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 from biodiesel
production
Maria Jerzykiewicz,a Irmina Cwielaga and Wojciech Jerzykiewiczb
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 transesterification 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. The results show new ways of utilising raw glycerol fraction without very costly and time-consuming purification.
c 2009 Society of Chemical Industry


Keywords: biodiesel; antioxidants; ESR spectroscopy; glycerol fraction; Herbert test

INTRODUCTION

1196

Biodiesel and glycerol fractions are both products of a transesterification reaction that can be manufactured from various oil
feedstocks; for example, oils from rapeseed, sunflowers, soybeans
and waste oil after thermal treatment.1 Because of the huge demand for a carbon-neutral and environmentally safe fuel, there
has been a worldwide increase in the production of biodiesel. This
results in a remarkable overproduction of 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 transesterification. Although there are many applications for pure glycerine,
especially of pharmaceutical grade, overproduction and a very
expensive purification process make its manufacture unprofitable.
Although the market is flooded with glycerol-containing products
such as cosmetics, soaps, toothpaste and sweeteners, there still
arises a question of finding new uses for the excess glycerol fraction
from biodiesel production. It is important to find a way to make this
glycerol fraction as valuable as biodiesel. Some chemical functionality of glycerol is also carried out in industry,2,3 including catalysed
synthesis of esters,4 glycerol carbonates5,6 and propanedioles.7 All
such attempts are focused on new uses of pure glycerol, but not on
the use of the crude unpurified 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

J Chem Technol Biotechnol 2009; 84: 11961201

antioxidant content as an important property has been measured

widely for different origins and qualities of oils8 10 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 transesterification process was one of the main
purposes of this study. The influence 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 AND METHODS

Glycerol fractions from different technologies
Glycerol fractions were investigated from seven different biodiesel
producers using different patented technologies. Fractions originated in processes on various technological scales, from industrial

Correspondence to: Maria Jerzykiewicz, Faculty of Chemistry, Wroclaw University, F JoliotCurie 14 St, 50-383 Wroclaw, Poland.
E-mail: mariaj@wchuwr.pl

a Faculty of Chemistry, Wroclaw University, F JoliotCurie 14 St, 50-383 Wroclaw,

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

Table 1. Labels of glycerol fraction samples

Catalyst
Alcohol

Scale

Oil type

KOH/CH3 OH

Industrial

Rapeseed

Producer B
B raw sample

NaOH/CH3 OH Industrial

Rapeseed

Producer C
C raw sample

KOH/CH3 OH

Medium-sized Rapeseed

KOH/CH3 OH

Medium-sized Rapeseed

KOH/CH3 OH

Medium-sized Rapeseed

Glycerol fraction
Producer A
A1technical
purity
A2 raw samples

Producer D
D1 raw sample
D2 raw samples
Producer E
E raw samples
Producer F
F1 raw sample

NaOH/CH3 OH Medium-sized Mixed used

oils

F2 after 1 stage
F3 after 2 stage
Producer G
G raw sample

KOH/C2 H5 OH Laboratory

Rapeseed

(100 000 t y1 biodiesel) to laboratory scale. Also, different catalysts and alcohols were used (Table 1). Samples from producer
D were differentiated between D1 and D2 owing to significant
differences in soap content.
Raw samples are defined 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; refined set
2; producer D) and its products. Additionally, anhydrous glycerol
(analytically pure, 99.5%) was investigated.
Saponification 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.

J Chem Technol Biotechnol 2009; 84: 11961201

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 field
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 G and 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 five 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
flame ionization detector and 11 m 0.22 mm 0.1 m hightemperature 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 flow rate of 2 mL min1 . The
column temperature was increased from 100 to 380 C at a rate of
15 C min1 . 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 according to norm PN-92/M-55 789. Unreacted glycerides contained
in the fractions were saponified using NaOH solution (Polish Patent
P-367 013). Pre-treated glycerol fractions were subjected to corrosivity 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 cm3 ) were kept in the investigated solution for 120 and
144 h at room temperature.

RESULTS AND DISCUSSION

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 purification.
This purification consisted of acidification, 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 collected from producer D are shown in Table 3. In particular, glycerol from set 1 differed from the others, with the saponification 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

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1197

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 dm3 solution in the respective solvent (hexane or
ethanol). Just before EPR measurement of the investigated sample free radical solution (200 L) was added and the EPR signal

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M Jerzykiewicz, I Cwielag, W Jerzykiewicz

Table 2. Composition of glycerol fractions from different producers

Glycerol fraction
Producer A
A1 technical purity
A2 raw samples
Producer B
B raw sample

Saponification
number
(mg KOH g1 )

Soap cont.
(%)

Glycerol
(%)

914
923

0
2225

97.8
6981

1520

8085

Producer C
C raw sample

32

23

61

Producer D
D1 raw sample
D2 raw samples

26
0

20
215

69
8496.5

Producer E
E raw samples

5059

2934

3650

Producer F
F1 raw sample
F2 after 1 stage
F3 after 2 stage

1
42
0

21
19
15

78
58
73

Producer G
G raw sample

35

17

69

345.0

345.5

346.0

346.5

347.0

347.5

348.0

348.5

349.0

[mT]

Table 3. Characteristics of glycerol fractions from producer D

Saponification number
(mg KOH g1 )
Soap content (%)
Glycerol content (%)

g=2.0045

Set 1
D1

Set2
D2

Set 3
D2

Set 4
D2

25.7

19.7
69

2.1
96.2

15.5
84.3

3.9
94.4

first esterification 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 saponification number. Samples from the first stage had a
much higher content of unreacted glycerides than that following
the second stage.
Gas chromatography of the ester fraction (biodiesel) from the
five sets showed EU14214 quality standard for C-18 : 3 ester content
(all samples below 12%), but slightly exceeded the limit for mono-,
di- and triglycerides. Monoglyceride content should be above 0.8%
and for the investigated samples was in the range 0.811.14%. The
norm for 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%.

1198

Free radicals in glycerol fractions

EPR spectra of raw glycerol fractions from producers A and D
show a typical radical line for phenolic/semiquinone (Fig. 1).11 The
calculated value of parameter g from the spectra at 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 hyperfine interaction, which could
be also associated with this kind of radical. Additionally, the
lack of hyperfine interaction and absence of nitrogen-containing

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Figure 1. EPR spectrum of glycerol fraction.

groups in the elemental analysis confirmed the absence of 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 saponification number do not have any influence. For
comparison, samples C and A2 have high and similar soap content
and high saponification number (free fatty acids and mono-, diand triglyceride 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 fractions were found
to have the strongest free radical scavenging activity, and most
of investigated glycerol samples quenched the galvinoxyl radical
in the first 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.12 15 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
reaction14
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 first minutes after preparing the sample was it
possible to observe the galvinoxyl radical signal; after 3 min the

c 2009 Society of Chemical Industry

J Chem Technol Biotechnol 2009; 84: 11961201

The antioxidant and anticorrosive properties of crude glycerol fraction from biodiesel production

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100
set 1
oil
glycerol
ester
set 2
oil
ester
glycerol - 0%

galvinoxyl signal remaining [%]

80

60

set 3
glycerol
ester
set 4
oil
glycerol
ester

ester
40

oils

set 5
(producer F)
oil
ester
glycerol

20
glycerol fractions
0
0

50

100
150
scavenging time [minutes]

200

Figure 2. Dependence of galvinoxyl radical scavenging in oils and its product versus time.

concentration of the transient radical is much higher regarding the

dilution and small volume of capillaries. In direct measurements
accumulation of scans was necessary. In addition, the radical was
formed in those samples where previously in direct measurements
of the glycerol fraction it was not found.

12 minutes

formed phenolic
radical

3 minutes

1 minute

galvinoxyl
lines

3450

3460

3470

3480

3490

[Gauss]
Figure 3. Illustration of the galvinoxylphenol reaction on EPR spectra.

J Chem Technol Biotechnol 2009; 84: 11961201

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

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1199

newly formed phenolic radical overlapping galvinoxyl and after

12 min only the phenolic radical was observed. The spectrum with
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
oxygen atom. Although the parameter is similar to those presented
before for the glycerol samples, the radicals are not the same. The

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 efficient
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 purified;
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 influence the free radical scavenging activity. Similarly,
it is difficult to find a relation between soap content or
saponification 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.

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M Jerzykiewicz, I Cwielag, W Jerzykiewicz

100
A2
A2
pure glycerol
A1
A2
C
G
F1
G
D1
D2
E
B

group III
galvinoxyl signal remaining [%]

80

60

40

group II

20
group I

0
0

50

100
150
scavenging time [minutes]

200

250

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
D2
E

Initial soap
content (%)

Final soap
content (%)

Corrosivity
(g m2 )

Radical
scavenging
(% of remained
radical)

15
34

0.75
0.73

0.28
0.72

2
60

glycerol fractions samples analysed by the Herbert method exhibited anticorrosive properties. Before measurement all fractions
were saponified, and corrosivity was analysed for solutions with
different initial and different final soap content. The corrosivity of
all saponified glycerol fractions decreases with increase of total
soap content, but what is more important is that the initial soap
content had an essential influence on the parameter. For comparison, in Table 4 are given two examples of fractions from different
producers with different initial soap content. After saponification
the soap content is comparable (about 0.75%) but corrosivity is
very different. The sample prepared from a glycerol fraction with
high soap content (E) has lower anticorrosive properties and lower
free radical scavenging abilities than samples with low initial soap
content (D2).

CONCLUSIONS

1200

The presented results show that most well-known phenolic

antioxidants14 16 occurring naturally in oils during esterification
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
purification 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 large enough to deal with the overproduction of glycerol.

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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 financially supported by EUREKA grant E! 3590
Use-Glycerol.

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