Note: Descriptions are shown in the official language in which they were submitted.
130~
"SEPARATION OF ClTRIC ACID FROM FERMENTATION BROTH
WITH A NON-ZEOLlTE POLYMERIC ADSORBENT'
Field of the Invention
The field of art to which this invention pertains is the solid bed
adsorptive separation of citric acid from fermentation broths containing citric acid,
carbohydrates, amino acids, proteins and salts. More specifically, the inventionrelates to a process for separating citric acid from fermentation broths containing
same which process employs a non-zeolite polymeric adsorbent, which selectively
adsorb citric acid, and is selected from the group consisting of a neutral, crosslinked
polystyrene polymer, a nonionic hydrophobic polyacrylic ester polymer, a weakly
basic anionic exchange resin possessing tertiary amine or pyridine functional groups,
and a strongly basic anionic exchange resin possessing quaternary amine functional
groups and mLxtures thereof
BACKGROUND OF THE INVENTION
Citric acid is used as a food acidulant, and in pharmaceutical,
industrial and detergent formulations. The increased popularity of liquid detergents
formulated with citric acid has been primarily responsible for growth of worldwide
production of citric acid to about 700 million pounds per year which is expected to
continue in the future.
Citric acid is produced by a submerged culture fermentation process
which employs molasses as feed and the microorganism, Aspergillus Niger. The
fermentation product will contain carbohydrates, amino acids, proteins and salts as
well as citric acid, which must be separated from the fermentation broth.
There are two technologies currently employed for the separation of
citric acid. The first involves calcium salt precipitation of citric acid. The resulting
calcium citrate is acidified with sulfuric acid. In the second process, citric acid is
extracted from the fermentation broth with a mixture of trilauryl-amine, n-octanol
and a Clo or Cll isoparaffin. Citric acid is reextracted from the solvent phase into
water with the addition of heat. Both technioues, however, are complex, expensive
and they generate a substantial amount of ~aste for disposal.
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The patent literature has suggested a possible third method for
separating citric acid from the fermentation broth, which involves mem~rane
filtration to remove raw materials or high molecular weight impurities and then
adsorption of contaminants onto a nonionic resin based on polystyrene or
polyacrylic resins and collection of the citric acid in the rejected phase or raffinate
and crystallization of the citric acid after concentrating the solution, or by
precipitating the citric acid as the calcium salts then acidifying with H2S04,
separating the CaS04 and contacting cation- and anion-exchangers. This method,
disclosed in European Published Application No. 151,470, August 14, 1985, is also a
rather complex and lengthy method for separating the citric acid. In contrast, the
present method makes it possible to separate the citric acid in a single adsorption
step and to recover the citric acid from the adsorbent to obtain the purified citric
acid using an easily separated desorbent.
SUMMARY OF THE INVENTION
This invention relates to a process for adsorbing citric acid from a
fermentation broth onto a polymeric adsorbent selected from the group consistingof a neutral, crosslinked polystyrene polymer, a nonionic hydrophobic polyacrylic
ester polymer, a weakly basic anionic exchange resin possessing tertiary amine or
pyridine functional groups, and a strongly basic anionic exchange resin possessing
quaternary amine functional groups and mixtures thereof and thereafter recovering
the citric acid by desorption thereof with a suitable desorbent under desorptionconditions. One aspect of the invention is in the discovery that highly selective
separation of citric acid from salts and carbohydrates is only achieved by adjusting
and maintaining the pH of the feed solution lower than the first ionization constant
(pKa1) of citric acid (3.13). The degree to which the pH must be lowered to
maintain adequate selectivity appears to be interdependent on the concentration of
citric acid in the feed mixture; the pH is inversely dependent on the concentration.
As concentrations are decreased below 13% to very low concentrations, the pH maybe near the pKa1 of citric acid of 3.13; at 13~o, the pH may range from 0.9 to 1.7;
however, at 40% citric acid feed concentration, the pH must be lowered to at least
about 1.2 or lower. At higher concentrations, the pH must be even lower; for
example, at 50% citric acid, the pH must be at or below 1Ø It is thus preferred to
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maintain the pH of the feed mixture in the range of O~S to 2.5 with a range of O~S to
2~2 giving best results~ Another aspect of the invention is the discovery that the
temperature of adsorption can be reduced for the polymeric adsorbent used hereinby the addition of acetone, or other low molecular weight ketone, to the desorbent;
the higher temperatures associated with adsorbent breakdown can thus be avoided~The invention also relates to a process for separating citric acid from
a feed mixture comprising a fermentation broth containing same, which process
employs a polymeric adsorbent selected from the group consisting of a neutral,
crosslinked polystyrene polymer, a nonionic hydrophobic polyacrylic ester polymer,
a weakly basic anionic exchange resin possessing tertiary amine or pyridine
functional groups, and a strongly basic anionic exchange resin possessing quaternary
amine functional groups and mixtures thereof which comprises the steps of:
(a) maintaining net fluid flow thorugh a column of said adsorbent in a
single direction, which column contains at least three zones having separate
operational functions occurring therein and being serially interconnected with the
terminal zones of said colurnn connected to provide a continuous connection of said
zones;
(b) maintaining an adsorption zone in said column, said zone defined
by the adsorbent located between a feed input stream at an upstream boundary of
said zone and a raffinate output stream at a downstream boundary of said zone;
(c) maintaining a purification zone immediately upstream from said
adsorption zone, said purification zone defined by the adsorbent located between an
extract output stream at an upstream boundary of said purification zone and saidfeed input stream at a downstream boundary of said purification zone;
(d) maintaining a desorption zone irnmediately upstream from said
purification zone, said desorption zone defined by the adsorbent located between a
desorbent input stream at an upstream boundary of said zone and said extract
output stream at a downstream boundary of said zone;
(e) passing said feed mixture into said adsorption zone at adsorption
conditions to effect the selective adsorption of said citric acid by said adsorbent in
said adsorption zone and withdrawing a ra~fi`nate output stream comprising the
nonadsorbed components of said ferrnentation broth from said adsorption zone;
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(f) passing a desorbent material into said desorpdon zone at
desorption conditions to effect the displacement of said citric acid from the
adsorbent in said desorption zone;
(g) withdrawing an extract output stream comprising said citric acid
and desorbent material from said desorption zone;
(h) passing at least a portion of said extract output strearn to a
separation means and therein separating at separation conditions at least a portion
of said desorbent material; and,
(i) periodically advancing through said column of adsorbent in a
downstream direction with respect to fluid flow in said adsorption zone the feedinput strearn, raffinate output stream, desorbent input stream, and extract output
stream to effect the shifting of zones through said adsorbent and the production of
extract output and raffinate output streams to produce a raffinate product having a
reduced concentration of desorbent material. Further, a buffer zone may be
maintained immediately upstream from said desorption zone, said buffer zone
defined as the adsorbent located between the desorbent input stream at a
downstream boundary of said buffer zone and the raffinate output stream at an
upstream boundary of said buffer zone.
Other aspects of the invention encompass details of feed mixtures,
adsorbents, desorbents and operating conditions which are hereinafter disclosed.
BRIEF DESCRTmON OF THE DR~WINGS
Figure 1 is a plot of concentration of various citric acid species versus
the pH of citric acid dissociation which shows the shifting of the equilibrium point of
the citric acid dissociation by varying the concentraffon of citric acid, citrate anions
and the hydrogen ion.
Figure 2 is a static plot to determine the effect of pH on amount of
citric acid that can be adsorbed by the adsorbent.
Figures 3A, 3B and 3C are the plots of the pulse tests in Example I
using XAD~ to separate citric acid from a feed containing 13% citric acid at pH's of
2.4,1.7 and 0.9, respectively.
Figures 4A, 4B, 4C, 4D and 4E are plots of the pulse tests of Example
II at pH's of 2.4,1.7, 0.9, 2.8 and 1.4, respectively, run on different adsorbent samples.
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Figures SA and 5B are plots of the pulse tests of E~ample III at pH'
of 2.8 and 1.4, respectively, and temperatures of 93 C.
Figures 6A, 6B and 6C are plots of the pulse tests of Example IV at
pH's of 1.94,1.13 and 0.5, respectively.
Figures 7A, 7B and 7C are plots of the pulse tests of Example V at
pH's of 1.82, 0.5 and 0.3, respectively.
Figures 8A and 8B are plots of the pulse tests of Example VI at pH's
of 1.5 and 1.0, respectively.
Figure 9 is a plot of the pulse test in Example YII showing the
adsorption achieved at lower temperatures (93C versus 4SC) through the
incorporation of 10% acetone in the desorbent water.
Figure 10 is the plot of the pulse test in Example VIII using a weakly
basic anionic exchange resin having a tertiary amine functionality in a cross-linked
acrylic resin matrix to separate citric acid from a feed containing 40% citric acid at a
pH of 1.6, desorbed with water.
Figures llA, llB and llC are plots of the pulse tests of Example IX at
pH's of 7.0, 3.5 and 2.4, respectively.
Figures 12,13A and 13B are the plots of the pulse test of Example X at
a pH of 1.6 run on several different adsorbent samples of weakly basic anionic
exchange resin possessing pyridine functionality in a cross-linked polystyrene resin
matrix. The citric acid is desorbed with 0.05N sulfuric acid or water.
Figure 14 is a plot of the pulse test of Example XIII at a pH of 1.6.
Figure lS is the plot of the pulse test of Example XIV at a pH of 2.2
run on a different adsorbent sample of a less strongly basic anionic exchange resin
possessing quaternary ammonium functionality in a cross-linked polystyrene resinmatrix, desorbed with dilute sulfuric acid.
DESCRIPIION OF THE INVENllON
At the outset the definitions of various terms used throughout the
specification will be useful in making clear the operation, objects and advantages of
the instant process.
A "feed mixture" is a mixture containing one or more extract
components and one or more raffinate components to be separated by the present
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process. The term "feed stream" indicates a stream of a feed mixture which passes
to the adsorbent used in the process.
An "extract component" is a compound or type of compound that is
more selectively adsorbed by the adsorbent while a "raffinate component" is a
compound or type of compound that is less selectively adsorbed. In this process,citric acid is an extract component and saltg and carbohydrates are raffinate
components~ The term "desorbent material" shall mean generally a material
capable of desorbing an extract component~ The term "desorbent stream" or
"desorbent input stream" indicates the stream through which desorbent mateAal
passes to the adsorbent. The term "raffinate stream" or "raffinate output stream"
means a stream through which a raffinate component is removed from the
adsorbent~ The composition of the raffinate stream can vary from essentially 100%
desorbent material to essentially 100~ raffmate components~ The term "extract
stream" or "extract output stream" shall mean a stream through which an extract
material which has been desorbed by a desorbent material is removed from the
adsorbent. The composition of the extract stream, likewise, can vary from
essentially 100% desorbent mateAal to essentially 100~ extract components. At
least a portion of the extract stream and preferably at least a portion of the raffinate
stream from the separation process are passed to separation means, typically
fractionators, where at least a portion of desorbent material is separated to produce
an extract product and a raffinate product. The terms "extract product" and
"raffinate product" mean products produced by the process containing, respectively,
an extract component and a raf~mate component in higher concentrations than
those found in the extract stream and the raffinate stream. Although it is possible
by the process of this invention to produce a high puAty, citric acid product at high
recoveries, it will be appreciated that an extract component is never completelyadsorbed by the adsorbent. Likewise, a raffinate component is completely
nonadsorbed by the adsorbent. Therefore, varying amounts of a raffinate
component can appear in the extract stream and, likewise, varying amounts of an
extract component can appear in the raffinate stream. The extract and raffinate
streams then are further distinguished from each other and from the feed mixture by
the ratio of the concentrations of an extract component and a raffinate component
appearing in the particular stream. More specifically, the ratio of the concentration
of citAc acid to that of the less selectively adsorbed components will be lowest in the
1;~0306~
raffinate stream, next highest in the feed mixture, and the highest in the extract
stream. ~ikewise, the ratio of the concentration of the less selectively adsorbed
components to that of the more selectively adsorbed citric acid will be highest in the
raffinate stream, next highest in the feed mixture, and the lowest in the extract
stream.
The term "selective pore volume" of the adsorbent is defined as the
volume of the adsorbent which selectively adsorbs an extract component from the
feed mixture. The term "nonselective void volume" of the adsorbent is the volumeof the adsorbent which does not selectively retain an extract component from thefeed mixture. This volume includes the cavities of the adsorbent which contain no
adsorptive sites and the interstitial void spaces between adsorbent particles. The
selective pore volume and the nonselective void volume are generally expressed in
volumetric quantities and are of importance in determining the proper flow rates of
fluid required to be passed into an operational zone for efficient operations to take
place for a given quantity of adsorbent. When adsorbent "passes" into an
operational zone (hereinafter defined and described) employed in one embodiment
of this process its nonselective void volume together with its selective pore volume
carries fluid into that zone. The nonselective void volume is utilized in determining
the amount of fluid which should pass into the same zone in a countercurrent
direction to the adsorbent to displace the fluid present in the nonselective void
volume. If the fluid flow rate passing into a zone is smaller than the nonselective
void volume rate of adsorbent material passing into that zone, there is a net
entrainment of liquid into the zone by the adsorbent. Since this net entrainment is a
fluid present in nonselective void volume of the adsorbent, it in most instancescomprises less selectively retained feed components. The selective pore volume of
an adsorbent can in cenain instances adsorb portions of raffinate material from the
fluid surrounding the adsorbent since in certain instances there is competition
between extract material and raffinate material for adsorptive sites within the
selective pore volume. If a large quantity of raffinate material with respect toextract material surrounds the adsorbent, raffinate material can be competitive
enough to be adsorbed by the adsorbent.
The feed material contemplated in this invention is the fermentation
product obtained from the submerged culture fermentation of molasses by the
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microorganism, Aspergillus Niger. The fermentation product will have a
composition exemplified by the following:
Citricacid 12.9~o + 3
Salts 6,000 ppm
Carbohydrates (sugars) l~o
Others (proteins and amino acids) 5%
The salts will be K, Na, Ca, Mg and Fe. The carbohydrates are sugars including
glucose, xylose, mannose, oligosaccharides of DP2 and DP3 plus as many as 12 or
more unidentified saccharides. The composition of the feedstock may vary from
that given above and still be used in the invention. However, juices such as citrus
fruit juices, are not acceptable or contemplated because other materials contained
therein will be adsorbed at the same time rather than citric acid alone. Johnson, J.
Sci. Food Agric., Vol 33 (3) pp 287-93.
It has now been discovered that the separation of citric acid can be
enhanced significantly by adjusting the pH of the feed to a level below the first
ionization constant of citric acid. The first ionization constant (pKa1) of citric acid
is 3.13, Handbook of Chemistrv & Physics, 53rd Edition, 1972-3, CRC Press, and
therefore, the pH of the citric acid feed should be below 3.13. When the pH for a
13% concentrated solution of citric acid is 2.4 or greater, for example, as in Figure
3A (Example I), citric acid "breaks through" (is desorbed) with the salts and
carbohydrates at the beginning of the cycle, indicating that all the citric acid is not
adsorbed. In contrast, less '~reak through" of citric acid is observed when the pH is
1.7 and no '~reak through" when the pH is 0.9 at the 13% level, for example as in
Figures 3B and 3C, respectively.
In aqueous solution, unionized citric acid exists in equilibrium with
the several citra~eanions and hydrogen ions. This is shown in the following
equations, where the acid dissociation constants, pKal, pKa2 and pKa3 of citric acid
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at 25C are 3~, 4.74 and 5.40, respectively.
~q~
PKal ~ 3.13 PKa2 ' 4 74 pl~a3 ~ 5.40
H3CA --- H2CA;l ~ HCA;2 ~ CA-3
The equilibrium point of citric acid dissociation can be shifted by var~ring theconcentrations of atric acid, the citrate anion or the hydrogen ion. This is
demonstrated in Figure 1, for the concentrat;on o~the several citric acid species in
solution versus pH at 90C The result shows a higher percent of noruonized citric
acid (H3CA) at a higher hydrogen ion concentration (lower pH). Decreasing the
pH (raising the H+ ion concentration) will introduce more nonionized dtric add
while reducing the citrate anionic species (H2CA l, HCA-2 and CA 33in the
solution.
Based on the citric acid equilibrium and the resin propert;es
mentioned above, nonionizcd dtric acid vill be separated from other ionic species
(inc1uding citrate anions) in the fermentation broths using the resin adsorbentsdescribed. However, for a higher citric acid recovery, a lower pH solution is
required. The static adsorption isotherm of a particular resin falling within the
invention, Amberlite XAD~, for citric acid was carried out at room temperature
about 25C, as a function of feed pH. Figure 2 shows the results of the study. The
results show adsorpdon of the nonionic citric acid as the pH is lowered. Without the
intention of being lirnited by this explaDation, it appears that the nonionic citric acid
species in the soludon is preferentially adsorbed on the adsorbents of the present
invention either through an acid-base interacdon mechanism or a hydrogen bondingmechanism or a mechanism based on a strong affinity for relatively hydrophobic
species or a combination of these mechanisms.
Desorbent materials used in various prior art adsorptive separation
processes vary depending upon such factors as the type of operation employed. Inthe SWiDg bed system, in which the selectively adsorbed feed component is removed
from the adsorbent by a purge stream, desorbent sdection is not as critical and
desorbent materials comprising gaseous hydrocarbons sucb as methane, ctbane, ctc.,
or other types of gases such as nitrogen or hydrogen may be used at elcvatcd
* trade mark
30306
temperatures or reduced pressures or both to effectively purge the adsorbed feedcomponent from the adsorbent. However, in adsorptive separation processes which
are generally operated continuously at substantially constant pressures and
temperatures to ensure liquid phase, the desorbent material must be judiciously
selected to satisfy many criteria. First, the desorbent material should displace an
extract component from the adsorbent with reasonable mass flow rates without itself
being so strongly adsorbed as to unduly prevent an extract component from
displacing the desorbent material in a following adsorption cycle. Expressed in
terms of the selectivity (hereinafter discussed in more detail), it is preferred that the
adsorbent be more selective for all of the extract components with respect to a
raffinate component than it is for the desorbent material with respect to a raffinate
component. Secondly, desorbent materials must be compatible with the particular
adsorbent and the particular feed mixture. More specifically, they must not reduce
or destroy the critical selectivity of the adsorbent for an extract component with
respect to a raffinate component. Desorbent materials should additionally be
substances which are easily separable from the feed mixture that is passed into the
process. Both the raffmate stream and the extract stream are removed from the
adsorbent in admixture with desorbent material and without a method of separating
at least a portion of the desorbent material the purity of the extract product and the
raffinate product would not be very high, nor would the desorbent material be
available for reuse in the process. It is therefore contemplated that any desorbent
material used in this process will preferably have a substantially different average
boiling point than that of the feed mixture to allow separation of at least a portion
of the desorbent material from feed components in the extract and raffinate streams
by simple fractional distillation thereby permitting reuse of desorbent material in
the process. The term "substantially different" as used herein shall mean that the
difference between the average boiling points between the desorbent material andthe feed mixture shall be at least about 5C. The boiling range of the desorbentmaterial may be higher or lower than that of the feed mixture. Finally, desorbent
materials should also be materials which are readily available and therefore
reasonable in cost. In the preferred isothermal, isobaric, liquid phase operation of
the process of the present invention, it has been found that water is a particularly
effective desorbent material. Also, it has been determined that acetone and other
low molecular weight ketones, such as methylethyl ketone and diethyl ketone to be
~30306~
11
effective in admixture with water in small amounts, up to 15 wt.~o. The key to their
usefulness lies in their solubility in water. Their advantage, however, lies in their
ability to reduce the temperature at which the desorption can take place. With
some adsorbates and water as desorbent, the temperature must be raised to aid the
desorption step. Increased temperatures can cause premature deactivation of the
adsorbent. A solution to that problem in this particular separation is to add acetone
in the amount of I to 15 wt.% of the desorbent, preferably, 1 to 10 wt.æ with the most
preferred range of 5 to 10 wt.%. The low molecular weight ketone may also affectthe adsorbent stability in possibb two ways, by removing solubilizing componentswhich cause deactivation or by effecting regeneration, i.e., by removing the
deactivating agent or reversing its effect. For example, a reduction of the
desorption temperature of this separation by approximately 50C has been achieved
by adding 10 wt.æ acetone to the desorbent. A reduction of from about 5C to
about 70C can be achieved by the addition of 1 to 15 wt.æ acetone to the water
desorbent. Dilute inorganic acids have also been found to give good results whenused as desorbents. Aqueous solutions of sulfuric acid, nitric acid, hydrochloric
acid, phosphoric acid and rnixtures thereof can be used in amounts corresponding to
0.01 to l.ON (normal), with best results obtained with dilute sulfuric acid at 0.01 to
l.ON.
The prior art has also recognized that certain characteristics of
adsorbents are highly desirable, if not absolutely necessary, to the successful
operation of a selective adsorption process. Such characteristics are equally
important to this process. Among such characteristics are: (1) adsorptive capacity
for some volume of an extract component per volume of adsorbent; (2) the
selective adsorption of an extract component with respect to a raffinate component
and the desorbent material; and (3) sufficiently fast rates of adsorption and
desorption of an extract component to and from the adsorbent. Capacity of the
adscrbent for adsorbing a specific volume of an extract component is, of course, a
necessity; without such capacity the adsorbent is useless for adsorptive separation.
Furthermore, the higher the adsorbent's capacity for an extract component the
better is the adsorbent. Increased capacity of a particular adsorbent makes it
possible to reduce the amount of adsorbent needed to separate an extract
component of known concentration contained in a particular charge rate of feed
mixture. A reduction in the amount of adsorbent required for a specific adsorptive
i30306~
12
separation reduces the cost of the separation process. It is important that the good
initial capacity of the adsorbent be maintained during actual use in the separation
process over some economically desirable life. The second necessary adsorbent
characteristic is the ability of the adsorbent to separate components of the feed; or,
in other words, that the adsorbent possess adsorptive selectivity, (B), for one
component as compared to another component. Relative selectivity can be
expressed not only for one feed component as compared to another but can also beexpressed between any feed mixture component and the desorbent material. The
selectivity, (B), as used throughout this specification is defined as the ratio of the
two components of the adsorbed phase over the ratio of the same two components
in the unadsorbed phase at equilibrium conditions. Relative selectivity is shown as
Equation 2 below:
Equation 2 .
Selectivity = (B) = rVOl. percent C/vol. percent D~ A
~vol. percent C/vol. percent D~ U
where C and D are two components of the feed represented in volume percent and
the subscripts A and U represent the adsorbed and unadsorbed phases respectively.
The equilibrium conditions were determined when the feed passing over a bed of
adsorbent did not change composition after contacting the bed of adsorbent. In
other words, there was no net transfer of material occurring between the
unadsorbed and adsorbed phases. Where se1ectivity of two components approaches
l.O there is no preferential adsorption of one component by the adsorbent with
respect to the other; they are both adsorbed (or nonadsorbed) to about the same
degree with respect to each other. As the (B) becomes less than or greater than l.O
there is a preferential adsorption by the adsorbent for one component with respect
to the other. When comparing the selectivity by the adsorbent of one component Cover eomponent D, a (B) larger than l.O indicates preferential adsorption of
component C within the adsorbent. A (B) }ess than 1.0 would indicate that
component D is preferential~ adsorbed leaving an unadsorbed phase richer in
component C and an adsorbed phase richer in component D. Idealb desorbent
materials should have a selectivity equal to about 1 or slightly less than l with
`" 1303061
13
respect to all extract components so that all of the extract components can be
desorbed as a class with reasonable flow rates of desorbent material and so thatextract components can displace desorbent material in a subsequent adsorption
step. While separation of an extract component from a raffinate component is
theoretically possible when the selectivity of the adsorbent for the extract
component with respect to the raffinate component is greater than 1, it is preferred
that such selectiv;ty approach a value of 2. Iike relative volatility, the higher the
selectivity, the easier the separation is to perform. Higher selectivities permit a
smaller amount of adsorbent to be used. The third important characteristic is the
rate of exchange of the extract component of the feed mixture material or, in other
words, the relative rate of desorption of the extract component. This characteristic
relates directly to the amount of desorbent material that must be employed in the
process to recover the extract component from the adsorbent; faster rates of
exchange reduce the amount of desorbent material needed to remove the extract
component and therefore permit a reduction in the operating cost of the process.With faster rates of exchange, less desorbent material has to be pumped through the
process and separated from the extract stream for reuse in the process.
Resolution is a measure of the degree of separation of a two-
component system, and can assist in quantifying the effectiveness of a particular
combination of adsorbent, desorbent, conditions, etc. for a particular separation.
Resolution for purposes of this application is de~med as the distance between the
two peak centers divided by the average width of the peaks at 1/2 the peak height as
determined by the pulse tests described hereinafter. The equation for calculating
resolution is thus:
Equation 3
R = 1/ 2 ( Wl I W2 J
where Ll and 1~ are the distance, in ml, respectively, from a reference point, e.g.,
zero to the centers of the peaks and W1 and W2 are the widths of the peaks at 1/2
the height of the peaks.
A dynamic testing apparatus is employed to test various adsorbents
with a particular feed mixture and desorbent material to measure the adsorbent
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characteristics of adsorptive capacity, selectivity and exchange rate. The apparatus
consists of an adsorbent chamber comprising a helical column of approximately 70cc volume having inlet and outlet portions at opposite ends of the chamber. The
chamber is contained within a temperature control means and, in addition, pressure
control equipment is used to operate the chamber at a constant predetermined
pressure. Quantitative and qualitative analytical equipment such as refractometers,
polarimeters and chromatographs can be attached to the outlet line of the chamber
and used to detect quantitatively or determine qualitatively one or more
components in the effluent stream leaving the adsorbent chamber. A pulse test,
performed using this apparatus and the following general procedure, is used to
determine selectivities and other data for various adsorbent systems. The adsorbent
is filled to equilibrium with a particular desorbent material by passing the desorbent
material through the adsorbent chamber. At a convenient time, a pulse of feed
containing known concentrations of a tracer and of a particular extract component
or of a raffinate component or both, all diluted in desorbent, is injected for aduration of several minutes. Desorbent flow is resumed, and the tracer and the
extract component or the raffinate component (or both) are eluted as in a liquid-
solid chromatographic operation. The effluent can be analyzed onstream or,
alternatively, effluent samples can be collected periodically and later analyzedseparately by analytical equipment and traces of the envelopes of corresponding
component peaks developed.
From information derived from the test adsorbent, performance can
be in terms of void volume, retention volume for an extract or a raffinate
component, selectivity for one component with respect to the other, and the rate of
desorption of an extract component by the desorbent. The retention volume of an
extract or a raffinate component may be characterized by the distance between the
center of the peak envelope of an extract or a raffinate component and the peak
envelope of the tracer component or some other known reference point. It is
expressed in terms of the volume in cubic centimeters of desorbent pumped duringthis time interval represented by the distance between the peak envelopes.
Selectivity, (B), for an extract component with respect to a raffinate component may
be characterized by the ratio of the distance between the center of the extract
component peak envelope and the tracer peak envelope (or other reference point)
to the corresponding distance between the center of the raffinate component peak
30306
envelope and the tracer peak envelope. The rate of exchange of an extract
component with the desorbent can generally be characterized by the width of the
peak envelopes at half intensity. The narrower the peak width, the faster the
desorption rate. The desorption rate can also be characterized by the distance
between the center of the tracer peak envelope and the disappearance of an extract
component which has just been desorbed. This distance is again the volume of
desorbent pumped during this time interval.
To further evaluate promising adsorbent systems and to translate this
type of data into a practical separation process requires actual testing of the best
system in a continuous countercurrent liquid-solid contacting device. The general
operating principles of such a device have been previously described and are found
in Broughton U.S. Patent 2,98S,S89. A specific laboratory size apparatus utilizing
these principles is described in deRosset et al., U.S. 3,706,812. The equipment
comprises multip]e adsorbent beds with a number of access lines attached to
distributors v.1ithin the beds and terminating at a rotaly distributing valve. At a
given valve position, feed and desorbent are being introduced through two of thelines and the raffinate and extract streams are being withdrawn through two more.
All remaining access lines are inactive and when the position of the distnbutingvalve is advanced by one index, all active positions will be advanced by one bed.
This simulates a condition in which the adsorbent physically moves in a direction
countercurrent to the liquid flow. Additional detaUs on the above-mentioned
nonionic adsorbent testing apparatus and adsorbent evaluation techniques may be
found in the paper "Separation of C8 Aromatics by Adsorption~ by A. J. deRosset,R. W. Neuzil, D. J. Korous, and D. H. Rosback presented at the American ChemicalSociety, Los Angeles, California, March 28 through April 2, 1971.
One class of adsorbents to be used in the process of this invention will
comprise nor~ionogenic, hydrophobic, water-insoluble, crosslinked styrene-
poly(vinyl)benzene copolymers and copolymers thereof with monoethylenically
unsaturated compounds or polyethylenically unsaturated monomers other than
poly(vinyl)benzenes, including the ac~ylic esters, such as those described in
Gustafson U.S. Patent Nos. 3,531,463 and 3,663,467.
~ s stated in Patent No. 3,531,463, the
polymers may be made by techniques disclosed in U.S. Serial No. 749,526, filed July
18,1958, now Patent Nos. 4,221,871; 4,224,415; 4,256,840; 4,297,220; 4,382,124 and
~ .
~ 30306~
16
4,501,826 to Meitzner et al~
Adsorbents such as just described are manufactured by the Rohm and Haas
Company, and sold under the trade name "Amberlite." The types of Amberlite
polymers known to be effective for use by this invention are referred to in Rohmand Haas Company literature as Amberlite adsorbents XAD-1, XAD 2, XAD 1,
XAD-7 and XAD-8, and described in the literature as ~hard, insoluble spheres of
high surface, porous polymer." The various types of Amberlite polymeric
adsorbents differ somewhat in physical properties such as porosity volume percent,
skeletal density and nominal mesh sizes, but more so in surface area, average pore
diameter and dipole moment. The preferred adsorbents u ill have a surface area of
1~2000 square meters per gram and preferably from 100-1000 m2/g. These
properties are listed in the following table:
.-,. 'p~, .
,~
130306i
TABLE 1
Properties of Amberlite Polymeric Adsorbents
XAD-l XAD-7 XAD^8
Poly- XAD-2 XAD-4 Acryl~c Acrylic
Chem~cal Nature styrene Polystyrene Polystyrene Ester Ester
Porosity 37 42 51 55 52
True Wet Dens~ty
grams/cc 1.02 1.02 1.02 1.05 1.09
Sur~;ce Area 100 300 780 450 160
Average Pore D~ameter 225
Angstroms 200 90 50 9
Skeleta/cDcens Y 1.07 1.07 1.08 1.24 1.23
Nominal Mesh Size 20-50 20-50 20-50 20-50 25-50
Dipole Moment of
Functional Groups0.3 0.3 0.3 1.8 1.8
130306~
18
Applications for Amberlite polymeric adsorbents suggested in the Rohm and Haas
Company literature include decolorizing pùlp rnill bleaching effluent, decolorizing
dye wastes and removing pesticides from waste effluent. There is, of course, no hint
in the literature of my surprising discovery of the effectiveness of Amberlite
polymeric adsorbents in the separation of citric acid from Aspergillus-Niger
fermentation broths.
A second class of adsorbents to be used in the process of this
invention will comprise wealdy basic anion exchange resins possessing tertiaIy
amine or pyridine functionality in a cross-linked polymeric matrix, e.g., acrylic or
styrene. They are especially suitable when produced in bead form, have a high
degree of ur~iform polymeric porosity, exhibit chemical and physical stability and
good resistance to attrition (not common to macroreticular resins).
Adsorbents such as just described are manufactured by the Rohm and
Haas Company, and sold under the trademæk "Amberlite." The types of
Amberlite*polymers known to be effective for use by this invention are referred to in
Rohm and Haas Company literature as Amberlite*adsorbents XE-275 (IRA-35),
IRA-68, and described in the literature as "insoluble in all common solvents andhaving open structure for effective adsorption and desorption of large moleculeswi~hout loss of capacity, due to organic fouling." Also, suitable are AG3-X4A and
AG4-X4 manufactured by Bic Rad and comparable resins sold by Dow Chernical
Co., such as Dowex 66, and Dow experimental resins made in accordance with U.S.
Patents 4,031,038 and 4,098,867.
l~e various types of polymeric adsorbents of these classes available,
will differ somewhat in physical properties such as porosity volume percent, skeletal
density and nominal mesh sizes, and perhaps more so in surface area, average pore
diameter and dipole moment. The preferred adsorbents will have a surface area of10-2000 square meters per gram and preferably from 100-1000 m2/g. Specific
properties of the materials listed above can be found in company l;terature and
technical brochures, such as those in the following Table 2 which are incorporated
hercin by reference. Others of the general class are also available.
* trade mark
1303Q61
-
19
TAB~ F 2
Adsorbent MatrLl~ Type Reference to Company Literature
AG34A Polystyrene Chromatography Electrophoresis
(Bio Rad) Immunochemistry Molecular
BioloEy - HPLC - Price List M
April 1987 (Bio-Rad)
AG4-X4 Acrylic Chromatography Electrophoresis
Immunochemistry Molecular
Biology - HPLC - Price List M
April 1987 (Bio-Rad)
Dow Polystyrene U.S. Patent Nos. 4,031,038 and
Experimental 4,098,867
Resins
Dowex 66 Polystyrene Material SafetY Data Sheet
Printed 2/17/87
(Dow Chemical USA)
lRA-35 Acrylic Amberlite lon Exchange Resins
(XI~275) (~275) Rohm & Haas Co. 1975
lRA-68 Acrylic Arnberlite Ion Exchange Resins -
Amberlite IRA-68
Rohm & Haas Co. April 1977
Applications for Amberlite polymeric adsorbents suggested in the Rohm and Haas
Company literature include decolorizing pulp mill bleaching effluent, decolorizing
dye wastes and removing pesticides from waste effluent. There is, of course, no hint
in the literature of my surprising discovery of the effectiveness of Amberlite
polymeric adsorbents in the separation of citric acid from Aspergillus-Niger
fermentation broths.
A third class of adsorbents to be used in the process of this invention
will comprise strongly basic anion exchange resins possessing quaternary arnmonium
functionality in a cross-linked polymeric matrix, e.g., divinylbenzene cross-linked
acrylic or styrene resins. They are especially suitable when produced in bead form
and have a high degree of uniform polymeric porosity and exhibit chemical and
physical stability. In the instant case, the resins can be gelular (or "gel-typen) or
"macroreticular" as the term is used in some recent literature, namely Kunin and
......
. . .
i303061
Hetherington, A Progress Report on the Removal of Colloids From Water bv
Macroreticular Ion Exchange Resins, paper presented at the International Water
Conference, Pittsburg, PA, October 1969, reprinted by Rohm & Haas Co. In recent
adsorption technology, "the term microreticular refers to the gel structure per se,
size of the pores which are of atornic dimensions and depend upon the swelling
properties of the gel" while "macroreticular pores and true porosity refer to
structures in which the pores are larger than atomic distances and are not part of the
gel structure. Their size and shape are not greatly influenced by changes in theenvironmental conditions such as those that result in osmotic pressure variations"
while the dimensions of gel structure are "markedly dependent upon the
environmental conditions." In "classical adsorptionn nthe terrns microporous andmacroporous norrnally refer to those pores less than 20 A and greater than 200 A,
respectively. Pores of diameters between 20 A and 200 A are referred to as
transitional pores.~ The authors selected the term "macrore~icularn, instead, to apply
to the new ion exchange resins used in this invention, which "have both a
microreticular as well as a macroreticular pore structure. The forrner refers to the
distances between the chains and crosslinks of the swollen gel structure and thelatter to the pores that are not part of the actual chernical structure. The
macroreticular portion of structure may actually consist of micro-, macro-, and
transitional-pores depending upon the pore size distribution." (Quotes are from
page 1 of the Kunin et al. article). The macroreticular structured adsorbents also
have good resistance to attrition (not comrnon to conventional macroreticular
resins). In this application, therefore, all reference to "macroreticular~ indicates
adsorbent of the types described above having the dual porosity defined by Kunin
and I~etherington. "Gel" and "gel-tvpe" are used in thelr conventional sense.
Looking at both the quaternary ammonium function~ntaining ion
exchange resins of the invention, the quaternary amine has a positive charge and can
form an ionic bond with the sulfate ion. The sulfate form of quaterna~y ammonium
~303061
21
anion exchange resin has a weakly basic proper~r, which in turD, can adsorb ci~ic
acid throu~h an acid-base interactiorL
p N+ (R) or P - N - (R)2 (C2H4~H)
.. I ................... .. p .,
:0 ~ S ~ 0: 0 ~ ,S ~ 0:
~ - H+ - C.A. ~ - H~-C.A.
where P ~ res~nous mo1ety
R ~ lower ~lkyl, Cl 3
C.A. ~ cltr~te lon
Adsorbents such as just descnbed are manufactured by the Rohm and
Haas Company, and sold under the trade ~a~l "Amberlite.~ The types of
Amberlite*polymers known to be effective for use by this invention are referred to in
Rohm and Haas Company literature as Arnberlite*IRA 400 and 900 series
adsorbents described in the literaturc as "insoluble in all comrnon solvents, open
structure for effective adsorption and desorption of large molecules without loss of
capacity, due to organic fouling." Also suitable are AG1, AG2 and AGMP-1 resins
manufactured by Bio Rad and comparable resins sold by Dow Chemical Co7 such as
Dowex 1, 2, 11, MSA-1 and MSA-2, etc. Also useful in this invention are the so-
called intermediate base ion exchange which are mixtures of strong and weak baseexchange resins. Among these are the following commercially available resins: Bio-
Rex S (Bio-Rad 1); Amberlite IRA47 and Duolite*A-340 (both Rohm & Haas).
For exarnple, they may be useful where a basic ion exchange resin is needed which is
not as basic as the strong base resins, or one which is more basic than the wealdy
basic resins.
The various types of polymeric adsorbents of these classes available
will differ somewhat in physical properties such as porosity volume percent, skeletal
deDsity and non~inal mesh sizcs, and perhaps more so in surface area, average pore
diameter and dipole moment. The preferred adsorbents will have a surface area of1~2000 square meters per gram and preferably from 100 1000 m2/g. Specific
properties of thc materials listed above can be found in company literature and
technical brochures, such as those mentioned in the following Table 3,
" ~30306~
22
TABLE 1
PROPERTIES OF ADSORBENTS
Adsorbent Matrix Resin Type Reference to Company L~terature
IRA 458 Acryl~c Amberlite Ion Exchange Resins
(Rohm & Haas) gel-type 1986 & Technical Bulletin
IE-207-74 84
IRA 958 Acrylic Technical Bulletin and Material
macroporous Safety Data Sheet are available
IRA 900 Polystyrene Technical Bulletin is available
macroporous and Amberlite Ion Exchange
Res~ns, IE-100-66.
IRA 904 Polystyrene Technical Bulletin, 1979 and
macroporous IE-208/74, Jan. 1974
IRA 910 Polystyrene Technical Bullet~n, 1979 and
macroporous IE-101-66, May 1972
IRA 400, 402 Polystyrene Amberlite Ion Exchange Resins,
macroporus Oct., Sept. 1976, April 1972 and
IE-69-62, ~ctober 1976
IRA 410 Polystyrene Amberlite Ion Exchange Resins
gel-type IE-72-63, August 1970
AG 1 Polystyrene Chromatography Electrophoresis
(Bio Rad) gel-type Immunochemistry Molecular
Biology HPLC, Pr~ce List M
April 1987
A& 2 Polystyrene Chromatography Electrophores~s
gel-type Immunochemistry Molecular
B~ology HP~C, Price List M
April 1987
AG-MP-1 Polystyrene Chromatography Electrophores~s
macroporous Immunochem~stry Molecular
Biology HPLC, Pr~ce L~st M
April 1987
B~o Rex 5 Mixture of strong Chromatography Electrophoresls
(Bio Rad) base and weak base Immunochemistry Molecular
resins (e.g. AG-2 Biology HPLC, Pr~ce L~st M
and AG-3 or AG-4 April 1987
30306
23
The adsorbent may be employed in the form of a dense compact fixed
bed which is alternadvely contacted with the feed mixture and desorbent materials.
In the simplest embodiment of the invention the adsorbent is employed in the form
of a single static bed in which case the process is only semicontinuous. In another
cmbodiment a set of two or more static beds may be employed in fixed bed
contacdng with appropriate Yalving so that the feed mixture is passed through one
or more adsorbent beds while the desorbent materials can be passed through one or
more of the other beds in the set. The flow of feed mLsture and desorbent materials
may be either up or down through the desorbent. Any of the conventional
apparatus employed in static bed fluid-solid contacting may be used.
Countercurrent moving bed or simulated moving bed countercurrent
flow systerns, however, have a much greater separation efficiency than fixed
adsorbent bed systems and are therefore preferred. In the moving bed or simulated
moving bed processes the adsorption and desorption operations are continuously
taking place which allows both continuous production of an extract and a raffinate
stream and the continual use of feed and desorbent streams. One preferred
cmbodiment of this process utilizes what is known in the art as the simulated
moving bed countercurrent flow system. The operating principles and sequence of
such a flow system are described in U.S. Patent 2,985,589.
In such a system it is the progressive movement of multiple liquid
access points down an adsorbent chamber that simulates the upward movement of
adsorbent contained in the chamber. Only four of the access lines are active at any
one time; tbe feed input stream, desorbent inlet stream, raffinate outlet stream, and
extract outlet stream access lines. Coincident with this simulated upward movement
of the solid adsorbent is the movement of the liquid occupying the void volume of
the packed bed of adsorbent. So that countercurrent contact is maintained, a liquid
aOw down the adsorbent chamber may be provided by a pump. As an active liquid
access point moves through a cycle, that is, from the top of the chamber to the
bouosn, the chamber circulation pump moves through different zones which requiredifferent flow rates. A programmed flow controller may be provided to set and
regulate these flow rates.
The active liquid access points effectively divided the adsorbent
chamber into separate zones, each of which has a different function. In this
embodiment of my process it is generally necessary that three separate operational
~'
~3a306~
24
zones be present in order for the process to take place although in some instances
an optional fourth zone may be used.
The adsorption zone, zone 1, is defined as the adsorbent located
between the feed inlet stream and the rafBnate outlet stream. In this zone, the
feedstock contacts the adsorbent, extract component is adsorbed, and a raffinatestream is withdrawn. Since the general flow through zone 1 is from the feed stream
which passes into the zone to the raffinate stream which passes out of the zone, the
flow in this zone is considered to be a downstream direction when proceeding from
the feed inlet to the raffinate outlet streams.
Imrnediately upstream with respect to fluid flow in zone 1 is the
purification zone, zone 2. The purification zone is defined as the adsorbent between
the extract outlet stream and the feed inlet strearn. l he basic operations taking
place in zone 2 are the displacement from the nonselective void volume of the
adsorbent of any raffinate material carried into zone 2 by shifting of adsorbent into
this zone and the desorption of any raffinate material adsorbed within the selective
pore volume of the adsorbent or adsorbed on the surfaces of the adsorbent particles.
Purification is achieved by passing a portion of extract stream material leaving zone
3 into zone 2 at zone 2's upstream boundary, the extract outlet stream, to effect the
displacement of raffinate material. The flow of material in zone 2 is in a
downstream direction from the extract outlet stream to the feed inlet stream.
Immediately upstream of zone 2 with respect to the fluid flowing in
zone 2 is the desorption zone or zone 3. The desorption zone is de~med as the
adsorbent between the desorbent inlet and the extract outlet stream. The function
of the desorption zone is to allow a desorbent material which passes into this zone
to displace the extract component which was adsorbed upon the adsorbent during aprevious contact with feed in zone 1 in a prior cycle of operation. The flow of fluid
in zone 3 is essentially in the same direction as that of zones 1 and 2.
In some instances an optional buffer zone, zone 4, may be utilized.
This zone, de~med as the adsorbent between the raffinate outlet stream and the
desorbent inlet stream, if used, is located immediately upstream with respeci to the
fluid flow to zone 3. Zone 4 would be utilized to conserve the amount of desorbent
utilized in the desorption step since a portion of the raffinate stream which isremoved from zone 1 can be passed into zone 4 to displace desorbent material
present in that zone out of that zone into the desorption zone. Zone 4 will contain
1303061
enough adsorbent so that raffinate material present in the raffinate stream passing
out of zone 1 and into zone 4 can be prevented from passing into zone 3 thereby
contaminating extract stream removed from zone 3. In the instances which the
fourth operational zone is not utilized the raffinate stream passed from zone 1 to
zone 4 must be carefully monitored in order that the flow directly from zone 1 to
zone 3 can be stopped when there is an appreciable quantity of raffinate material
present in the raffinate stream passing from zone 1 into zone 3 so that the extract
outlet stream is not contaminated.
A cyclic advancemen~ of the input and output streams through the
fixed bed of adsorbent can be accomplished by utilizing a manifold system in which
the valves in the manifold are operated in a sequential manner to effect the shifting
of the input and output streams thereby allowing a flow of fluid with respect to solid
adsorbent in a countercurrent manner. Another mode of operation which can effectthe countercurrent flow of solid adsorbent with respect to fluid involves the use of a
rotating disc valve in which the input and output streams are connected to the valve
and the lines through which feed input, extract output, desorbent input and raffinate
output streams pass are advanced in the same direction through the adsorbent bed.
Both the manifold arrangement and disc valve are known in the art. Specifically
rotary disc valves which can be utilized in this operation can be found in U.S.
Patents 3,040,777 and 3,422,848. Both of the aforementioned patents disclose a
rotary type connection valve in which the suitable advancement of the various input
and output streams from fixed sources can be achieved without difficulty.
In many instances, one operational zone will contain a much larger
quantity of adsorbent than some other operational zone. For instance, in some
operations the buffer zone can contain a minor amount of adsorbent as compared to
the adsorbent required for the adsorption and purification zones. It can also beseen that in instances in which desorbent is used which can easily desorb extract
material from the adsorbent that a relatively small amount of adsorbent will be
needed in a desorption zone as compared to the adsorbent needed in the buffer
zone or adsorption zone or purification zone or all of them. Since it is not required
that the adsorbent be located in a single column, the use of multiple chambers or a
series of columns is within the scope of the invention.
It is not necessary that all of the input or output streams be
simultaneously used, and in fact, in many instances one of the strearns can be shut
30306
26
off while others effect an input or output of material. The apparatus which can be
utilized to effect the process of this invention can also contain a series of individual
beds connected by connecting conduits upon which are placed input or output tapsto which the various input or output streams can be attached and alternately andperiodically shifted to effect continuous operation. In some instances, the
connecting conduits can be connected to transfer taps which during the normal
operations do not function as a conduit through which material passes into or out of
the process.
It is contemplated that at least a portion of the e%tract output stream
will pass into a separation means wherein at least a portion of the desorbent
material can be separated to produce an extract product containing a reduced
concentration of desorbent material. Preferably, but not necessary to the operation
of the process, at least a portion of the raffinate output stream will also be passed to
a separation means wherein at least a portion of the desorbent material can be
separated to produce a desorbent stream whicb can be reused in the process and araffinate product containing a reduced concentration of desorbent material. The
separation means will typically be a fractionation column, the design and operation
of which is well-known to the separation art.
Reference can be made to D. B. Broughton U.S. Patent 2,985,589, and
to a paper entitled "Continuous Adsorptive Processing--A New Separation
Technique" by D. B. Broughton presented at the 34th Annual Meeting of the Society
of Chemical Engineers at Tokyo, Japan on April 2, 1969,
for further explanation of the simulated moving bed countercurrent
process flow scheme.
Although both liquid and vapor phase operations can be used in many
adsorptive separation processes, liquid-phase operation is preferred for this process
because of the lower temperature requirements and because of the higher yields of
extract product than can be obtained with liquid-phase operation over those
obtained with vapor-phase operation. Adsorption conditions will include a
temperature range of from about 20C to about 200C with about 65C to about
100C being more preferred and a pressure range of from about atmospheric to
about 500 psig (3450 kPa gauge) being more preferred to ensure liquid phase.
Desorption conditions will include the same range of temperatures and pressures as
used for adsolption conditions.
r
30306:
27
The size of the units which can utilize the process of this invention can
~ary anywhere from those of pilot plant scale (see for example our assignee's U.S.
Patent 3,706,81~ to those of cornmercial scale
and can range in ~ow rates from as little as a few cc an hour up to many thousands
of gallons per hour.
The following examples are presented to illustrate the selectivity
relationship that makes the process of my invention possible. The example is notintended to unduly restrict the scope and spi~it of claims attached hereto.
EXAMPLE I
In this example, three pulse tests were run with a neutral styrene
divinylbenzene polymeric adsorbent (XAD~ made by Rohm ~ Haas Company) to
determine the ability of the adsorbent to separate citric acid, at di~erent pH's, from
its fermentation rnn~ture of carbohydrates (DP1, DP2, DP3, including glucose,
~ylose, arabinose and rafEinose) and ions of salts, including Na+, K+, Mg+ +,
Ca+ +,Fe+ + +, Cl-, SO4=, PO4-- and NO3; amino acids and proteins. The first
test was run at a pH of 2.4 and 45C. Two further tests were run at a pH of 1.7 and
0.9. Citric acid was desorbed with water. The fermentation feed mixture had the
following composition:
Feed Composition Amount
Citric Acid 12.9%
Salts (K+, Na+, Ca+ +, Mg+ + Fe+ + +) 0.60% (6000 ppm)
Carbohydrates (Sugars) 1%
Others (SO =, Cl; PO"-, NO3-, 5~o
proteins an~} amino acids)
Water 815%
Retendon volumes and resolution were obtained using the pulse test
apparatus and procedure previously described. Specifically, the adsorbent was
tested in a 70 cc straight column using the following sequence of operadons for the
pulse test. Desorbent material was continuously run upwardly through the column
, ~
130306i
28
containing the adsorbent at a nominal liquid hourly space velocity (LHSV) of about
1Ø A void volume was determined by observing the volume of desorbent required
to fill the packed dry column. At a conver~ient time the flow of desorbent material
was stopped, and a 10 cc sample of feed mixture was injected into the column via a
sample loop and the flow of desorbent material was resumed. Samples of the
effluent were automatically collected in an automatic sample collector and lateranalyzed for salts and citric acid by chromatographic analysis. Some later samples
were also analyzed for carbohydrates, but since they were eluted at approximately
the same rate as the carbohydrates, they were not analyzed in these examples norwere other minor ingredients, amino acids and proteins. From the analysis of these
samples, peak envelope concentrations were developed for the feed mixture
components. The retention volume for the citric acid was calculated by measuringthe distance from the midpoint of the net retention volume of the salt envelope as
the reference point to the midpoint of the citric acid envelope. The resolution, R, is
calculated from Equation 3, given earlier. The separation factor, B, was calculated
as previously indicated.
The results for these pulse tests are shown in the follov~ing Table 4.
30306
29
TABLE 4
Test A - pH - 2.4
Net retention Peak ~idth Resolution
Component Volume at 0.5 Height (O.S He~ght)
Salts 0 14.4 1.39
C~tric ac~d 44.4 49.5 Reference
Test B - pH - 1.7
Net retention Peak W~dth Resolution
Component Volume at 0.5 Height (0.5 He~ght)
Salts 0 11.6 1.49
Citric ac~d 42.2 4~.1 Reference
Test C - pH - 0.9
Net retent10n Peak W1dth Resolut~on
Component Volume at 0.5 He~ght (0.5 He~ght)
SJlts 0 13.3 1.4
Citr~c ac~d 40.9 45.1 Reference
The results are also shown in Figure 3A in which it is clear that while
citric acid is more strongly adsorbed than the other components, there is a
substantial loss of citric acid which is unadsorbed and removed with the salts and
carbohydrates (not shown). Citric acid is satisfactorily separated in the process in
Figure 3B where the results are judged good and in Figure 3C where the results are
judged excellent. The process clearly will have commercial feasibility at a pH of 1.7
and lower. At a pH of 2.4 (Figure 3A), however, it is noted that a substantial
amount of the citric acid will be recovered in the form of the citrate, H2CA-l, in the
raffinate with the salts and carbohydrates. From this, it is evident that the ionized,
i3Q306
soluble species should be reduced, as explained previously, by maintaining a lower
pH in the feed, thereby driving the equilibrium in Equation 1 to the left.
LXAMPLE II
This example presents the results of using a neutral crosslinked
styrene divinylbenzene (XAD-4) and a neutral crosslinked polyacrylic ester
copolymer (XAD-8) with the same separation feed mixture as Example I at
different pHs to demonstrate the poor separation when the pH is 2.4 or higher, or
above the first ionization constant, pKal = 3.13, of citric acid. The same procedure
and apparatus previously described in Example I were used in the separation, except
the temperature was 60C and 5 ml of feed mixture was used.
Figures 4A, 4B and 4C are, respectively, graphical presentations of the
results of the pulse tests using XAD-4 at pHs, respectively, of 2.4, 1.7 and 0.~.
Figure 4A shows that citric acid "breaks through" with the salts (and carbohydrates).
This problem can be partially alleviated by lowering the pH to 1.7 as in Figure 4B.
An excellent separation can be achieved by lowering the pH further to 0.9 as in
Figure 4C. This separation, with adjustment of the pH, again, clearly has
commercial utility.
Figures 4D and 4E are, respectively, graphical representations of the
results of pulse tests, run under the same conditions as above, using XAD-8 at pHs
of 2.8 and 1.4 and temperatures of 65C. Figure 4D, which was made at a pH of 2.8,
shows no separation, but rather the salts, carbohydrates and citric acid elutingtogether initially. After about 67 ml, after most of the carbohydrates and salts and
some of the citric acid have been recovered, some relatively pure citric acid can be
obtained, but recovery is low. Figure 4E, which was made at a pH of 1.4, shows aselectivity between citric acid and carbohydrates and salts which results in a
satisfactory separation and recovery of the citric acid.
EX~MPLE III
This exarnple presents the results of using a neutral crosslinked
styrene divinylbenzene copolymer (XAD-4) with the same separation feed mixture
as E~xample I at two different pHs to demonstrate the poor separation when the pH
~30306
31
is 2.4 or higher. The same procedure and apparatus previously described in
Example I were used, except the temperature was 93C in Figures SA and SB and
the amount of feed mixture was 10 ml.
Figures SA and 5B are, respectively, graphical presentations of the
results of pulse tests using ~ 4 at pHs, respectively, of 2.8 and 1.4. Figure SAshows that citric acid "breaks through" with the salts and carbohydrates. This
problem can be alleviated by lowering the pH to 1.4 as in Figure 5B. This
separation, with adjustment of the pH, again, clearly has comme}cial utility.
EXAMPLE IV
The procedure and apparatus previously described in Example I was
used on the samples of this example. The temperature was 60C and S ml of feed
mixture was used. The feed composition was similar to that previously used except
that citric acid has been concentrated to 40% in the feed mixture. The effect ofconcentration on the pH will be seen. In Figure 6A, even with the temperature at60C, the pH of 1.9 is too high to separate the citric acid at 40% concentration. By
adjusting the pH downward as in Figures 6B and 6C, the citric acid is preferentially
adsorbed and excellent separation is achieved at a pH of 1.13 and at a pH of 0.5. In
each of these samples, carbohydrates were not analyzed, but it can be assumed that
the carbohydrates closely followed the salts in the separation.
EXAMPLE V
The procedure and apparatus previously described in Example I was
used on the three samples of this example. The temperature was 93C and the
amount of feed mixture was 5 ml. The feed composition was similar to that
previously used except that citric acid has been concentrated to 40% in the feedmixture to demonstrate the further effect of concentration on the pH. In Figure 7A,
even with the temperature at 93C, the pH of 1.8 is too high to separate the citric
acid at 40% concentration. By adjusting the pH downward as in Figures 7B and 7C,the citric acid is preferentially adsorbed and exce]lent separation is achieved.Again, carbohydrates were not analyzed, but it can be assumed that the
ca~bohydrates closely followed the salts in the separation.
1303061
32
EXAMPLE VI
The pulse test of Example I was repeated on two 50% citric acid
samples using XAD-4 adsorbent. The desorbent in both cases was water. The
composition of the feed used was the same as used in Example I except that citric
acid has been concentrated to 50%. The temperature was 93C. In the first sample,
the pH was 1.5. As shown in Figure 8A, citric acid was not separated. After
reducing the pH to 1.0 in the second sample, citric acid was readily separated as
seen in Figure 8B. Again, carbohydrates were not analyzed, but assumed to closely
follow the salts. The separation in Figure 8B was judged good.
EXAMPLE VII
The separation example represented by Figures 7B and 7C required
high temperatures, e.g., 93C to achieve the separation of 40~o citric acid due to the
difficulty in desorbing citric acid from the XAD-4 adsorbent. In this example, high
temperatures, which adversely affect the adsorbent life and the cost to operate, are
eliminated and the separation is readily achieved at 45C through the use of a
desorbent mixture of 10% (by wt.) acetone and 90% water. Referring to Figure 9, a
feed comprising 40% (wt.) citric acid, 4% carbohydrates and 2% salts of the
following elements: K+, Na+, Mg+ +, Fe+ + +, Ca+ + plus proteins and amino
acids, was introduced into the pulse test apparatus as set forth previously and the
test ran as before except that the temperature was 45C. In this test, the pH was
maintained at 0.5, but the desorbent contained acetone as mentioned above. The
net retention volume for citric acid was lQ7 ml, and the resolution was Q61 and,therefore, the separation was easily made.
EXAMPLE VIII
In this example, four pulse tests were run with a weakly basic anion
exchange resin having a teniary amine function hydrogen bonded to a sulfate ion, in
a cross-linked gel-type acrylic resin matrix (AG4-X4 made by Bio Rad Laboratories,
Richmond, California) having a tertiary amine function hydrogen bonded to a
1:~0306~.
33
sulfate ion, in a cross-linked acrylic resin matrix to determine the ability of the
adsorbent to separate citric acid from its fermentation mixture of carbohydrates(DP1, DP2, DP3, including glucose, xylose, arabinose and raf~nose) and ions of
salts, including Na+, K+, Mg+ +, Ca+ +,Fe+ + +, Cl~, SO4=, PO4= and NO3-,
amino acids and proteins at a pH of 1.6. The first test was run at a temperature of
75C. The remaining tests were run at 60C. Citric acid was desorbed with water
in Pulse Test No. I (Figure 10) and sulfuric acid in two concentrations: 0.05 (Pulse
Test No. 2) and 0.25N (Pulse Test No. 3). Pulse Test No. 4 was like Pulse Test No.
2 except that it was made after the adsorbent was used with 24 bed volDes of feed.
The fermentation feed mixture had the following composition:
Feed Composition Per Cent
Citric Acid 40%
Salts (K+, Na+, Ca+ +, Mg+ + Fe+ + +) 1.5%
Carbohydrates (Sugars) 4%
Others (SO , Cl, POd4 , NO3; 5%
proteins an amino acl s)
Water 49.5%
Retention volumes and separation factor (O were obtained using the
pulse test apparatus and procedure previously described in Example I except that a
5cc sample was used. The net retention volume (NRV) for the citric acid was
calculated by measuring the distance from the midpoint of the salt envelope as the
reference point to the midpoint of the citric acid envelope. The separation factor,B,
is calculated from the ratio of the retention volumes of the components to be
separated to the retention volume for the first salt componet (i.e. Salts l).
The results for these pulse tests are shown in the following Table No.
5.
1303~6i
34
TABLE N0. 5
Pulse Feed
Test Resin/Desorbent Component NRV B
1 AG4-X4/Water Salts 1 1.6 34.25
Citric Acid 54.8 Reference
Unknowns A 0 Tracer
Unknowns B 6.6 8.30
Salts 2 54.6 1.00
2 AG4-X4/0.05N H2S04 Salts 3.2 11.87
Citric Acid 38.0 Reference
Unknown A 0 Tracer
Unknown B 2.7 14.07
3 AG4-X4/0.25N H2S04 Unknowns A 0 Tracer
Citric Acid 26.9 Reference
Salts 2.3 11.70
Unknowns B 7.6 3.54
4 AG4-X4/0.05N H2S04 Unknowns A 0 Tracer
Citric Acid 38,0 Reference
Salts 2.4 15.8
Unknowns B 7.2 5.28
,
13~306~
The results of Pulse Tests 2~ are similar to Figure lO. From Table 5,
it is clear that while citric acid is satisfactori1y separated in the process, in highly
purified form, with water, desorption with water is slower than with dilute sulfuric
acid as evidenced by larger net retention volume. After aging the adsorbent with 24
bed volumes of feed, the adsorbent shows no signs of deactivation.
EXAMPLE IX
The first pulse test of Example VIII was repeated using the same
procedure and apparatus except that the temperature was 65C The desorbent was
water. This exarnple presents the results of using a macroporous weakly basic
anionic exchange resin possessing a cross linked polystyrene matrix (Dowex 66) with
the same separation feed mixture as Example VII~ (40% citric acid) in the first two
pulse tests at a pH of 7.0 and 3.5 (Figures llA and llB, respectively) to demonstrate
the failure to accomplish the desired separation when the pH is above the first
ionization constant, pKal = 3.13, of citric acid, and more specificaDy in these two
samples, where the concentration of citric acid is 40~o, when the pH is above 1.7. In
the third part of the example (represented by Figure llC), the feed was diluted to
13% citric acid and the pH reduced to 2.4. While there is evident improvement, it is
apparent that the pH and/or concentration will have to be reduced further to
prevent "breakout" of the citric acid. For example, at 13% concentration, it is
estimated that the pH must be lowered to about 1.6 to 2.2
Figures lL~ and llB are, respectively, graphical presentation of the
results of the pulse test using Dowex 66 at pHs, respectively, of 7.0 and 3.5. Figures
IL~ and llB show that citric acid '~reaks through" with the salts (and carbohydrates)
at the higher pHs. This problem can be partially alleviated by reducin~g the
concentration to 13% and lowering the pH to 2.4 as in Figure DC, where it is shown
that only a small amount of citric acid is not adsorbed and "breaks through" in the
raffinate while most is adsorbed onto the adsorbent resin (but not desorbed in this
F;gure). This separation, w~th adjustment of the concentration and pH to optimumlevels, clearly will have commercial utility.
B
13~306~
36
EXAMPLE X
Three additional pulse tests under the same conditions as Example
vm, except as Doted, were made on citric acid samples of the same feed
composition, but with two different adsorbents. The desorbent in the first two
samples was 0.05 N H2S04 (Figures 12 and 13A) while water was used in the third
sample (Figure 13B). The composition of the feed used was the same as used in
Example VIII. The temperature was 60C and the pH was 1.6. The adsorbent #1
in the first test was a macroporous pyridine function-containing divinylbenzene
cross-linked resin of the followiDg formula:
CH3 ~
H+50;
where P is the polystyrene moiety forming tbe resin. The second adsorbent
(#2), used in the second and third samples, is a tertiary amine, also with a pyridine
~unctional group, having the following formula:
P - CH2 - I - CH2 ~
CH2 H+SO4'
CHOH
I
CH3
where P is as def~ned above. Both resins are cross-linked with
divinylbenzene. In some cases, while water is an effective desorbent, with excellent
separation, it is not strong enough to recover the adsorbed citric acid quickly enough
to make the process commercially attractive. See Figure 13B, in which the
conditions are the same as above, using adsorbent #2, where water is the desorbent.
In this case citric acid does not elute until about 9S ml of desorbent have passed
through the adsorbent. Dilute sulfuric acid is, therefore, the preferred desorbent, as
will be apparent from the results shown in Figures 12 and 13A. Also, from Fig~res
12,13A and 13B, it will be seen that an excellent separation of citlic acid is obtained.
E3~
The procedure, conditions and apparatus previously described in
Example VIII were used to separate four samples of citric ac~d from the same feed
with two different resins of the same class of adsorbent as Example vm, (except
that in the first and fourth samples, the column temperature was S0C and the
desorbent was 0.05N H2SO4; in the second and third samples the pH was 2.2 and
the desorbent was dilute sulfuric acid at 0.15 N concentration). Both resins, IR~-68
and IRA-35, obtained from Rohm and Haas, have an amine function and the
following structural forrnula:
,R'
p - CH2 N: H+S04
R"
where P is the polya~ylic matrix, and
R'andR" = CH3.
Amberlite IRA-68 (Sarnple Nos. 1, 2 and 3) is a gel-type resin. IRA-35 (Sample No.
4) is a macroreticular-type resin. Sample No. 3 was identical to Sample No. 2,
except that the adsorbent had previously been used to separate 69 bed columns ofthe feed. Sample Nos. 1 and 2 are both excellent adsorbents for separating citric
acid from its fermentation broth within the pH range of 1.6 to 2.2. Sample No. 3,
after aging the adsorbent with 69 bed volumes of feed, demonstrates the stability of
the resin (little or no deactivation has taken place) in this separation. Net retention
volume (NRV) and selectivity (E~) are shown in the following Table 6.
130~
38
~LE 6
Sample No. Re Component NRV ``~
Amberlite Salts ~.5 8.24
IRA-68 Citric Acid45.3 Reference
Unknown A 0 Tracer
Unknown B 9.3 4.87
2 Amberlite Salts 2.3 12.61
IRA-68 Citric Acid29.0 Reference
Unknown A 0 Tracer
UnknDwn B 6.5 4.46
3 Amberlite Salts 2.85 10.32
IRA-68 Citric Acid29.4 Reference
Unknown A 0 Tracer
Unknown B 7.0 4.2
4 Amberlite Salts 1 .3 27.38
IRA-35 Citric Acid36.9 Reference
Unknown A 0 Tracer
Unknown B 5.9 6.25
In a furtber comparison of the claimed adsorbents of Examples I
through VII with ~xamples VIII through Xl, several sarnples of the e~tract were
analyzed for readily carbonizable impurities (RCS) (Food & Chemical Codex
(FCC) Monograph #3) and potassium level. RCS is determined in the following
manner: a 1 gm sample of the extract (actual concentration of citric acid is
determined) is carbonized at 90C with 10 ml of 95% H2S04. The carbonized
substance is spectrophatometrically measured at 500 nm using a 2-cm cell with a 05
inch diameter tube and the amount of RCS is calculated for 50% citric acid solution.
The number arrived at can be compared with that obtained by using this procedureon the cobalt standard solution of the FCC test mentioned above. Potassium is
detemuned by atomic adsorption spectroscopy. For comparison, the same
analytical determinations were made on a sample of the same feed and RCS
calculated for 50~o citric acid with XAD4, AG4-X4, and adsorbents No. I and No. 2
of Example m. The results are shown in Table 7.
.....
.
l3b306l
39
TABl E 7
EXI~ACI` OUALlTY (RCS/POTASSIUM) BY PULSE TEST
RCS
Calculated CA. Net
Adsorbent Desorbent (for 50~ C.~.) ppmK Ret. Vol.
XAD~ H2O 6.86,8.98 59, 137 13.0
AG4-X4 .05N.H2sO4 1.77, 1.42 24, 81 34.8
#2 (Ex. X) .05N.H2SO4 3.17, 3.33 24, 54 30.8
#1 (Ex. X) .05N.H2SO4 2.17 62 31.0
An improvement in both reduction of levels of RCS and K for the weakly basic
resins compared to the neutral resins is indicated by this data. In all samples, RCS
was reduced by at least 50% and in two samples, K was reduced by over 50%. It isnoted from the net retention volume that both classes of adsorbents have good
resolution, but the strong base adsorbents suffer somewhat from increased cycle
times. The cycle times can be reduced by using higher concentrations of sulfuricacid, e.g., up to about 0.2N, in the preferred range of 0.1 to 0.2N.
In another embodiment, citric acid adsorbed on the adsorbent may be
converted in situ to a citrate before being desorbed, for example, by reaction with an
alkaline earth metal or alkali metal hydroxide or ammonium hydroxide and then
immediately eluted using a metal hydroxide, ammonium hydroxide or water as the
desorbent. Deactivation of the adsorbent by the unknown impurities may take place
in time, but the adsorbent may be regenerated by flushing with a stronger desorbent,
e.g., a higher concentration of sulfuric acid than the desorbent, an alkali metal
hydroxide or NH40H, or an organic solvent, e.g., acetone or alcohol.
EXAMPLE XII
In this example, two pulse tests were run with a gel-type strongly basic
anion exchange resin (IRA 458 made by Rohm & Haas Co.) having the structural
formula like (1) on page .1 ~ above, substituted with three methyl groups, to
i303~6i
determine the ability of the adsorbent to separate citric acid from its fermentation
rnil~ture of carbohydrates (DP1, DP2, DP3, including glucose, xylose, arabinose and
raffinose) and ions of salts, including Na+, K+, Mg+ +, Ca+ +,Fe+ + +, Cl~,
SO4=, PO4- and NO3-, amino acids and proteins at a pH of 2.2. P is acrylic cross-
linked with divinylbenzene. Pulse test Sample No. I was run at a temperature of
50C. Pulse test Sample No. 2 was run at 60C, but after the bed had been aged
with 33 bed volumes of feed. Further runs to 62 bed volumes have been made with
no signs of deactivation of the adsorbent. Citric acid was desorbed with 0.1N
solution of sulfuric acid in both samples. The fermentation feed mixture had thefollowing composition:
Feed Composition Per Cent
Citric Acid 40
Salts (K+, Na+, Ca+ +, Mg+ + Fe+ + +) 1.5
Carbohydrates (Sugars) 4
Others (SO4=, Cl, PO4=, NO3-, 5
proteins and amino acids)
Water 49.5
Retention volumes and separation factor were obtained using the
pulse test apparatus and procedure previously described in Example I.
The results for these pulse tests are shown in the following Table No.
8.
1303061
T~BIJE N 0.8
Feed
Sample No. Resin Component NRY B
1 IRA-458 Salts 1.0 38.9 `
Citric Acid 38.9 Reference
Unknowns A O Tracer
Unknowns B 6.6 5.89
2 IRA-458 Salts 0.9 43.3
Citric Acid 39.0 Reference
Unknown A O Tracer
Unknown B 7.1 5.49
It is clear that citric acid is satisfactorily separated in the process, and
after aging the adsorbent with 33 bed volumes of feed, the adsorbent shows no signs
of deactivation, which is substantially identical to the results under closely identical
conditions with fresh adsorbent.
EX~MP~ XIII
The pulse test of Example XII was repeated on additional citric acid
samples using the same feed, but a different, macroporous, strongly basic anionic
exchange adsorbents, IRA-958, possessing quaternary ammonium functions and an
acrylic resin matrix cross-linlced with divinylbenzene matmL The desorbent was 0.05
N H2S04. The composition of the feed used was the same as used in EKample XII.
The temperature was 60C and the pH was 1.6. The adsorbent in this test was a
resin obtained from Rohm & Haas having the structure (1) shown on page~ 1,
where R is methyl.
As shown in Figure 14, citric acid starts eluting after 45 ml of
desorbent have passed through the adsorbent and is very effectively separated from
the fermentation mixture in high purity with excellent recovery.
~30306i
42
EXAMPLE XIV
The pulse test of Example XII was repeated on an additional citric
add sample using the same feed, but a different, strongly basic anionic exchangeresin adsorbent, AG2-X8 (obtained from Bio Rad Company) having a structure like
formula (2) above, (page~l ~ where R is methyl, with a cross-linked polystyrene gel-
type resin matrix having quaternaly ammonium functional groups thereon. The
desorbent was 0.15N H2SO4. The composition of the feed used was the same as
used in Example XI. The temperature was 50C and the pH was 2.2.
As shown in Figure lS, citric acid starts eluting at about 43 ml of
desorbent have passed through the adsorbent and is very effectively separated from
the fermentation mixture in high purity with excellent recovery.
In a further comparison of adsorbents of Exarnples I through VII with
Examples XII through XIV, several samples of the extract were analyzed for readily
carbonizable impurities (RCS) (Food & Chemical Codex (FCC) Monograph #3)
and potassium level as described above. The results for each of the adsorbents,
XAD-4, IRA 458, IRA 959 and AG 2 - X4 with the indicated desorbent are shown
in the following Table 9.
TABLE 9
EXTRACT QUAL~TY (RCS/POTASSIUM) BY PULSE TEST
RCS
(CalculatedppmK CA Net
at 50% CA)(Calculated Retention
Adsorbent Desorbent (Units!at 50% C.A.) Volume
6 86 59
XAD-4 H20 8 98 . 137 13.0
IRA 458 0.1N H2S04 1.5 80 37.9
IRA 958 o.oSN H2SO4 2.73 82 32
AG2-X4 0.15N H2SO4 5.3 131 43
An improvement in both reduction of levels of RCS and K for the strongly basic
resins compared to the neutral resins is indicated by this data. In all sarnples, RCS
1303061
43
was reduced by between 40-85~ and K was reduced between
0-20%. It is no~ed from Examples XII, XIII (Fig. 14) and
XIV (Fig. 15), that both classes of adsorbents have
good separation, but the present adsorbents suffer somewhat
from increased cycle times. The cycle times can be reduced by
using higher concentrations of sulfuric acid, e.g., up to
about 0.2N in the preferred range of 0.1 to 0.2N.
Deactivation of the adsorbent by the unknown
impurities may take place in time, but the adsorbent may be
regenerated by flushing with a stronger desorbent, e.g., a
high concentration of sulfuric acid than the desorbent, an
alkali metal hydroxide or NH40H, or an organic solvent, e.g.,
acetone or alcohol.
D
