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Research Article
Received: 13 October 2008 Revised: 21 January 2009 Accepted: 26 January 2009 Published online in Wiley Interscience: 13 March 2009
(www.interscience.wiley.com) DOI 10.1002/jctb.2157
The antioxidant and anticorrosive properties
of crude glycerol fraction frombiodiesel
production
Maria Jerzykiewicz,
a
Irmina Cwielag
a
and Wojciech Jerzykiewicz
b
Abstract
BACKGROUND: Raw glycerol fractions as the waste material from biodiesel production were investigated for its antioxidant
and anticorrosive properties. For comparison raw glycerol fractions and biodiesel fuels together with different oil feedstocks
(as an input material for the transesterication process) were measured by means of electron paramagnetic resonance (EPR)
spectroscopy. Additionally glycerol fractions were subjected to corrosivity testing (Herbert method).
RESULTS: 1,2,3-Propanetriol-containing fractions exhibited high free radical scavenging properties, in comparison with
biodiesel and oil material. Additionally, the Herbert test proved the anticorrosive ability of the crude glycerol fractions
investigated. Radical scavenging (antioxidant) properties obtained from EPR spectroscopy paralleled the anticorrosive
investigations, with samples having the highest quenching abilities giving the best results in the Herbert tests.
CONCLUSION: The anticorrosive activity of glycerol fractions examined in this paper generates a new application in lubricant
manufacturing. Theresultsshownewwaysof utilisingrawglycerol fractionwithout verycostlyandtime-consumingpurication.
c 2009 Society of Chemical Industry
Keywords: biodiesel; antioxidants; ESR spectroscopy; glycerol fraction; Herbert test
INTRODUCTION
Biodiesel and glycerol fractions are both products of a transes-
terication reaction that can be manufactured from various oil
feedstocks; for example, oils fromrapeseed, sunowers, soybeans
and waste oil after thermal treatment.
1
Because of the huge de-
mand for a carbon-neutral and environmentally safe fuel, there
has been a worldwide increase in the production of biodiesel. This
results ina remarkable overproductionof the glycerol fraction. The
mixture of 1,2,3-propanetriol and some minor constituents was
for a long time treated as an unwanted by-product of transesteri-
cation. Although there are many applications for pure glycerine,
especially of pharmaceutical grade, overproduction and a very
expensive purication process make its manufacture unprotable.
Although the market is ooded with glycerol-containing products
such as cosmetics, soaps, toothpaste and sweeteners, there still
arises aquestionof ndingnewuses for theexcess glycerol fraction
frombiodiesel production. It is important tonda way tomakethis
glycerol fraction as valuable as biodiesel. Some chemical function-
ality of glycerol is alsocarriedout inindustry,
2,3
includingcatalysed
synthesis of esters,
4
glycerol carbonates
5,6
andpropanedioles.
7
All
suchattempts arefocusedonnewuses of pureglycerol, but not on
the use of the crude unpuried glycerol fraction. Theoretically the
yield of glycerine production accompanying biodiesel should be
10% but in practice it varies depending on the technology and is
generally around 30%. Apart from 1,2,3-propanetriol, the glycerol
fraction also contains methyl/ethyl esters, soaps, unreacted fatty
acids and glycerides. Also other natural compounds originated
in the oils can be found; for example, phenolic antioxidants. The
antioxidant content as an important property has been measured
widely for different origins and qualities of oils
810
but has never
been investigated in glycerol fractions.
Glycerol fractions from various biodiesel technologies were
found to be a very interesting subject for antioxidant studies by
electron paramagnetic resonance (EPR). Finding the antioxidant
routes in the transesterication process was one of the main
purposes of this study. The inuence of the technology on
antioxidant content was also investigated. As the main method of
investigation, EPR spectroscopy (scavenging radical method) was
used and for comparison Herbert corrosivity measurements were
performed.
MATERIALS ANDMETHODS
Glycerol fractions fromdifferent technologies
Glycerol fractions were investigatedfromsevendifferent biodiesel
producers using different patented technologies. Fractions origi-
nated in processes on various technological scales, fromindustrial

Correspondence to: Maria Jerzykiewicz, Faculty of Chemistry, WroclawUniver-

sity, F JoliotCurie 14 St, 50-383 Wroclaw, Poland.
E-mail: mariaj@wchuwr.pl
a Faculty of Chemistry, WroclawUniversity, FJoliotCurie14St, 50-383Wroclaw,
Poland
b Institute of Heavy Organic Synthesis Blachownia, Energetykow 9 St, 47-225
Kedzierzyn-Kozle, Poland
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The antioxidant and anticorrosive properties of crude glycerol fraction from biodiesel production www.soci.org
Table 1. Labels of glycerol fraction samples
Glycerol fraction
Catalyst
Alcohol Scale Oil type
Producer A
A1technical
purity
KOH/CH
3
OH Industrial Rapeseed
A2 raw samples
Producer B
B raw sample NaOH/CH
3
OH Industrial Rapeseed
Producer C
C raw sample KOH/CH
3
OH Medium-sized Rapeseed
Producer D
D1 raw sample KOH/CH
3
OH Medium-sized Rapeseed
D2 raw samples
Producer E
E raw samples KOH/CH
3
OH Medium-sized Rapeseed
Producer F
F1 raw sample NaOH/CH
3
OH Medium-sized Mixed used
oils
F2 after 1 stage
F3 after 2 stage
Producer G
G raw sample KOH/C
2
H
5
OH Laboratory Rapeseed
(100 000 t y
1
biodiesel) to laboratory scale. Also, different cat-
alysts and alcohols were used (Table 1). Samples from producer
D were differentiated between D1 and D2 owing to signicant
differences in soap content.
Raw samples are dened as crude glycerol fractions before
the removal of soap and neutralization. Metal ions (sodium
or potassium respectively) are only in the form of soaps.
Standardization methods were necessary owing to the different
alcohol and water contents of the glycerol fractions. All glycerol
fractions were pre-treated by removing the water and alcohol by
distillation.
Oils, glycerol fractions and ester sample sets
Five sets of oils, their methyl esters and glycerol fractions from
two different producers (D and F) were studied. One set (set 5,
producer F) contained a mixture of different waste oils from the
thermal treatment of food and its products; other sets consisted
of fresh rapeseed oil (cold-pressed sets 1, 3 and 4; rened set
2; producer D) and its products. Additionally, anhydrous glycerol
(analytically pure, 99.5%) was investigated.
Saponication number and soap content were determined
using standard titration methods according to Polish and EN
norms PN-86/C-45 051/05 and PN-87/C-04 821, respectively.
Electron paramagnetic resonance investigations
EPR analysis was conducted using the free radical scavenging
method with the free radical galvinoxyl (Sigma-Aldrich, St Louis,
MO, USA). Measurements of the oil and ester samples were
performed in hexane, with glycerol samples in ethyl alcohol,
using samples (19 mg) diluted in solvent (200 L).
The galvinoxyl free radical solution consisted of a
1.1 mmol dm
3
solution in the respective solvent (hexane or
ethanol). Just before EPR measurement of the investigated sam-
ple free radical solution (200 L) was added and the EPR signal
was recorded immediately. As a reference solution (blank sample)
200 L solvent and 200 L galvinoxyl solution were used.
The prepared samples were measured in glass capillaries
(0.8 mm i.d.), which were kept in a quartz EPR test tube. EPR
spectra were obtained using a Bruker ESP300E spectrometer
(Wroclaw University, Poland) with a 100 kHz magnetic eld
modulation equipped with a Bruker NMR gaussmeter ER 035M
and a microwave frequency counter (HP 5350B, Hewlett-Packard,
Palo Alto, CA, USA) operating at X-band frequencies at room
temperature. Microwave power of 20 mW, modulation amplitude
of 1 Gand double resonance cavities were used. The blank sample
was located in one cavity, and the analysed sample in the second
one. Measurements were performed for both cavities over the
same period of time with at least ve replications.
For calculations of the EPR spectra WINEPR version 2.11 of the
Bruker program was used.
Gas chromatography
The composition of the oil, glycerol fractions and esters was
determined using gas chromatography. The chromatograph
(model 5890 GLC II, Hewlett-Packard) was equipped with a
ame ionization detector and 11 m 0.22 mm 0.1 m high-
temperature capillary (Ultra 2, HP). Separation of esters C-18: 1, 2,
3 was achieved using a HP-FFAP capillary column.
Argon was used as a carrier gas at a owrate of 2 mL min
1
. The
column temperature was increased from100 to 380

C at a rate of
15

C min
1
. This last temperature was maintained for 2 min.
Data obtained from chromatography concerns only glycerol,
traces of different esters and between fractions and were
recalculated according to soap content.
Corrosivity
Corrosivity was measured using the standard Herbert method ac-
cording to normPN-92/M-55 789. Unreacted glycerides contained
inthefractions weresaponiedusingNaOHsolution(PolishPatent
P-367 013). Pre-treated glycerol fractions were subjected to cor-
rosivity with different concentration of the solution (0.55%) and
soap content (0.141.84%). Additionally, for comparison in some
tests the commercial water softeners Ergon 3 and Dequest were
used. As a blank, sample water or pure soap was used. Steel plates
(25 and 43 cm
3
) were kept in the investigated solution for 120 and
144 h at room temperature.
RESULTS ANDDISCUSSION
Diversity of the glycerol fractions
The composition of the glycerol fractions obtained from different
producers (Table 2) show some differences between them. Raw
samples from producer A had high level of impurities, but the
technical purity A2 had zero soap content due to its purication.
This purication consisted of acidication, which helps separate
the glycerol from fatty acids and glycerides. The highest content
of glycerol in the raw glycerol fractions was found for samples
from producer B.
Even samples from the same producer varied in soap and fatty
acid contents. As an example, compositions of different glycerol
fractions collectedfromproducer Dare shown in Table 3. In partic-
ular, glycerol from set 1 differed from the others, with the saponi-
cation number and percentage soap being especially high here.
Interesting results were found for producer F. The technology
used here requires two steps of biodiesel production. After the
J ChemTechnol Biotechnol 2009; 84: 11961201 c 2009 Society of Chemical Industry www.interscience.wiley.com/jctb
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www.soci.org M Jerzykiewicz, I Cwielag, W Jerzykiewicz
Table 2. Composition of glycerol fractions from different producers
Glycerol fraction
Saponication
number
(mg KOH g
1
)
Soap cont.
(%)
Glycerol
(%)
Producer A
A1 technical purity 914 0 97.8
A2 raw samples 923 2225 6981
Producer B
B raw sample 0 1520 8085
Producer C
C raw sample 32 23 61
Producer D
D1 raw sample 26 20 69
D2 raw samples 0 215 8496.5
Producer E
E raw samples 5059 2934 3650
Producer F
F1 raw sample 1 21 78
F2 after 1 stage 42 19 58
F3 after 2 stage 0 15 73
Producer G
G raw sample 35 17 69
Table 3. Characteristics of glycerol fractions from producer D
Set 1
D1
Set2
D2
Set 3
D2
Set 4
D2
Saponication number
(mg KOH g
1
)
25.7 0 0 0
Soap content (%) 19.7 2.1 15.5 3.9
Glycerol content (%) 69 96.2 84.3 94.4
rst esterication the separated glycerol fraction is removed and
another portion of alcohol and catalyst added to the reactor tank.
Fractions received from both stages (F2 and F3) varied, especially
in the saponication number. Samples from the rst stage had a
much higher content of unreacted glycerides than that following
the second stage.
Gas chromatography of the ester fraction (biodiesel) from the
vesetsshowedEU14214qualitystandardfor C-18 : 3ester content
(all samples below12%), but slightly exceeded the limit for mono-,
di- andtriglycerides. Monoglyceridecontent shouldbeabove0.8%
andfor the investigatedsamples was inthe range 0.811.14%. The
normfor di- and triglycerides is 0.2%and the sets studied indicate
the limits: 0.08, 0.18 (for sets 2 and 3) and higher, up to 0.7%.
Free radicals in glycerol fractions
EPR spectra of raw glycerol fractions from producers A and D
showa typical radical line for phenolic/semiquinone (Fig. 1).
11
The
calculatedvalueof parameter g fromthespectraat 2.00432.0046
is characteristic for radicals with an unpaired electron centred on
an oxygen atom substituted to an aromatic ring. The discussed
spectra are not resolved by hyperne interaction, which could
be also associated with this kind of radical. Additionally, the
lack of hyperne interaction and absence of nitrogen-containing
345.0 345.5 346.0 346.5 347.0 347.5 348.0 348.5 349.0
[mT]
g=2.0045
Figure 1. EPR spectrumof glycerol fraction.
groups intheelemental analysis conrmedtheabsenceof Maillard
polymers, which would be found in products of natural origin.
As stability is one of the most important features of radicals,
the signal was measured several times. The oldest samples of the
glycerol fractions were about 3 years old and the EPR parameters
remained unchanged. Sample A1 (the same producer as A2) does
not exhibit a spectrum even after accumulating 10 spectral scans,
which can be easily explained by the higher purity (technical
grade) of the glycerol fraction. Although the purity of the glycerol
fraction is essential for the presence of the radical, the soap
content and saponication number do not have any inuence. For
comparison, samples C and A2 have high and similar soap content
and high saponication number (free fatty acids and mono-, di-
andtriglyceride contents) but only A2 exhibits the phenolic radical
spectrum.
Free radical scavenging results for esters, oils and glycerol
fractions
Methyl esters, oils and glycerols show varying effectiveness of
galvinoxyl radical scavenging(Fig. 2). Glycerol fractionswerefound
to have the strongest free radical scavenging activity, and most
of investigated glycerol samples quenched the galvinoxyl radical
in the rst few minutes of measurement. As shown in Fig. 2,
oil samples have medium scavenging properties, and esters the
smallest effect. Samples from set 5, producer F, differed from the
others. The general dependence remained the same, with the
glycerol fraction having the strongest radical quenching ability
and the ester the smallest, but scavenging abilities of all fractions
from producer F were smaller than from producer D. This change
in behaviour is described below.
Galvinoxyl radical is known as a good indicator of phenolic
systems in different natural mixtures.
1215
During the scavenging
reaction protons are transferred to galvinoxyl radicals from the
phenolic systems, which become paramagnetic species with
an unpaired electron on the oxygen atom according to the
reaction
14
Galv-O +phenol-OH Galv-OH +phenol-O
Although these new-formed radicals are not stable, for samples
with the highest phenol content (glycerol from Sets 24) an ESR
signal was observed for several minutes. As an example, spectra
of the investigated glycerol fraction from set 3 are shown in
Fig. 3. The scales of the presented spectra are the same for better
quantitative comparison of changes in radical concentration with
time. Only in the rst minutes after preparing the sample was it
possible to observe the galvinoxyl radical signal; after 3 min the
www.interscience.wiley.com/jctb c 2009 Society of Chemical Industry J ChemTechnol Biotechnol 2009; 84: 11961201
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The antioxidant and anticorrosive properties of crude glycerol fraction from biodiesel production www.soci.org
0 50 100 150 200
0
20
40
60
80
100
g
a
l
v
i
n
o
x
y
l

s
i
g
n
a
l

r
e
m
a
i
n
i
n
g

[
%
]
scavenging time [minutes]
set 1
oil
glycerol
ester
set 2
oil
ester
glycerol - 0%
set 3
glycerol
ester
set 4
oil
glycerol
ester
set 5
oil
ester
glycerol
ester
oils
glycerol fractions
(producer F)
Figure 2. Dependence of galvinoxyl radical scavenging in oils and its product versus time.
3450 3460 3470 3480 3490
1 minute
[Gauss]
galvinoxyl
lines
3 minutes
formed phenolic
radical
12 minutes
Figure 3. Illustration of the galvinoxyl phenol reaction on EPR spectra.
newly formed phenolic radical overlapping galvinoxyl and after
12 min only the phenolic radical was observed. The spectrumwith
no signal was taken about 20 min from mixing. The g parameter
of the newly formed and short-lived radical was 2.0046, typical
for an aromatic system with an unpaired electron centred on an
oxygenatom. Althoughtheparameter is similar tothosepresented
before for the glycerol samples, the radicals are not the same. The
concentrationof the transient radical is muchhigher regardingthe
dilution and small volume of capillaries. In direct measurements
accumulation of scans was necessary. In addition, the radical was
formedinthose samples where previously indirect measurements
of the glycerol fraction it was not found.
Free radical scavenging results: glycerol fractions from
different producers
Scavenging properties of the investigated glycerol fraction from
seven different biodiesel technologies were compared. Results
(Fig. 4) show differences between ability of scavenging galvinoxyl
radicals. The glycerol fractions investigated could be divided
conventionally into three groups depending on their scavenging
capability.
Group I consists of glycerol fractions having the highest radical
scavenging capability. This group is represented by glycerol
fractions A2, D1 and D2. In these fractions the most efcient
quenching of galvinoxyl radical is observed.
Medium concentrations of phenols, which refer to lower
quenching activity, are found for the rest of the producers.
No scavenging (group III) was observed for analytical-grade
glycerol and a technical-grade sample (laboratory puried;
data not shown). Figure 4 shows that the sensitivity of the
galvinoxyl radical (its quenching) was related to the chemical
composition of the glycerol fractions and their different origin.
The scale of production, type of catalyst and type of alcohol
did not inuence the free radical scavenging activity. Similarly,
it is difcult to nd a relation between soap content or
saponication number and free radical scavenging ability. It is
worth pointing out that samples with the highest scavenging
properties are those with the free radical line in the EPR
spectrum. The same samples also showed transient radical during
scavenging.
Anticorrosive investigations
High concentration of phenolic compounds, known from their
antioxidant behaviour in the glycerol fractions, encouraged the
investigation of the anticorrosive properties of the samples. All
J ChemTechnol Biotechnol 2009; 84: 11961201 c 2009 Society of Chemical Industry www.interscience.wiley.com/jctb
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www.soci.org M Jerzykiewicz, I Cwielag, W Jerzykiewicz
0 50 100 150 200 250
0
20
40
60
80
100
g
a
l
v
i
n
o
x
y
l

s
i
g
n
a
l

r
e
m
a
i
n
i
n
g

[
%
]
scavenging time [minutes]
A2
A2
pure glycerol
A1
A2
C
G
F1
G
D1
D2
E
B
group I
group II
group III
Figure 4. Dependence of galvinoxyl radical scavenging in glycerol fractions from different producers versus time.
Table 4. Comparison of Herbert test with free scavenging results
Glycerol
fraction
Initial soap
content (%)
Final soap
content (%)
Corrosivity
(g m
2
)
Radical
scavenging
(% of remained
radical)
D2 15 0.75 0.28 2
E 34 0.73 0.72 60
glycerol fractions samples analysed by the Herbert method ex-
hibited anticorrosive properties. Before measurement all fractions
were saponied, and corrosivity was analysed for solutions with
different initial and different nal soap content. The corrosivity of
all saponied glycerol fractions decreases with increase of total
soap content, but what is more important is that the initial soap
content had an essential inuence on the parameter. For compar-
ison, in Table 4 are given two examples of fractions from different
producers with different initial soap content. After saponication
the soap content is comparable (about 0.75%) but corrosivity is
very different. The sample prepared from a glycerol fraction with
highsoapcontent (E) has lower anticorrosive properties andlower
free radical scavenging abilities than samples with lowinitial soap
content (D2).
CONCLUSIONS
The presented results show that most well-known phenolic
antioxidants
1416
occurring naturally in oils during esterication
reaction are transferred to the glycerol fraction. The proven high
content of phenolic antioxidants in the glycerol fraction indicates
new possibilities of using the fraction without very expensive
purication processes. The mixture of antioxidants with glycerol,
although not pure, could be used as an anticorrosive soap with
glycerol. This kind of use could only be possible for technical
demands washing of cars, tanks, trucks, trains, etc. but this
market is largeenoughtodeal withtheoverproductionof glycerol.
Electron spin resonance spectroscopy was found to be a very
useful and suitable tool for studying the antioxidant properties of
glycerol fractions. These measurements could be carried out in a
few minutes and this may lead to the use of this technique in the
future utilization of glycerol fractions.
Standard gas chromatographic methods, used for establishing
glycerol content, esters and fatty acids, is ineffective in the
investigation of phenolic antioxidants without pre-treatment and
separation of the investigated samples.
ACKNOWLEDGEMENT
The research was nancially supported by EUREKA grant E! 3590
Use-Glycerol.
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