第27巻 第8号 (1978) 507

Effect of Citric Acid on the Oxidative and

Flavor Stability of Soybean Oil

Kojl MIYAKOSHI, Mamoru KOMODA, and

Shlgezo MATSUBARA

Sugiyama Chemical and Industrial Laboratory

(11, Kagetori-cho, Totsuka-ku, Yokohama)

Citric acid was added into the deodorized soybean oil. Citric acid content in the oil was determined
by GLC wtih 5% Silicone OV-17 and the effect of citric acid on the oxidative and flavor stability of
the soybean oil was tested. Citric acid did not protect the oil against fluorescent light. On the other
hand, citric acid markedly improved flavor and oxidative stability when the oils were deteriorated under
spontaneous autoxidation conditions. However, its efficiency was independent of its concentration.
Citric acid solubility in the soybean oil was measured. At least 50ppm citric acid could be dissolved
in the oil after 4 weeks of storage.

serve poor quality for flavor and oxidative

1 Introduction stability.

For many years, citric acid has been used as When soybean and other oils containing

synergist of antioxidants for edible oils and linolenic acid are exposed to air for long

fats. Many workers reported studies of the periods, off-flavors are developed.1) A large

use of citric acid in edible oils and fats. amount of soybean oil is used for salad and

Cowan and Evans1) insisted that citric acid acts frying oils. Therefore, clarification of flavor

entirely as a metal-chelating agent. On the reversion mechanism and its inhibition is

other hand, Lundberg2) and Lemon et al.3) necessary. Thus, we have made the laboratory

described that citric acid functions not only experiments to investigate the effect of citric

as a metal scavenger but also as a socalled acid on the oxidative and flavor stability of

acidic synergist like ascorbic acid. Privett soybean oil.

and Quackenbush4) considered that citric and

2 Experimental procedure
ascorbic acids function as synergists in natural
fats and oils by inhibiting the antioxidant 2.1 Deodorization

catalysis of peroxide decomposition. However, General features of the bleached soybean oil,

these published papers do not refer to the manufactured commercially, are shown in

citric acid content in the oils. Table-1. Tocopherol and iron contents in the

The authors reported in the previous paper5) oil were determined by the method of Nelson

that thermally decomposed products, of citric and Milun6), and Evans et al.,7) respectively.

acid i.e., aconitic, itaconic, and citraconic acid The bleached soybean oil was deodorized in

are not formed in the commercial edible oils. the glass laboratory deodorizer. The deodori-

Therefore, citric acid alone can be expected to zation conditions were as follows: temperature,

act as a synergist. We also reported that citric 260•Ž; vacuum, 5mmHg; time, 1h; and steam,

acid is decomposed when added during deodo- 3%. Citric acid aqueous solution was added

rization. Therefore, the relationship between into the oil at the beginning or during cooling

citric acid content and oxidative stability of phase of deodorization.

fully refined edible oils is very important. 2•22 Determination of citric acid by GLC

Because the oils which contain no citric acid Soybean oil (50g) containing citric acid was

31
 508 油 化 学

taken into a 500ml separatory funnel. Hexane was used for judging quality and intensity of
(100ml) and 50ml of distilled water were flavor. t-Test was used to test the sample
added into the funnel to extract the citric acid. means for differences.
The funnel was automatically shaken for 1h. Oxidative stability of the soybean oil was
The water layer containing citric acid was also measured by the Active Oxygen Method,
collected through a filter paper. The aqueous i.e., peroxide value was determined after hold-
solution (25ml) was pipetted to a 50ml round ing the oil 8h under the AOM conditions, and
bottom flask and was neutralized with 0.1N the time required for the oil to reach peroxide
sodium hydroxide. value 100 was measured.
After the water was evaporated, the citric The effect of fluorescent light on the flavor
acid in the flask was esterified with 2ml of deterioration of the soybean oil was examined
1-butanol, 0.2ml of sulfuric acid, and 2g of according to the method of Moser et al.10)
anhydrous sodium sulfate. Then, tributyl citrate Soybean oil (150g) was taken into the 250ml
was collected into a 5ml test tube using transparent wide-mouthed bottles, and they
hexane. The details of this paragraph are were placed in the fluorescent light exposure
described in published paper8). apparatus (Fig.-1). After the bottles were
Hexane solution (2ml) containing 0.2mg of exposed to the light of 7000 luxes for 10h,
o-terphenyl internal standard was added to the organoleptic evaluation of the oil was carried
tube and filled up to 5ml with hexane. Two out as described above. Peroxide value was
microliters of the sample solution were then

chromatographed two times for each sample.

The following GLC conditions were used:

Instrument: Shimazu GC-5A Gas Chromato-

graph with a dual flame ioniza-

tion detector.

Column: Glass, 3mm ID, 2m long, packed

with 5% Silicone OV-17, Chro-

mosorb W DMCS AW, 60•`80

mesh.

Flow Rate: Nitrogen carrier gas, 60ml/min

to each column, hydrogen 50ml/

min to each flame, air 100ml/

min.

Temperature: Inlet and detector temperature

were 250•Ž. Column oven was

220•Ž.

Sensitivity: 103.

Range: 8.

Chart speed: 10mm/min.

2•E3 Oxidative stability test

Oxidative stability was assayed by determin-

ing the peroxide of the soybean oil after

holding it in a 60•Ž oven in the dark for 3 or

5d. The samples were also organoleptically

A: Rotating table on which sample.
evaluated following the procedure of Moser et
bottles are placed.
al..9) After the soybean oil (150g) taken into
B: Fluorescent lamp (10W•~12)
the 250ml wide-mouthed bottles was stored at
C: Motor
the above conditions, 10 trained panels evalu- D: Ventilator to keep the tempera-

ated the oil. The paired comparison method ture constant.

was employed and a 10-point scoring system Fig.-1 Fluorescent light exposure apparatus.

32
 第27巻 第8号 (1978) 509

also determined. A: Control.

B: Citric acid was added at the beginning

3 Results and discussion
of deodorization.

3• 1 Gas chromatography C: Citric acid was added at 130•Ž during

The retention time of o-terphenyl and tribu- cooling phase of deodorization.

tyl citrate ester were 7 and 15min, respectively. D: Citric acid was added when the oil

The calibration curve of citric acid is shown was cooled to 200•Ž after deodoriza-

in Fig-2.8) Esterification rate of citric acid t ion. The temperature was maintained

at 200•Ž for 10min, and then the oil

was rapidly cooled to 30•Ž

Citric acid (29.2ppm) was found in the oil

C. However, citric acid could not be detected

in the oils B and D. Citric acid was decom-

posed by heat during deodorization.5)

The results of 60•Ž oven test and AOM test

indicate that citric acid is highly effective on

flavor and oxidative stability of soybean oil,

which means that citric acid protects the oil

against spontaneous autoxidation. Citric acid

addition during cooling phase of deodorization

markedly improved flavor and oxidative stabil-

ity, and the stability of the oil C containing

a: Standard ester, b: Esterified, c: Recovery
29.2ppm citric acid was most improved.
Fig.-2 Calibration curves of citric acid.
Although citric acid could not be detected in

was 88.4%. Recovery of citric acid from the the oil D, stability was greatly improved. It

oil was 94.0%. seems most likely that trace amount of citric

3•E2 Deodorization and citric acid addi- acid which is formed metal complexes still

tion remained in the oil D. This result suggests

Citric acid (50ppm) was added into the that citric acid does not function as an acidic

soybean oil at the beginning or during cooling synergist such as ascorbic acid, for it is difficult

phase of deodorization, and its effect on the to think that such trace amounts of citric acid

oxidative and flavor stability of the soybean oil can act as an acidic synergist. Meanwhile addi-

was tested. Results are given in Table-1 and- tion of citric acid at the beginning of deodori-

2, where A, B, C, and D were as follows: zation slightly improved oxidative stability.

Table-1 General characteristics of the soybean oil.

*1 Fatty acid compositions were palmitic 10 .0%, stearic4.1%, oleic 25.7%, linoleic52.8%, and linolenic
7.4%.
*2 Citricacid (50ppm) was added into B , C, and D.

33
510 油 化 学

Table-2 Effect of citric acid upon the oxidative Dutton et al11). reported similar results. How-
and flavor stability of the deodorized ever, as citric acid is completely decomposed
soybean oil.*1
during deodorization at 260•Ž5), Cowan's metal-

chelating theory cannot interprete this result.

Further clarification is necessary.

On the other hand, citric acid exhibited no

Citric acid content (ppm)
synergistic effect on flavor stability when the

soybean oil was exposed to fluorescent light.

Moser et al.12) reported that the addition of

Flavor score at 0 time
citric acid to soybean oil do not protect the oil

against photooxidation. Sattar and Deman13)

Fluorescent light exposure for 10h observed the deterioration of oils and fats by

fluorescent light, and discussed that spontaneous

autoxidation and photooxidation may proceed

by different mechanism. Present data support

their idea. First, citric acid was ineffective

against photooxidation as described above.

Second, the photooxidized soybean oil was

really tasted •glight struck•h as Moser et al.10)

After 3d storage at 60℃
expressing, which was completely different

from spontaneously autoxidized soybean oil.

Third, the soybean oil was extremely deterio-

rated by fluorescent light in spite of its small

After 5d storage at 60℃ peroxide increment. Inhibition of photooxida-

tion is now a subject of importance since

soybean salad and frying oils are usually sold in

clear glass or plastic bottles. Therefore, further

study of this problem is necessary.

3.3 Citric acid solubility

Since it was proved that citric acid improved

the stability of soybean oil, solubility of citric

AOM (peroxide values after 8h)
acid in the soybean oil was then determined.

In general, it is said that a maximum of 50ppm

citric acid can be dissolved in the oil.14) How-

ever, the authors have been unable to find

any report in which citric acid solubility in the

*1 Duplicate deodorizationexperiments.
oils and fats are determined. So we have
*2 Key to symbols: * Significant difference (5%
estimated the citric acid solubility in the soy-
level), ** highly significant difference (1%
level),+ no significantdifference. bean oil (Table-3). Citric acid, 30, 80, and
*3 Peroxide value at time of organolepticevaluation. 500ppm, was added into each soybean oil at

Table-3 Solubility of citric acid in the soybean oil (ppm).

*1 Citric acid was determined before and after filtration through a filter paper
.
*2 Citric acid in the oil was determined after filtration through a filter paper
.
*3 Peroxide value .

34
 第27巻 第8号 (1978) 511

130•Ž during cooling phase of deodorization. Table-4 Effect of citric acid concentration on
These oils were stored in the dark at ambient the oxidative and flavor stability of
soybean oil.
temperature for 4 weeks.

Considerable amounts of citric acid were

deposited on the glass deodorization flask,

therefore, e.g., only 196ppm of citric acid

Flavor score at 0 time
could be found despite 500ppm addition. When

the citric acid content was determined immedi-

ately after the deodorization, 22.2, 57.5. and Fluorescent light exposure for 10h
196.0ppm had been found in each oil. When

the oils were filtered through a filter paper,

19.2 43.7 and 142.7ppm of citric acid was

found in each oil. Small amount of citric acid

was adsorbed by the filter paper and most part

of it had passed through the filter paper.

When less than 60ppm citric acid was dissolved

in the soybean oil, it had not precipitated after

4 weeks of storage. Although 196ppm was

After 3d storage at 60℃
dissolved in the oil at the initial stage, only

29.2ppm could be found after 4 weeks of

storage. Therefore, it was confirmed that at

least 50ppm of citric acid can de dissolved in

the soybean oil after 4 weeks of storage.

The authors, however, consider from the

After 5d storage at 60℃
results above that citric acid is not dissolved

but just dispersed as a fine particle in the oil.

Citric acid is expected to precipitate out after

a long period of storage.

3•E4 Optimum citric acid concentration

Since a maximum of 50ppm citric acid can

be dispersed in the soybean oil, investigations

were made to clarify the optimum citric acid

concentration for oxidative and flavor stability.

Citric acid was added into the soybean oil at

130•Ž during cooling phase of deodorization.

Citric acid (8.5, 22.1, and 49.0ppm) was found

AOM (peroxide value after 8h)

in each deodorized oil (Table-4). Similar to

the results in Table-2, citric acid was ineffec-

tive towards fluorescent light deterioration.

The results of 60•Ž oven and AOM test indi-

cated that although citric acid is effective on *1 Citric acid found in the oil.
flavor and oxidative stability its efficiency is *2 For explanation of symbols, see Table-2.
independent of its concentration. Conclusion is *3 Peroxide value and citric acid content at time
therefore that existence of small amount of of organoleptic evaluation.
citric acid in the soybean oil improved flavor

and oxidative stability. Little citric acid was as an acidic synergist. Although a very small
decomposed by fluorescent light exposure or amount of iron was found in the soybean oil,
60•Ž of storage. citric acid may act as a metal-inactivating agent
These results, together with the results in as Cowan et al.1) insist. On the oher hand,
Table-2, suggest that citric acid do not function citric acid may inhibit the antioxidant catalysis
35
 512 油 化 学

of peroxide decomposition as privett et al.4) 9) H.A. Moser, C.M. Jaeger, J.C. Cowan, and
described. So research is needed to elucidate H.J. Dutton, J. Am. Oil Chem. Soc., 24, 291
the mechanism of citric acid's synergisms. (1947).
10) H.A. Moser, C.D. Evans, J.C. Cowan, and
Acknowledgment
W.F. Kwolek, J. Am. Oil Chem. Soc., 42, 30
The authors are grateful to Mr. Noriji Onuki of (1965).
Hohnen Oil Co. Ltd, for his helpful advice in connec- 11) H.J Dutton, A.W. Schwab, H.A. Moser, and
tion with GLC; to Miss Emiko Seki for invaluable J.C. Cowan, J. Am. Oil Chem. Soc., 26, 441
assistance and to the members of the taste panel for (1949).
their continuing interest and participation. 12) H.A. Moser, C.D. Evans, G. Mustakas, and
(Received March 11, 1978) J.C. Cowan, J. Am. Oil Chem. Soc., 42, 811
References (1965).
13) A. Sattar and J.M. Deman, J. Am. Oil Chem.
1) J.C. Cowan and C.D. Evans, •gAutoxidation
Soc., 53, 473 (1976).
and Antioxidants•h, ed. by W.O. Lundberg,
14) H.W. Vahlteich, C.M. Gooding, C.F. Brown,
Vol. 2, John Wiley and Sons Inc., New York
and D. Melnick, Food Tech., p. 6 Jan. (1954).
(1962) p. 593.

2) W.O. Lundberg,•h Symposium on Foods: Lipids

大 豆 油 の 風 味 の も ど りに 及 ぼ す ク エ ン 酸 の 効 果
and Their Oxidation•h, ed. by H.W. Schultz,

Avi Pub. Co., Westport (1962) p. 31.

宮 腰 宏 治 ・菰 田 衛 ・松 原 繁 左 右

3) H.W. Lemon, R.M. Knapp, and A.H. Allman, 大 豆 油 を脱 臭 した とき クエ ン酸 を添 加 した 。大 豆 油 中

Can. J. Research, F 28, 453 (1950). の ク エ ン酸 含 最 を5%シ リニー ンOV-17 GLCに て定
4) O.S. Privett and F.W. Quackenbush, J. Am. 量 し,ま た 大 豆 油 の 風 味 及 び 酸 化 安 定性 に 及 ぼす ク エ ン
Oil Chem. Sac., 31, 321 (1954). 酸 の 影 響 につ い て 検 討 した 。 大 豆 油 をけ い 光燈 照 射 にて
5) K. Miyakoshi, and M. Komoda, Yukagaku, 劣 化 させ た と き,ク エ ン酸 は風 味 の劣 化 を防止 す る こ と
27, 381 (1978). は で きな か った 。 一 方,大 豆 油 を 自動酸 化 的 に劣 化 させ
6) J.P. Nelson and A.J. Milun, J. Am. Oil Chem. た と き,ク エ ン酸 は大 豆 油 の酸 化 及 び 風 味 安定 性 を著 し
Soc., 45, 848 (1968). く改 善 した 。 し か し な が ら,そ の 効果 は ク エ ン酸 濃 度
7) C.D. Evans, G.R. List, and L.T. Black, J. に は 関 係 が な か った 。 大 豆 油 へ の クエ ン酸 の溶 解 度 を測
Am. Oil Chem. Soc., 48, 840 (1971). 定 した 。4週 間 の 保 存 試 験 の結 果 か ら,少 な くと も50
8) K. Miyakoshi and M. Komoda, J. Am. Oil
ppmの ク エ ン酸 が 大 豆 油 中 に溶 解 で き るこ とが わ か っ
Chem. Soc., 54, 331 (1977). た。

36