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International Journal of Food Properties

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Effect of operational parameters on determination of

oxidative stability measured by rancimat method
a a a a a
Nilkanth Pawar , Akash Purohit , Kamal Gandhi , Sumit Arora & R R B Singh
a
Dairy Chemistry Division, National Dairy Research Institute , Haryana , India , Karnal-132
001
Accepted author version posted online: 11 Jul 2013.

To cite this article: International Journal of Food Properties (2013): Effect of operational parameters on
determination of oxidative stability measured by rancimat method, International Journal of Food Properties, DOI:
10.1080/10942912.2012.680220

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 ACCEPTED MANUSCRIPT

Effect of operational parameters on determination of oxidative

stability measured by rancimat method
Nilkanth Pawara, Akash Purohita, Kamal Gandhia,Sumit Aroraa* and R R B Singha
a
Dairy Chemistry Division, National Dairy Research Institute, Karnal-132 001, Haryana, India

* Corresponding author

E-mail: sumitak123@yahoo.com

Tel: 91-184-2259156
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Fax: 91-184-2250042

Running title- Rancimat and shelflife prediction of ghee

ABSTRACT

Operational parameters of Rancimat, including sample weight (3, 6, 9 g), airflow rate (10, 15, 20

L/h) and temperature (110,120,130°C) were evaluated to determine their effects on oxidative

stability index (OSI), temperature coefficient, Q10 number and shelf life prediction of ghee

(anhydrous milk fat). These parameters showed statistically significant effects on the OSI. It was

observed that when the sample weight and airflow rate at a given temperature were at saturated

air condition, the OSIs showed no significant differences (p<0). As the temperature increased,

OSI decreased and average coefficient of variation was minimal at 120°C. The conditions under

which the ghee sample was saturated with air and had a relatively lower OSI, was with sample

weight of 6 g, temperature at 120°C and an airflow rate of 15 L/h. Temperature coefficient and

Q10 number were independent of sample weight and air flow rate, however, they had a

significant effect on shelf life prediction of ghee.

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Keywords: Oxidative stability index; Q10 number; Rancimat; Ghee; Temperature coefficient.

INTRODUCTION

Oxidative stability is an important parameter for the quality assessment of milk fat and vegetable

fats and oils. Lipids undergo oxidative degradation during processing and storage, resulting in

an alteration of major quality parameters such as colour, flavour, aroma and nutritive value,
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affecting their suitability for consumption. Majority of these changes are initiated by reactive

oxygen species, which lead to the formation of various primary and secondary products like
[1, 2]
carbonyls, peroxides and hydroperoxides .Therefore, assessing the extent of oxidative

degradation of fats, oils and foods containing lipids is essential to the food industry [3].

Rapid methods are required to predict the stability and shelf life of edible oils and fats.

The determination of oxidative stability with the Active Oxygen Method (AOM; AOCS Method

Cd 12-57) and Schaal oven method, involves the use of elevated temperatures to increase the
[4]
reaction rate of lipid oxidation. However, both are costly and labour intensive . Rancimat
[5]
method developed by Hadorn and Zurcher is used to assess oil stability index (OSI) and it

agrees well with AOM method. It has gained acceptance, owing to its ease of use,

reproducibility, continuous measurement requiring no periodic analytical determinations and

[6, 7]
uses no organic solvents for titrations . This method is based on the measurement of

conductance, produced by volatile organic acids collected in deionised water. In this test, the fat

sample is placed in a vessel, subjected to an above-ambient temperature while oxygen is bubbled

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through it, to accelerate the oxidative process until short-chain volatile acids are produced. In a

separate vessel, the acids formed are immediately dissolved in distilled water, and the

conductivity of this solution is monitored at ambient temperature [7]. End point of rancimat test is

determined by induction period (time before rapid deterioration of fat occurs), which is used to

establish relative stability of fats. The oxidative stability index (OSI) obtained from the rancimat

correlated well with stability of fats under various conditions of lipid oxidation and also with

data obtained from independent sensory and/or analytical methods [8, 9, 10].
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This method may also provide some other useful information regarding oxidative

stability of edible fats. Plotting the logarithms of OSIs versus elevated temperatures and

extrapolating it to room temperature, the shelf-life of the fat sample at ambient conditions can be
[6]
predicted. The slope of the curves represents the temperature coefficients for fat samples .

Temperature acceleration factor, known as Q10 number, which predicts the increase in oxidation

rate with a 10°C increase in temperature, can also be calculated [11].

Sample weight, air flow rate and temperature are the operational parameters that can be

adjusted easily in the rancimat method and may affect the determination of the OSI, temperature

coefficient and Q10 number (oxidative stability measures) as well as the shelf-life prediction of

ghee. A number of studies have investigated the effect of these parameters independently and/or

in combination for vegetable oils and fish oils. However, no studies have been reported till date

regarding the effect of these three operating parameters of rancimat on oxidative stability of ghee

(AMF). Therefore, the aim of the present work was to investigate the effect of operational

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parameters on the determination of the oxidative stability and to predict the shelf-life of ghee

(AMF) using rancimat.

MATERIALS AND METHODS

Materials

Freshly prepared ghee was procured from experimental dairy plant, National Dairy Research

Institute, Karnal and stored at 4°C. The peroxide value of ghee was determined according to IS:
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3508[12].

Apparatus

A Metrohm rancimat model 743 (Herisau, Switzerland) capable of operating over a temperature

range of 50 to 220°C was used. The glassware were rigorously cleaned between each run to

avoid any contamination that would catalyze peroxidation. The tubes were soaked in hot water

detergent overnight followed by cleaning with hot water, distilled water and then thoroughly

dried in an oven. Measuring vessels, electrodes, and connecting tubes were cleaned with

detergent, washed off by running warm water and were rinsed several times with alcohol and

distilled water before use.

OSI Measurement

A stream of air was bubbled into ghee (AMF) samples (3, 6 and 9 g) in a reaction vessel placed

in an electric heating block. Effluent air containing volatile organic acids from the ghee (AMF)

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sample were collected in a measuring vessel containing double distilled water (60 mL). The

conductivity of the water was measured automatically as oxidation proceeded. Filtered, cleaned

and dried air was allowed to bubble through the hot ghee at the rates of 10, 15, and 20 L/h. The

OSIs of the ghee samples were recorded at 110, 120 and 130°C. In each run, eight samples were

accommodated in the equipment and analyzed simultaneously.

Shelf-life prediction, temperature coefficient and Q10 number calculation

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The logarithms of OSIs versus elevated temperatures (110, 120, and 130°C) were plotted and

lines were fitted to the data. Equations for each line were also determined. Slopes of the lines

yielded temperature coefficients. The shelf-lives as OSIs at 37°C (OSI37) were calculated from

the corresponding equations. Q10 numbers were calculated as the OSIs at T/OSI time at T+10°C
[6, 7, 11].

Statistical Analysis

All determinations were carried out in triplicate and data was subjected to analysis of variance.

Analysis of variance was performed using the ANOVA, according to the SYSTAT software. P

values less than 0.05 were considered statistically significant.

RESULTS AND DISCUSSION

The OSIs and coefficient of variation (CV) of treatment combinations (ghee sample

weight × air flow rate) at different temperatures are presented in table 1. All ghee samples had

zero peroxide values prior to the start of the rancimat test.

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At 110°C with sample weight of 3, 6 and 9 g, the air flow rates of 10, 15 and 20 L/h gave

statistically different OSIs of ghee. It is evident from table 1 that at 110°C, lowest OSI was

observed at 6 g sample weight and air flow rate of 15 L/h, whereas highest OSI was observed at

3 g sample weight and air flow rate of 20 L/h at same temperature. Hill and Perkins [13] reported

significantly different OSIs for an oil sample size of 2.5 g at air flow rates of 12–20 L/h. It was

interesting to find that the OSI of the ghee sample at an air flow rate of 20 L/h was significantly
[14]
higher than that of low air flow rates. Jebe et al. reported that temperature stabilization at a
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sample weight of 2.5 g was difficult when using the rancimat method and thus recommended a

5g sample weight. However, it seems that air-saturated conditions cannot be maintained for low

ghee sample weights exposed to high air flow rates. The rigorously turbulent status of the ghee
[15]
sample resulted in more air escaping from the ghee sample than used for lipid oxidation . At

110 and 120°C, for all air flow rates and sample weight of 6 g ghee the OSI was minimal. These

relatively low OSIs for 6 g sample size might be because of the creation of sufficient air

saturated conditions which was necessary for lipid oxidation as against 3 and 9g sample weights,

whereas the above trend was not observed for 130°C temperature as the OSIs for 9 g sample size

were higher from that of 6g sample size. This suggests that the concentration of air in the ghee

sample was not sufficient for the creation of air-saturated conditions essential for oxidation. It

can, therefore, be interpreted that treatment combinations 1, 3, 4, 6, 7, 9 in Table 1 cannot create

the air-saturated conditions whereas, a relatively large coefficient of variation (CV) for treatment

combinations 2 and 8 suggests that the conditions under which oxidation occurred varied more

than the others. Hence, the preferred conditions at 110°C are 6 g sample weight with an air flow

rate of 15L/h.

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As expected, OSIs significantly decreased with temperature (Table 1). As shown in Fig. 1, at 110

and 120°C, a nearly similar trend for the OSIs was observed, but not at 130°C. For all treatment

combinations, the OSIs at 110°C were higher than corresponding OSIs at 120 and 130°C. The

average CV of all the treatment combinations varied considerably amongst all the three

temperatures and was minimal at 120°C (0.023). These observations indicated that the conditions

under which the sample oxidizes varies with temperature. From the table 1, it can be interpreted

that the optimum condition for determining OSI, where the sample was saturated with air (OSI
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is minimum) with a relatively lower CV, was sample weight of 6 g, air flow rate of 15 L/h and

the temperature of 120°C.

Table 2 represents the data calculated from the linear relationship between the natural

logarithm of OSI and the temperature for the treatment combinations in Table 1. There were non-

significant differences among the treatment combinations in terms of the temperature

coefficients and Q10 numbers. Thus, these two parameters can be determined for ghee

independent of the sample weight and air flow rate. The results obtained revealed that the

temperature coefficient for ghee had a mean value of -3.24×10-2 °C-1. The temperature

coefficient for soybean oil was found to be -3.12 × 10-2°C-1[15]. A mean value of 2.07 for Q10

number indicated that an increase in temperature by 10°C approximately halves the OSI of ghee.

Statistically significant differences between the calculated OSIs for the treatment combinations

at 37°C (OSI 37) showed that sample weight of ghee and air flow rate have a definite effect on
[11]
the shelf-life prediction for ghee. Frankel stated that the extrapolation of the stability results

obtained by the rancimat test at ambient storage leads to either over prediction or under

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prediction of the actual shelf life depending on the type of oil. Mendez et al. [7] attributed this to

the different mechanism of peroxidation and peroxide decomposition kinetics under accelerated

conditions of the rancimat test from the corresponding mechanisms at ambient storage

conditions. Nevertheless, the rancimat method correlated highly (r = 0.966; P = 0.000) with
[16]
rapeseed oil stability measured by peroxide development . Variations between shelf life

predictions from long-term storage studies and the OSI test can be minimized by choosing the

correct levels of these operational parameters in the rancimat method depending on the type of
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the product. Thus, by using a rancimat, we can rapidly assess the quality of fats, oils and fat

containing dairy foods which can save a lot of time and is very precious in quality control as well

as research and development laboratories.

CONCLUSION

Temperature, sample weight and air flow rate showed a significant effect on shelf-life prediction

of ghee. Therefore, choosing the right levels of these operational parameters in the rancimat

method may produce the least possible difference between predictions from long-term storage

studies and the OSI test.

ACKNOWLEDGEMENTS

This work was supported by the National Agricultural Innovation Project (component 4: Basic

and strategic research C30029), Indian Council of Agricultural Research. New Delhi, India.

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REFERENCES

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8. Anwar, F.; Bhanger, M.; Kazi, T. Relationship between Rancimat and active oxygen

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Chemists Society 2003, 80,151-155.

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9. Kowalski, B.; Ratusz, K.; Kowalska, D.; Bekas, W. Determination of the oxidative

stability of vegetable oils by differential scanning calorimetry and Rancimat measurements.

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11. Frankel, E. 1998. Lipid oxidation. The Only Press, Dundee, pp 99–114.

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Bureau of Indian Standards, New Delhi.

13. Hill, S.E.; Perkins, E.G. Determination of oxidation stability of soybean oil with the

oxidative stability instrument: Operation parameter effects. Journal of American Oil Chemists

Society 1995, 72, 741–743.

14. Jebe, T.A.; Matlock, M.G.; Sleeter, R.T. Collaborative study of the oil stability index

analysis. Journal of American Oil Chemists Society 1993, 70, 1055–1061.

15. Farhoosh, R. Effect of operational parameters of the Rancimat method on determination

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Oil Chemists Society 2007, 84, 205-209.

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with storage at 200°C. Journal of American Oil Chemists Society 1994, 71(6), 649-651.

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Table 1 The oxidative stability index (OSI) and coefficient of variation (CV) of the treatment
combinations at different temperatures

Treatm Sample Air Temperature (oC)

ent size (g) flow

rate(l/h 110 120 130

)
OSI CV OSI CV OSI CV
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1 3 10 21.98aA 0.08 10.72aB 0.035 4.72aC 0.026

2 3 15 19.63bA 0.08 9.97aB 0.046 4.64aC 0.091

3 3 20 24.35cA 0.00 10.57aB 0.017 4.31aC 0.027

4 6 10 20.43abdA 0.09 10.18aB 0.049 4.52aC 0.050

5 6 15 18.57beA 0.07 9.38aB 0.010 4.59aC 0.033

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6 6 20 22.37afA 0.00 10.28aB 0.006 4.00aC 0.040

7 9 10 21.28adfA 0.12 10.28aB 0.022 4.79aC 0.006

8 9 15 18.76begA 0.10 9.77aB 0.003 4.93aC 0.025

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9 9 20 22.84afhA 0.00 10.44aB 0.022 4.10aC 0.046

Avg. 0.06 0.023 0.038

CV 4

Different uppercase as well as lowercase letters show significant difference within a row and a

column, respectively at p<0.05. CV = (SD/average) ×10. SD standard deviation

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Table 2 The results calculated from the linear relationship between the natural logarithm of the
OSI assessed by the rancimat test and the temperature for the treatment combinations in Table 1

Treatment Log OSI = A(T) + B Tcoeff(×10-2) OSI37(h) Q10

A ± SE B ± SE R2

1 -0.0330 ± 0.0015 5.0160 ± 0.1139 0.994 -3.30a 5948a 2.05a

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2 -0.0320 ± 0.0017 4.9150 ± 0.1486 0.998 -3.20a 3729b 2.00a

3 -0.0320 ± 0.0017 4.8790 ± 0.1334 0.999 -3.20a 12530c 2.07a

4 -0.310 ± 0.0023 4.7420 ± 0.1890 0.997 -3.10a 3845bd 1.968a

5 -0.0296 ± 0.0014 4.6030 ± 0.1175 0.999 -2.96a 3057bd 1.979a

6 -0.0286 ± 0.0014 4.4553 ± 0.3626 0.999 -2.86a 12595ce 1.92a

7 -0.0330 ± 0.0035 5.527 ± 0.0905 0.999 -3.30a 4934bdf 2.303a

8 -0.0366 ± 0.0003 5.4406 ± 0.1110 0.996 -3.66a 2480g 2.17a

9 -0.0366 ± 0.0003 5.4360 ± 0.0914 o.997 -3.66a 12162cei 2.18a

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Different lowercase letters show significant difference within a column at p<0.05. SE standard

error

Tcoeff temperature coefficient, OSI37 OSI at 37°C, Q10 OSI at T/OSI at T + 10°C
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Fig 1 The variation trend of the oxidative stability index (OSI) for different treatment
combinations (ghee sample sizexair flow rate) at 110°C (a), 120°C (b) and 130°C (c)
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