Latest Developments on Characterization of Pure Biodiesel (B100) and

Finished Biodiesel Blends by Conventional GC, Ultra Fast GC and CGxCG

Daniela
1.

1
Cavagnino ,

Fausto

1
Munari ,

Danilo

2
Pierone

Thermo Fisher Scientific, Milan, Italy 2. Nova Analtica, So Paulo (SP), Brazil (E-mail for contact: danilo.pierone@novanalitica.com.br)

Overview

Here below the curves obtained for glycerin and triolein are reported

Purpose:

Glycerin

Triolein

To describe the characterization of Biodiesel B100 in compliance

with GC-based ASTM and EN methods regulating the analysis of free
and total glycerin, residual methanol, and total FAME content.
All methods can be applied using a single assembly with two GCs
automated by a single autosampler for utmost productivity. In addition,
the need to speed up the analysis time when high sample throughput
is required, led to a deeper investigation into the application of an
Ultra Fast GC technique for the determination of Free and Total
Glycerin content. Application of GCxGC for the characterization of
finished Biodiesel blends is also shown.

Methanol
EN 14110 requires a headspace GC method, based on either polar
or non-polar columns. In this work, the analysis was performed using
an external calibration procedure, which is preferred over internal
standard calibrations when automatic headspace equipment is used.
A Thermo Scientific TRACE GC Ultra equipped with a split/splitless
(SSL) inlet and a flame ionization detector (FID), automated by a
TriPlus Autosampler configured for headspace was used. The
analytical column is a non-polar Thermo Scientific TRACE TRBIODIESEL(M), 30 m, 0.32 mm I.D. 3.0 m f.t. The linearity of the
Methanol curve, the repeatability tested on different sample types and
the analytical parameters are reported in Figure 4.

Results:
The reported data demonstrate the suitability of the optimized
hardware solutions for the analysis of Biodiesel in full compliance with
the official standards. In addition, the Ultra Fast GC approach
generated promising results in terms of separation, linearity and
repeatability, offering an additional analytical tool for fast sample
screening. The advantages of GCxGC for the analysis of finished
blends are clearly stated.

FIGURE 4. Analytical parameters, linearity and repeatability obtained for EN 14110

EN 14105/ ASTM D6584 Optimized

Introduction
Interest in Biodiesel as a clean-burning alternative fuel produced
from renewable sources such as vegetable oils has increased
tremendously over the last few years, mainly because of its reduced
environmental impact in comparison with conventional mineral diesel.
In order to be commercialized as pure biofuel or blending stock,
biodiesel must meet a set of requirements defined in ASTM D6751 and
EN 14214 standards. Gas Chromatography is commonly adopted to
determine free and total glycerin (EN 14105/ ASTM D6584), total
FAME (EN 14103) and methanol (EN 14110) in pure biodiesel.

for Ultra Fast Analysis

To achieve Ultra Fast GC separation,

a narrow bore column is coiled along
with a heating element and a properly
insulated temperature sensor. It is held
in an easy to handle metal cage
(UltraFast Module - UFM) installed into a
TRACE GC Ultra oven.
For this application, an SE54 phase 0.05 m thickness on 3 m of
capillary 0.32 mm ID has been used and integrated in an UFM.
In order to minimize the possible discrimination of heavier glycerides
during the injection, a PTV (Programmable Temperature Vaporizing)
injector was used. A deactivated glass liner with glass wool showed
better performance with a suitable transfer of both volatile and nonvolatile compounds (Figure 2).

As a further possibility, methanol content can be successfully

determined by direct liquid injection of Biodiesel diluted in n-Butanol,
exploiting the PTV Backflush system to vent out the heavier fraction
without letting it enter the capillary column.
This solution overcomes the time-consuming sample preparation
procedure for Headspace analysis and the possible critical availability
of reference FAMEs mixture for calibration (Figure 5).
In addition, the same hardware configuration used for the method
EN 14103 for FAMEs analysis can be used.
FIGURE 5. Methanol determination by direct liquid injection and
backflush of heavier components
R2 = 0.9999 with external calibration

B100 in n-Butanol spiked

with Methanol and Ethanol

BKF at 3 minutes

Name

Avrg

SD

% RSD

MeOH

110133.8

2074.5

1.9

Concentration range
0.001% - 0.5% w/w

Peak Area Repeatability (10 runs)

FIGURE 2. Ultra fast separation of the calibration mixture and typical Biodiesel samples

Instrumentation
Glycerin
R2 = 0.9996

All the methods are achieved using two

Thermo Scientific TRACE GC Ultra and
the TriPlus Autosampler, able to uniquely
automate sequences on two GCs.

5 min

Sample
Biodiesel #1

Triolein
R2 = 0.9989

Biodiesel Blends
A very common use of biodiesel is its blending with ultra low sulfur
petrodiesel as a lubricity additive.
GCxGC has been effectively applied to get comprehensive
compositional characterization of biodiesel blends, providing triple
information in a single analysis: the petrodiesel composition, the
percentage of biodiesel in the blend, the FAMEs detailed composition
(Figs. 6 and 7).
FIGURE 6. FAMEs chraracterization in petrodiesel at 5% (v/v) (BD5 blend)

Sample
Biodiesel #2

The UltraFast GC technique has been accomplished with the

Thermo Scientific UltraFast Module, which directly heats the capillary
column for a faster temperature programming rate and a much shorter
analysis time.
The GCxGC analyses were performed with the Thermo Scientific
TRACE GCxGC equipped with a cryogenic (CO2 liquid) Dual Jet
modulator and a Fast FID detector.

Free and Total Glycerin

EN 14105/ ASTM D6584
The analysis of glycerin, mono, di and triglycerides by GC requires
a non-discriminative injection system. The true cold On-column
injector used on the TRACE GC Ultra is a permanently cold system,
able to prevent discrimination of the heavier fraction and to eliminate
any risk of degradation of labile components like triglycerides.

Total FAME and Linolenic Acid Methyl Ester

EN 14103
A PTV injector with Back Flush option was used, with a polar
capillary column Thermo Scientific TRACE TR-BIODIESEL(F), 30 m,
0.25 mm ID, 0.25 m f.t.

FIGURE 7. Comparison of typical compositional data (% m/m) in various vegetable oils

When FAMEs are analyzed, the heavier fraction present in biodiesel

samples (like di- and tri-glycerides) enters the column, getting stacked
onto the polar phase. This means that a few nanograms of heavy
compounds will accumulate inside the column, which increases the risk
of compromised chromatographic performance after a number of
sequences and reduces the column lifetime.

PTV Back flush (reverse flow device):

The glycerides fraction is vented out without entering the column.

The analytical column used is a non-polar Thermo Scientific

TRACE TR-BIODIESEL(G), 10 m, 0.32 mm ID, 0.1 m f.t. This
column is designed for this high temperature GC method, featuring
enhanced mechanical robustness at high oven temperature, and thus
prolonged lifetime. A 1m x 0.53 mm ID pre-column is connected to the
column by a high temperature gas-purged metal tee. This low dead
volume leak-free metal tee has been specifically conceived for a
reliable connection between the guard column and the analytical
column at high temperature operation, removing the need for normal
glass press-fit unions. This connector remains leak-free even with
extremely large and frequent oven temperature variations.

Conclusions
A single dual-GC system equipped with cold On-Column and PTVBKF injectors is proposed, which includes the versatility of the Liquid
and HS Triplus autosampler for easiest operation and maximum
sample throughput. The PTV injector with backflush capabilities
reduces the analysis time and extend the polar column lifetime. By
using the PTV injector with backflush, it is possible to avoid the use of
the Headspace technique for the determination of Methanol and
Ethanol, increasing even more the sample throughput.
In addition, an Ultra Fast method has been optimized for screening
of Glycerides content in B100 in only 5 minutes.

By incorporating the Back Flush Option into the PTV, heavy compounds
are vented out of the inlet system, preventing column contamination while
still allowing efficient transfer of compounds of interest (Figure 3).

GCxGC has proven to be a powerful technique for the

characterization of finished biodiesel blends, delivering added-value
detailed information in a single analysis.

FIGURE 3. FAMEs chromatogram of an unknown Biodiesel, along with analytical parameters.

References
[1] Comprehensive two-dimensional gas-chromatography applied to
diesel and biodiesel analysis D. Cavagnino, A. Cadoppi, G. Zilioli, F.
Pigozzo Pittcon Conference 2006.
[2] Thermo Application Notes 10212, 10215, 10216 - Determination of
Total FAME and Linolenic Acid Methyl Ester, Free and Total Glycerin and
Methanol Content in Pure Biodiesel (B100).

The system is calibrated by analyzing a 4-component calibration

mix at four different concentrations, which generates four calibration
curves for glycerin, monoolein, diolein, triolein. The linear correlation
coefficients (R2) exceed the specification of 0.9 as requested in EN
14105 for each curve, demonstrating very good linearity of the true
cold On-column injection technique.

Additional Information
For additional information, please browse:
www.analiticaweb.com.br/biodiesel

www.thermo.com