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Influence of fat content on physico chemical and oxidative stability of foal liver pâté

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The effect of fat content on physicochemical properties and lipid and protein stability of foal liver pâté was studied. For this purpose, two batches (10 units per batch) of foal liver pâté with different pork back fat content 30% (30F) and 40% (40F) were manufactured. 30F foal liver pâté was darker (lower L value, P > 0.05), redder (higher a, Pb0.001) and harder (higher hardness value; Pb0.001) compared to those with 40F. Fat level was closely related to the calorific value of foal liver pâté, being more calorific in those with higher fat contents (352 kcal100 g; Pb0.001). Regarding total Fe content, 30F foal liver pâté showed the higher value (4.19 mg100 g; Pb0.01). Oxidative stability of foal liver pâté was influenced by fat level. 40F foal liver pâté presented higher TBARS and lower carbonyl contents compared to 30F ones (Pb0.001). Finally, foal pâtés with the two different fat contents had significantly (Pb0.001) different n−6n−3 ratios, foal liver pâtés with 30F showed the lowest values (9.97) compared to those with 40F content

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In fluence of fat content on physico-chemical and oxidative stability

of foal liver pâté

Centro Tecnológico de la Carne de Galicia, Rúa Galicia Nº 4, Parque Tecnológico de Galicia, San Cibrán das Viñas, 32900 Ourense, Spain

a b s t r a c t

a r t i c l e i n f o

Article history:

Received 7 February 2013

Received in revised form 10 April 2013

Accepted 13 April 2013

Keywords:

Fat content

Foal liver pâté

Lipid and protein oxidation

Physico-chemical properties

The effect of fat content on physico-chemical properties and lipid and protein stability of foal liver pâté was stud-ied For this purpose, two batches (10 units per batch) of foal liver pâté with different pork back fat content [30% (30F) and 40% (40F)] were manufactured

30F foal liver pâté was darker (lower L* value, P > 0.05), redder (higher a*, Pb 0.001) and harder (higher hard-ness value; Pb 0.001) compared to those with 40F Fat level was closely related to the calorific value of foal liver pâté, being more calorific in those with higher fat contents (352 kcal/100 g; P b 0.001) Regarding total Fe con-tent, 30F foal liver pâté showed the higher value (4.19 mg/100 g; Pb 0.01) Oxidative stability of foal liver pâté was influenced by fat level 40F foal liver pâté presented higher TBARS and lower carbonyl contents compared to 30F ones (Pb 0.001) Finally, foal pâtés with the two different fat contents had significantly (P b 0.001) different

n−6/n−3 ratios, foal liver pâtés with 30F showed the lowest values (9.97) compared to those with 40F content (13.41)

© 2013 Elsevier Ltd All rights reserved

1 Introduction

Liver pâté, a traditional cooked meat product consumed in many

countries, particularly in Europe (Denmark, France, Germany, Spain),

is usually considered to be of high quality (Le Ba & Zuber, 1996) It

con-sists of minced liver, fat and meat mixed with water and different

addi-tives, and is packed in glass containers and thermally treated This

product is characterized by its high iron content, in some cases

provid-ing up to 40% of daily requirements (Mataix & Aranceta, 2002) Due to

its chemical composition (high amounts of fat and non-heme iron,

and low content of natural antioxidants) and its manufacturing

process, this product is susceptible to lipid oxidation (Estévez,

Ramírez, Ventanas, & Cava, 2007; Russell, Lynch, Lynch, & Kerry, 2003)

Horse meat is characterized by low fat (6.63 g/100 g) and

cholester-ol contents (61 mg/100 g), and is rich in iron (3.89 mg/100 g) and

vita-min of the B group (Badiani, Nanni, Gatta, Tolomelli, & Manfredini,

1997) This meat has a favorable dietetic fatty acid profile, with a high

content of unsaturated fatty acids relative to saturated acids and

con-tains a greater proportion of components from theα-linolenic fatty

acid family (Lorenzo, Fuciños, Purriños, & Franco, 2010; Sarriés,

Murray, Troy, & Beriain, 2006; Tateo, De Palo, Ceci, & Centoducati,

2008) These nutritional characteristics mean that this type of meat

Martin-Rosset, Jussiaux, & Trillaud-Geyl, 1984) Consumption has

in-creased in recent years, with Spain being the fourth major producer of

horse meat in the U.E in 2009 with 6400 tons (FAOSTAT, 2009), but is

still not comparable to the consumption of other meats such as beef, chicken or pork (Franco et al., 2011; Lombardi-Boccia, Lanzi, & Aguzzi,

2005) These increases might be due to changes in attitude towards this type of meat and the wish of consumers to taste new meat products (Hoffman & Wiklund, 2006; Sarriés et al., 2006) The meat is mainly consumed as fresh meat however, it is starting to be used in the

Temperán, Bermúdez, Cobas, & Purriños, 2012) Although, foal meat represents a good alternative for these products, some manufactures add pork fat to compensate for its low fat content

Several studies have evaluated the meat quality of foal fresh meat (Franco et al., 2011; Juárez et al., 2009; Lanza, Landi, Scerra, Galofaro,

& Pennisi, 2009; Lorenzo et al., 2010; Sarriés & Beriain, 2005, 2006; Sarriés et al., 2006; Tateo et al., 2008), but there is little information about the physico-chemical and nutritional quality of products made with this meat (Lorenzo et al., 2012) These types of products would uti-lize the parts of the carcass that have less value for fresh consumption Some studies concerning the physico-chemical characteristics of pork, duck, goat and ostrich liver pâté (Dalmás, Bezerra, Morgano, Milani, & Madruga, 2011; Delgado-Pando, Cofrades, Rodríguez-Salas, & Jiménez-Colmenero, 2011; Estévez, Morcuende, Ramírez, Ventanas, & Cava, 2004; Estévez, Ventanas, & Cava, 2005; Estévez, Ventanas, Cava,

& Puolanne, 2005; Fernández-López, Sayas-Barberá, Sendra, & Pérez-Álvarez, 2004; Russell et al., 2003) have been carried out

Howev-er, foal meat has never been used in the production of these products Furthermore, the level of fat has been demonstrated to influence the nutritional and sensory characteristics of these products (M Estévez,

S Ventanas et al., 2005; M Estévez, J Ventanas et al., 2005) The aim

of this work is to develop a new value-added foal pâté and study the

⁎ Corresponding author Tel.: +34 988 548 277; fax: +34 988 548 276.

E-mail address: jmlorenzo@ceteca.net (J.M Lorenzo).

0309-1740/$ – see front matter © 2013 Elsevier Ltd All rights reserved.

Contents lists available atSciVerse ScienceDirect

Meat Science

j o u r n a l h o m e p a g e : w w w e l s e v i e r c o m / l o c a t e / m e a t s c i

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effect of different fat levels on the physico-chemical and oxidation

sta-bility characteristics of this product

2 Material and methods

2.1 Manufacture of the foal liver pâté

Two different formulations of foal liver pâté were considered,

differ-entiated in terms of fat content [30% fat (30F) and 40% fat (40F)] The

pâtés were prepared in the pilot plant of the Meat Technology Center

of Galicia The composition of each sample is presented in theTable 1

Foal liver, foal meat (from the hind quarter, composed principally of

gluteus medius, semitendinosus and semimembranosus muscles) and

subcutaneous fat from commercial slaughter pigs was used as the

main ingredients Foal meat used to the manufacture of pâté had a

chemical composition of 76.3% moisture, 20.8% protein and 1.5% fat,

while foal liver had values of 69.9%, 23.6% and 1.0% for moisture, protein

and fat, respectively The fatty acid profile of liver, muscle and adipose

tissue is shown inTable 2 The day before the preparation, liver and

foal meat were ground through 10 mm diameter mincing plate in a

cooled chopped (La Minerva, Bologna, Italy) at 4 °C and mixed with

the nitrificant ingredients (sodium chloride, sodium nitrite and sodium

ascorbate) This blend was kept in darkness and refrigerated until the

following day On the day of manufacture, the fat was chopped using

the same conditions used for the meat and liver, and heated in water

to 65 °C Then, the remaining ingredients were added, sodium caseinate

to the heated fat, and water, milk powder and potassium phosphates to

the meat mixture Finally, both mixtures were blended to obtain a

ho-mogeneous raw paste The liver pâtés were packed in glass containers

prior to thermal treatment (80 °C/30′) The samples were cooled in a

blast chiller (−21 °C/30′) and then analyzed

2.2 Analytical methods

2.2.1 Physico-chemical analysis

The pH of samples was measured using a digital pH-meter (Thermo

Orion 710 A +, Cambridgeshire, UK) equipped with a penetration

probe Color measurements were carried out using a CR-600

color-imeter (Minolta Chroma Meter Measuring Head, Osaka, Japan)

Three measurements were performance for each sample CIELAB

space (CIE, 1976): lightness, (L*); redness, (a*); yellowness, (b*)

were obtained Before each series of measurements, the instrument

was calibrated using a white ceramic tile Hue (hab) and chroma (C*)

were calculated from the a* and b* values according to the formula:

C¼qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffið Þa2þ bð Þ 2

and hab¼ acr tanba: Moisture, fat, protein (Kjeldahl N × 6.25) and ash were

1997), 1443:1973 (ISO, 1973), 937:1978 (ISO, 1978), and 936:1998

(ISO, 1998), respectively Briefly, moisture percentage was calculated

by weight loss of the sample (5 g) maintained in an oven (Memmert

UFP 600, Schwabach, Germany) at 105 °C, until constant weight Ash percentage was calculated by weight loss of the sample (5 g) in a muffle furnace (Carbolite RWF 1200, Hope Valley, England) in a porcelain capsule at 600 °C until constant weight For fat content samples (2 g) were subjected to a liquid–solid extraction using petroleum

USA) for 90 min Fat content was calculated by gravimetric differ-ence Protein content was determined according to Kjeldahl total ni-trogen method, multiplying the total nini-trogen content by 6.25 A sample (1 g) was reacted with sulfuric acid (cupric sulfate was employed as a catalyst) in a digester (Gerhardt Kjeldatherm KB, Bonn, Germany) Organic nitrogen was transformed to ammonium sulfate, which was distilled in alkali conditions (Gerhardt Vapodest

50 Carrousel, Bonn, Germany)

2.2.2 Total Fe content Five gram (5.000 ± 0.001 g) samples were weighed into porce-lain crucibles for total Fe analysis The foal pâté samples were incin-erated in a furnace at 450 °C for 12 h The ash was dissolved in 10 mL

directly

induc-tively coupled plasma-optical emission spectroscopy (ICP-OES), using a Thermo-Fisher ICAP 6000 plasma emission spectrometer (Thermo-Fisher, Cambridge, UK), equipped with a radio frequency source of 27.12 MHz, a peristaltic pump, a spraying chamber and a concentric spray nebulizer The system was totally controlled by ICP software using 99.996% liquid argon plasma gas (Praxair, Madrid, Spain) Operating conditions of the ICP-OES equipment were: reflected

0.5 L/min; main argonflow, 12 L/min; background correction, 2 points;

Table 1

Recipe used for the preparation of foal liver pâtés with different fat contents.

Table 2 Fatty acid composition (means ± SD) of liver, muscle and adipose tissue.

Liver (from foal) Muscle

(from foal)

Adipose tissue (from pork)

P value SEM

C14:0 0.45 ± 0.03 a

2.29 ± 0.09 c

1.24 ± 0.01 b

0.000 0.33 C15:0 0.21 ± 0.02 b

0.31 ± 0.02 c

0.00 ± 0.00 a

0.001 0.06 C16:0 16.58 ± 0.27 a

25.07 ± 0.21 c

23.24 ± 0.12 b

0.000 1.63 C16:1cis-9 1.28 ± 0.03 a 2.55 ± 0.07 b 2.84 ± 0.01 c 0.000 0.30 C17:0 0.63 ± 0.03 c

0.51 ± 0.01 b

0.25 ± 0.03 a

0.002 0.07 C17:1cis-9 0.20 ± 0.01 0.51 ± 0.19 0.26 ± 0.01 0.124 0.07 C18:0 20.88 ± 0.09 c

6.20 ± 0.08 a

11.36 ± 0.07 b

0.000 2.72 C18:1cis-9 9.85 ± 0.92 a

15.94 ± 0.30 b

43.40 ± 0.15 c

0.000 6.52 C18:2n−6 31.64 ± 0.06 c

16.88 ± 0.75 b

14.64 ± 0.13 a

0.000 3.37 C20:0 0.12 ± 0.02 b

0.02 ± 0.02 a

0.09 ± 0.02 ab

0.045 0.02 C20:1 0.17 ± 0.01 a 0.21 ± 0.02 a 0.80 ± 0.01 b 0.000 0.12 C18:3n−3 12.05 ± 0.11 b

25.31 ± 0.46 c

0.86 ± 0.02 a

0.000 4.47 C20:2 0.29 ± 0.01 b

0.24 ± 0.01 a

0.60 ± 0.01 c

0.000 0.07 C20:3n−6 0.44 ± 0.01 c

0.28 ± 0.02 b

0.05 ± 0.02 a

0.001 0.07 C20:3n−3 0.49 ± 0.03 b

0.68 ± 0.01 c

0.10 ± 0.01 a

0.000 0.11 C20:4n−6 3.84 ± 0.09 c

1.82 ± 0.22 b

0.24 ± 0.01 a

0.000 0.66 C20:5n−3 0.12 ± 0.16 0.13 ± 0.09 0.00 ± 0.00 0.505 0.04 C22:6n−3 0.47 ± 0.03 c 0.27 ± 0.03 b 0.00 ± 0.00 a 0.001 0.08 SFA 38.89 ± 0.48 c

34.78 ± 0.24 a

36.18 ± 0.23 b

0.003 0.77 MUFA 11.59 ± 0.95 a

19.36 ± 0.41 b

47.30 ± 0.14 c

0.000 6.86 PUFA 49.51 ± 0.47 c

45.85 ± 0.66 b

16.51 ± 0.09 a

0.000 6.60 TUFA 61.10 ± 0.48 a

65.21 ± 0.24 c

63.81 ± 0.23 b

0.003 0.77 Σn−6 36.38 ± 0.13 c

19.45 ± 0.99 b

15.54 ± 0.11 a

0.000 4.05 Σn−3 13.12 ± 0.33 b 26.39 ± 0.33 c 0.97 ± 0.02 a 0.000 4.64 n−6/n−3 2.77 ± 0.06 b 0.73 ± 0.05 a 16.05 ± 0.28 c 0.000 3.03 SFA/TUFA 0.64 ± 0.01 b

0.53 ± 0.01 a

0.56 ± 0.05 a

0.004 0.02 Results expressed as percentage of total fatty acid analyzed.

SEM: Standard error of mean.

PUFA = Σ (C18:2n−6 + C18:3n−3 + C20:2 + C20:3n6 + C20:3n3 + C20:4n−

6 + C20:5n3 + C22:6n3).

MUFA = Σ (C16:1cis-9 + C17:1cis-9 + C18:1cis-9 + C20:1).

TUFA = Σ MUFA + PUFA.

SFA = Σ (C14:0 + C15:0 + C16:0 + C17:0 + C18:0 + C20:0).

Σn−6 = Σ(C18:2n−6 + C20:3n−6 + C20:4n−6).

Σn−3 = Σ(C18:3n−3 + C20:3n−3 + C20:5n3 + C22:6n−3).

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integration and reading time, 5 s; replicate number, 3; height of vertical

observation, 19 mm; nebulizer pressure, bar and radial torch con

figura-tion The operating wavelength was 259.940 nm Stock solution

(1000 mg/L; SCP-SCIENCE, Countaboeuf, France) was used for

prepar-ing the standard solutions in 4% HNO3, v/v The concentration range

was 0.01 to 50 mg/kg of Fe Thefinal value was the average of three

de-terminations The results were expressed in mg/100 g of pâté

2.2.3 Analysis of heme iron

Total heme pigments in the samples were determined as hemin

after extraction with acidified acetone solution (Hornsey, 1956) Briefly,

fresh meat samples (5 g) were dissolved in 1 mL deionized water,

0.5 mL concentrated HCl (37%) and 20 mL of acetone in glass test

tubes The tubes were sealed to reduce evaporation, held at room

tem-perature in darkness to minimize pigment fading during the 24 h

ex-traction and then filtered through 0.45 μm (Filter Lab, Spain) The

absorbance was measured (Agilent 8453, Waldbronn, Germany) at

512 nm The heme iron content was calculated with the factor

0.0882μg iron/μg hematin All samples were assayed in duplicate The

results were expressed as mg/100 g of pâté

2.2.4 Analysis of non-heme iron

The non-heme iron was determined by the ferrozine method

(Purchas, Simcock, Knight, & Wilkinson, 2003) Briefly, dry samples

of meat (500 mg) were ground using a mortar and pestle, dissolved

in a mixture of 3 mL of 0.1 M citrate phosphate buffer (pH 5.5) and

1 mL of 2% ascorbic acid (as reducing agent) in 0.2 M HCl and left to

stand at room temperature for 15 min before adding 2 mL of 11.3%

trichloroacetic acid After centrifugation at 3000 g for 10 min the

su-pernatant was removed To 2 mL of the susu-pernatant, 0.8 mL of 10%

ammonium acetate and 0.2 mL ferrozine reagent (40 mM) were

added and the absorbance was measured at 562 nm Concentrations

were obtained using a standard curve from 0 to 5 mg of iron/L

made with ferrous sulfate heptahydrate (Panreac Química S.L.U.,

Bar-celona, Spain) All samples were assayed in duplicate The results

were expressed in mg/100 g of pâté

2.2.5 Gross energy determination

The gross energy (the heats of combustion of protein, fat and

car-bohydrate) was determined in duplicate using an automatic adiabatic

bomb calorimeter (Parr 6100, Parr Instrument Company, USA),

cali-brated with benzoic acid The process essentially involves measuring

the energy evolved on total combustion of the sample in a stream of

oxygen The results were expressed in kcal/100 g of pâtés

2.2.6 Texture measurement

The Texture Analyzer (TA-XT.plus, Stable Micro Systems, Vienna

Court, UK) was used (Bourne, 1978) The penetration test was carried

out at room temperature (22 °C) and performed with a 6 mm

diam-eter penetration probe linked to a 5 kg cell at a velocity of 0.8 mm/s

and for a distance of 8 mm Hardness (kg/cm2), cohesiveness,

the available computer software (TEE32 Exponent 4.0.12 Stable Micro

Systems, Vienna Court, UK)

2.2.7 Lipid oxidation

Lipid stability was evaluated using the method proposed by

Vyncke (1975) Briefly, a meat sample (2 g) was dispersed in 5%

tri-chloroacetic acid (10 mL) and homogenized in an Ultra-Turrax (Ika

T25 basic, Staufen, Germany) for 2 min The homogenate was

solu-tion (5 mL) and incubated in a water bath at 96 °C for 40 min

The absorbance was measured at 532 nm Thiobarbituric acid

reac-tive substances (TBARS) values were calculated from a standard

curve of malonaldehyde with 1,1-3,3 tetraetoxipropane (TEP) and expressed as mg MDA/kg sample

2.2.8 Protein oxidation Protein carbonyls, as measured by the total carbonyl content,

and Stadtman (1987) Meat samples were homogenized in 20 mL

of 0.15 M KCl buffer for 60 s using an Ultra-Turrax (Ika T25 basic, Staufen, Germany) Two aliquots of homogenate (0.1 mL) were transferred to Eppendorf vials Then, proteins were precipitated in both aliquots by 10% trichloroacetic acid (TCA) (1 mL) and centri-fuged for 5 min at 5000 g One pellet was treated with 1 mL of 2 N

containing 0.2% 2,4-dinitrophenyl hydrazine (DNPH) (carbonyl con-tent) Both samples were incubated for 1 h at room temperature

(shak-en every 20 min) After incubation, 10% TCA was added (0.8 mL) The samples were vortexed for 30 s, centrifuged for 5 min at 5000 g and the supernatant removed The pellet was washed three times with

1 mL of ethanol-ethyl acetate (1:1 v/v) and then was dried under N2 gas Finally the pellet was dissolved in 2 mL of 6 M guanidine HCl in

20 mM sodium phosphate buffer (final pH 6.5), stirred and centrifuged for 2 min at 5000 g to remove insoluble fragments Protein concentra-tion was calculated from the absorbance at 280 nm using bovine serum albumin (BSA) as standard The amount of carbonyls was expressed as nmol of carbonyl per milligram of protein using an adsorp-tion coefficient of 21.0 mM−1cm−1at 370 nm for protein hydrazones 2.2.9 Analysis of fatty acid methyl esters

Fat was extracted from 5 g of foal pâté, according toFolch, Lees, and Stanley (1957) Lipid extracts were evaporated to dryness

methanol, as described byCarreau and Dubacq (1978) Fifty

stored at−80 °C until chromatographic analysis

Separation and quantification of FAMEs was carried out using a gas chromatograph, GC-Agilent 6890N (Agilent Technologies Spain, S.L., Madrid, Spain) equipped with aflame ionization detector and an auto-matic sample injector HP 7683, and using a Supelco SPTM-2560 fused silica capillary column (100 m, 0.25 mm i.d., 0.2μm film thickness, Supelco Inc., Bellafonte, PA, USA) Chromatographic conditions were as follows: initial oven temperature 120 °C (held for 5 min),first ramp at

2 °C/min to 170 °C (held for 15 min), second ramp at 5 °C/min to

200 °C (held for 5 min) and third ramp at 2 °C/min to afinal tempera-ture of 235 °C (held for 10 min) The injector and detector were maintained at 260 and 280 °C respectively Helium was used as carrier gas at a constantflow-rate of 1.1 mL/min, with the column head pres-sure set at 35.56 psi 1μL of solution was injected in split mode (1:50) The fatty acids were quantified using nonadecanoic acid methyl ester

at 0.3 mg/mL, as internal standard, it was added to samples prior to fat extraction and methylation Identification of fatty acids was performed

by comparison of the retention times with those of known fatty acids and the results expressed as a percentage of total fatty acids identified The proportion of polyunsaturated (PUFA), monounsaturated (MUFA), total unsaturated (TUFA) and saturated (SFA) fatty acid contents, and SFA/TUFA, n−6/n−3 and nutritional ratio were calculated

2.3 Statistical analysis

An analysis of variance (ANOVA) of one way using SPSS package (SPSS 19.0, Chicago, IL, USA) was performed for all variables in the study The least squares mean (LSM) were separated using Duncan's t-test All statistical tests of LSM were performed for a significance

correlation analyses using Pearson's linear correlation coefficients using the above statistical software package

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3 Results and discussion

3.1 Effect of fat content on chemical composition of foal liver pâtés

The results for the foal pâtés manufactured with different fat levels

are presented inTable 3 As expected, the differences in formulation

(Table 1) produced significant changes in the proximate composition

of the pâtés 30F foal pâtés had a higher water content than the 40F

batch (54.04 vs 52.48%, P = 0.055) Pearson correlation indicated that

moisture contents were positively related to the color attribute, a*

(r = 0.53, Pb 0.01), in agreement withM Estévez, S Ventanas et al

(2005)andDelgado-Pando et al (2011)who found higher water levels

in low-fat pâtés Fat and protein content were also significantly affected

by the fat level showing the opposite behavior Protein content

followed the decreasing order: 30F > 40F (Pb 0.001) In contrast,

40F foal pâtés had a higher fat content (26.33 g/100 g foal pâtés)

than 30F (23.20 g/100 g foal pâtés) (Pb 0.001) These results agree

withM Estévez, S Ventanas et al (2005)who found higher protein

content and lower fat contents in low-fat pâtés Regarding ash content,

fat level did not significantly affect the batches (3.25–3.26 g/100 g foal

pâtés; P = 0.932).Pinho, Ferreira, Casal, Oliveira, and Ferreira (1998)

found in15 brands of bovine liver pâtés on sale in Portugal, a moisture

content of 53.4%, similar to the present study The same authors found

11.8% protein, 29.4% fat and 2.6% ash In comparison to bovine pâtés,

the 30F foal pâtés had higher ash and protein contents, and lower values

for lipids, showing that the 30F foal pâtés could have consumer appeal,

being healthier, low-fat meat products

With reference to the chemical forms of iron, significant

differ-ences (P≤ 0.001) between groups were found (Table 3) The total

Fe content was higher in pâtés with 30F (4.19 mg/100 g) compared

to those with high-fat content (3.61 mg/100 g) (P = 0.001) The

re-sults agree withM Estévez, S Ventanas et al (2005), who found

higher levels of total Fe in pâtés samples with low-fat content The

foal pâtés samples (2.50 mg/100 g) as compared to those with

high-fat content (2.22 mg/100 g) On the other hand, the levels of

pâtés samples (1.14 mg/100 g) than the other one (1.01 mg/100 g)

These results are similar to those reported byM Estévez, S Ventanas

et al (2005), who observed that fat reduction causes increased heme

iron and decreased non-heme iron Fe deficiency is the most prevalent

nutritional disorder in the world, especially in developing countries

Knowledge of the levels of total, heminic and non-heminic Fe in meat

is of importance, because of the differences in bioavailability of these

forms of Fe (Lombardi-Boccia, Martínez-Domínguez, & Aguzii, 2002)

Foal pâtés from the two different fat content presented signi

fi-cantly different caloric values, being higher in those with high-fat

content (40F: 352.55 kcal/100 g pâtés and 30F: 315.88 kcal/100 g

pâtés, P = 0.000) These results are logical in view of the different

fat contents and agree with those reported by (Delgado-Pando et al.,

2011; M Estévez, S Ventanas et al., 2005) who found lower energy

con-tents in the low-fat pâtés The caloric values were positively related

(Pb 0.01) to fat level (r = 0.63) and negatively related (P b 0.01) to protein content (r =−0.67)

3.2 Effect of fat content on physical properties of foal liver pâtés Table 4shows the physical properties of foal liver pâtés with the two different fat contents No significant differences were found in pH 40F foal pâtés samples presented higher pH values and are in agreement with those reported byM Estévez, S Ventanas et al (2005) The values obtained (6.68–6.69) were slightly higher to those described for this type of product (Delgado-Pando et al., 2011; M Estévez, S Ventanas

et al., 2005; Fernández-López et al., 2004; Hong, Lee, & Min, 2004) pH was positively correlated with fat content (r = 0.44; Pb 0.05) Color parameters were affected by fat content (Table 4) These re-sults were expected because the color of pâtés is closely related to the color properties of raw material used for the formulation (M Estévez, J Ventanas et al., 2005) and therefore, changes in the pro-portions of the ingredients might lead to different colors The results indicated that higher contents of fat and lower contents of meat, in-creased lightness and reduced redness in the samples The 40F foal pâtés samples were lighter than the 30F samples (54.21 vs 52.97;

P > 0.05, respectively) and was positively correlated with fat con-tent (r = 0.39; Pb 0.05) and negatively related with protein

positively correlated with protein content (r = 0.58) Consequently, 30F foal pâtés samples had a more intense color (Chroma values: 16.33 vs 15.55; Pb 0.01) with lower values of hue (55.37 vs 58.78;

Pb 0.001) compared to foal pâtés with higher fat levels

As expected, the manufacture of foal pâtés with increasing fat levels resulted in products with different textural properties (Table 4) Fat re-duction increased hardness (0.22 vs 0.37 kg; Pb 0.001 for 40F and 30F groups, respectively), chewiness (0.11 vs 0.19 kg mm; Pb 0.001 for 40F and 30F groups, respectively), gumminess (0.11 vs 0.21 kg;

(0.49 vs 0.54; Pb 0.01 for 40F and 30F groups, respectively) Fat con-tent was negatively correlated (Pb 0.01) with hardness (r = −0.54),

content was positively related (Pb 0.01) with hardness (r = 0.85), chewiness (r = 0.82) and gumminess (r = 0.83)

This behavior is in agreement withM Estévez, S Ventanas et al (2005)in liver pâtés, who observed that fat reduction increased

Viana, Silva, Delvivo, Bizzotto, & Silvestre, 2005) found that fat re-duction decreased (Pb 0.05) penetration force, or had no effect on

Table 4

pH, color parameters and instrumental texture of foal liver pâtés from foal with differ-ent fat levels (means ± SD).

Color parameters Luminosity (L*) 52.97 ± 2.01 54.21 ± 1.37 0.40 0.127

Yellowness (b*) 13.43 ± 0.48 13.30 ± 0.54 0.11 0.564 Chroma (C*) 16.33 ± 0.51 15.55 ± 0.59 0.15 0.006

TPA test Hardness (kg) 0.37 ± 0.07 0.22 ± 0.05 0.02 0.000 Springiness (mm) 0.91 ± 0.03 0.94 ± 0.04 0.01 0.171 Chewiness (kg mm) 0.19 ± 0.05 0.11 ± 0.03 0.01 0.000 Gumminess (kg) 0.21 ± 0.05 0.11 ± 0.03 0.01 0.000

Table 3

Chemical composition and energy content of foal liver pâtés with different fat levels

(means ± SD).

Moisture (%) 54.04 ± 2.24 52.48 ± 0.86 0.41 0.055

Protein (%) 16.16 ± 0.58 14.99 ± 0.59 0.18 0.000

Total iron (mg/100 g) 4.19 ± 0.28 3.61 ± 0.39 0.10 0.001

Heme iron (mg/100 g) 2.50 ± 0.18 2.22 ± 0.09 0.04 0.000

Non-heme iron (mg/100 g) 1.01 ± 0.11 1.14 ± 0.03 0.02 0.001

Energy content (kcal/100 g) 315.88 ± 12.48 352.55 ± 3.60 4.65 0.000

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the textural parameters (Lurueña-Martínez, Vivar-Quintana, &

Revilla, 2004; Ordónez, Rovira, & Jaime, 2001)

3.3 Effect of fat content on oxidative stability of foal liver pâtés

The oxidative stability of foal liver pâtés, as measured by TBARS

from lipid oxidation and carbonyl content from protein oxidation, is

shown in Fig 1 The fat level presented significant differences in

TBARS values, compared to pâtés with lower fat content (0.54 vs

be-cause TBARS are derived from lipid oxidation and thus pâtés with

higher fat content would yield higher amounts of oxidation products

TBARS values were positively correlated (Pb 0.01) with fat content

(r = 0.64), agreeing withM Estévez, S Ventanas et al (2005)

Foal pâtés with higher TBARS values had lower contents of carbonyls

from protein oxidation (Fig 1) The carbonyl contents were higher in

pâtés with 30F (14.70 nmol carbonyls/mg protein) as compared to

those with high-fat content (11.48 nmol carbonyls/mg protein)

(Pb 0.001) Among amino acids, cysteine, tyrosine, phenylalanine,

tryptophan, histidine, proline, arginine, lysine and methionine have

been described as particularly susceptible to ROS (reactive oxygen

species) (Davies & Dean, 2003) The nature of the Pox products

formed is highly dependent on the amino acids involved and how

the oxidation process is initiated The side chains of some particular

amino acids such as arginine, lysine and proline are oxidized through

metal-catalyzed reactions into carbonyl residues while others such as

cysteine or methionine are involved in cross-linking or yield

sulfur-containing derivatives (Lund, Heinonen, Baron, & Estévez, 2011)

Car-bonyl content was positively related (Pb 0.01) to protein content

(r = 0.67) and negatively related (Pb 0.01) to fat level (r = −0.57)

These results disagree withM Estévez, S Ventanas et al (2005)who

reported higher carbonyl contents in pâtés manufactured with

high-fat levels

3.4 Effect of fat content on fatty acid composition of foal liver pâtés

The fatty acid composition of foal liver pâtés of two different fat

contents is shown inTable 5 The fatty acid profiles were dominated

“approximately 18.5% of total methyl esters” (Table 5) These results

are in agreement with other authors for liver pâtés (Estévez et al.,

2004; M Estévez, J Ventanas et al., 2005; Ordóñez, D'Arrigo,

Cambero, Pinc, & de la Hoz, 2003) since MUFA were the most

abun-dant of fatty acids As expected, the fatty acid profile of the foal

liver pâtés reflected the fatty acid composition of the porcine adipose

tissue (Table 2) as the proportion of lard in the recipe was the highest

of all ingredients The higher MUFA contents observed in foal liver pâtés were very significantly (r = 0.97, P b 0.01) correlated with

C18:1cis-9content and significantly (r = 0.58, P b 0.01) with C16:1cis-9

content The fat level showed significant differences in MUFA content (44.30 vs 44.86%, P = 0.000; for 30F and 40F groups, respectively) and in PUFA (18.82 vs 18.23%, P = 0.001; for 30F and 40F groups, respectively)

differences between batches (40.64 vs 41.03%, P = 0.003; for 30F and 40F groups, respectively), followed by palmitoleic acid These re-sults are in agreement with those ofEstévez et al (2004)who found that oleic acid was the predominant fatty acid in liver pâtés Within SFA, the main fatty acid was palmitic, which did not differ signi fi-cantly between groups (22.86 vs 22.96%, P = 0.188; for 30F and 40F batches, respectively), in agreement withEstévez et al (2004),

M Estévez, J Ventanas et al (2005)who observed similar percent-ages in liver pâtés Finally, within PUFA, linoleic acid was

15.82%, P = 0.451; for 30F and 40F batches, respectively), followed

by linolenic acid, which was higher in pâtés with 30F (1.59%) com-pared to those of higher fat content (1.26%) (Pb 0.001)

The nutritional ratio (C14:0+ C16:0) / (C18:1cis-9+ C18:2n−6), which indicates the healthiness of the diet with regard its lipid content, is im-portant (Estévez et al., 2004) In the present study, fat content did not affect (P > 0.05) this ratio Finally, foal pâtés with the two different fat contents presented significantly different n−6/n−3 ratios, those

of low-fat content showed the lowest values (9.97) compared to those with higher fat content (13.41) (P = 0.000) These values were higher

Health (1994)andFAO (2010)for the human diet, it should not exceed 4.0

40F 30F

Fig 1 Lipid and protein oxidative stability of foal liver pâté with different fat content as

assessed by TBARS (mg MDA/kg foal pâté) and carbonyl (nmol carbonyls/mg protein)

Table 5 Fatty acid composition (means ± SD) of foal liver pâtés from foal with different fat levels.

Nutritional ratio 0.43 ± 0.00 0.43 ± 0.00 0.00 0.673 Results expressed as percentage of total fatty acid analyzed.

SEM: Standard error of mean.

PUFA = Σ (C18:2n−6 + C18:3n−3 + C20:2 + C20:3n6 + C20:3n3 + C20:4n−6) MUFA = Σ (C16:1cis-9 + C17:1cis-9 + C18:1cis-9 + C20:1).

TUFA = Σ MUFA + PUFA.

SFA = Σ (C14:0 + C16:0 + C17:0 + C18:0 + C20:0).

Σn−6 = Σ (C18:2n−6 + C20:3n−6 + C20:4n−6).

Σn−3 = Σ (C18:3n−3 + C20:3n−3).

Nutritional ratio = (C14:0 + C16:0)/(C18:1cis-9 + C18:2n−6).

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4 Conclusions

As expected, fat level affected most of the physico-chemical

proper-ties Higher fat levels in foal liver pâtés, decreased redness and hardness

Concerning protein and lipid oxidation increased fat level encouraged

the production of TBARS and decreased carbonyl contents On the

other hand, 30F foal pâtés had a better n−6/n−3 ratio compared to

pâtés of higher fat levels Using meat and liver from foals and back fat

from pigs for the manufacture of pâtés results in a high quality product

in which the iron is highly bioavailable

Acknowledgments

Authors are grateful to Xunta de Galicia (Conselleria de Medio Rural)

for thefinancial support Special thanks to Monte Cabalar (A Estrada,

Pontevedra) for the foal samples supplied for this research

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