Biblid: 1821-4487 (2011) 15; 3; p.

169-172 Original Scientific Paper

UDK: 664.1:546.17 Originalni naučni rad

NITROGEN COMPOUNDS IN THE MOLASSES

AZOTNE MATERIJE U MELASI
Rada JEVTIĆ-MUČIBABIĆ*, Jasna GRBIĆ*, Nevena MIŠLJENOVIĆ**,
Gordana KOPRIVICA**, Tatjana KULJANIN**, Stevan RADIVOJEVIĆ*
* University of Novi Sad, Institute for Food Technology, Bul. Cara Lazara 1, 21000 Novi Sad, Serbia
**
University of Novi Sad, Faculty of Technology, Bul. Cara Lazara 1, 21000 Novi Sad, Serbia
e-mail: [email protected]

ABSTRACT
Composition of the molasses depends on the sugar beet composition and applied processing technology. Molasses, obtained un-
der adequate conditions of sugar beet processing, has about 80% of dry matter content (about 50% is saccharose and about 30%
non-saccharose matter). The molasses containes concentrated non-sugar components that have not been removed in the purification
process, non-sugar components that are formed during the evaporation and crystallization, as well as the remains of the auxiliary
materials used in the sugar beet processing. The non-sugar components of the molasses are divided into three major groups: inor-
ganic (mineral) compounds, organic compounds with nitrogen and organic compounds without nitrogen. This paper examined the
composition of organic nitrogen compounds in the molasses and the impact of sugar beet processing on the content of mentioned
compounds. The research was comprehended within six local sugar factories by use of average campaign samples of molasses.
Key words: molasses, nitrogen compounds, sugar beet.

REZIME
Melasa, nusproizvod prerade šećerne repe, je polikomponentni sistem koji predstavlja vrlo vrednu sirovinu za čitav niz
tehnologija. Razlog za to leži u činjenici da melasa sadrži preko dvesta organskih i neorganskih jedinjenja, neophodnih za njenu
primenu u prerađivačkim i fermentativnim tehnologijama. Sastav melase zavisi od šećerne repe i primenjene tehnologije prerade.
Pod adekvatnim uslovima prerade šećerne repe dobija se melasa sa oko 80% suve materije, od čega oko 50% čini saharoza i oko
30% nesaharozne materije. U melasi se koncentrišu nešećeri koji nisu uklonjeni postupkom čišćenja, nešećeri koji se formiraju tokom
uparavanja i kristalizacije kao i ostaci pomoćnih materijala koji se koriste u peradi repe. Nesaharozne komponente melase podeljene
su u tri velike grupe: neorganske (mineralne), organske sa azotom i organske bez azota. Organske nešećerne materije bez azota se
sastoje od produkata razlaganja hemiceluloze, produkata toplotnog razlaganja saharoze, neisparljivih i isparljivih organskih
kiselina, pektinskih materija i produkata njihovog razlaganja, kao i čitavog niza drugih jedinjenja. Sadržaj azotnih jedinjenja u
melasi zavisi od istih u šećernoj repi i transformacije nekih jedinjenja u procesu prerade repe. Oko jedne trećine azotnih materija
melase čini betain, a ostalo amino-kiseline, belančevine, amidi, melanoidini i u neznatnoj količini soli amonijaka. Za fermentativne
tehnologije, a naročito za proizvodnju pekarskog kvasca, melasa koja ne sadrži dovoljnu količinu azotnih jedinjenja je nekompletna
sirovina. Kriterijum za ocenu melase u fermentativnim tehnologijama nije sadržaj ukupnog azota, nego sadržaj azota koji proizvodni
mikroorganizam može lako da asimiluje. U radu je ispitan sastav organskih jedinjenja azota u melasi kao i uticaj uslova prerade
šećerne repe na sadržaj istih. Istraživanja su obuhvatila prosečne, kampanjske uzorke melase iz šest domaćih fabrika šećera.
Ključne reči: melasa, azotne materije, šećerna repa.
ess, non-sugar components that are formed during the evapora-
INTRODUCTION tion and crystallization, as well as the remains of the auxiliary
Molasses is a byproduct of sugar production from sugar beet materials used in the sugar beet processing: antifoams, disinfec-
or sugar cane, which is no longer possible to get the sugar by tants, flocculants, substances for pH correction and prevention of
crystallization, and that production has an economic justifica- the incrustation in the plant (Van der Poel et al., 1998, Hoffmann
tion. et al., 2009).
The main components of the molasses are carbohydrates, pri- The non-sugar components of the molasses are divided into
marily sucrose, but it also contains a significant percentage of three major groups: inorganic compounds (~33%), organic com-
non-sugar compounds. Qualitative and quantitative composition pounds with nitrogen (~ 40%) and organic compounds without
of nonsugar compounds in the molasses is a main criterion for nitrogen (~ 20%).
evaluation of the molasses as raw material in processing and The content of nitrogen compounds in the molasses depends
fermentative industries (Mahn et al., 2002, Lević et al., 2008). on their content in sugar beet and transformation of some com-
Sugar beet molasses is a good source of many micronutrients pounds during the sugar beet processing and varies in the range
(vitamins and minerals), especially potassium, calcium, sodium of 1.4 to 2.1%. The composition of nitrogen compounds in the
and magnesium. Due to such a good nutritive composition, mo- sugar beet and their behavior during the different phases of the
lasses is used as a supplement in feed production and also beet processing (up to sugar and molasses) was the subject of
showed good behavior as hypertonic solution in osmotic dehy- research of many authors. Common conclusion was that is a
dration (Koprivica et al., 2009) . strong dependence between soil fertilization with nitrogen and
The composition of molasses depends on sugar beet compo- nitrogen compounds in beet and molasses. At the same time, the
sition and technology that has been applied in the sugar beet changes and transformations of nitrogen compounds from beet to
processing. The molasses containes concentrated non-sugar molasses were followed. About one-third of the nitrogen com-
components that have not been removed in the purification proc- pounds in molasses is betaine and the rest are amino acids, pro-

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 Jevtić-Mučibabić, Rada et al. / Nitrogen Compounds in the Molasses

teins, amides, melanoidines and a negligible amount of ammo- biological material whose quality varies depending on the proc-
nium salts. (Schiweck, 1995). essing conditions. Variations of this group of indicators are in-
Earlier researches show that total nitrogen content varies fluenced by characteristics of the processing conditions, used in
from year to year, but it is also noted that the content of the be- some factories.Coefficients of variation, calculated as the ratio of
taine is approximately constant and that nitrogen from betaine is standard deviation and average values for all factories involved
33-43% of the total nitrogen content in the molasses. in the study, were significantly higher for the content of reducing
For fermentative technology, especially for the production of substances, which indicate on significant deviation of this ratio
baker's yeast, molasses, which does not contain sufficient in function of the conditions of production.
amounts of nitrogen compounds is incomplete raw material. A
criterion for evaluation of the quality of the molasses in fermen- Table 1. Methods of analysis and used instruments
tative technologies is not the content of total nitrogen, than con-
Instruments
tent of nitrogen which can be easily assimilated by microorgan- Components Methods
Type Producer
isms (Roehr, 2001).
Refractometer,
The proportion of nitrogen, of total nitrogen content, that can Dry matter Refractrometry Schmidt & Haensch
DUR-S
be assimilated, according to the literature from 1952 (Weber and
Sucrose Polarimetry Saccharomat IV Schmidt & Haensch
Becker) is 39-42%, while Olbrich (1956) reported values of 35- Reducing
55%, indicating that nitrogen content that can be assimilated de- Berlin Institute - -
substances
pends on amino acids content. Plevako (1970) was found that the Kjelfoss,
amount of nitrogen that can be assimilated was in range from Total nitrogen Kjeldahl Foss - Electric
automatic 16200
0.15 to 0.35%, calculated on the weight of molasses, which Atomic absorption
Flame
represents 15-25% of total nitrogen in the molasses. Potassium
photometry
spectrophotometer, Carl Zeiss
Research of Schiwecka et al. (1993) was shown that the con- AAS 30
tent of free amino acids was only 10-12% of total nitrogen con- Atomic absorption
Flame
Sodium spectrophotometer, Carl Zeiss
tent and indicated that yeast can also assimilate nitrogen from photometry
AAS 30
other nitrogen compounds. Primarily, oligo and polypeptides, as pH meter,
well as polydicarbonic acid and even some inorganic nitrogen pH value Potentiometry Iskra
MA 5706
compounds (nitrate, ammonium). Otherwise, betaine and nitro- Alfa-amino Spectrophotometer
Spectrophotometry Carl Zeiss
gen compounds that were not precisely identified can not be as- nitrogen Specord M 40
similated. That not identified nitrogen compounds are probably Ammonium and
colored substances, preproducts of colored substances, etc. Distillation - -
amide nitrogen
The aim of the study was examination of composition of the Spectrophotometer
organic nitrogen compounds in domestic sugar factories and the Betaine Spectrophotometry Carl Zeiss
Specord M 40
effect of sugar beet processing on the content of mentioned Betaine
Calculation - -
compunds. nitrogen
Assimilable
Microbial - -
MATERIAL AND METHOD nitrogen

The study was conducted using the average samples of mo- Table 2. Basic quality parameters of the molasses
lasses, taken from six local sugar factories (I, II, III, IV, V and Parameter Standard Coefficient
VI). The average samples for the campaign were prepared from Minimum Maximum Average
% deviation of variation
the heptade samples taken during the production of sugar from Dry matter 80.12 86.00 82.41 2.93 3.56
sugar beet. Polarization 48.90 51.60 50.02 1.19 2.37
The basic quality parameters of molasses, as well as nitrogen Purity
components, were determinated according to the methods given 59.86 61.56 60.72 0.82 1.35
quotient
in the laboratory manuals for the control of the process of sugar pH value 7.30 9.50 8.05 0.94 11.65
production (Milić et al., 1992, 1993). The methods are in accor- Reducing
0.279 0.677 0.448 0.19 42.09
dance with the regulations given by the International Commis- substances
sion for Uniform Methods of Sugar Analysis (ICUMSA, 2003) Potassium 2.62 3.82 3.36 0.43 12.92
and shown in Table 1, together with the instruments used for that Sodium 0.71 1.93 1.06 0.23 22.13
purpose.
Statistical data analysis was performed using statistical soft- Statistically determined, the least significant differences for
ware Costat (Cohort) in order to calculate the basic descriptive the basic quality indicators of the molasses (dry matter, polariza-
statistical indicators, as well as to identify the significance of tion, reducing substances and pH value), depending on process-
differences in quality indicators of molasses (Hadživuković and ing conditions, are shown in Table 3, with indications of differ-
Čobanović,1994). ences among individual factories.
Depending on the processing conditions, the values of the
RESULTS AND DISCUSSION basic indicators of molasses quality had statistically significant
variations and that means that factories in different ways and
In order to obtain more realistic picture of quality of the ex- with different effects led the production process.
amined samples of molasses, the average values and statistical Factories V and VI had a significantly higher content ofdry
analysis of the studied quality parameters of molasses samples matter in the molasses and consequently also had a significantly
from six sugar factories, are shown in Table 2. higher content of sucrose, while the content of the reducing sub-
The results show differences in the values of exam- stances and pH values, as indicators of possible hydrolysis proc-
ined parameters, which supports the fact that the sugar beet is a

170 Journal on Processing and Energy in Agriculture 15 (2011) 3

 Jevtić-Mučibabić, Rada et al. / Nitrogen Compounds in the Molasses

ess in molasses, did not show a statistically significant variations - acids, which means that production microorganisms, except
depending on the conditions of production. amino acids, can accept some other nitrogen compounds. Hun-
It is emphasized that, the content of nitrogen compounds in garian authors, who have examined the composition of the mo-
molasses depends on their content in sugar beet and transforma- lasses produced in sugar factories in Hungary (Hajdu, 1994) as
tion of some compounds in the sugar beet processing. Table 4 well as the composition of molasses in neighboring countries
shows the content of some determinated nitrogen compounds. (Romania, Ukraine, Slovakia), come to the similar results.
Table 3. Significant differences of the basic indicators of the Table 5.The share of nitrogen components in the total nitro-
quality of molasses, depending on processing conditions (p = gen in molasses
0.05)
Betaine Alfa-amino Ammonium Assimilable
Least significant difference* Factory nitrogen nitrogen, and amide nitrogen,
Reducing % % nitrogen, % %
Dry matter, 0Bx Polarization, % pH value
substances, % I 31.79 17.34 2.89 49.13
LSD=2.209 LSD=1.687 LSD=1.105 LSD=0.378 II 26.19 18.45 2.98 53.57
Lable of Lable of Lable of Lable of III 26.97 17.76 2.63 57.89
the Ratings the Ratings the Ratings the Ratings
factory factory factory factory IV 27.32 18.02 3.49 49.42
VI a VI a VI a II a V 26.67 19.44 3.33 53.33
V a V a I a IV a VI 27.96 18.23 3.23 53.23
I b III b V a III a
IV b IV b III a I a All conclusions indicate that sugar beet, from this area, has
III b I b IV a V a such a composition of nitrogen components that by transferring
II b II b II a VI a into molasses become easy "prey" for the yeasts.
* The differences ranked with the same letter (a, b, c) is not Non-sugar fraction of molasses with nitrogen is the greatest.
statistically significant For our molasses samples, average value of "crude protein" is
over 32% compared to non-sugar fraction. The value is obtained
Table 4. Nitrogen compounds in molasses by multiplying the total nitrogen content, determinate by the
Kjeldahl method, with a 6.25 factor. According to the studies of
Parameters Standard Coefficient Schiwecka et al. (1994, 1995), conversion factor 6.25 seems too
Minimum Maximum Average
% deviation of variation low for the molasses, which means that the non-sugar fraction
Total with nitrogen is even higher.
1.34 1.97 1.72 0.15 8.72
nitrogen The average values of nitrogen compounds in molasses, ex-
Betaine 2.74 6.22 4.01 0.91 22.69 pressed in g/100 g of non-sugars and interval of dispersion of
Betaine values are given in Table 6. It shows that the percentage of al-
0.33 0.74 0.48 0.11 22.92
nitrogen
pha-amino nitrogen is low and that covers only about 35% of
Alfa-amino
0.19 0.45 0.31 0.07 22.58 assimilable nitrogen.
nitrogen
Ammonium Table 6. Average values of nitrogen compounds in molasses,
and amide 0.03 0.07 0.06 0.01 16.67 interval of dispersion of the values and percentage of nitrogen
nitrogen components in total nitrogen
Assimilable
0.46 1.36 0.86 0.33 38.37 Average value, Interval of
nitrogen Nitrogen Share,
g/100 g dispersion,
compounds %
For tested molasses samples is characteristic that they have of nonsugars g/100 g of nonsugars
enhanced content of nitrogen compounds, as a result of intensive Total nitrogen 5.23 5.03 – 5.40 100
soil fertilization, planted with sugar beet, with nitrogen fertiliz-
Betaine nitrogen 1.45 1.10 – 1.70 27.72
ers. Good adoption of nitrogen from the soil is caused by
drought in the growing season. Alfa-amino nitrogen 0.94 0.71 – 1.19 17.97
Total nitrogen content in molasses varies from 1.34 to Ammonium
0.18 0.15 – 0.19 3.44
1.97%. It was found that, from the total nitrogen content, 49.7% and amide nitrogen
is betain nitrogen, alpha-amino nitrogen, ammonium and amide Assimilable nitrogen 2.77 1.91 – 3.71 52.96
nitrogen.
The share of determinated nitrogen components in the total Variation of the nitrogen compounds in the molasses,
nitrogen, in each factorie, is shown in Table 5. depending on processing conditions are shown in the Table 7.
From the aspect of baker's yeast production, the content of Significant differences between nitrogen compounds,
assimilable nitrogen is very important because it is an amount of depending on processing conditions, were not found. Factory III
nitrogen, of total nitrogen content, that yeasts can accept. The with the lowest content of total and alpha-amino nitrogen had
results of these examinations show that the content of assimi- approximately the same and even higher content of assimilable
lable nitrogen is in the range of 0.46 to 1.36% and indicate that nitrogen, compared to the factories where the content of these
there is no direct, linear dependences between total nitrogen and components were higher. All this supports the claim that
assimilable nitrogen, but the content of assimilable nitrogen is in assimilable nitrogen does not depend linearly on the content of
function of participation of the component which yeasts may ac- total nitrogen and present amino-nitrogen, but depends on the
cept in the total nitrogen. At the same time, the results arisen presence and ratio of nitrogen components that microorganisms
from this research deny Olbrich’s claims (1956,1970) and coin- can easily assimilate.
cide with the Schiweck’s examinations (1993, 1995) that the con-
tent of assimilable nitrogen is greater than the content of amino

Journal on Processing and Energy in Agriculture 15 (2011) 3 171

 Jevtić-Mučibabić, Rada et al. / Nitrogen Compounds in the Molasses

Table 7. Variation of nitrogen compounds in molasses, Koprivica, G., Mišljenović, N., Lević, Lj., Kuljanin, T. (2009).
depending on the processing conditions Influence of nutrients present in sugar beet molasses and
saccharose solution on the quality of osmodehydrated carrot.
Nitrogen compounds % Journal on Processing and Energy in Agriculture, 13 (2), 178-
Factory Total Alfa- Ammonium 180.
Betaine Assimilable
nitrogen Betaine amino and amide Lević, Lj., Razmovski, R., Vučurović, V., Koprivica, G., Mišlje-
nitrogen nitrogen
nitrogen nitrogen nović, N. (2008) Appliance of sugar beet molasses after osmo-
I 1.73 4.61 0.55 0.30 0.05 0.85 tic dehydration of apple for bioethanol production. Journal on
II 1.68 3.67 0.44 0.31 0.05 0.90 Processing and Energy in Agriculture, 12 (4), 219-221.
III 1.52 3.46 0.41 0.27 0.04 0.88 Mahn, K., Hoffmann, C., Märländer, B. (2002). Distribution of
IV 1.72 3.94 0.47 0.31 0.06 0.85 quality components in different morphological sections of su-
V 1.80 3.99 0.48 0.35 0.06 0.96 gar beet (Beta vulgaris L.), European Juornal of Agronomy 17,
VI 1.86 4.38 0.52 0.34 0.06 0.99 29-39.
Milić, M., Karadžić, V., Obradović, S. (1992). Metode za
CONCLUSION laboratorijsku kontrolu procesa proizvodnje fabrika šećera,
Based on the obtained results, it can be concluded that the Tehnološki fakultet, Zavod za tehnologiju šećera, Novi Sad.
basic indicators of the quality of molasses, depending on Milić, M., Karadžić, V., Obradović, S. (1993). Specijalni deo
processing conditions, have statistically significant variation, analitičke kontrole pogona, sporednih proizvoda, pomoćnog
indicating that the factories, in different ways and with different materijala, voda za piće i otpadnih voda i voda u energetskom
effects were implemented technology of sugar beet processing. postrojenju, Tehnološki fakultet, Zavod za tehnologiju šećera,
In addition to the results of variability of the basic quality Novi Sad.
parameters, analysed molasses, produced in local sugar factories, Olbrich, H. (1956). Die Melasse, Berlin.
meet the requirements of the processing and fermentative Olbrich, H. (1970). Geschichte der Melasse, Berlin.
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confirms the fact that they are a function of sugar beet quality. Verhalten der stickstoffhaltigen Nichtzuckerstoffe von Rüben
während des Fabrikationsprozesses, Zuckerindustrie 118 (1),
ACKNOWLEDGEMENT: The research in this paper is 15-23.
funded by the Ministry of Science and Technological Schiweck, H., Kozianowska, G., Anderlei, J., Burba, M. (1994).
Development of Serbia, 2011-2014. (III 46005) Errechnung der Dicksaft - Nichtzuckermasse aus
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