, 356:60 (1945).
facturer of the product at present, basis. Calcium is determined volumet- (6) Bricker, L. G., and Proctor, K. L.,
largely because the method of production rically, using permanganate. Correc- Ind. Ene. Chem., Anal. Ed.,. 17,.
tions are made for moisture and ash- 511-12-(1945). .
affects somewhat the characteristics of ( 7 ) Graham, R. P., and Shepherd, A.
the product. For this reason, the char- minus-carbonate. KO difficulty has been D., Gel Dewatering Reel, to be
acteristics of low-methoxyl pectin pre- experienced in maintaining the calcium published.
pared by ammonia de-esterification in content below this value. (8) Graham, R. P., and Shepherd, A.
aqueous solution are described. The The Federal Food and Drug D., U. S. Patent 2,503,258
characteristics are those of the product Lead Administration has not es- ( 19 50).
which has been found most desirable Ontent tablished any tolerance for (9) Zbid.,2,548,895 (1951).

1000 AGRICULTURAL A N D FOOD C H E M I S T R Y


(10) Hall? H. H., and Fahs, F. J., and Maclay, W. D., Food Znds., (27) Owens, H. S., and Schultz, T. H.,
Confectioner (October 1946). 16, 794-6, 864-5, 906-8 (1944). U. S. Patent 2,517,595 (1950).
(11) Hills, C. H., Mottern, H . H., Nut- (20) McCready, R. M., Owens, H. S., (28) Pollari, V. E., Murray, W. G., and
ting, G. C., and Speiser, R., Food and Maclay, W. D., U. S. Patent Baker, G. L., Fruit Products J.,
Technol., 3, 90-4 (1949). 2,448,818 (1948). 25, 6-8 (1945).
(21) McCready, R. M., Owens, H. S.,
(12) Hills, C. H., White, J. W., Jr., Shepherd, A. D., and Maclay, (29) Schultz, T. H., Lotzkar, H., Owens,
and Baker, G. L., Proc. Inst. W. D.. Ind. Ene. H. S., and Maclay, W. D., J .
" Chem.. 38. 1254-
Food Technol., 3, 47-58 (1 942). 6 (1946).
I ,

Phys. Chem., 49, 554-63 (1945).


(13) Jeppesen, C. R., Eastmond, E. J., (22) Mottern, H. H., and Hills, C. H., (30) Shepherd, A. D., and Graham, R.
and Logan, H. G., J . Ojbt. SOC. Ibid.. 38. 1153-6 (19461. P., Food Technol., 6, 41 1-13
dmer., 34,313-18 (1944). (23) Olsen,'A. G., Stuewer, R, F., Fehl- (1 952).
(14) Kaufman, C. W., Fehlberg, E. R., berg, E. R., and Beach, N. M., (31) Shepherd, A. D., McCready, R.
and Olsen, A. G., Food Inds.. 14, Ibid., 31, 1015-20 (1939). M . , and Owens, H . S., Food Eng.,
57-8, 109 (1942); 15, 58-60 (24) Owens, H. S., McCreadv, R. SI., 23 (7), 44-5, 180 (1951).
(1943). and Maclav, W.D., Food Tprh-
nol., 3, 77-82 (1949). (32) LVard, W. H., Swenson, H. .4.,
(15) Kertesz, Zoltan, "The Pectic Sub- and Owens, H. S., J . Phys. @
stances," New York, Interscience (25) Owens, H. S., McCready, R. M.,
and Maclav, \Y. D., Ind. Eng. Colloid Chem., 51, 1137-43 (1947).
Publishers, 1951.
Chem., 36,936 -8(1944). (33) Woodmansee, C. W., and Baker,
(16) Lineweaver, H., J . '4m. Chem. SOC., (26) Owens, H. S., McCready, R. M , G. L., Food Technol. 3, 82, 85
67, 1292-3 (1943). Shepherd. A. D., Schultz, T. H . , (1949).
(17) Lineweaver, H., and McCready, Pippen, E. L., Swenson, H . A ,
R . M., U. S. Patent 2,386,323 Miers, J. C., Erlandsen, R. F.,
( 1 945). Received for review July 27, 1953. Acctpted
and Maclay, W.D., U. S. Dept. September 25, 1953. Presented before the Divi-
(18) Maclay. W. D., and McCready, Agr., Bur. Agr. Ind. Chem., Mim- sion of Agricultural and Food Chemistry at the
R . M., Ibid., 2,478$170 (1949). eographed Circ. Ser. AIG31.0 (June ? U r d Meeting of the AMERICAN CHEMICAL
(19) McCready, R. M ~ Obvens,
: H . S., 1952). SOCIETY, Los Angeles, Calif.

Factors in Destruction of Alfalfa Carotene Evaluated

CAROTENE
Factors Affecting Destruction in Alfalfa
KENNETH A. WALSH' and S. M. HAUGE
Department of Biochemistry, Purdue University Agricultural Experiment Station, Lafayette, Ind.

The rapid destruction of carotene in alfalfa during field curing has been attributed to
the summation of the losses by enzymatic and photochemical processes. Accurate
evaluation of the extent of destruction under field conditions is confused by continual
changes in temperature, moisture, light intensity, and physical state of the tissue. By
incubating aqueous suspensions of macerated alfalfa leaves under controlled conditions
it was possible to measure separately the effects of temperature, pH, heat treatments, and
cyanide upon the enzymatic, photochemical, and autoxidative losses of carotene. The
enzyme system was found to have a temperature optimum of about 43' C., to be more
active at pH 4 to 5 than at higher pH values, to be heat labile, and to be partially in-
hibited by cyanide. Photochemical destruction was not markedly affected by pH
changes between 4 and 8, by temperature changes between 10" and 45' C., or by pro-
longed heat treatments after enzyme inactivation. The loss of carotene in the absence of
enzymatic and photochemical destruction was attributed to autoxidation. This mech-
anism appears to be of minor importance below 40' C., and is little affected by pH
changes between 4 and 8. It is difficult to conclude that one mechanism of carotene
destruction predominates over the other during field curing. Both contribute to carotene
loss and both must be controlled to reduce the total loss.

A LFALFA IS POTENTIALLY
SOURCE O F CAROTENE for
A
livestock,
but 43 to 90% of the carotene is lost
RICH 73, 74). This loss is attributed chiefly
to the rapid destruction of carotene by
oxidative processes which are catalyzed
Attempts have been made to determine
the relative importance of enzymatic and
photochemical destruction of carotene
during the field curing process (3, 9, by enzymes (5, 6 ) and light ( 5 ) ,and to a during field curing by comparing the
slower destruction, probably by autoxi- losses of carotene in samples incubated
Present addrfss, *VationaI Research Council dation in the absence Of light and en- at similar temperatures in the light and
of Canada, OttaEa, Ontario, Canada. zyme activity. dark (I, 72). However, it is difficult

V O L . 1, NO. 16, O C T O B E R 2 8 , 1 9 5 3 1001