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Palatability and Stability of Shortbread Madewith Low Saturated Fat ContentOmbretta Marconi, Roberto Martini, Andrea Mangione, Caterina Falconi, Carolina Pepe, and Giuseppe Perretti

Abstract: High consumption of saturated fatty acids (SFA) is associated with increased risk of cardiovascular diseaseand the European Food Safety Agency has called for lower SFA intake. This study assessed the formulation of low SFAshortbreads by replacing 60% and 70% of the butter content with high oleic sunflower oil and water. The quality of thelow SFA shortbreads was evaluated through acidity, peroxide value, moisture, ash content, water activity, pH, protein,fat content, and fatty acid profiles. A sensory evaluation was performed to ascertain the effect on flavor. Stability of thenew formulations was assessed by conducting accelerated shelf-life studies. The high oleic sunflower oil replacement ofbutter at levels of 60% and 70% decreased the final SFA content by 52% and 61%, respectively. On the other hand,monounsaturated fat content increased 55% on average while polyunsaturated fat content increased by 40%. Furthermorethe new formulations possess quality parameters similar to those of traditional shortbreads (TSs). The study of the shelflife of the products showed that there are no significant variations in peroxide values, malondialdehyde content, or fattyacid profiles in biscuits over time, confirming their high stability. The quantitative descriptive analysis showed that theTS and low SFA shortbreads have similar sensory profiles, and the consumer tests indicated that the low SFA shortbreadswere well liked.

Keywords: fatty acids, food composition, food quality, omega-3 fatty acids, optimization

Practical Application: Consumers attention about obesity, diseases, and low fat foods is increasing. Therefore, oneof the aims of the food industry is the production of fat-reduced products characterized by the same quality of thefull-fat counterparts. Furthermore, the formulations of low saturated fatty acids shortbreads may be optimized by partiallyreplacing butter with high oleic sunflower oil. This study of the shelf-life stability and the sensory profiles of low saturatedfat shortbreads confirm that the new formulations possess quality parameters similar to those of traditional shortbreads.

IntroductionBiscuits are one of the most popular bakery products due to

their great variety, convenience, and long shelf life (Rababah andothers 2006; Caponio and others 2009). The main ingredients inbiscuits are flour, sugar, fat, and water, and the quality of theseingredients can all affect the overall quality of the product. Inparticular, fat plays an important role in biscuits as it influencesshelf life and contributes to the sensory experience (Zoulias andothers 2002; Laguna and others 2012; Taracon and others 2013).Shortbreads are biscuits characterized by being dense and brittle

as their relatively high quantities of fat and sugar create a glutennetwork (Manohar and Rao 1999). The quality of the fat usedin shortbread influences the flavor of the final product. Butter iscommonly used for its flavor in shortbread recipes, but it is rich insaturated fatty acids (SFA) especially myristic, palmitic, and stearicacids (McKevith 2005). Even though fat is an important source ofenergy and facilitates the absorption of fat-soluble vitamins, highfat consumption is generally associated with obesity and subse-quent health problems. In particular, high consumption of SFAand/or trans fatty acids (TFA) is associated with an increased riskfor cardiovascular disease (Micha andMozaffarian 2010). Recently,

MS 20130994 Submitted 7/18/2013, Accepted 1/9/2014. Authors Marconi,Mangione, Falconi, Pepe, and Perretti are with Dept. of Agricultural, Food andEnvironmental Science, Univ. of Perugia, Via san Costanzo, 06126, Perugia, Italy.Author Martini is with Colussi Group, Petrignano di Assisi, Perugia, Italy. Directinquiries to author Marconi (E-mail: ombretta.marconi@unipg.it).

the European Food Safety Agency (EFSA) published recommen-dations on the dietary reference values for fats including SFA,TFA, polyunsaturated fatty acids (PUFA), and monounsaturatedfatty acids (MUFA; European Food Safety Agency 2010). In par-ticular, the EFSA recommended that intakes of SFA and TFAshould be as low as possible, however they did not suggest an up-per limit for MUFA or PUFA intake. The EFSA also emphasizedthe relationship between dietary SFA intake and increased bloodcholesterol/low density lipoprotein (LDL) concentrations; the re-placement of SFA with MUFA and/or PUFA may help maintainnormal blood LDL concentrations (European Food Safety Agency2011).Some previous studies have focused on improving sensory and

nutritional profiles of biscuits (Maache-Rezzoug and others 1998;Chevallier and others 2000) with attention to proper diet. Addi-tional studies have focused on developing low-fat biscuit recipesusing emulsifiers, fat mimetics, and interesterified shorteningsmade from palm and cottonseed oils (Manohar and Rao 1999;Zoulias and others 2002; Dogan and others 2007; Handa andothers 2010; Forker and others 2012; Taracon and others 2013).One previous study (Regnicoli and others 2011) used 50% corn,soybean, and sunflower oils to replace SFA in shortbread and con-cluded that sunflower oil is a good SFA replacer because it had thebest sensory profile.Some vegetable oils (for example, sunflower, corn, and soybean)

are rich in MUFA and PUFA. Oleic acid (OA) and palmitoleicacid are the most common MUFA found in nature. Among thepolyunsaturated fats, the essential fatty acids linoleic acid (LA) and

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Table 1Shortbread recipes.

Ingredients (g) TS LSFS 60 LSFS 70

Flour 300 325 325Sugar 140 140 140Butter 100 40 30Sunflower oil 0 51 59.5Yolk 24 24 24Albumin 37 37 37(NH4)HCO3 1 1 1NaHCO3 1.5 1.5 1.5NaCl 1.25 1.25 1.25Water (mL) 25 34 35.5

TS: traditional shortbread.LSFS 60: low saturated fatty acids shortbread (60% high oleic sunflower oil).LSFS 70: low saturated fatty acids shortbread (70% high oleic sunflower oil).

linolenic acid (LNA) are of particular interest because of theirrequirement in the diet (McKevith 2005). Sunflower oil is charac-terized by a low content of SFA and high content of unsaturatedfatty acids, mainly OA and LA (Preeti and others 2007). OA isthe main fatty acid present in high oleic sunflower oil (Merrill andothers 2008).Studies have shown that diets rich in MUFA may play an im-

portant role in controlling cardiovascular risk factors such as hy-perlipidemia by lowering levels of LDL cholesterol and raisinghigh density lipoprotein cholesterol levels (Allman-Farinelli andothers 2005; Micha and Mozaffarian 2010). Although OA and LAcan have positive effects on health, they can also become oxidizedin the presence of light, moisture, and high temperatures therebyimparting peroxides and off-flavors in food products (Merril andothers 2008). In general, the more unsaturated a fat is, the moreunstable and susceptible to oxidation and rancidity it is. Therefore,the short shelf life of high oleic sunflower oil and all vegetable oilsrestricts their use in food formulations.The aim of the present study was to assess formulations of low

saturated fat shortbreads made by partially replacing butter with60% or 70% high oleic sunflower oil. The studies were conductedon both laboratory and pilot plant scales. Furthermore, acceleratedshelf-life studies were conducted in order to evaluate the stabil-ity of the new formulations through peroxide values, fatty acidprofiles, and malondialdehyde (MDA) content, a key product oflipid oxidation (Ray and Husain 2002). MDA has been used asa marker of oxidative damage in both biological samples (Kinter1995) and foods (St. Angelo 1996), but few studies have focusedon lipid oxidation in bakery products (Sakai and Kawahara 2005).Sensory evaluation of the new formulations was also performed inorder to ascertain the effects on the flavor of the low saturated fatshortbreads.

Materials and Methods

MaterialsBiscuit ingredients. Wheat flour (Triticum aestivum L.) that

was finely ground (00, with ash, protein, and moisture values of0.55%, 11.00%, and 15%, respectively, and a strength ofW = 146)was procured from Molini Popolari Riuniti di Ellera-Umbertide,Italy. The other shortbread ingredients were sucrose (Eridania,Bologna, Italy), butter (Grifolatte, Perugia, Italy), water, class Aeggs (Ovito, Perugia, Italy), high oleic sunflower oil with 79.2%of cis-OA, 4.5% of palmitic acid, 3.6% of stearic, and 11.2% ofLA (Oleificio Speroni, Fidenza, Italy), salt (Italkali, Palermo, Italy),sodium bicarbonate (E 500; Solvay, Milano, Italy), and ammoniumbicarbonate (E 503; Bertolini, Brescia, Italy).

Reagents. Sulfuric acid 96%, 0.1 N sulfuric acid solution(Fluka, Steinhem, Germany). Sodium hydroxide solution 40%,sodium hydroxide solution 0.02 N, potassium sulfate anhydrous,selenium, hydrogen peroxide 35%, petroleum ether, pumicestone, potassium hydroxide, methanol, sodium sulfate anhydrous(J. T. Baker, Deventer, Holland). 1,3-Diethyl-2-thiobarbituricacid, butyl hydroxyl toluene, sodium dodecyl sulfate, ethyl ac-etate, acetonitrile, boric acid solution 4% (Sigma Aldrich, St.Louis, Mo., U.S.A.). Kjeldahl tablet and sodium chloride solu-tion 2.5% (Carlo Erba, Milano, Italia). Hexane anhydrous (PanreacQuimica, Barcelona, Spain). All reagents were of analytical gradeand the highest purity available.

MethodsBiscuit preparation.

Laboratory scale. The quantities of ingredients used to producetraditional shortbread (TS) and the 2 levels of low saturated fattyacid shortbread (LSFS) are reported in Table 1. The replacementinvolved only the fat fraction of butter (85%).The shortbread in this study was made by the doughing up

method. In the first phase the butter and flour were mixed in alaboratory mixer (Kenwood Ltd., U.K.) for 5 min at 120 rpm and25 C. Meanwhile the sugar and the liquid ingredients (includingthe high oleic sunflower oil) were mixed separately in the mixer for3 min at 150 rpm. In the second phase, the 2 mixtures were com-bined, baking powder ((NH4)HCO3 and NaHCO3) was added,and the final dough was mixed for 5 min at 80 rpm. The doughwas allowed to rest at 4 C for 1 h after which it was manuallysheeted and shaped into pieces of 40 mm in diameter 5 mmin thickness. The biscuits were placed on a steel oven tray (length33 cm width 38 cm height 3 cm thickness 1 mm) andbaked in a laboratory ventilated oven (Binder, N.Y., U.S.A.) at180 C for 15 min. The biscuits were packaged and stored inpolypropylene bags after cooling.

Pilot plant scale. Once the LSFS recipe was optimized on the lab-oratory scale it was transferred to a pilot scale plant equipped witha vertical mixer IL30 Model (Dominici, Italy), a rotary press T280Model (Padovani, Italy) and a static oven C/2B Model (TMP,Italy).Biscuit evaluation. Moisture, pH, total nitrogen, protein, and

ash contents were determined according to their respective Asso-ciation of Official Analytical Chemists methods (Official Methodsof Analysis 1980). The water activity (aw) was determined usingan AquaLab series 3 (Decagon, Pullman, Wash., U.S.A.) calibratedwith lithium chloride solution (aw = 0.250 0.003).Determination of the acidity was conducted following the

method of Tateo (1969). Briefly, 4 g of ground biscuit and 100mL of an aqueous solution of neutral 50% ethanol were stirredfor 15 min at room temperature. After filtration, 50 mLof thesolution was titrated with NaOH 0.02 N using phenolphthaleinas the indicator. The peroxide values of the oil samples were de-termined according to the European Official Methods of Analysis(European Community 1991).The total fat content was determined according to the AOCS

method (American Oil Chemists Society 1984). Lipid extractionwas carried out by a Soxhlet extractor using petroleum ether assolvent for 6 h. The solvent was removed by vacuum evapora-tion. The fat content of the sample was expressed as a percentageof dry matter. The fat extract was used for the determinationof fatty acid profiles by gas chromatography. The lipid extractswere trans-esterified by treatment with methanol/KOH solution,

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Table 2Definitions of physical and flavor descriptors used inthe quantitative descriptive analysis.

Descriptor Definition

Color Burned, undercooked, or well-cookedHardness By steadily compressing the biscuit between

the molars, the force required forcompression.

Oiliness By gently rubbing both sides of the biscuitwith fingers, the greasy/oily sensation.

Sweetness Fundamental taste sensation associated withsugars, perceptible on the tip of thetongue.

Saltiness Taste of salt perceptible on the tip of thetongue and on the sides around it.

and the resulting fatty acid methyl esters were injected into theHRGC-FID system. The fatty acids were identified by comparingtheir retention times with those of commercial standards (SigmaAldrich; Bravi and others 2009).The MDA content of the biscuits was determined according to

the method of Sakai and Kawahara (2005) using HPLC coupledwith Flourimetric Detector DETBA-MDA adducts. The acceler-ated shelf-life test (ASLT), where storing the shortbreads for 1 d at55 C corresponds to 18 d at room temperature (20 C), was per-formed in order to evaluate the shelf life of the biscuits (Robertson1993). The shortbreads were kept in an oven at 55 C for 5, 10,and 20 d corresponding, respectively, to 90 (T5), 180 (T10), and360 (T20) days at room temperature and then subjected to chemicaland sensorial analysis.Sensory evaluation. Sensory evaluation was conducted to

evaluate the palatability of the cookies and to evaluate whetherthere was a statistically significant difference between the TS andLSFS at T0 and T20. In total 3 sensory tests were carried out:discrimination, quantitative descriptive analysis, and the hedonictest. For each test, panelists were presented with 2 biscuits inpolyethylene bags, a glass of water to cleanse the palate, and thescorecard.

Discrimination test. The triangle-discriminating test was performedby 30 untrained panelists of university students aging from 20 to30 of which 19 were female and 13 were male. The panelistswere self-reported traditional consumers of biscuits. Two tests werecarried out, one comparing LSFS 60 (LSFS with 60% of high oleicsunflower oil) with TS and the other comparing LSFS 70 (LSFSwith 70% of high oleic sunflower oil) with TS. Both tests wereperformed at T0.

Quantitative descriptive analysis. A total of 6 trained panelists agedfrom 30 to 45 consisting 4 males and 2 females evaluated thebiscuits using the quantitative descriptive analysis technique. Thepanelists were trained in 10 sessions to identify and determinedescriptors relating to smell, taste, and texture. The terms andtheir corresponding definitions (Table 2) were available to thepanelists during all sessions. The panelists belong to industrialbakery products and taste their products for the sensory analysis,almost daily. The evaluation of the biscuits was carried out over 2 din 2 sessions in which the LSFS 70 and TS at T0 and LSFS 70 andTS at T20 were evaluated. One replication of each treatment wasperformed by each panelist. A 7-point intensity scale was used;color: 0 = much too light, 1 = too light, 2 = slightly light, 3 =just about right, 4 = slightly too dark, 5 = too dark, 6 = much toodark; hardness: 0 = much too soft, 1 = too soft, 2 = slightly soft,3 = just about right, 4 = slightly hard, 5 = too hard, 6 = much

Table 3Chemical quality parameters of the different short-breads.

Quality parameter TS LSFS 60 LSFS 70

Acidity (g oleic ac./100 g product) 0.75a 0.66a 0.65a

Ash (%) 0.76a 0.80a 0.77a

PH 8.43a 8.44a 8.41a

Peroxide values (meq O2/g) 26.25a 23.96a 23.93a

Moisture (%) 3.25a 4.29b 4.40ab

aw 0.260a 0.310b 0.340b

Protein (%) 7.36a 7.94b 7.90b

Values with superscript letters, on the same row, that are different are significantlydifferent (P < 0.05).Mean reported of duplicate analyses from 3 technological replications.TS: traditional shortbread.LSFS 60: low saturated fatty acids shortbread (60% high oleic sunflower oil).LSFS 70: low saturated fatty acids shortbread (70% high oleic sunflower oil).

Table 4Fatty acid profiles of the shortbreads.

TS LSFS 60 LSFS 70

Total lipid (% of biscuit weight) 18.60a 16.35b 17.58ab

Fatty acids (% of total fat content)Saturated fatty acids (SFA)

Butyric (C4:0) 2.13a 0.95b 0.72c

Caproic (C6:0) 1.68a 0.76b 0.55c

Caprylic (C8:0) 1.13a 0.48b 0.35c

Capric (C10:0) 2.69a 1.12b 0.81c

Lauric (C12:0) 3.26a 1.31b 0.95c

Myristic (C14:0) 11.01a 4.28b 3.12c

Palmitic (C16:0) 31.08a 15.10b 12.42c

Stearic (C18:0) 9.39a 5.43b 4.81c

Behenic (C22:0) 0.06a 0.50b 0.57c

SFA 62.43a 29.93b 24.30c

Monounsaturated fatty acids (MUFA)Palmitoleic (C16:1) 1.79a 0.85b 0.69c

Oleic (C18:1) 25.55a 57.67b 63.22c

MUFA 27.34a 58.52b 63.91c

Polyunsaturated fatty acids (PUFA)t Linoleic (C18:2) 0.44a 0.19b 0.15c

cis-Linoleic (C18:2) 4.70a 8.74b 9.41c

-Linolenic (C18:3. n3) 0.77a 0.38b 0.33b

PUFA 5.91a 9.31b 9.89c

Values with superscript letters, on the same row, that are different are significantlydifferent (P < 0.05).Mean reported of duplicate analyses from 3 technological replications.TS: traditional shortbread.LSFS 60: low saturated fatty acids shortbread (60% high oleic sunflower oil).LSFS 70: low saturated fatty acids shortbread (70% high oleic sunflower oil).

too hard; oiliness, sweetness, saltiness, and flavor: 0 = much tooweak, 1 = too weak, 2 = slightly weak, 3 = just about right, 4 =slightly strong, 5 = too strong, 6 = much too strong. Open-endedcomments relative to secondary descriptors identified by trainedpanelists were also collected.

Consumer sensory analysis. A total of 114 consumers (54 female and60 male) aged from 19 to 59 y took part in the study to test theacceptance of the LSFS70 at T0. The consumer sample populationwas composed by students and employees of University of Perugiaand 52% of the participants reported frequent consumption of thistype of biscuit.Each consumer received a packet containing 2 pieces of LSFS

and tested the samples for acceptability of appearance, texture,fleste, and overall acceptance, reported in Table 3. Overall ac-ceptance was reported using a 5-point hedonic scale (where 1 =dislike extremely, 2 = dislike moderately; 3 = neither like nordislike, 4 = like moderately, and 5 = like extremely).

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Statistical analysisStatistical analyses were performed using the software STATIS-

TICA 8.0 (StatSoft Inc., Tulsa, Okla., U.S.A.) on data collectedfrom chemical and sensory evaluation of TS, LSFS 60, and LSFS70 during ASLT. Each biscuit sample was produced in triplicateand all the chemical analyses were performed in duplicate; so, thedata reported in tables are means of 6 values. Significant differ-ences (P 0.05) in the chemical parameters of TS, LSFS 60, andLSFS 70 after 0 d at 55 C (T0) and their fatty acid profiles werediscriminated using an unpaired Students t-tests, whereas a pairedt-test was used to discern the fatty acid profiles of the LSFS 60and LSFS 70 in ASLT. A paired t-test was used to discern signifi-cant differences of sensory evaluation tests. Values were consideredsignificantly different at P < 0.05.

Results and DiscussionThe present study tested the use of high oleic sunflower oil

as a replacement for 60% and 70% of the butter in shortbread.The shortbreads were prepared following the recipes reported inTable 1. The chemical properties of the LSFSs compared with thecontrol (TS) are reported in Table 3. The LSFS showed highermoisture, aw and protein values compared with the TS. The highmoisture value in the LSFS is likely to be related to the amountof water added to compensate for the butter replacement withvegetable oils as has been reported elsewhere (Forker and andothers 2012). The higher protein content of LSFS is probably dueto the greater use of flour during the kneading phase in order tofacilitate the workability of the product. The aw value is importantfor biscuits because it affects crispness. In fact, knowledge of justa products moisture content does not give information about itshydration (Arimi and others 2010). Biscuits typically have an awof about 0.25, and they will not be crisp if the aw is above 0.35(Manley 2000). Therefore, the aw values of 0.310 and 0.340 of theLSFS 60 and 70 were acceptable considering the aw limit value forthe crispness.Water activity and moisture content do not solely influence

rheological properties but also the microbiological and sensory(flavor) properties of foods. Foods with an aw < 0.60 are consid-ered microbiologically stable, although some of their constituentsmay undergo chemical reactions. A value of aw < 0.20 has beenshown to enhance lipid oxidization accompanied by pronouncedalterations of sensory qualities during storage (Labuza and De-

Table 5Fatty acid profiles of the LSFS 60 in the shelf-life study.

Fatty acids (% of total fat) T0 T5 T10 T20

Saturated fatty acids (SFA)Butyric (C4:0) 0.95a 0.98a 0.98a 0.93a

Caproic (C6:0) 0.77a 0.75a 0.75a 0.71a

Caprylic (C8:0) 0.48a 0.49a 0.49a 0.46a

Capric (C10:0) 1.11a 1.14a 1.13a 1.06b

Lauric (C12:0) 1.31a 1.34a 1.33a 1.24b

Myristic (C14:0) 4.29a 4.35a 4.31a 4.13a

Palmitic (C16:0) 15.11a 15.23a 15.16a 14.87a

Stearic (C18:0) 5.44a 5.45a 5.38a 5.84b

Behenic (C22:0) 0.50a 0.49a 0.57a 0.51a

SFA 29.95a 30.22a 30.13a 29.75a

Monounsaturated fatty acids (MUFA)Palmitoleic (C16:1) 0.84a 0.84a 0.86a 0.81a

Oleic (C18:1) 57.64a 57.67a 57.73a 58.33a

MUFA 58.48a 58.51a 58.59a 59.14a

Polyunsaturated fatty acids (PUFA)t-Linoleic (C18:2) 0.19a 0.19a 0.19a 0.19a

cis-Linoleic (C18:2) 8.76a 8.73ab 8.61b 8.44 c

-Linolenic (C18:3. n3) 0.38a 0.38a 0.37a 0.35b

PUFA 9.33a 9.30a 9.17b 8.98c

Mean reported of duplicate analyses from 3 technological replications.T0 = 0 d; T5 = 90 d; T10 = 180 d; T20 = 360 d.Values with superscript letters, on the same row, that are different are significantlydifferent (P < 0.05).

gun 1971; Reed and others 2002; Cervenka and others 2006).Thus, the aw values of our LSFSs which contain a high content ofunsaturated fatty acids known to readily oxidize, are acceptable.Total lipid content and fatty acid profiles of the shortbreads at

T0 are reported in Table 4. The lipid contents of the LSFS 60 and70 were lower than that of the TS. The fatty acid profiles werealso different between the groups. Among the SFA, palmitic acidwas the most abundant in all the samples that is mainly due tothe butter concentrations. OA was the most prevalent unsaturatedfatty acid followed by LA.Total SFA content was significantly decreased in the LSFS 60

and LSFS 70. Substituting high oleic sunflower oil for the butterreduced SFA content by about 52% in LSFS 60 and 61% in LSFS70. On the other hand, there was a significant increase in MUFAcontent by 53% and 57% for LSFS 60 and LSFS 70, respectively.PUFA content was increased by about 40% in both LSFS proto-types. The increases were the highest in OA and LA; this is due totheir relative abundance in high oleic sunflower oil. These results

Figure 1Peroxide values of LSFS 60 and LSFS 70during the accelerated aging tests.Number of replications: 3LSFS 60: low saturated fatty acids shortbread(60% high oleic sunflower oil)LSFS 70: low saturated fatty acids shortbread(70% high oleic sunflower oil)T0: 0 d; T5: 90 d; T10: 180 d; T20: 360 d.

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Table 6Fatty acid profiles of the LSFS 70 in the shelf-life study.

Fatty acids (% of total fat) T0 T5 T10 T20

Saturated fatty acids (SFA)Butyric acid (C4:0) 0.73a 0.68ab 0.65b 0.55c

Caproic (C6:0) 0.5a 0.54a 0.53a 0.45b

Caprylic (C8:0) 0.35a 0.35a 0.35a 0.29b

Capric (C10:0) 0.81a 0.83a 0.83a 0.67b

Lauric acid (C12:0) 0.96a 0.99b 0.99b 0.78c

Myristic acid (C14:0) 3.13a 3.23a 3.20a 2.64b

Palmitic (C16:0) 12.40a 12.60b 12.55b 11.22c

Stearic (C18:0) 4.80a 4.79a 4.80a 4.80a

Behenic (C22:0) 0.57a 0.56a 0.57a 0.62b

SFA 24.25a 24.57b 24.47c 22.02d

Monounsaturated fatty acids (MUFA)Palmitoleic (C16:1) 0.69a 0.68ab 0.68ab 0.62b

Oleic (C18:1) 63.34a 62.99a 63.09a 66.14b

MUFA 64.03a 63.67b 63.77b 66.76c

Polyunsaturated fatty acids (PUFA)t-Linoleic (C18:2) 0.15a 0.14a 0.14a 0.12b

cis-Linoleic (C18:2) 9.31a 9.39a 9.41a 9.12b

-Linolenic (C18:3. n3) 0.32a 0.32a 0.33a 0.27b

PUFA 9.78a 9.85a 9.88a 9.51b

Mean reported of duplicate analyses from 3 technological replications.T0 = 0 d; T5 = 90 d; T10 = 180 d; T20 = 360 d.Values with different superscript letters within the same row are significantly different(P < 0.05).

suggest that high oleic sunflower oil can be used to replace butterin shortbread dough to obtain LSFS.Alteration of the shortbread recipe can affect the workability

of the dough. In fact, the dough substituted with high oleic sun-flower oil was more oily and adhesive than the traditional dough(Baltsavias and others 1999). The workability was increased byadding a significant amount of flour during kneading and storingat a cool temperature before lamination. This suggests the watercontent should be reduced when the recipe is scaled up.Temperature is one of the main factors influencing lipid oxida-

tion that leads to the formation of off-flavor compounds that areunpalatable to the consumer (Calligaris and others 2007). There-fore, the LSFS were tested in an accelerated shelf-life test duringwhich fatty acid profiles were monitored. MDA content was alsoperformed to assess if MDA can be used as a marker for oxidativedamage in shortbread (Bergamo and others 1998).

Figure 1 shows the changes in peroxide values of the LSFS 60and the LSFS 70 stored at 55 C during the accelerated shelf-lifetest. Peroxide values increased during storage, as expected (Cal-ligaris and others 2007). At baseline (T0) the 2 samples showedequivalent peroxide values, which increased at T5 (which corre-sponds to 90 d at room temperature) and then it remained constantuntil T20 (corresponding to 360 d at room temperature). At T5,T10, and T20 the peroxide values of LSFS 70 were higher thanLSFS 60, as expected given the higher content of unsaturatedfatty acids. The fatty acid profiles of the 2 shortbreads did notvary during the test as reported in Tables 5 and 6. These tablesshow that, during the ASLT of the LSFS 60, most of the fattyacids did not change significantly (P < 0.05). A nonsignificantdecrease in lauric (C12:0), capric (C10:0), t-linoleic (C18:2), and-LNAs (C18:3, n3) was observed at T20. On the other hand,a nonsignificant increase in stearic acid (C18:0) was observed atT20. Similarly, LSFS 70 also showed nonsignificant decreases in allfatty acids except for stearic acid (C18:0) at T20.Figure 2 shows the changes in MDA content in the shortbreads

during the ASLT. The results showed no significant variations inthe amount of MDA during the storage time. While not signifi-cant, the higher MDA values at T20 in the LSFS 70 correspond asexpected to their higher peroxide value. Taken together the per-oxide values, fatty acid profiles and the MDA contents suggest thepartial replacement of butter with high oleic sunflower oil at thecurrent levels does not negatively influence the chemical stabilityof shortbread.Two discriminating tests were performed to compare the short-

breads. Panel members were asked to identify the correct identitiesof the TS compared with the substituted biscuits. The tests showedthat the LSFS 60 and LSFS 70 were perceived equal to the TS;only 13 panelists correctly discriminated between TS and LSFS 60,and only 7 between LSFS 70 and TS. Given that the sample of 33panel members had a fixed = 0.05, 17 positive responses wererequired to conclude that the samples are perceptibly different.Therefore, the replacement of butter with high oleic sunflower oildid not introduce a sensorial difference in this study.After observing that the new formulations lowered SFA content

while maintaining quality and storage parameters similar to thoseof TS, we decided to transfer the recipe to a pilot scale.The LSFS produced at the pilot plant was subjected to the

ASLT and sensory evaluation to compare the LSFS 70 with the

Figure 2MDA content in the LSFS 60 andLSFS 70 during the accelerated aging tests.Number of replications: 3LSFS 60: low saturated fatty acids shortbread(60% high oleic sunflower oil)LSFS 70: low saturated fatty acids shortbread(70% high oleic sunflower oil)T0: 0 d; T5: 90 d; T10: 180 d; T20: 360 d.

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Table 7Sensory evaluation scores of TS and LSFS 70 at T0 andT20.

ColorA HardnessB OilinessC SweetnessC SaltinessC FlavorC

TS T0 2.95a 2.43a 2.68a 2.88a 2.33a 3.45a

LSFS 70 T0 3.40b 3.33b 3.12a 3.23a 1.93a 1.85b

TS T20 2.57c 2.95c 2.92c 3.45c 2.78c 3.23c

LSFS 70 T20 2.72c 3.85d 2.93c 3.37c 2.27c 2.63d

n = 6; T0 = 0 d; T20 = 360 d.Pairs (TS T0 compared with LSFS 70 T0 and TS T20 compared with LSFS 70 T20),with the different letters within the same column are significantly different (P < 0.05).TS: traditional shortbread.LSFS 60: low saturated fatty acids shortbread (60% high oleic sunflower oil).LSFS 70: low saturated fatty acids shortbread (70% high oleic sunflower oil).AColor: 0 = much too light, 1 = too light, 2 = slightly light, 3 = just about right, 4 =slightly too dark, 5 = too dark, 6 = much too dark.BHardness: 0 = much too soft, 1 = too soft, 2 = slightly soft, 3 = just about right, 4 =slightly hard, 5 = too hard, 6 = much too hard.COiliness, sweetness, saltiness and flavor: 0 = much too weak, 1 = too weak, 2 = slightlyweak, 3 = just about right, 4 = slightly strong, 5 = too strong, 6 = much too strong.

Table 8Consumer evaluation scores for LSFS 70 at T0.

OverallAppearancea Texturea Flavora Acceptancea

LSFS 70 at T0 3.82 0.70 3.89 0.83 3.67 0.63 3.95 0.58Values are presented as means standard deviations.LSFS 70: low saturated fatty acids shortbread (70% high oleic sunflower oil). T0: 0 d.aAppearance, texture, flavor, overall acceptance: 1 = dislike extremely, 2 = dislikemoderately, 3 = neither like nor dislike, 4 = like moderately, and 5 = like extremely.

TS at T0 and T20. Table 7 shows the results of the quantitativedescriptive analysis for LSFS 70 compared with TS at T0. Studentst-tests indicated that the LSFS 70 was significantly different fromTS (P < 0.05) with respect to color, hardness, and flavor, howeverthey were not significantly different in terms of oiliness, sweetness,and saltiness. At the end of the accelerated shelf-life test, T20, theLSFS 70 was significantly different compared to TS in hardness andflavor. While both shortbreads showed normal chromatic decay atT20, the LSFS 70 was reported by some panelists to exhibit afry flavor. Other panelist comments described the butter flavorof TS more positively and pleasurable than the LSFS 70 flavor,but this result is acceptable for a new formulation and has beenpreviously reported (Forker and others 2012; Tarancon and others2013). Our data indicate that 60% butter replacement with higholeic sunflower oil and water could be better for the flavor ofshortbread. Regardless, the consumer sensory analysis of the LSFS70 resulted in scores ranging from 3.67 to 3.95 for all descriptors,with an overall acceptance of 3.95 (Table 8). The results indicatedthat the LSFS 70 could be accepted by the consumers as a newformulation for shortbread biscuits.

ConclusionsHigh oleic sunflower oil can be used to produce shortbreads

with low levels of SFA, high levels of PUFA and MUFA, andgood stability. The proposed formulations allow the use of thefollowing claim to be included on the label per EFSA regulations:low or reduced saturated fat (hard fat) or replacement of saturatedfat with MUFA, PUFA (soft fat) low cholesterol.Another benefit is that because butter contains TFA, the use

of high oleic sunflower oil to replace butter can also reduce TFAcontent in shortbread. One drawback of the oil substitution wasthat it produced a less consistent, more oily and adhesive doughthan traditional dough, but this issue was resolved by adding ahigher concentration of flour during kneading and storing at a

cool temperature before lamination. The results of sensory analysisshowed that the TS and the LSFS 70 have similar sensory profiles,and the consumer tests indicated that the LSFS 70 was palatable.

AcknowledgmentsWe thank the ItalianMinistry of Agricultural, Food and Forestry

Policies for financial support of the project CERSUOM, and Dr.Valerio Colasante for his collaboration on the laboratory activity.

ReferencesAllman-Farinelli MA, Gomes K, Favaloro EJ, Petocz P. 2005. A diet rich in high-oleic acid

sunflower oil favorably alters low-density lipoprotein cholesterol, triglycerides and factor VIIcoagulant activity. J Am Diet Assoc 105(7):10719.

American Oil Chemists Society 1984. Official methods and recommended practices of theAOCS. In: Walker RC, editor. Champain, Ill.: AOCS.

Arimi JM, Duggan E, OSullivan M, Lyng JG, ORiordan ED. 2010. Effect of water activity onthe crispiness of a biscuit (crackerbread): mechanical and acoustic evaluation. Food Res Int43:16505.

Association of Official Analytical Chemists(AOAC). 1980. Official methods of analysis. In:Horwitz E, editor. Washington, DC: AOAC.

Baltsavias A, Jurgens A, Van Vliet T. 1999. Fracture properties of short-dough biscuits: effect ofcomposition. J Cereal Sci 29:23544.

Bergamo P, Fedele E, Balestrieri M, Abrescia P, Ferrara L. 1998. Measurement of malondialde-hyde levels in food by high-performance liquid chromatography with fluorometric detection.J Agric Food Chem 46(6):21716.

Bravi E, Perretti G, Buzzini P, Della Sera R, Fantozzi P. 2009. Technological steps and yeastbiomass as factors affecting the lipid content of beer during the brewing process. J Agric FoodChem 57:627984.

Calligaris S, Manzocco L, Kravina G, Nicoli MC. 2007. Shelf-life modeling of bakery productsby using oxidation indices. J Agric Food Chem 55(5):20049.

Caponio F, Summo C, Paradiso VM, Pasqualone A, Gomes T. 2009. Evolution of the oxidativeand hydrolytic degradation of biscuits fatty fraction during storage. J Sci Food Agric 89:13926.

Cervenka L, Brozkova I, Vytrasova J. 2006. Effects of the principal ingredients of biscuits uponwater activity. J Food Nutr Res 45:3943.

Chevallier S, Colonna P, Della Valle G, Lourdin D. 2000. Contribution of major ingredientsduring baking of biscuit dough system. J Cereal Sci 31:24152.

Dogan IS, Javidipour I, Akan T. 2007. Effect of interesterified palm and cottonseed oil blendson cookie quality. Int J Food Sci Technol 42(2):15764.

European Community, 1991, Regulation 2568/91. Official Journal of the European Commu-nities 1991, L 248.

European Food Safety Authority. 2010. Scientific opinion on dietary reference values for fats,including saturated fatty acids, polyunsaturated fatty acids, monounsaturated fatty acids, transfatty acids, and cholesterol. EFSA J 8(3):1461568.

European Food Safety Authority. 2011. Scientific Opinion on the substantiation of healthclaims related to the replacement of mixtures of saturated fatty acids (SFAs) as present infoods or diets with mixtures of monounsaturated fatty acids (MUFAs) and/or mixtures ofpolyunsaturated fatty acids (PUFAs), and maintenance of normal blood LDL-cholesterolconcentrations (ID 621, 1190, 1203, 2906, 2910, 3065) pursuant to Article 13(1) of Regulation(EC) No 1924/2006. EFSA J 9(4):206987.

Forker A, Zahn S, Rohm H. 2012. A combination of fat replacers enables the production of fat-reduced shortdough biscuits with high-sensory quality. Food Bioprocess Technol 5(6):2497505.

Handa C, Goomer S, Siddhu A. 2010. Performance and fatty acid profiling of interesterifiedtrans free bakery shortening in short dough biscuits. Int J Food Sci Technol 45(5):10028.

Kinter M. 1995. Analytical technologies for lipid oxidation product analysis. J Chromatogr B671:22336.

Labuza TP, Dugan L. 1971. Kinetics of lipid oxidation in foods. CRC Crit Rev Food Technol2(3):355405.

Laguna L, Varela P, Salvador A, Sanz T, Fiszman SM. 2012. Balancing texture and others sensoryfeatures in reduced fat short-dough biscuits. J Texture Stud 43:23545.

Maache-Rezzoug Z, Bouvier JM, Allaf K, Patras C. 1998. Effect of principal ingredients onreological behavior of biscuit dough and on quality of biscuit. Food Eng 35:2342.

Manley D. 2000. Secondary processing. Technology of biscuits, crackers and cookies. 3rd ed.Cambridge: Woodhead Publishing Limited, CRC Press. p 42756.

Manohar RS, Rao PH. 1999. Effect of emulsifiers, fat level and type on the rheological charac-teristics of biscuit dough and quality of biscuits. J Sci Food Agric 79(10):122331.

Mckevith B. 2005. Nutritional aspects of oilseeds. Nutr Bull 30:1326.Merrill LI, Pike OA, Ogden LV, Dunn ML. 2008. Oxidative stability of conventional and

high-oleic vegetable oils with added antioxidants. J Am Oil Chem Soc 85:7716.Micha R, Mozaffarian D. 2010. Saturated fat and cardiometabolic risk factors, coronary heart

disease, stroke, and diabetes: a fresh look at the evidence. Lipids 45(10):893905.Preeti NK, Sudesh J, Rajni G. 2007. Fatty acid composition and physico-chemical characteristics

of cooking oils and their blends. J Dairy Foods Home Sci 26(3/4):2028.Rababah TM, Al-Mahasneh MA, Ereifej KI. 2006. Effect of chickpea, broad bean, or isolated

soy protein additions on the physicochemical and sensory properties of biscuits. J Food Sci 71(6):43842.

Ray G, Husain SA. 2002. Oxidants, antioxidants and carcinogenesis. Ind J Exp Biol 40(11):121332.

Reed KA, Sims CA, Gorbet DW, OKeefe SF. 2002. Storage water activity affects flavor fade inhigh and normal oleic peanuts. Food Res Int 35:76974.

Regnicoli GF, Falconi C, Perretti G. 2011. 10 Congresso Italiano di Scienza e Tecnologia degliAlimenti (Ciseta) Sostituzione parziale di grassi saturi nei biscotti. 910 Maggio 2011. Milan,Italy.

Robertson Gordon L. 1993. Food packaging: principles and practices, chapter 13. New YorkN.Y.: Marcel Dekker, Inc.

C474 Journal of Food Science Vol. 79, Nr. 4, 2014

C:Fo

odCh

emist

ry

Low saturated fatty acid shortbread . . .

Sakai T, Kawahara S. 2005. Malonaldehyde content in some breads and its change during storage.J Food Hyg Soc Jpn 46(3):1213.

St. Angelo A J. 1996. Lipid oxidation in foods. Crit Rev Food Sci 36(3):175221.Taracon P, Salvador A, Sanz T. 2013. Sunflower oil-water-cellulose ether emulsion as trans-fatty

acid-free fat replacers in biscuits: texture and acceptability study. Food Bioprocess Technol6(9):238998.

Tateo F. 1969. Gli sfarinati di frumento. In: Chiriotti, editor. Analisi dei prodotti alimentari.Vol. 1, Torino: Giuseppini Publishing. p 24771

Zoulias E, Kounalaki E, Oreopoulou V. 2002. Effect of fat and sugar replacement on cookieproperties. J Sci Food Agric 82:163744.

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