Citric Acid 1

Citric Acid
Frank H. Verhoff, Miles Laboratories, Elkhart, Indiana 46515, United States

1. Introduction . . . . . .. . . . . . . . . 1 4.3.1. Surface Fermentation . . . . . . . . . . 4

2. Physical Properties . .. . . . . . . . . 2 4.3.2. Submerged Fermentation . . . . . . . . 5
3. Chemical Properties .. . . . . . . . . 3 4.3.3. Other Fermentation Methods . . . . . 5
4. Production . . . . . . .. . . . . . . . . 4 4.4. Recovery . . . . . . . . . . . . . . . . . . 5
4.1. Laboratory Synthesis . . . . . . . . . 4 5. Quality Specifications and Uses . . . 6
4.2. Early Production . . .. . . . . . . . . 4 6. Economic Aspects . . . . . . . . . . . . 6
4.3. Fermentation . . . . . .. . . . . . . . . 4 7. References . . . . . . . . . . . . . . . . . 6

1. Introduction cycle, also known as the Krebs cycle or tricar-

boxylic acid cycle (see Fig. 1), is pivotal in the
Citric acid [77-92-9], C6 H8 O7 , M r 192.13, oxidation of sugars and acetates to carbon diox-
is a tricarboxylic acid. According to IUPAC- ide and water, releasing energy for physiological
nomenclature, citric acid is 2-hydroxypropane- functions [5], [6].
1,2,3-tricarboxylic acid; it is also known as β-
hydroxytricarballylic acid.

Citric acid was isolated and crystallized in

1784 by Scheele, who precipitated calcium cit-
rate by adding lime to lemon juice. The presence
of three carboxyl and one hydroxyl groups was
recognized by Liebig in 1838. Citric acid was
first prepared from calcium citrate in 1860 in
the United Kingdom. By 1880 citric acid was
being produced from calcium citrate in France,
Germany, and the United States.
Citrus fruits contain citric acid in large quan-
tities, from 5 % in the fruit [1] to 9 % in the
juice [2]. Citric acid is also found in many other Figure 1. Citric acid cycle of plants and animals [5], [6]
fruits. The citric acid content (in mg/100 mL) of
About 100 years after citric acid had been iso-
various common fruits is as follows [3], [4]:
lated from citrus fruits, it was synthesized from
glycerol. The finding, in 1893, that certain mi-
Lime 7000
Lemon 5630 croorganisms secrete citric acid when grown on
Raspberry 2480 sugar media suggested the possibility of produc-
Black currant 1170 ing the acid commercially by fermentation. The
Tomato 1018
Pineapple 605 first successful operation of this type began 1923
Strawberry 580 in New York; later large-scale fermentation pro-
Cranberry 202
Apple 14 cesses were developed in the former Czechoslo-
vakia, the United Kingdom, Belgium, and Ger-
In addition to fruits citric acid is found in al- many. Submerged fermentation has been used
most all plant and animal species. The citric acid since the middle of this century.

c 2005 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

10.1002/14356007.a07 103
2 Citric Acid

Citric acid fermentation, whether surface or

submerged, used carbohydrate substrates with
certain strains of the mold Aspergillus niger.
Since the early 1970s, other microorganisms
have been introduced, including molds, yeasts,
and bacteria, and other substrates, e.g., various
carbohydrates and n-alkanes from crude oil.

2. Physical Properties
Citric acid is a colorless, odorless substance with
an agreeable, though strongly acid taste. The
monohydrate crystallizes from a saturated so-
lution below 36.6 ◦ C and the anhydrous form Figure 2. Solubility of citric acid in water as a function of
at higher temperature. The monohydrate, M r temperature
210.13, has a relative density of 1.542 and a heat
of combustion at 30 ◦ C of 1973 kJ/mol; it softens
at 75 ◦ C and melts at 100 ◦ C. Its translucent, col-
orless crystals belong to the orthorhombic sys-
tem. Anhydrous citric acid, mp 153 ◦ C, has a rel-
ative density of 1.665 and a heat of combustion
at 30 ◦ C of 1990 kJ/mol. The crystals are translu-
cent and colorless, belonging to the holohedral
class of the monoclinic system. Both forms exist
under conditions of normal humidity; dehydra-
tion of the monohydrate occurs in very dry air
and is more rapid when it is carried out under
vacuum in the presence of concentrated sulfuric
acid. The anhydrous crystals gradually absorb
water in moist air; both crystalline forms clump
and harden in moist air.
The solubility of citric acid in water as a func-
tion of temperature is shown in Figure 2; the Figure 3. Concentration of citric acid in water vs. pH
temperature of the solution drops as the acid dis- ∗ Concentration is in (grams per 100 mL solution) × 100 %
solves. The pH of citric acid solutions as a func-
tion of concentration is shown in Figure 3. The Table 2. Solubility of hydrated and anhydrous citric acid in organic
freezing point depression and boiling point el- solvents at 25◦ C and density of solutions
evation of citric acid solutions are given in Ta- Form Solubility,∗ d 25
ble 1. The dissociation constants at 18 ◦ C are K 1 Solvent g/100 g
8.2 × 10−4 , K 2 1.8 × 10−5 , and K 3 4.0 × 10−6 . Monohydrate
Amyl acetate 5.980 0.8917
Table 1. Freezing point depression and boiling point elevation of Amyl alcohol 15.430 0.8774
aqueous citric acid solutions Ethyl acetate 5.276 0.9175
Ether 2.174 0.7228
Concentration, Freezing point Boiling point
Chloroform 0.007 1.4850
mol/kg H2 O depression, ◦ C elevation, ◦ C
Anhydrous
Amyl acetate 4.22 0.8861
0.01 0.023 –
Ether (absolute) 1.05 0.7160
0.05 0.042 –
0.10 0.203 –
∗ Saturated solution.
0.50 0.965 0.284
1.00 1.940 0.577
2.00 1.000 1.214
5.00 – 3.512
The solubility of citric acid in organic sol-
10.00 – 8.390 vents is listed in Table 2, along with the relative
20.00 – 16.600
 Citric Acid 3

density of the solutions. The distribution bet- Citric acid forms stable complexes with many
ween ether and water is shown in Table 3. metals; some, such as ferroammonium citrates,
Table 3. Distribution of citric acid between water and ether
can be crystallized. Copper gives a complex re-
sembling the copper – tartaric acid complex of
Concentration, mol/L Distribution Fehling’s solution. These complexes have been
coefficient
studied by various classical methods.
In water In ether Citric acid can chelate many metal ions in
At 15 ◦ C
solution by forming bonds between the metal
0.902 0.0077 117 and the carboxyl or hydroxyl groups of the cit-
0.460 0.0036 128 ric acid molecule. Sometimes two or more citric
0.220 0.0017 129
0.297 0.0023 129 acid molecules are involved in the interaction
At 25 ◦ C with the metal ion. This property is valuable for
0.9175 0.0063 114 preventing precipitation (e.g., bathtub ring from
0.487 0.0031 155
0.241 0.00155 155 soap), changing chemical potential, and altering
0.315 0.0020 158 other chemical properties.
Citric acid is easily esterified with many alco-
hols under the usual conditions in the presence
of a catalyst, such as sulfuric acid, p-toluene-
3. Chemical Properties sulfonic acid, or an acid ion-exchange resin; es-
terification with alcohols boiling above 150 ◦ C
Citric acid loses water at 175 ◦ C to form aconitic
requires no catalyst. Benzyl chloride and sodium
acid, HOOCCH=C(COOH) (CH2 COOH),
citrate yield di- or tribenzyl esters. Trimethyl, tri-
which loses carbon dioxide to yield itaconic
ethyl, and tributyl citrate are used as plasticizers
anhydride [2170-03-8]. Itaconic anhydride re-
in food-packaging materials.
arranges to citraconic anhydride [616-02-4] or
Dihydric alcohols, dihydric phenols, and
adds water to form itaconic acid [97-65-4],
polyhydric alcohols, such as mannitol, sorbitol,
(HOOCCH2 ) (HOOC)C=CH2 .
and glycerol, form polyesters with citric acid.
In some cases the esterification reaction can be
stopped before completion, leaving at least one
of the carboxylic acid moieties free to form a
salt; the resulting polyester may be soluble in
water.
Acid chlorides and anhydrides react with the
Addition of water to citraconic anhy- hydroxyl group of citric acid; usually the ester
dride gives citraconic acid [498-23-7], cis- of citric acid is used and the allyl ester is formed.
HOOCCH=C(CH3 ) (COOH). Evaporation of a Dicarboxylic acid chlorides give bis(citric acid
citraconicacid solution in the presence of nitric esters).
acid yields mesaconic acid [498-24-8], the trans Epoxides, such as ethylene oxide, propylene
isomer of citraconic acid. oxide, and styrene oxide, form polymers by re-
Potassium permanganate oxidation of cit- acting with citric acid or its esters at the available
ric acid at 35 ◦ C yields 1,3-acetonedicarboxyl- hydroxyl and carboxyl groups.
ic acid [542-05-2], HOOCCH2 COCH2 COOH; Ammonia, amines, amides, and carbamides
at 85 ◦ C the productis oxalic acid [144-62-7], react with citric acid as they do with sim-
HOOCCOOH. Fusion with potassiumhydroxide ple carboxylic acids. Aldehydes form adducts
produces oxalic and acetic [64-19-7] acids. with citric acid; e.g., formaldehyde gives
Citric acid forms crystalline mono-, di-, anhydromethylenecitric acid [144-16-1] 5-oxo-
and tribasic salts with manycations. The de- 1,3-dioxolane-4,4-diaceticacid:
gree of hydration of these salts varies; thus,
trisodium citrate can crystallize with 2 or 5.5
molecules of water. Mixtures of metal cations
give complex salts, such as ZnNa3 H(C6 H5 O7 )2 ,
ZnNa4 (C6 H5 O7 )2 , and Zn(Na3 C6 H4 O7 )2 .
4 Citric Acid

4. Production which employs a strain of Aspergillus niger to

convert sugar to citric acid. Both surface fermen-
4.1. Laboratory Synthesis tation and submerged fermentation have been
used.
The classic laboratory synthesis of citric acid
beginning with acetone has not been used on an
industrial scale: 4.3.1. Surface Fermentation

In surface fermentation, A. niger is grown on a

liquid substrate in pans stacked vertically in a
chamber. The chamber and pans are sterilized
either before or after introduction of the sub-
Reaction conditions at each stage are critical strate. The pans are filled manually or automat-
and use of bromine and hydrogen cyanide is ically. The chamber is warmed by the introduc-
hazardous. A simplified version of this synthe- tion of moist, sterile air at a controlled tempera-
sis has been patented [7]. ture. The liquid and the surface microorganisms
Another synthesis is as follows: are removed manually or automatically from the
pans. The pans are cleaned before the next batch
is introduced.
The substrate for the fermentation is a car-
bohydrate, usually molasses or a sugar, such as
raw beet, refined beet, or cane sugars, or a syrup.
Glucose syrups can be prepared from wheat,
The key step in this route is the catalytic con-
corn, potato, or other starch. The sugar content
densation of ketene with oxalacetic anhydride,
of the syrup can vary from about 10 to 25 wt %.
which can be obtained by catalytic oxidation of
Certain inorganic nutrients, such as ammonium
malic or fumaric acid.
nitrate, potassium phosphate, magnesium sul-
Other syntheses are found in the patent and
fate, zinc sulfate, and potassium ferrocyanide,
scientific literature, but apparently none have
are added. The pH is adjusted to between 3 and
been commercialized [8], [9].
7, depending on the carbohydrate source. Ster-
ilization may be batchwise or continuous; the
latter uses less energy and is usually faster. Af-
4.2. Early Production ter sterilization, the temperature is adjusted as
required.
For about 50 years, citric acid was obtained ex- The surface of the sterile substrate in the pans
clusively from unripe citrus fruit. In southern is inoculated with A. niger spores, which germi-
Italy, an entire industry with orchards and pro- nate and cover the surface of the liquid with a
cessing facilities was devoted to this purpose. mat of mold. After two to three days the surface
The juice was pressed from unripe fruit and is completely covered and citric acid production
mixed with lime (CaO) to precipitate calcium begins, continuing at almost a constant rate until
citrate. The solid was treated with sulfuric acid 80 – 90 % of the sugar is consumed. Fermenta-
to precipitate calcium sulfate and form a solu- tion then continues more slowly for an additional
tion from which citric acid was crystallized. The six to ten days.
yield of purified product was about 2 – 3 wt % of The theoretical yield from 100 kg of sucrose
the fruit; citrus oils were also obtained. This pro- is 123 kg of citric acid monohydrate or 112 kg
cedure has been displaced by fermentation. of anhydrous acid. However, the A. niger uses
some sugar for growth and respiration, and the
actual yield varies between 57 and 77 % of the-
4.3. Fermentation oretical, depending on such factors as substrate
purity, the particular strain of organism, and the
The most economical method for producing cit- control of fermentation.
ric acid since the 1930s has been fermentation,
 Citric Acid 5

4.3.2. Submerged Fermentation First, biological solids usually are removed by

filtration using a rotary vacuum filter or the more
Submerged fermentation is similar to surface recent belt-press filter, or by centrifugation. The
fermentation, but takes place in large fermenta- solids are washed to improve recovery of citric
tion tanks. This method is used more frequently acid.
because labor costs are lower with large tanks The dissolved citric acid must then be sep-
than with small pans; equipment costs are also arated from residual sugars, proteins generated
lower. by the fermentation, and other soluble impuri-
The fermentation vessel can be short and ties. This has traditionally been accomplished
wide or tall and narrow, and equipped with mix- by precipitation and crystallization. Addition of
ing devices, such as top-entering or side-entering lime precipitates calcium citrate, which is fil-
agitators of the turbine or propeller type. Agita- tered and stirred in dilute sulfuric acid to form a
tion can be increased by use of a draft tube, a precipitate of calcium sulfate; filtration yields a
recirculation loop, or a nozzle through which air purified citric acid solution. Control of pH and
and recirculated substrate is pumped. Spargers temperature in these operations helps to opti-
located at the bottom of the vessel or under the mize the results. Citric acid is then crystallized
stirrer supply air, which may be enriched with from solution and recrystallized from water; the
oxygen. Oxygen is usually recovered from the mother liquors are recycled to remove accumu-
exhaust gas. The air is supplied by a compres- lated impurities.
sor and passes through a sterile filter; if neces- Other purification techniques can be used, in-
sary, the air is cooled. Because the process is cluding ion exchange, carbon adsorption, mem-
exothermic, the vessel must be equipped with brane filtration, chromatography, and liquid ex-
heat-exchange surfaces, which can be the out- traction.
side walls or internal coils. Ports are provided After filtration or centrifugation, the citric
for introducing substrate, inoculum, and steam acid crystals are dried, classified, and sold in
or other sterilizing agents; sampling and exhaust bags or in bulk. Crystals of the wrong size are
ports are also provided. recycled. Citric acid is also sold as an aqueous
The substrate is prepared in a separate tank solution of varying degrees of purity.
and its pH adjusted; the micronutrients may be Byproducts of these purification processes in-
added to this tank or directly to the fermen- clude biomass from the primary filtration, which
tor. The substrate is sterilized by a batchwise can be sold for feed, and calcium sulfate, which
or, more commonly, by a continuous operation. can be used for landfill or for industrial purposes,
The fermentor is sterilized, charged with sub- such as manufacture of gypsum board. Soluble
strate, and inoculated. Fermentation requires 3 – organic impurities are given normal wastewater
14 days. After it is completed, the air supply treatment.
is stopped to prevent the microorganisms from Few changes in production methods have oc-
consuming the citric acid. curred in recent years, but research has been
done on the fermentation process [10] and on
new separation techniques [11].
4.3.3. Other Fermentation Methods
Citric acid can also be produced from hydrocar-
bons using yeast. The process resembles sub- 5. Quality Specifications and Uses
merged fermentation, but requires more air. It
is a four-phase process, i.e., air, organic liquid, Specifications. The Food Chemical Code
aqueous liquid, and solid, and some operational specifications for citric acid are as follows [12]:
details are different.
Description: colorless, translucent crystals
or white, granular to fine crystalline powder;
4.4. Recovery odorless, strongly acid taste; hydrate is efflo-
rescent in dry air
The citric acid broth from the surface or sub-
merged fermentation processes must be purified.
6 Citric Acid

Solubility: 1 g of anhydrous acid dissolves in jor producers; however, capacity has recently in-
0.5 mL of water, in 2 mL of alcohol, and in creased in the Far East and in Eastern Europe.
30 mL of ether The location of new facilities depends on access
Identification: 1-in-10 solution gives posi- to raw materials and markets.
tive tests for citrate
Assay: not less than 99.5 % C6 H8 O7 , calcu-
lation based on the anhydrous form 7. References
Water: not more than 0.5 % (anhydrous
form) and not more than 8.8 % (hydrate) 1. H. Moellering, W. Gruber, Anal. Biochem. 17
Limits of impurities: arsenic (as As): not (1966) 369 – 376.
more than 3 ppm (0.0003 %) 2. H. Fricke, S. B. Hensen, Food Process. Ind.
heavy metals (as Pb): not more than 10 ppm 1975, 38 – 44.
(0.001 %) 3. J. J. Ryan, J. A. Du Pont, J. Agri. Food Chem.
oxalate: passes test; no turbidity produced 21 (1973) 45 – 49.
readily carbonizable substances: passes test 4. K. N. Paulson, M. A. Stevens, J. Food Sci. 39
residue on ignition: not more than 0.05 % (1974) 354 – 357.
For specific test methods, see [12]. 5. H. A. Krebs, Angew. Chem. 66 (1954)
313 – 319.
Uses. Citric acid and its sodium or potassium 6. A. L. Lehninger: A Short Course in
salts are used in many food products, includ- Biochemistry, Worth Publ., New York 1973,
ing carbonated beverages, dry-packaged drinks, p. 217.
7. Montecatini Edison, DE-OS 2 245 892, 1972.
fruit drinks, jams, jellies, and canned fruits. They
8. Chevron Research Corp., DE-OS 2 225 986,
are also important in producing vegetable oils
1972.
and in preserving the color, flavor, and vitamin
9. Ethyl Corp., DE-OS 2 240 723, 1972.
content of fresh and frozen vegetables and fruit. 10. M. Rohr, C. P. Kubicek, J. Kominek: “Citric
Citric acid is widely used in cleaning, e.g., for Acid” in H. Dellweg (ed.): Biotechnology
boilers and heat exchangers. Its chelating ability Microbiology Products, Biomass, and Primary
assists in removal of scale. Citric acid is em- Products, vol. 3, Verlag Chemie, Weinheim
ployed as a detergent builder, especially in liq- 1983, pp. 456 – 465.
uid formulations. Its solutions can remove sulfur 11. D. T. Friesen et al.: “Separation of Citric Acid
dioxide from gases and chelate micronutrients from Fermentation Beer Using Supported
in fertilizer. Citric acid esters are plasticizers for Liquid Membranes”, Paper no. 194, Am.
food-grade plastic containers. Chem. Soc. Meeting, New York, April 1984.
12. Food Chemicals Codex, 3rd. ed., National
Academy Press, Washington, D.C., 1981.
6. Economic Aspects
Citric acid is manufactured worldwide. The
United States and Western Europe are the ma-

Citronellal → Flavors and Fragrances

Citrus Juice → Beverages, Nonalcoholic
Cladding of Metals → Metals, Surface Treatment
Claus Process → Sulfur