EFFECTS OF USING ROSEMARY EXTRACT AND ONION JUICE ON OXIDATIVE STABILITY OF SARDINE (SARDINA PILCHARDUS) MINCE

Journal of Food Quality

28

(2005) 109–120.

All Rights Reserved.

©

Copyright 2005, Blackwell Publishing

109

EFFECTS OF USING ROSEMARY EXTRACT AND ONION JUICE ON OXIDATIVE STABILITY OF SARDINE

(

SARDINA PILCHARDUS

) MINCE

MELTEM SERDARO LU

1

and ELVAN FELEKO LU

Ege UniversityEngineering Faculty

Food Engineering Department 35

1

00, Bornova, zmirTurkey

Accepted for Publication December 20, 2003

ABSTRACT

Sardine

(Sardina pilchardus)

mince was treated with rosemary extract(RE – 300 ppm) and onion juice (OJ – 1 mL/100 g) then stored at

-

20C for5 months. Proximate composition, thiobarbutiric acid (TBA), free fatty acids(FFA) and peroxide value (PV) were determined on 0 and 15 days and 1, 2,3, 4 and 5 months of storage. Fatty acid composition was also determined on0 and 5 months of frozen storage. TBA, PV and FFA levels increased on allexperimental groups due to the lipid oxidation. RE showed antioxidative effecton sardine mince during frozen storage as indicated by TBA, PV and FFAlevels. Oxidation was delayed for 3 months by OJ treatment. At the end of5 months’ storage, the TBA values in OJ treatment and control (C) treatmentwere out of consumable limits. After frozen storage of 5 months polyunsatu-rated fatty acid level decreased and saturated fatty acid level increased in thecontrol treatment. No significant change was observed in fatty acid composi-tion in samples of RE and OJ treatments.

INTRODUCTION

Sardine is an important species in Turkey; total catch was 20,500 tons in1999 (Kasımo lu

et al.

2003). It is generally consumed as fresh, canned orsalted and also utilized as fish meal and oil. Lipid oxidation is one of the most

G (

G (

I

g (

Blackwell Science, LtdOxford, UKJFQJournal of Food Quality0146-9428Copyright 2005 by Food & Nutrition Press, Inc., Trumbu ll, Connecticut.2005282109120Original Article

ROSEMARY EXTRACTS AND ONION JUICE EFFECTS ON

SARDINE MINCEM. SERDAROLU and E. FELEKOLU

1

Corresponding author. TEL:

+

90 232 3882395; FAX:

+

90 232 3427592; EMAIL: [email protected]

110 M. SERDAROG LU and E. FELEKOG LU

important factors responsible for quality deterioration of fish during bothrefrigerated and frozen storage. Lipid oxidation in muscle foods can be initi-ated by nonenzymic and enzymic reactions (Akhtar

et al.

1998). Reactionsbetween the by-products that are derived from lipid oxidation and proteinscause undesirable changes of food properties including protein denaturation,loss of protein solubility, alteration of texture and functional properties ofprotein and destruction of nutrient components (Verma

et al.

1995; Akhtar

et al.

1998). Frozen storage inhibits microbial spoilage and helps to slow downlipid oxidation. However, it does not inhibit lipid oxidation. Fish mince is lessstable than whole fish due to the disruption of cellular membranes, whichincreases the rate of enzymatic and chemical reactions (Pastoriza

et al.

1994).The rate and extent of oxidative deterioration depends on factors such as thestorage period and temperature, saturation degree of fatty acids, presence ofantioxidants or prooxidants and availability of oxygen (Gögü and Kolsarıcı1992).

The highly unsaturated fatty acids commonly found in seafood are par-ticularly sensitive to oxidative changes during storage. Antioxidants are usedby the food industry to delay the oxidation process (Brand-Williams

et al.

1995). The most commonly used antioxidants are butylated hydroxytoluene(BHT) and butylated hydroxyanisole (BHA). However, they are volatile andeasily decompose at high processing temperatures and are synthetic chemi-cals. The possible toxicity of the synthetic chemicals used as antioxidants hasbeen a subject of study for many years (Chang

et al.

1977).Recently the food industry focused on the use of natural antioxidants,

such as tocopherols, various spices and herbs, vegetable extracts and ascorbicacid. The antioxidant properties of spices and herbs are attributed to theirphenolic contents (Akhtar

et al.

1998). Many studies reported the effective-ness of these additives in retarding lipid oxidation (Inatani

et al.

1983;Akhtar

et al.

1998; Serdaro lu and Yıldız-Turp 2001; Kamil

et al.

2002). Theuse of rosemary as an antioxidant was reported by Rac and Ostric in 1955.More recently a patent was issued to Brener and Jobson in 1973 for theextraction of rosemary with oil (Chang

et al.

1977). The antioxidative effectof rosemary is based on its phenolic diterpenes, carnosol and carnosinic acidas well as rosmanol, epirosmanol and iso rosmanol (Inatani

et al.

1983;Schwarz and Ternes 1992). Many studies have demonstrated the effective-ness of rosemary extracts in retarding lipid oxidation (Cavoski

et al.

1991;Liu

et al.

1992; Boyd

et al.

1993; Akhtar

et al.

1998; Yıldız-Turp andSerdaro lu 2002). Onions are often used as an additive in meat and fishpatties. Younathan

et al.

(1983) reported that the presence of flavonoids,ascorbic acid and sulfur compounds in onion extract probably contributes tothe antioxidant effect of onion extract. The objective of this study was todetermine the effect of frozen storage on oxidative quality of sardine mince

s

g (

g (

ROSEMARY EXTRACTS AND ONION JUICE EFFECTS ON SARDINE MINCE 111

(

Sardina pilchardus

) and to measure the antioxidant effectiveness of rose-mary extract and onion juice.

MATERIALS AND METHODS

Material

The samples of sardine (

Sardina pilchardus

) each approximately inweight 60–80 g and 14–16 cm in length were obtained from a local fish marketin Izmir (in the Aegean Region of Turkey). Fish were iced and transported tothe Food Engineering Department, Ege University, in Izmir and were filletedon the same day. The fillets were minced using a meat grinder with a 3 mmhole plate.

The following treatment groups were prepared: 300 ppm rosemaryextract (Dragoco 9/037174), 1 mL/100 g onion juice (prepared by pressingwhite large onions with food processor in laboratory conditions), control batch([C] without antioxidant). Batches were ground twice by using the meatgrinder to ensure an even distribution of rosemary extract (RE) or onion juice(OJ) with the fish mince. All samples weighed 250 g each and were packedin polyamide/polyethylene bags and than stored at

-

20C

±

2C for 5 months.Samples were randomly drawn for analysis at the evaluation periods. Thiobar-buturic acid value (TBA), peroxide value (PV) and free fatty acids (FFA) weretested after 0 and 15 days and 1, 2, 3, 4, 5 months of storage. Fatty acid profileof samples was evaluated on 0 and 5 months of storage. Zero time (day 0)samples were taken after 24 h of mincing, monthly evaluated samples weretaken on the last day of each month. All analysis were done on three packages.

METHODS

Proximate Composition

Moisture content was measured using the oven-drying procedure accord-ing to the AOAC (1990). Fat content was determined by the chloroform-methanol extraction according to Flynn and Bramblett (1975). Protein contentwas determined (Anon 1979). Ash content was measured according to theAOAC (1990) procedure.

Lipid Oxidation Parameters

Thiobarbuturic acid values were determined according to Tarladgis

et al.

(1960); the results were expressed as mg malonaldehyde equivalents/kg of


sample. Free fatty acids and peroxide value were assessed by the AOAC(1990) method.

Fatty Acid Composition

Fatty acid composition was determined according to the method outlinedby Morrison and Smith (1964). Sardine lipids were converted to their fattyacid methyl esters by heating the lipids in a mixture of benzene and borontrifluoride methanol complex solution at 85C for 30 min. Methyl esters wereanalyzed by gas liquid chromatography. As outlined by Wada and Fang (1992)a decline in the ration of eicosapentaenoic acid (EPA)

+

docosahexaenoic acid(DHA)/16 : 0 fatty acid was measured to elucidate oxidative deterioration ofpolyunsaturated fatty acid in fish lipids.

Statistical Analysis

The trial was performed twice and all analysis was done on three pack-ages at each trial. Data were analyzed by ANOVA using general linear model(GLM) procedure of SPSS (1997) V.8 with a significance level of (

P

<

0.05).Differences among means were determined by Least Significance Differences(

P

<

0.05).

RESULTS AND DISCUSSION

The average proximate composition of the sardine mince for protein, fat,moisture and ash was 16.2, 5.2, 77.2 and 1.2%, respectively. Those valueswere generally in line with the results reported by Kilinç (2003) and Gokoglu

et al.

(1998).Changes in TBA values of treatment groups stored at

-

20C are given inFig. 1. Initial TBA values of RE, OJ and C treatments were found to be 0.95,1.02 and 1.26 mgma/kg, respectively. All samples showed an increased TBAvalue with storage (

P

<

0.05) period. No differences were found in TBAvalues of treatment groups at the first (initial) and second evaluation period(

P

>

0.05). The TBA values of RE treatment and OJ treatment were signifi-cantly lower than that of the control after 1 month of storage. The mean TBAvalue was 4.17 mgma/kg for the control samples at 1 month. After 3 monthsof storage no differences were found in TBA values between the control andOJ treatments. TBA values of the control samples increased sharply in frozenstorage from day 15 to 30. Samples treated with RE showed a sharp increasein TBA values between month 2 and 3. However, at the end of the storage thelowest TBA value was recorded as 5.97 mgma/kg for the RE treatment. TBAvalues indicated that control samples and samples with added OJ were more


rancid than samples treated with RE throughout the storage time at

-

20C. OJtreated samples had TBA values in acceptable limits after 4 months of storage;however, there were no differences in TBA values between the OJ-treatedsamples and control samples at the end of the storage period. Similar to ourfindings Younathan

et al.

(1983) reported the antioxidative effect of onionjuice for frozen shark and mackerel muscle. Antioxidative activity of onionjuice may be attributed to the high level of ascorbic acid, sulfur and flavonoidcompounds. At the end of the storage period the TBA values in OJ and Cgroup were quite high, which Sinnuber and Yu (1977) reported as acceptablefor frozen seafood.

Changes in peroxide values of sardine minces likewise occurred. At zeroand 15 days of storage there were no significant differences in peroxide valuesof the treatment groups. Peroxide values of RE, OJ and C treatments were10.94, 11.96 and 14.24 meqPO/kg, respectively. High PV levels of lipids fromsardine during the initial storage stages may be attributed to the high degreeof unsaturation of fatty acids and the mincing process. It has been reportedthat mincing fish muscle creates a larger surface area and then the lipids areeasily oxidized (Pastoriza

et al.

1994). The mincing process disturbs themuscle membrane system, thereby exposing the lipid components to oxygen,or causes other reactions (Love and Pearson 1976). The PV for all samplesincreased during frozen storage (

P

<

0.05). Treating with RE or OJ showedthe same effect on peroxide value at each storage step. The control sampleshad the highest peroxide values at each storage time (

P

<

0.05). The meanlevel of PV for the RE treatment was 19.15 meqPO/kg after 5 months ofstorage. The PV value was lower in the OJ added samples than the controlsamples at each evaluation period. PV levels of samples stored at

-

40C

FIG. 1. CHANGES IN TBA VALUES OF SARDINE MINCES STORED AT

-

20C

0

2

4

6

8

10

0 1 2 3 4 5

Storage (month)

TB

A (

mg

ma/

kg)

RE

OJ

C


increased more rapidly than the sample stored at 0C (Hwang and Regenstein1996).

Similar to our results Wada and Fang (1992) reported that PV levels ofrosemary extract added to sardine oil were lower than

μ

-tocoferol addedsamples. In our research after 5 months of storage, the lowest PV wasobserved for sardine mince treated with RE. Ackman and Gunnlakgsdattir(1992) reported that PV of mackerel fillets ranged from 20 to 30 meqPO/kgafter 5 months of storage at

-

15C. In our study peroxide value for all treatmentgroups did not exceed 30 meqPO/kg.

FFA formation as a result of lipid hydrolysis (triglyseride and phospho-lipid classes) has provided a suitable means for assessment of fish damageduring frozen storage (de Koning and Mol 1991; Hwang and Regenstein1996). FFA values of samples stored at

-

20C are given in Fig. 2. FFA valuesincreased significantly as a function of time (

P

<

0.05). At zero time the meanFFA level was recorded 5.15 for RE treated sample, 5.92 for OJ treated sampleand 7.57

m

mole/g for the control sample. Treatment with RE or OJ signifi-cantly affected FFA levels of samples during storage. At zero time and on day15 there were no significant differences among treatment groups (Fig. 3);however, FFA level had a sharp increase in control samples after 1 month ofstorage. No differences were found in FFA levels between OJ treated andcontrol samples after 3 months of storage (

P

> 0.05).At the end of storage period the lowest FFA level was found in samples

treated with RE. FFA develops in fish even at -29C although at a very slowrate (Olley et al. 1969). The increase is due to the hydrolysis of phospholipidsand triglyserides by the action of lipases and phospholypases (Oshima et al.

FIG. 2. CHANGES IN PV (meqPO/kg) VALUES OF SARDINE MINCES STORED AT -20C

0

5

10

15

20

25

30

0 1 2 3 4 5

Storage (month)

PV

(m

eqP

O/k

g)

RE

OJ

C


1984; Fazal and Srikar 1989). Abdel-aal (2001) reported that Nile karmoutfish (Claries lazera) mince treated with antioxidants and stored at -18Cshowed significant increment in FFA levels and no effect of added antioxi-dants on FFA levels was observed. Hwang and Regenstein (1996) did notdetect any significant changes in FFA levels of mackerel mince patties storedunder vacuum at -40C. Accumulation of FFA in frozen fish is related to someextent with lack of acceptability, because FFA are known to cause deteriora-tion by interacting with proteins, strongly interrelated with lipid oxidation(Han and Liston 1989; Auburg 1999) and affect taste or odor.

Fatty acid profiles of sardine mince treatments are given in Table 1. Therewas similarity in fatty acid profiles among the treatment groups. The majorfatty acids in the total lipids of sardines were palmitic acid (16 : 0), stearicacid (18 : 0), palmitoleic acid (16 : 1), eicosatrienoic acid (20 : 3 n-3), eicos-apentaenoic acid (20 : 5 n-3) and docosahexaenoic acid (22 : 6 n-3). Signifi-cant differences were observed in fatty acid profiles of control treatmentbefore and after storage whereas no significant differences were recorded forthe other treatment groups (RE or OJ) in any class of fatty acid at month 5.In control samples saturated fatty acid (SFA) percent increased from 32.17 to42.1 whereas polyunsaturated fatty acid (PUFA) percent decreased from 43.81to 32.78. The decrement in PUFA percent reflects enzymatic hydrolysis ofsardine lipids. Similar to our results, Kundakçı (1979) reported that SFApercent increased from 31.75 to 37.48 in mullet fillets after 18 months storageat -18C. Sant’Ana and Manchini-Filho (2000) showed that the use of antiox-idants altered fatty acid composition of fish fillets. Fish muscle containscharacteristic high amounts of EPA and DHA. Since EPA and DHA are veryeasily oxidized because of their high unsaturation, a decline in the ratio ofEPA + DHA/16 : 0 fatty acid has been used to elucidate oxidative deteriora-tion of polyunsaturated fatty acid in fish lipids (Wada and Fang 1992). In

FIG. 3. CHANGES IN FFA VALUES OF SARDINE MINCES STORED AT -20C

0

5

10

15

20

25

0 2 4 51 3

Storage (month)

FF

A (

mm

ole

/g)

RE

OJ

C


TAB

LE

1.

FAT

TY

AC

ID P

RO

FIL

E O

F SA

RD

INE

MIN

CE

S ST

OR

ED

AT

-20

C

Fatty

aci

d (g

/100

g t

otal

fat

ty a

cids

)*R

EO

JC

0 da

y5t

h m

onth

0 da

y5t

h m

onth

0 da

y5t

h m

onth

14 :

04.

99 ±

0.1

33.

99 ±

0.2

14.

34 ±

0.2

15.

99 ±

1.2

14.

56 ±

0.7

83.

67 ±

1.9

616

: 0

19.1

1 ±

0.11

19.7

6 ±

0.13

21.1

7 ±

1.54

20.6

5 ±

1.08

19.1

1 ±

1.76

24.6

7 ±

0.58

18 :

06.

51 ±

0.2

37.

68 ±

0.0

98.

09 ±

1.1

18.

02 ±

0.9

28.

50 ±

1.2

213

.76

± 0.

0116

: 1

13.8

6 ±

0.32

13.2

2 ±

0.12

12.0

6 ±

0.14

11.7

9 ±

0.13

12.5

4 ±

1.54

10.3

4 ±

0.32

18 :

12.

61 ±

0.4

31.

95 ±

0.2

12.

22 ±

0.2

12.

33 ±

0.8

72.

33 ±

1.2

21.

45 ±

0.4

618

: 2

3.48

± 0

.12

3.0

± 1.

213.

59 ±

0.1

73.

42 ±

0.7

83.

60 ±

1.0

92.

56 ±

0.3

718

: 3

n-3

3.73

± 0

.18

3.22

± 0

.43

2.24

± 0

.34

2.18

± 0

.45

2.27

± 0

.45

1.87

± 0

.34

20 :

20.

49 ±

0.2

10.

35 ±

0.2

10.

28 ±

1.2

20.

23 ±

1.9

80.

31 ±

0.6

90.

26 ±

0.1

120

: 3

n-3

10.2

0 ±

0.14

8.02

± 0

.32

10.5

3 ±

1.13

8.45

± 0

.78

10.4

2 ±

1.09

8.98

± 0

.23

20 :

5 n-

3 (E

PA)

10.4

4 ±

0.09

10.3

1 ±

0.21

11.3

8 ±

1.01

11.3

0 ±

0.56

10.4

5 ±

1.09

6.36

± 0

.42

22 :

5 n-

30.

51 ±

0.0

80.

35 ±

0.2

10.

49 ±

1.1

20.

42 ±

0.9

90.

55 ±

1.0

20.

33 ±

1.0

322

: 6

n-3

(DH

A)

17.1

3 ±

0.12

16.3

4 ±

0.12

16.3

± 0

.99

16.6

5 ±

0.56

16.2

1 ±

1.76

12.4

2 ±

1.04

SFA

30.1

1 ±

0.13

31.4

3 ±

0.13

33.6

± 1

.32

34.6

6 ±

1.22

32.1

7 ±

0.45

42.1

± 1

.09

MU

FA16

.47

± 0.

1315

.17

± 0.

2314

.28

± 2.

0114

.12

± 1.

7614

.87

± 0.

9911

.79

± 2.

13PU

FA45

.98

± 0.

1141

.59

± 0.

1244

.88

± 1.

1142

.65

± 0.

0343

.81

± 0.

0432

.78

± 0.

21T

UFA

62.4

5 ±

0.12

56.7

6 ±

0.11

59.1

6 ±

0.23

56.7

7 ±

0.98

58.6

8 ±

1.02

44.5

7 ±

0.23

Oth

ers

7.44

± 0

.05

11.8

1 ±

0.09

7.24

± 0

.45

8.57

± 0

.92

9.15

± 0

.78

13.3

3 ±

0.92

EPA

+ D

HA

/16

: 01.

44 ±

0.1

21.

34 ±

0.1

21.

31 ±

0.5

31.

35 ±

0.4

51.

39 ±

0.2

20.

76 ±

0.1

4

*M

ean

± SD

.R

E, r

osem

ary

extr

act;

OJ,

oni

on j

uice

; C

, con

trol

; E

PA, e

icos

apen

taen

oic

acid

; D

HA

, doc

osah

exsa

enoi

c ac

id;

SFA

, sat

urat

ed f

atty

aci

d; M

UFA

, mon

oun-

satu

rate

d fa

tty a

cid;

PU

FA, p

olyu

nsat

urat

ed f

atty

aci

d; T

UFA

, tot

al u

nsat

urat

ed f

atty

aci

d.


control samples mean EPA + DHA/16 : 0 value decreased from 1.39% to0.76% at the end of storage; no changes were observed in EPA + DHA/16 : 0value for RE or OJ treatments after 5 months of storage. Beltran and Moral(1990) observed an increment in EDA and DHA levels for fish fillets storedat -18C for 180 days. Younathan et al. (1983) showed the effect of addingonion extract on the stability of PUFA of shark mince. Dissimilar to our resultsaccording to Boyd et al. (1993), fatty acid composition of RE treated cookedfish fillets was not different than control fillets after storage at -20C. Thus,differences may be attributed to cooking of sardine mince in their research.

CONCLUSIONS

Results of our investigation revealed that 300 ppm rosemary extractretarded oxidative changes in frozen sardine mince. Onion juice (1 mL/100 g)was not as effective as rosemary extract on oxidative stability. Adding onionjuice retarded rancidity development for 3 months of storage. Further researchis necessary to study the effects of different levels of onion extract on oxidativestability of frozen fish mince.

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EFFECTS OF USING ROSEMARY EXTRACT AND ONION JUICE ON OXIDATIVE STABILITY OF SARDINE (SARDINA PILCHARDUS) MINCE