Food Hydrocolloids Vol.S nO.1 pp.

9-17, 1994

On the production of low esterified pectins by acid maceration of

pectic raw materials with ultrasound treatment

I.N.Panchev, N.A.Kirtchev and Chr.G.Kratchanov

Higher Institute of Food and Flavour Industry, blvd. Maritza 26, Plovdiv 4002,
Bulgaria

Abstract. The effect of temperature, time and nitric acid concentration on the yield and quality of
low esterified pectins obtained from apple pressings by preliminary acid maceration of the raw
materials was studied. Regression equations describing the changes in the degree of esterification,
the yield, the polyuronide content and the characteristic viscosity of the pectins obtained depending
on the temperature and time of maceration of apple pressings at various nitric acid concentrations
were derived. The opportunity of intensifying the process of pectin deesterification by combining the
acid maceration of raw materials with ultrasound treatment was investigated. An additional decrease
in the degree of esterification of pectin (about 6%) and an important increase in the yield of pectin
(about 18%), as compared to the reference sample, were established. The optimum ultrasound
treatment proved to be within 24-30 min. A more prolonged ultrasound treatment resulted in
obtaining pectins of lower gel strength.

Introduction
Pectic substances find a vast and various application as hydrocolloids in food
industry.
A determining index of the pectin macromolecule properties appears to be the
distribution of electric charge density on its skeleton, finding a quantitative
expression in the degree of esterification (DE) (1,2). The degree of esterification
was found to exceed 70% for the major part of industrial pectic substances, i.e.
ester groups prevailed over free carboxyl groups. During the process of pectin
extraction from the plant raw material, the storage and the application, a
decrease in DE changing the behaviour of pectin macromolecules occurred (3).
An important pursuit of the production and application of pectins of low
degree of esterification (LM pectins) for their interesting properties was
observed. Their production under laboratory and industrial conditions was
carried out by acid, alkaline or enzymic hydrolysis of ester groups in either
homogeneous or heterogeneous media. The deesterification of pectin macro-
molecules could be completed directly in the pectic raw material during
extraction in diluted or concentrated acidic solutions, and in the raw or ready
pectin preparations as well. A process of depolymerization of pectin macro-
molecules occurred in parallel with the deesterification.
In industry, the most widely used method is acid-catalysed deesterification by
strong mineral acids for the easier control of the process and the parallel
hydrolysis of ballast substances (mainly hemicellulose) leading to an increase in
the purity of the LM pectins obtained.
Acid deesterification in a homogeneous medium was most widely studied. It
also found a practical application under laboratory and industrial conditions
(3,4). Acid deesterification of pectin in diluted ethanol or isopropanol
9
I.N.Panchev, N.A.Kirtchev and Chr.G.Kratchanov

suspension was studied on a small scale (5,6,7,8,9). Kratchanov and Balakireva

(7,8) found that the efficiency of the process depended on the degree of swelling
of the pectin-the higher moisture content of the mixture resulted in
accelerating the reaction and increasing the gel strength.
Studies of acid deesterification in a heterogeneous medium achieved by a
continuous maceration of pectic substances appeared to be rather rare.
Wallerstein (10) patented a method for the production on LM pectins from
apple pressings, citrus peels, sugar beet slices and others using aqueous solutions
of hydrochloric, sulphuric and nitric acids and revealed that the LM pectins
obtained were free of starch and protein substances.
Francois and Arnaud (11) patented a method for the production of LM
pectins from citrus peels, also using maceration of raw materials in hydrochloric
and sulphuric acids at a temperature of 100°e. In a patent for the production of
LM pectins as stated by Winkler (12) the use of sulfaminic acid was suggested.
After the method patented, apple pressings were allowed to swell first in water
at a 1:10 hydromodule for 1 h at 40°C, then sulfaminic acid at a ratio of 1:10 (pH
2.2) was added and hydrolysis for 16 h at 75°C was carried out.
Woodmansee and Baker (13) suggested a method for the production of
calcium pectinates of LM pectins from apple pressings using simultaneous acid
maceration and extraction followed by precipitating the LM pectins obtained by
calcium chloride solution.
Joseph (14) advised that maceration of pectic raw material should be
conducted for 6-36 h at a temperature of 25-45°C using a mixture of ethanol or
acetone in water containing hydrochloric acid or sulphur dioxide.
Georgieva and Kratchanov (15) studied the production of LM pectin by
macerating the plant raw materials with mineral acids at a concentration interval
of 3-6%. According to them, raising the temperature of maceration to 70°C
resulted in some decreasing the yield of pectin. It was found that when the
maceration was carried out at a lower hydromodule (the ratio of dry apple
pressings to water acid solution), the deesterification occurred faster at a lower
acid consumption and the pectin obtained was of higher quality.
Granchev (16) attempted to describe the changes in the degree of esterifi-
cation (DE), the gel strength (GS), the polyuronide content (PUC) and the yield
of pectin under various conditions of acid maceration of dry apple and carrot
pressings. Out of the regression equations suggested, however, the one
describing the change in DE depending on the time, the temperature of
maceration and the amount of nitric acid might be accepted as reliable.
The aim of the present work was to study the effect of temperature, time and
nitric acid concentration on the yield and quality of the LM pectins obtained
while investigating the opportunities for intensifying the acid deesterification
process by ultrasound action on the raw materials.

Materials and methods

Deesterification of pectin by acid maceration of pectic raw materials was
conducted according to the following experimental set-up. As independent
10
 Production of low esterified pectins by acid maceration with ultrasound

factors affecting the process , X, (temperature of running the acid maceration) ,

X z (time of carrying out the maceration) and X3 concentration of mineral acid
(HN0 3 ) used, were chosen . The tests were conducted as 100 g of dry apple
pressings were put in a glass vessel of 3 drn' working volume and 700 ern? of
definite nitric acid aqueous solution were added. After wetting the mixture, the
vessel was placed in a thermostat at a constant temperature for a defined time.
After that 1 drn' of water at 60°C was poured in while NazCO ] was added with
gentle stirring to correct pH to 4.0-4 .5. Hot water was added up to 3 drrr' and
extraction at 80°C for 30 min with continuous stirring was performed. The
treatment of the extract and the production of dry pectin were completed after
procedures as described in (17).
The ultrasound treatment of the raw materials subjected to acid maceration
while studying the opportunities for intensifying the deesterification of pectin
was performed using an ultrasound apparatus GU6-0.63 , complete with an
ultrasound tub V2-25A, made in the 'Promishlena Electronika' Plant, Gabrovo,
Bulgaria. The average value of the ultrasound field intensity emitted by the
apparatus at a working frequency of 22 kHz ranged within 1-1.2 W cm- z.
The pectic substances (PS) obtained were characterized by amount (yield) of
pectin in the extract, DE, PUC, intrinsic viscosity [ll] and gel strength (GS),
"TB. The degree of esterification and PUC of the pectins were determined after
the method described in (18) . The gel strength was established after the Tarr-
Baker method as shown in (19) and the intrinsic viscosity [1')] of pectin aqueous
solutions in 0.1 M NaCI was found by the method described in (20). Processing
of the experimental results obtained by the Ubellode type viscosimeter and the
Huggins equation 11sp = [11] + [11j2.K H .C was made using a REO computer
program, FORTRAN 77 on a Pravetz-16, Bulgaria computer. Development of
stochastic regression models based on data obtained for the effect of the
maceration time , the temperature and the concentration of acid on DE, yield of
pectin, [11] and PUC, was achieved using a PALMR program realizing an
algorithm for finding a multiple regression model. The program was taken as
stated in (21) and adapted to PC Pravetz-16 , Bulgaria.

Results and discussion

Table I shows data obtained for pectin extraction using a preliminary acid
maceration of apple pressings in nitric acid. As independent factors X, affecting
the pectin extraction, the temperature of maceration X, changed within the
range of 400:s X, :s 60°C, time of maceration X z varied at interval of
2 -s X z :s 6 h and nitric acid concentration X3 varied within the limits of
1.7 :s X3 :s 7% .
In parallel with the experimental results for DE, yield, PUC, [11] of the pectins
obtained , Table I reveals the values calculated for the same quantities Y, on the
basis of stochastic regression models describing the effect of the independent
factors Xj ' The particular equations for the values stated such as DE, yield of
pectic substance, PUC and intrinsic viscosity [11] as a function of temperature of
11
.....
N -
Z
:.0
~

'":='"'
II>
.'"
Z
~
~
:;"
Table I. Production of pectin by preliminary acid maceration of apple pressings using nitric acid at various temperatures ::;
:='
II>
No. T(°C) t(h) q%) DE(%) Y\(%) Yield(g) Y 2(g) puq%) Y,(%)
-_
["l](dl/g) Yidllg)
--_._-
'"
0>

"r'"
.. _--
0-
1 40 4 3 72.58 74.59 7.12 6.34 61.21 63.06 5.93 6.15
2 40 6 3 71.00 70.02 7.98 7.25 65.84 63.07 6.07 5.86 ...
:='

3 40 2 5 74.62 74.71 6.71 6.50 62.34 59.45 6.68 6.52 C'l

4
5
40
40
4
4
5
7
71.17
6556
70.14
65.68
7.28
8.62
7.41
8.50
58.30
54.64
58.27
53.46
5.96
5.59
5.96
5.76
...~
~

40
::;
6 6 7 61.10 61.11 9.02 9.41 51.81 51.12 5.09 4.93 :='
~
7 40 6 5 63.80 65.56 7.59 8.32 54.57 5.29 5.39
8 50 2 5 67.68 66.24 8.00 7.88 54.94
57.09
60.73 5.73 5.84
=0
-e
9 50 4 5 60.52 60.82 8.29 8.79 60.27 61.39 5.41 5.02
10 50 6 5 56.44 55.40 10.02 9.71 60.66 62.06 3.84 4.15
11 60 4 3 60.10 63.21 8.84 9.10 63.22 60.54 4.67 5.04
12 60 6 3 51.79 56.94 9.46 10.01 65.83 62.74 4.36 4.20
13 60 2 5 57.88 57.77 8.64 9.26 63.52 62.01 5.56 5.16
14 60 4 5 53.10 51.50 10.53 10.18 66.76 64.52 3.97 4.04
15 60 6 1.7 71.55 64.55 9.01 9.31 57.16 59.94 5.36 5.02
16 60 6 5 40.42 45.22 12.66 11.09 63.26 67.03 2.15 2.92
17 60 4 7 39.81 39.79 11.45 11.25 68.14 68.50 2.63 3.04
18 60 6 7 36.77 33.51 11.82 12.17 72.88 71.33 2.41 1.65
 Production of 10\\ esterified pectins b~ acid ma ceration with ultrasound

maceration XI , time of ma cerat ion X z and nitric acid concentrati on X,. arc
mentioned below.
The variation of DE as affe ct ed by the factors stud ied is de scribed by the
equation:

Y I = 84 .59 + 0.1457X I - O.5842 X 2 + 5.113IX, - 0.04257X 1X ,< - () .1 ~15X IXI

( 1)

where the multiple coefficient of correlation R.\t = (I. ~72. the e xpe rime nt a l
coefficient of Fisher F"" = 44.43 a nd the theoretical coefficient of Fisher P" =
3.02 for the degrees of freedom VI = 5, \':, = 12 and the significance level IX -rr-
0.05 according to (22) .
The analysis of the changes in the yields of PS obtained wh ile varying the same
independent factors enabled us to derive the following equ ation:

Y2 = -2.633 + O.1382X 1 + 0.4568X 2 + 0.5387X, (2)

(R M = 0.939; Fe" = 37.46; P" (3.15; IX = (1.05) = 3.2~),

The statistical analysis of the changes in PUC of PS obt ained gave us no
sufficiently satisfactory re sults. The regression equation obtained (3) de scribes
77% of dispersions from the experimental results, which appe a red to be
considerably less as compared to equations (I) and (2) whi ch re vealed ~4 and
88% of dispersions in Y, and Y2 , respectivel y.

Y, = 98.3 - 0.7866X, + O.8656X 2 - 7.085X-, + O.146IX \X,

- 1.029X 2X -, + 0.01844X 1X :,X , (3)

(R M = 0.883; F"k = 7.1; P" (6.12; IX = (1.05) = 2.996).

An indirect indication of the depol ymerization processes o f pectin macro-

molecules during the acid maceration was given hy th e changes in the intrinsic
viscosity (1)] depending on temperature , time and acid con centration. As a result
of the processing of the experimentally calculated valu es for [1']], the following
regression equation was developed :

Y 4 = 3.739 I O.06116X, + 0.6249X 2 + O.9863X,

- O.01403X 1X2 - 0,(l2015X\X-, - O.06903X:,X-, (4)

(R M = 0.962; F"" = 22 .64 ; f\ (6, II; 0: = (1.05) = 2.(9).

Equation (4) describes 95 .2% of dispersions from the expe rim ent al result s and
shows the presence of complex mutual influence of the independent factors X i
on the [1)] values, hence of the pectin molecular mass as well.
It is evident from the results obtained that the depolymerization of pectin
macromolecules was mo st slightly expressed when maceration of plant raw
13
I.:"/.Panchev. ~.A.Kirtchev and Chr.G.Kratchanov

materials was completed at low temperatures (40aC) for a short time irrespective
of the high acid concentration-7%.
Trying to find a way of intensifying the process of deesterification using acid
maceration of pectic substances directed our attention to the application of the
ultrasound action. Our idea was based on the evidence that the ultrasound field
exerted an important positive effect on the capillary soaking of liquid in the
porous structure of the plant raw material (23). The acceleration of the mase-
exchange processes in combination with the expected enhancement of the tissue
hydrolysis would intensify the deesterification of pectin in theend.
The ultrasound action was applied in the course of the maceration while the
macerated raw material was sound treated for a definite time at intervals of 30
min. Data obtained from the tests (preparatory screenings) and presented in
Table II show that the periodic ultrasound treatment accelerated the acid
deesterification of pectin.
In general. the three kinds of apple pressings used revealed an additional
decrease in DE of -4-6% and a significant increase in the yield of pectin (16-
18%) respectively. as compared to the untreated sample. The optimum
ultrasound treatment proved to be within 24-30 min. A prolonged action (test 3)
resulted in obtaining a pectin of lower gel strength.
The effect of the temperature and the time of maceration in combination with
a 30 min ultrasound action in the yield of pectin was followed in a subsequent
series of tests. The extraction was carried out at a temperture of 80a C after the
aforesaid method. The experimental results are given in Table III.
The regression equation describing the changes in DE (Y I) in connection with
the variations of the temperature (X]) and the time of maceration (X 2 ) is as
follows:

YI = -0.8621 + 3.377X I + 6.656X 2 - 0.1151X 1X2 - 0.03948X I 2 - 0.5575X/

(5)

(R M = 0.966; F"k = 65.85; P k (2,6; Ci = 0.(5) = 5.14).

The changes in the yield of PS (Y 2) under the influence of the variations of the
temperature (XI) and the time of maceration (X 2 ) are revealed by the equation:

Y2 = -2.796 + 0.1993X] + 0.5875X 2 (6)

(R M = 0.978; F"k = 65.85; P k (2,6; Ci = 0.(5) = 5.14).

The changes in PUC of the pectins obtained within the range of XI and X 2
studied are described by the equation:

Y.. = 53.66 - 0.195X 1 - 2.04X 2 - 0.OO8625X IX2 + 0.OO4617X 12 + 0.5742X/

(7)

(R M = 0.992; F"k = 36.16; P k (5,3; Ci = 0.05) = 9.01).

14
 Table II. Ex pe rime nta l data of the acid mace ration (6 h a t 50°C and hydrom odul 7: I) of a pple pressings co mbined with peri odi cal ultrasound treatm ent ,
subseq ue nt ex trac tion (45 min at 80°C) a nd cha racte rsitics of the pectins o bta ined
"Q

Kind of apple pressings No . test Macer ati on with Extraction dat a Pectin characte riza tion 8-e
(% ) ultr asound treatment pH hydrornodu l Yield PUC DE GS ~
(min) (o,{, ) 0'
( %) (% ) (OTB) :::l
...,
Q

1 co ntro l 1.9 20: I 8. 1 7 1.6 76.8 220 0-

PU C = 14.1 2 - 4.5 20: I 9.0 74.8 57.8 2 18 :Ii
DE = 83. 1 3
4
60
30
4.5
4. 5
20: I
20: 1
11.0
10.8
75.6
76. 1
52.1
53.0
180
2 12
.
l4..,
5
"'Co
5 co ntro l 1.9 30: I R.6 5R.6 72.R 220 ~
PU C = 14.1 ~
DE = 83. 1
6 - 4.5 30: I 9.7 60.1 55.6 222 :i"
7 24 4.5 30: 1 12.0 61. 1 50.0 2 16 '"
r::r
-e
~
8 cont rol 2.0 30: 1 9.0 58.4 72.2 245 "Q:
PU C = 14.8
9 - 4.4 30: 1 10.5 58.4 58.1 255
DE = 76.2 3
10 24 4.5 30: 1 12.6 61. 1 52.1 250
~
"'
~
is·
:::l

:1.
;.
=..
~
~
....
VI
c
:::l
Co
I.IIi.Panchev. N.A.Kirtchev and Chr.G.Kratchanov

Table III. Experimental data of 5'i'0 HNO, acid maceration combined with ultrasound treatment (30
min) of apple prcssings of IH.7% PUC and H2.1 % DE

T t CE YI Yield Y, PUC Y, [Til Y..

(0C) (h) (%) (0Ia) (%) ('Yo) (%) (%) (dllg) (dl/g)

40 2 74.49 72.85 5.90 6.35 50.75 50.76 6.20 6.30

40 4 70.58 70.07 800 7S, 52.30 52.89 5.04 5.34
40 11 60.68 112.83 9.16 H.70 60.21 59.60 3.7H 3.99
50 2 65.99 h8.86 H.33 8.35 5.'.07 5280 5.18 5.26
50 4 61.93 63.77 8.98 9.52 55.47 54.75 4.54 4.61
50 0 58.94 54.D 10.18 111.70 60.30 6U9 3.72 3.57
60 2 58.21 56.98 10.79 11)]4 55.51 55.76 3.05 2.88
60 4 50.93 49.60 11.56 11.51 57.40 57.53 2.30 2.54
60 (j 35.19 37.75 12.73 12.69 M.28 63.90 1.87 1.89

As a first approximation. the dependence of [ll] on the temperature XI and

the time X 2 could be expressed by the equation:

Y4 = 12.72 - O.14X] - 0.4217X 2 (8)

(R M = 0.953; F"k = 29.4; F\ (2,6; ex = 0.05) = 5.14).

It is seen from the equation that when the temperature and the time of
maceration are increased, lnl (the molecular mass, respectively) decreases due
to accelerating the process of depolymerization of the pectin macromolecule.
The better approximation of the dependence of [ll] on the temperature XI and
the time X 2 could be expressed by the equation:

Y4 = -1.211 + 0.4713X I - 0.81X 2 + 0.0155X\X 2 - 0.006733X\2 - 0.04833X 22

(9)

(R M = 0.992; F\ = 38.91; F\ (5,3; ex = 0.05) c- 9.(1).

The regression models obtained from the experimental data shown in Table
III afford an opportunity to make satisfactory predictions about the process of
production of LM pectins using acid maceration of apple pressirigs intensified by
ultrasound treatment for 30 min.
The validity of the equations proved to be statistically reliable within the range
of changes in the temperature of maceration from 40-60°C and in the time of
acid maceration from 2-6 h. The rectilinear character of the equations allwos
their application to practical purposes within the investigated intervals of
temperature and time of acid maceration of dry apple pressings. It is of interest
to investigate the influence on the pectin extraction and deesterification in other
pectic raw materials and with other acid extragents.

Conclusions
During comparative studies of the production of low esterified apple pectin
16
 Production of low esterified pectins by acid maceration with ultrasound

using acid maceration of raw mat erials , the ultrasound treatm ent (30 min) was
found to int ensify the deesterification and the extraction of pectin.
Regression models of the yield and the degree of ester ification allowing
predictions about acid mac er ati on of apple pressings in the temperature range of
40-60°C were derived. The equations could be also applied to practical
purpose s.

References
1. Nelson ,D .B. (1977) Proc. 1111. Soc. Citriculture 3, 739-742 .
2. BeMiller ,J .N. (1986) ACS Symposium , 310, 2- 12.
3. Kert esz,Z .1. ( 195 1) The pectic substances, Int erscience Publishers, New York .
4. May,C.D . ( 199;) Carbohydrat e polymers 12, 79- 99.
5. Smit,C.J .B. and Bryant ,E.F . (1968) J. Food Sci. , 33, 262.
6. Wiles,R.R . and Smit ,C.J .B. (1971) USA Patent No 3622559.
7. Kratchanov ,Chr. and Balakireva.B , (1972) Travaux Scientifiqu es , Institute technologique ,
Superiour des industries alimentaires Plovdiv ; Bulgaria, 19(1), 197-209.
8. Kratchanov,Chr. and Balakir eva,B . (1972) Travaux Scientifiq ues , Institute Technologique
Superiour des industries alimentair es, Plo vdiv , Bulgaria, 19(3) ,81-90.
9. Kratchanov, Chr. (1977) Die Lebensmittel Industrie . 24(9), 406-408.
10. Wallerst ein.L. (1935) USA Patent No 2008999.
11 . Francois.R. and Arnaud ,M . (1963) France Patent No. 1343687.
12. Winkler ,G. (1954) FRG Patent No . 917709.
13. Woodm ansee ,c.W. and Baker ,G .L. ( 1954) Del. A gr. Expt . State Bull . No . 305.
14. Jose ph,G .H., Kieser ,A.H . and Bryant ,E .F. (1949) Food Techn ol. ; 3, 85.
15. Georgi eva.Iv . and Krat chan ov,Chr. ( 1978) Scientific Works. Ca nning Research Institute ,
Plovdiv , vol. XIV , pp . 77- 88.
16. G ranchev ,D . (1988) A study of some basic factor s in the technological flow diagram for
industria l production of pectin. Dissert at ion , Plovdiv, Bulgari a.
17. Kratchan ov.Chr., Mare v,K., Kirtchev,N. and Bratanoff,A. ( 1986) J. Food Techn ol. , 21, 75 1-
761.
18. O wens,U .S., McCre ad y,R.M . and Sheph erd ,A .D. (1952) Meth ods used at western region al
research laboratory for extrac tion and analysis of pectic mat erial s, A .1. C. Report 340, Western
Region al Research Laboratory, Alban y, CA .
19. Bend er ,W .A . (1949) A nalyt. Chern. 21,408-411.
20. Mor awetz ,H . (1967) Macrom olecules in solutio n , Interscience Publi shers, New York .
21. IBM (1970) System 1360 Scient ific Subrout ine Package (360 A-CM-03X ), Version Ill , Techni cal
Publicati ons Department , New York .
22. Draper ,N .R . and Srnith.H . ( 1981) Applied regression analyses, John Wiley & Sons, New York.
23. Panch ev,l. , Kirtchev ,N. and Kratchano v,Chr. (1988) J. Food Sci. Tee/mol. 23, 337-341.

17