ORNL/TM-2004/192

Direct Capture of Organic Acids from

Fermentation Media Using Ionic Liquids

November 2004

Prepared by
K. Thomas Klasson
Whitney B. Ridenour
Brian H. Davison
Joanna McFarlane
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 ORNL/TM-2004/192

Nuclear Science and Technology Division

DIRECT CAPTURE OF ORGANIC ACIDS FROM FERMENTATION

MEDIA USING IONIC LIQUIDS

K. Thomas Klasson*
Whitney B. Ridenour†
Brian H. Davison‡
Joanna McFarlane

Date Published: November 2004

Prepared by
OAK RIDGE NATIONAL LABORATORY
P.O. Box 2008
Oak Ridge, Tennessee 37831-6283
managed by
UT-Battelle, LLC
for the
U.S. DEPARTMENT OF ENERGY
under contract DE-AC05-00OR22725

__________________________
*U.S. Department of Agriculture, New Orleans, La.
†The University of Tennessee, Knoxville (student)
‡Life Sciences Division, ORNL
 CONTENTS

Page

LIST OF FIGURES .....................................................................................................................................iv

LIST OF TABLES.......................................................................................................................................iv

ACKNOWLEDGMENTS ..........................................................................................................................vii

EXECUTIVE SUMMARY........................................................................................................................... v

1. INTRODUCTION ................................................................................................................................... 1

2. MATERIALS AND METHODS............................................................................................................. 2

2.1 IONIC LIQUIDS ............................................................................................................................2
2.2 PROCEDURES ..............................................................................................................................3

3. RESULTS ................................................................................................................................................ 4
3.1 EXTRACTION OF SUCCINIC ACID FROM FERMENTATION MEDIUM ............................8
3.2 ESTIMATION OF CONTINUOUS LIQUID-LIQUID EXTRACTION REQUIREMENTS .......9

4. CONCLUSIONS ................................................................................................................................... 11

5. REFERENCES ...................................................................................................................................... 11

iii
 LIST OF FIGURES

Figure Page

1 Trihexyltetradecylphosphonium methanesulfonate. ............................................................................. 2

2 Trihexyltetradecylphosphonium dodecylbenzenesulfonates................................................................. 3

3 Tributyltetradecylphosphonium dodecylbenzenesulfonates. ................................................................ 3

4 Schematic of experimental procedure. .................................................................................................. 4

5 Results from scoping studies with ionic liquids from Merck................................................................ 5

6 Scoping studies with ionic liquid containing sulfonate(s) anions. ........................................................ 6

7 Succinic acid distribution coefficient as a function of pH at room temperature for different

TOA/ionic liquid/nonanol mixtures. ..................................................................................................... 6

8 Succinic acid distribution coefficient for different temperatures with

TOA/trihexyltetradecylphosphonium methanesulfonate/nonanol. ....................................................... 7

9 Results from comparison study with extraction solvent suggested by Hong........................................ 8

10 Distribution coefficient as a function of succinic acid concentration. .................................................. 9

11 Equilibrium diagram and working graph for determining theoretical extraction stages..................... 10

12 Schematic of extraction stages for succinic acid extraction from aqueous to ionic liquids. ............... 10

LIST OF TABLES

Table Page

1 Fermentation medium for production of succinic acid. ........................................................................ 9

iv
 ACKNOWLEDGMENTS

This work was supported by the Oak Ridge National Laboratory’s State Partnership Program and
administered by the U.S. Department of Energy’s Energy Efficiency and Renewable Energy Program.

Several of the ionic liquids used in this study were kindly supplied by Dr. Rex Ren of Wesleyan
University (Middleton, CT) and Dr. Huimin Luo in the Nuclear Science and Technology Division at Oak
Ridge National Laboratory.

v
 EXECUTIVE SUMMARY

Several ionic liquids have been investigated for the extraction of organic acids from fermentation broth.
Partitioning of representative organic acids (lactic, acetic, and succinic) between aqueous solution and
nine hydrophobic ionic liquids was measured. The extraction efficiencies were strongly dependent on pH
of the aqueous phase. Distribution coefficient was very good (approximately 60) at low succinic acid
concentrations for one of the ionic liquids (trihexyltetradecylphosphonium methanesulfonate) at neutral
pH. However, this ionic liquid had to be diluted with nonanol due to its high viscosity in order to be
useful. A diluent (trioctylamine) was also added to this mixture. The results suggest that an extraction
system based on ionic liquids may be feasible for succinic acid recovery from fermentation broth and that
two ideal extraction stages are needed to reduce the concentration from 33 g/L to 1 g/L of succinic acid.
Further studies are needed to evaluate other issues related to practical applications, including ionic liquid
loss in the process, toxicity effects of ionic liquids during simultaneous fermentation and extractions.

vi
 1. INTRODUCTION

Ionic liquids are a new class of solvents that may promise reduced energy usage and minimized emissions
in replacing conventional solvents in industrial chemical processes (1). Typically, these liquids are
composed of combinations of two large ions, with at least one of them organic in nature. Ionic liquids
offer several potential advantages, including new chemistries with improved kinetics, selectivities, and
yields (2). They support greater, longer-lasting catalytic activity at milder processing conditions (lower
temperatures and pressures). Their extremely low vapor pressure could be employed in improved
processes for greatly reduced solvent loss, less air pollution, and easier separations that could result in
large energy savings compared to traditional separations processes. Despite the attractive promise of new
“green” processes based on ionic liquids, development of industrial technologies utilizing the unique
properties of these fluids as solvents or in separations processes has been hampered by a lack of process
data upon which industry can evaluate technical and economic feasibility.

Liquid-liquid extraction is a mature technology and extraction of chemicals produced via fermentation
has been the topic of many research activities and patents. Baniel et al. described extraction and back
extraction of carboxylic acids such as lactic or citric acid in an amine-based solvent system (3, 4, 5, 6, 7).
Replacement of filtration and crystallization in fermentation-based succinic acid production with low-cost
liquid-liquid extraction was suggested as a method for improving process economics (8). Several aspects
of extractive fermentation as well as general approaches to two-phase bioreactor systems have been
discussed in a recent review paper (9). Computer-aided design of solvents in extractive fermentation have
been proposed by Wang and Achenie, addressing issues such as biocompatibility, easy of separation,
stability, and inertness to reactions (10). Often, solvents are not used in the pure form—Hong et al. found
that when tri-alkylamines were added to octanol the extraction of succinc acid from water was improved
(11).

Extractive fermentation or extraction of fermentation products with ionic liquids has been suggested and
recently reported in the literature. Ionic liquids, such as 1-butyl-3-methylimidazolium
hexafluorophosphate, have been used to extract fermentation chemicals (e.g., erythromycin) and have
shown good biocompatibility in fermentation experiments with Rhodococcus R312 (12). Amino acids
produced via fermentation have also been recovered by the same ionic liquid containing a small amount
of crown ether (13). However, investigators tested 1-butyl- and 1-octylmethylimidazolium
hexafluorophosphate for extraction of butanol from fermentation medium and expressed concerns about
toxicity (14). Results obtained by Matsumoto et al. from lactic acid fermentation experiments suggest
that imidazolium-based ionic liquids containing tributyl phosphate were not toxic to Lactobacillus
rhamnosus and performed the same as conventional organic solvents (15).

The focus of this study was on the collection of preliminary data for extraction of organic acids from
fermentation broth. The recovery of fermentation products such as succinic and lactic acids is often
costly and technically challenging. The U.S. Department of Energy Vision 2020 Advanced Separations
Team has evaluated the potential impact of biobased products and identified the “direct capture of
products from biotransformations” as the most significant barrier to large-scale implementation (16).

1
 2. MATERIALS AND METHODS

Liquid extraction experiments were carried out with different ionic liquids and ionic liquid mixtures to
determine if they could be used for the recovery of fermentation products. Several aqueous stock
solutions were prepared at different organic acid concentrations with either brine or deionized water with
a mixture of lactic, acetic, hexanoic, and succinic acids. Single-component solutions of succinic acid in
water were also prepared. The solutions were prepared in volumetric flasks by adding organic acids at
several concentrations and adjusting to a desired pH. The stock solution, containing 1–100 g/L of organic
acid, was then contacted with several different ionic liquid solvents. After contact, the aqueous phases
were analyzed by high-performance liquid chromatography (HPLC) for organic acids.

2.1 IONIC LIQUIDS

Twelve ionic liquids and ionic-liquid/solvent mixtures were tested for their extraction potential of organic
acids from aqueous phases:
1. butylmethylimidazolium bis(trifluoromethanesulfonyl)imide
2. octylmethylimidazolium bis(trifluoromethanesulfonyl)imide
3. butylmethylimidazolium hexafluorophosphate
4. trihexyl(tetradecyl)phosphonium bis(trifluoromethanesulfonyl)imide
5. 1-hexyl-3-methyl-imidazolium bis(trifluoromethanesulfonyl)imide
6. 1-butyl-1-methyl-pyrrolidinium bis(trifluoromethanesulfonyl)imide
7. trihexyltetradecylphosphonium methanesulfonate (Fig. 1)
8. trihexyltetradecylphosphonium dodecylbenzenesulfonates ( Fig. 2)
9. tributyltetradecylphosphonium dodecylbenzenesulfonates ( Fig. 3)

Ionic liquids 1 through 3 were produced and obtained from Dr. Huimin Luo in the Nuclear Science and
Technology Division at Oak Ridge National Laboratory. These ionic liquids were chosen because of
their low viscosities, their hydrophobicity, and their success with other organic extractions from aqueous
phases such as toluene and complex organic acids (17). Some of these ionic liquids were also tested with
the diluent nonanol. Ionic liquids 4 through 6 were obtained from Merck KGaA (Darmstatd, Germany)
and were selected due to their hydrophobic properties and because they were either soluble or miscible
with several organics. Ionic liquids 7 through 9 were kindly supplied by Dr. Rex Ren of Wesleyan
University (Middleton, CT). These ionic liquids were chosen based on their extreme hydrophobicity.
The ionic liquids obtained from Dr. Ren were very viscous and were diluted with nonanol to a 50 vol %
mixture with stirring for 2 hours.

H3C

CH3

H3C

+
P

O -
S O
O CH3
H3C

Fig. 1. Trihexyltetradecylphosphonium methanesulfonate.

2
 H3C

CH3

H3C

+
P

O - Rn
S
H3C
O O

CH3

Fig. 2. Trihexyltetradecylphosphonium dodecylbenzenesulfonates. The ‘Rn’

bond indicates different ring bonding sites of the alkane chain.

CH3
H3C

H3C

+
P

H3C O -
S Rn

O O

CH3

Fig. 3. Tributyltetradecylphosphonium dodecylbenzenesulfonates. The ‘Rn’

bond indicates different ring bonding sites of the alkane chain.

2.2 PROCEDURES

The ionic liquids were tested under different conditions of temperature, pH, salinity, and contact time. A
typical experiment used a contact ratio of 1:10 (solvent-to-aqueous). The mixture was then shaken hard
by hand for 3 min, centrifuged for 15 min at 1000 rpm, and left overnight to help separate the phases (as
the aqueous phase was sometimes cloudy) and reach equilibrium. After contact and separation, the pH of
the aqueous phase was measured, followed by filtration through a 0.2-µm-pore-size filter, and finally
injected into a high-performance liquid chromatograph (HPLC). The mobile phase (filtered 5 mM
H2SO4) of the HPLC was pumped at 0.6 mL/min through a 300 × 7.8 mm (8-µm-particle-size) RHM
Monosaccharide column (Phenomenex, Torrance, CA) held at a temperature of 65°C to a refractive index
detector (Model 2410, Waters Corporation, Miltford, MA) held at a temperature of 40°C. The sample
injection volume was 10 µL, and the resulting chromatograms were compared with injections of acid
standards or controls not in contact with the ionic liquid to compare the amount of organic acid extracted
into the ionic liquid. A schematic of the procedure is shown in Fig. 4.

3
 Aqueous organic acid Mix and centrifuge Analyze on HPLC
solution of known weight (Wa) (control solution) (result = Ai)

Calculate Kd

Ionic liquid solution Mix and centrifuge Analyze on HPLC

of known weight (Wo) to separate phases (result = Af)

Fig. 4. Schematic of experimental procedure.

The ionic liquids trihexyltetradecylphosphonium methanesulfonate, trihexyltetradecylphosphonium

dodecylbenzenesulfonates, and tributyltetradecylphosphonium dodecylbenzenesulfonate (each with 50%
nonanol) were also mixed with trioctylamine (TOA) as a diluent in at 1 mol TOA/kg of ionic
liquid/nonanol mixture. Trioctylamine was chosen as the diluent based on results in the literature (11).
These three ionic liquids were the only ones able to dissolve trioctylamine after stirring for 24 h.

One of the most important variables measured was the distribution coefficient, Kd. It was defined as the
ratio between the concentrations of the solute (e.g., succinic acid) in the organic phase and in the aqueous
phase after contact. As the organic phase concentration could not be measured directly using the HPLC,
it was calculated from the aqueous-phase concentration according Eq.(1) and the schematic shown in
Fig. 4:

Wa (Ai − A f )
Kd = , (1)
Wo A f

where

Wa = weight of the aqueous phase,

Wo = weight of the organic (ionic liquid) phase,
Ai = initial concentration of solute in the aqueous phase = concentration of solute in the control
sample,
Af = final (equilibrium) concentration of solute in the aqueous phase.

3. RESULTS

Scoping tests with butylmethylimidazolium bis(trifluoromethanesulfonyl)imide did not produce very

promising results for either acidic, lactic, or succinic acid (data not shown). These results were in
agreement with those of Matsumoto et al. (15), who tested some of the same imidazolium-based ionic
liquids for extraction of carboxylic acids from fermentation medium and obtained Kd value of 1 or less.
In fact, we speculate that this ionic liquid interfered with the HPLC analysis of succinic acid, often
producing unreasonable results. Other research has shown that this ionic liquid has a nonnegligible
solubility in water, 8 g/L (18). Tests with octylmethylimidazolium bis(trifluoromethanesulfonyl)imide
and butylmethylimidazolium hexafluorophosphate also gave disappointing results. This was true when
nonanol was added as a diluent as well, often producing calculated values of Kd that were less than zero
(data not shown). Negative values of Kd arise when the measured concentration in the aqueous phase
after equilibrium exceeds the concentration in control samples (see Fig. 4). These unreasonable results
may be caused by interferences in the analyses of organic acids present at low concentrations (~ l g/L)
because of increased background levels by ionic liquids with aqueous solubilities of a few weight percent.
(19)

4
Scoping studies with the ionic liquids purchased from Merck did not produce very promising results
( Fig. 5). These ionic liquids were not able to extract much of the organic acids when contacted with
the extractant at approximate ratio of 10 (aqueous solution to organic extractant mixture). This was true
for any of the pHs studied: low, medium, and high.

25
Succinic Acid
Lactic Acid
20
Acetic Acid

15

K d 10 1-hexyl-3-methyl-imidazolium
bis(trifluoromethanesulfonyl)imide

5 trihexyl(tetradecyl)phosphonium 1-butyl-1-methyl-pyrrolidinium
bis(trifluoromethanesulfonyl)imide bis(trifluoromethanesulfonyl)imide

0
pH 3

pH 5

pH 12

pH 3

pH 5

pH 12

pH 3

pH 5

pH 12
-5

Fig. 5. Results from scoping studies with ionic liquids from Merck. The x-axis is
labeled with the initial pH of the aqueous phase. The final pH did not differ much for the
lower pHs. For the high-pH case, the pH dropped 1–2 units. The initial concentrations
of organic acids were 1 g/L.

Scoping studies with the ionic liquids with sulphonate(s)-based anions showed some promise. The
greatest extraction efficiency was noted for succinic acid. Each of the organic acids was present at
~ 1 g/L and contacted with the extractant at approximate ratio of 10 (aqueous solution to organic
extractant mixture [ Fig. 6]). The best-performing ionic liquid was trihexyltetradecylphosphonium
methanesulfonate/nonanol, which worked well for all the organic acids, especially when the initial
aqueous pH was high.

The efficiency of extraction of succinic acid was studied further at additional pH ranges. The TOA/ionic
liquid/nonanol mixtures were contacted with approximately 1 g/L succinic acid solution at pH 3, pH 5,
and pH 6.5. This range of pHs bracketed the pKa values of succinic acid of 4.2 and 5.6 (20). The
contacts consisted of 0.5 mL of solvent and 5 mL of succinic acid stock solution. The control was
succinic acid stock solution not in contact with the solvent mixture. The contacts were then left in the
refrigerator overnight or allowed to remain at room temperature. The results for room temperature
conditions are shown in Fig. 7. As is noted, the use of trihexyltetradecylphosphonium
methanesulfonate as the ionic liquid constituted a superior extraction solvent. In Fig. 8, the results for
extractions systems refrigerated overnight are shown. As is noted, aqueous-organic phase equilibrium for
succinic acid established at refrigeration temperature shows lower Kd values. All the experiments
indicated that a pH below the lowest pKa (4.2) did not give the best results. These results are different
from some of the results obtained by Huddleston et al., who found the noncharged form of organic acids
are more prone to extraction by butylmethylimidazolium hexafluorophosphate (21).

5
 25
trihexyltetradecylphosphonium
methanesulfonate
Succinic Acid
Lactic Acid
20 Acetic Acid

15 tributyltetradecylphosphonium
dodecylbenzenesulfonates
Kd
10 trihexyltetradecylphosphonium
dodecylbenzenesulfonates

0
pH 3 pH 11 pH 3 pH 11 pH 3 pH 11

Fig. 6. Scoping studies with ionic liquid containing sulfonate(s) anions. All these
ionic liquids were diluted with about 50 vol % nonanol. The x-axis is labeled with the
initial pH of the aqueous phase. The final pH was 2.5–4.6 for the low-pH and 6.6–9.5 for
the high-pH case.

70
Trihexyltetradecylphosphonium
methanesulfonate
60
Trihexyltetradecylphosphonium
dodecylbenzenesulfonates
50 Tributyltetradecylphosphonium
dodecylbenzenesulfonates

40
Kd
30

20

10

0
pH 3 pH 5 pH 6.5

Fig. 7. Succinic acid distribution coefficient as a function of pH at room

temperature for different TOA/ionic liquid/nonanol mixtures.

6
 70

refrigerated
60
room temperature

50

40
Kd
30

20

10

0
pH 3 pH 5 pH 6.5

Fig. 8. Succinic acid distribution coefficient for different temperatures with

TOA/trihexyltetradecylphosphonium methanesulfonate/nonanol.

To investigate if the extraction system consisting of ionic liquids with sulfonate(s) anions (each with 50
vol % nonanol and TOA) performed better than other traditional solvent mixtures, contacts were also
made to duplicate the experiment of Hong (11) for comparison. These contacts consisted of 3 mL of 1
mol TOA/kg of octanol mixture (made by stirring two hours) and 3 mL of stock solution with 1 g/L
succinic acid. Control experiment contacts were also made with 0.5 mL of TOA/kg octanol mixture and
deionized water. In parallel experiments, the three ionic liquids with sulphonate(s) anions were used. As
noted in Fig. 9, the TOA/octanol performed as poorly as the some of the ionic liquids. However, the
capacity of the TOA/octanol for succinic acid may be greater—Hong reported that a distribution
coefficient of approximately 11 was obtained when an aqueous solution with 33 g/L of succinic acid was
contacted with an equal volume of octanol containing 1 mol TOA/kg. In our project, the opposite
correlation was observed—higher Kd values were noted when the extraction of succinic acid from water
was carried out at very low concentration (e.g., 1 g/L) than at higher values.

The effect of succinic acid concentration on the distribution coefficient is shown in Fig. 10. As is
noted, there is a strong dependence between Kd and the succinic acid concentration. At low concentration
of succinic acid in the aqueous phase, the Kd is large, which by definition means a preference of the
succinic acid to distribute into the ionic liquid phase.

7
 100

90
trihexyltetradecylphosphonium
80 metanesulfonate

70

60 trihexyltetradecylphosphonium
dodecylbenzenesulfonates
K d 50
40
trihbutyltetradecylphosphonium
dodecylbenzenesulfonates
30

20

10

0
Octanol

Fig. 9. Results from comparison study with extraction solvent suggested by

Hong. Each of the ionic liquids was diluted with 50 vol % nonanol, and all extraction
solvent mixtures contained 1 mol TOA/kg of solvent. The final pH was 4.7–5.3.

3.1 EXTRACTION OF SUCCINIC ACID FROM FERMENTATION MEDIUM

The 1 mol TOA/kg of 50 vol % trihexyltetradecylphosphonium methanesulfonate/nonanol solvent

mixture was tested with the succinic acid fermentation medium to determine if the medium would affect
the extraction coefficient of the solvent. The medium used is shown in Table 1. The pH of the medium
was adjusted to 6.5 with concentrated phosphoric acid. The medium was then contacted with extraction
solvent in a 1:10 ratio. The aqueous phase was analyzed in the HPLC along with medium without
succinic acid (to determine if the medium had any peaks inhibiting the peak of succinic acid) and medium
with succinic acid (which was a control).

Extraction of succinic acid from this medium did not appear much different than extraction of the same
acid from distilled water. The value of Kd was 1.5 when the fermentation medium was contacted with the
mixture of trihexyltetradecylphosphonium methanesulfonate/nonanol/TOA. In the case of extraction
from distilled water (under the same conditions), the Kd was 1.0. Although the Kd for the fermentation
medium was 60% higher, these low Kd’s were prone to experimental error. In practice, it is appropriate to
say that the Kd’s were essentially the same.

8
 70

60

50

40
Kd
30

20

10

0
0 5 10 15 20 25 30 35
Aqueous Phase Concentration (g/kg)

Fig. 10. Distribution coefficient as a function of succinic acid concentration. Kd

was measured between a 13 ± 3 mass ratio of succinic acid aqueous solution and a
mixture of 50 vol % trihexylxtetradecylphosphonium methanesulfonate-50% 1 mol
TOA/kg octanol.

Table 1. Fermentation medium for production of succinic acid

Chemical Concentration (g/L)

Yeast extract 2
K2HPO4 1.4
KH2PO4 0.6
(NH4)2SO4 2
MgSO4 0.2
Tryptone peptone 5
Succinic acid 33
Glucose 50 wt % 2

3.2 ESTIMATION OF CONTINUOUS LIQUID-LIQUID EXTRACTION REQUIREMENTS

In order to predict the number of unit operations needed to extract succinic acid from an aqueous phase
with an ionic liquid solvent, Fig. 11 was generated. Data from several experiments were plotted, and
an approximate equilibrium curve was drawn through the points. The working line, corresponding to an
assumed ratio of 0.7 kg/kg aqueous-to-ionic liquid flows, was drawn for ideal extraction stages (22). As
is noted in Fig. 11, two ideal stages are needed to reduce the succinic aqueous phase from 33 g/kg to 1
g/kg. The equivalent flow chart showing the unit operation with two extraction stages is shown in Fig.
12.

9
 35

Organic Phase Concentration (g/kg)

30

25

1
20

15

2
10

5 slope= 0.7

0
3
0 5 10 15 20 25 30 35
Aqueous Phase Concentration (g/kg)

Fig. 11. Equilibrium diagram and working graph for determining theoretical
extraction stages.

Product

Solvent
Regeneration

Extract
Solvent (ionic liquid effluent with
(fresh ionic liquid) succinic acid, 22 g/kg)
2
Extraction Extraction
3 1
Stage 2 Stage 1

Raffiate Feed
(aqueous phase with (aqueous phase with
succinic acid, 1 g/kg) succinic acid, 33 g/kg)

Fig. 12. Schematic of extraction stages for succinic acid extraction from aqueous
to ionic liquids.

10
 4. CONCLUSIONS

Several ionic liquids have been investigated for the extraction of organic acids from fermentation broth.
Most of the tests with ionic liquids failed to give promising results, and further experiments were not
pursued. The extraction efficiencies were (not surprisingly) strongly dependent on the pH of the aqueous
phase. Distribution coefficients were very good (approximately 60) at low succinic acid concentrations
for one of the ionic liquids (trihexyltetradecylphosphonium methanesulfonate) at neutral pH. However,
this ionic liquid had to be diluted with nonanol due to its high viscosity in order to be useful. A diluent
(trioctylamine) was also added to this mixture to improve performance. The results suggest that an
extraction system based on ionic liquids may be feasible for succinic acid recovery from fermentation
broth and that two ideal extraction stages are needed to reduce the concentration from 33 g/L to 1 g/L of
succinic acid.

It should be noted that this report contains preliminary results. Further studies are needed in the
following areas:
1. Studies with controlled pH: The pH was not “controlled” in the present studies. Effort was made
in many cases to adjust pH during contact if it had drifted too far from the desired value.
2. Measurement of ionic liquid losses: No effort was made in the present studies to estimate the loss
of ionic liquid in the extraction sequence.
3. Recovery of organic acid from the ionic liquid: No effort was made to recover the organic acid
(e.g., succinic acid) from ionic liquid after extraction. Some of the results suggest that both
temperature swing and pH swing may be useful methods.
4. Toxicity studies of ionic liquid: The present study was performed based on the idea that the acids
would be extracted after fermentation. If extraction is simultaneous to fermentation, the toxicity
of the ionic liquids on the microbial culture must be determined.

5. REFERENCES

1. K. N. Marsh, A. Deev, A. C.-T. Wu, E. Tran, and A. Klamt, “Room Temperature Ionic Liquids as
Replacements for Conventional Solvents―A Review,” Korean J. Chem. Eng. 19(3), 357–362
(2002).
2. K. N. Marsh, J. A. Boxall, and R. Lichtenthaler, “Room Temperature Ionic Liquids and Their
Mixtures—A Review,” Fluid Phase Equilib. 219, 93–98 (2004).
3. A. M. Baniel and A. M. Eyal, “Citric Acid Extraction,” U.S. Patent 5,426,220 (June 20, 1995).
4. A. M. Baniel et al., “Lactic Acid Production, Separation and/or Recovery Process,” U.S. Patent
5,510,526 (April 23, 1996).
5. A. M. Baniel et al., “Lactic Acid Production, Separation and/or Recovery Process,” U.S. Patent
5,780,678 (July 14, 1998).
6. A. M. Baniel, “Recovery of Carboxylic Acid from Organic Solution that Contains an Amine and an
Extraction Enhancer,” U.S. Patent 5,780,276 (July 14, 1998).
7. A. M. Baniel, “Recovery of Carboxylic Acid from Organic Solution that Contains an Amine and an
Extraction Enhancer,” U.S. Patent 5,773,653 (June 30, 1998).
8. J. G. Zeikus, M. K. Jain, and P. Elankovan, “Biotechnology of Succinic Acid Production and
Markets for Derived Industrial Products,” Appl. Microbiol. Biotechnol. 51, 545–552 (1999).

11
9. J. J. Malinowski, “Two-Phase Partitioning Bioreactors in Fermentation Technology,” Biotechnol.
Adv. 19, 525–538 (2001).
10. Y. Wang and L. E. K. Achenie, “Computer Aided Solvent Design for Extractive Fermentation,”
Fluid Phase Equilib. 201(1), 1-18 (2002).
11. Y. K. Hong, W. H. Hong, and H. N. Chang, “Selective Extraction of Succinic Acid from Binary
Mixture of Succinic Acid and Acetic Acid,” Biotechnol. Lett. 22, 871–874 (2000).
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18.

12
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2. M. A. Brown 8. H. Luo
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5. B. H. Davison 11. C. Tsouris
6. D. W. DePaoli 12. ORNL OTIC, OSTI, CRL

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13. R. E. Baltus, Department of Chemical Engineering, Clarkson University, Potsdam, NY 13699

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of Tennessee, Knoxville, TN 37996-08400