ORIGINAL PAPER

Effect of Formulation on Physicochemical Propertiesand Water Status of Nutritionally Enriched Fresh Pasta

Eleonora Carini & Elena Curti & Elisabetta Spotti &Elena Vittadini

Received: 6 July 2010 /Accepted: 14 November 2010 /Published online: 1 December 2010# Springer Science+Business Media, LLC 2010

Abstract A standard fresh pasta formulation (STD, thecontrol sample) was modified by introducing soy and carrotingredients both in dry and in liquid forms (soy and carrotflour and soy milk and carrot juice) to obtain eightnutritionally enriched fresh pasta samples with differentformulations. The effect of formulation on selected phys-icochemical properties and water status of fresh pasta werestudied. Colour, texture (force at rupture and extensibility),and cooking loss were found significantly affected by theformulation. Soy and carrot decreased the force at ruptureand extensibility of fresh pasta and increased the solids lossduring cooking. Improper gluten network development dueto either a steric hindrance of soy and carrot solids orimproper water availability for gluten hydration due todifferent water–solid interaction developed were hypothe-sized. Soy and carrot ingredients significantly altered thewater dynamics in the pasta matrix at different space-timelevels (macroscopic, moisture content and water activity;

macromolecular, frozen water content; molecular, protonnuclear magnetic resonance relaxometry) of fresh pasta in amanner dependent upon the physical state of the addedingredient. Soy flour increased both the frozen watercontent and the overall proton mobility (1H FID, 1H T1and T2) of fresh pasta while these parameters did notmarkedly differed from STD when soy milk was used. Thepresence of both carrot flour and carrot juice decreasedsignificantly the frozen water content of fresh pasta but, at amolecular level, carrot flour altered the proton molecularmobility, while carrot juice did not.

Keywords Fresh pasta . Functional ingredients .Waterstatus . Physicochemical properties . Formulation . 1H NMRmobility

Introduction

Durum wheat semolina and water are the two basicingredients of traditional Italian pasta. The variety of pastashape and size, the ease of manufacture and simplicity ofcooking make pasta a global food commodity. Moreover,the relatively simple formulation and process of pastamanufacturing make it an excellent model food system tovehicle specific “nutrients” (e.g. dietary fibres, antioxidantmolecules) to the diet of a large number of individuals(Brennan 2008). Pasta is, therefore, a good candidate tobecame a “functional food”, products that affect beneficial-ly one or more target functions in the body, beyondadequate nutritional effects, in a way relevant to improvestate of health and well-being, reduction of risk of diseases,or both (Riccardi et al. 2005).

Soy and carrot, may, therefore, be of interest to enhance thenutritional value of pasta to the basic pasta recipe. Recent

Part of this work was presented at the 9th International Conference onthe Applications of Magnetic Resonance in Food Science, Reykjavik,Iceland, September 2008.

E. CariniUniversità Telematica San Raffaele Roma,Via Val Cannuta 247,00166, Rome, Italy

E. Carini : E. Curti : E. Vittadini (*)Food Technology, Department of Industrial Engineering,University of Parma,Viale G.P. Usberti 181/a,Parma 43124, Italye-mail: elena.vittadini@unipr.it

E. SpottiStazione Sperimentale per l’Industria delle conserve Alimentari,Viale Tanara, 31/a,43100, Parma, Italy

Food Bioprocess Technol (2012) 5:1642–1652DOI 10.1007/s11947-010-0476-4

experimental and epidemiological studies have providedconvincing evidence for a variety of health benefits derivedfrom soy consumption (Valachovicova et al. 2004; Hasler1998). The reasons of these evidence are due to the presenceof isoflavones (Brynin 2002), the high nutritional value ofsoy proteins (amino acid composition and digestibility,Mariotti et al. 1999), and the lower low-density lipoproteincholesterol intake when soy is used to replace dairy oranimal proteins (Hoie et al. 2007; Wilson et al. 2007). Theuse of carrot may be encouraged as a good source ofcarotenoids, α- and β-carotene in particular. Carrot con-sumption has been shown to positively influence antioxidantstatus in healthy human subjects (Bub et al. 2000).

Changes in pasta formulation due to the addition ofingredients have been associated with an alteration ofcooking and textural properties. Edwards et al. (1995)reported an increase in pasta firmness when xanthan gumwas added at levels of 1% and 2%. Tudorica et al. (2002)found increased cooking loss when pea fibre and inulinwere substituted (7.5–15%) to durum wheat flour andobserved a progressive reduction in pasta firmness withincreasing fibre concentration. Sensory analysis of soy-enriched spaghetti containing up to 35% soy flour indicatedno significant difference in flavour and texture comparedwith a control without soy while spaghetti with 50% soyflour had slightly higher beany and bitter flavours ascompared to the control (Shogren et al. 2006).

The texture of both uncooked and cooked pasta isstrongly dependent on protein content and protein quality(Autran et al. 1987; D’Egidio et al. 1990; Matsuo et al.1982). In particular, the strength of the gluten network(related to protein composition and processing conditions)is universally acknowledged to be an important conditionfor high pasta quality (Ames et al. 1999). Water is known toplay a crucial role in defining the stability and quality ofpasta: the development of a correct gluten matrix is strictlyrelated to the way water interacts with other molecules atmultiple levels of structure (Zardetto and Dalla Rosa 2006;Carini et al. 2009a; Rahman et al. 2010).

It has been previously reported that the inclusion of otheringredients (e.g. soy) in other cereal-based products (e.g.bread, tortillas) affects not only macroscopic properties ofthe product (textural and sensory) but also shorter-rangemacromolecular (observed by differential scanning calo-rimetry, DSC) and molecular (observed by nuclear mag-netic resonance, NMR) properties. In particular, a differentwater–solids interaction was reported in bread and tortillascontaining soy, with an increased proton (1H) molecularmobility (Vittadini and Vodovotz 2003; Vittadini et al.2004; Serventi et al. 2009). It is, therefore, hypothesizedthat the addition of soy and/or carrot may affect the water–solids interaction in pasta, and, consequently, the macro-scopic properties of the product.

The objective of this work was to produce nutritionallyenriched fresh pasta by incorporating ingredients with welldocumented nutritional value (soy and carrot both in solidflour and liquid milk or juice forms) in a standard pastaformulation and to study the effect of formulation onphysicochemical properties and water status of fresh pasta.

Materials and Methods

Pasta Formulation

A standard fresh pasta (STD, semolina:water = 75:25)formulation was taken as control and was then modified byintroducing ingredients with documented functional prop-erties. Eight enriched fresh pasta samples were producedand are listed below:

– Soy flour enriched (SF), soy milk enriched (SM), soyingredients enriched (S-M) fresh pastas;

– Carrot flour enriched (CF), carrot juice enriched (CJ),carrot ingredients enriched (C-J) fresh pastas;

– Soy flour and carrot juice enriched (SF-CJ) and carrotflour and soy milk enriched (CF-SM) fresh pastas.

A different/altered development of the continuous phasewas expected in the formulation in which semolina waspartially substituted with other ingredients (not containinggluten forming proteins) and therefore, wheat gluten wasadded in SF, S-M, CF, C-J, SF-CJ and CF-SM formula-tions. The pasta formulations are reported in Table 1. Carrotflour was provided by Macor Mia Prada Company (Milan,Italy) and all other ingredients were obtained from a localsupermarket.

Fresh pasta samples were produced by a domestic doughmixer (1 kg flour capacity) with planetary action (KitchenAid, St. Joseph, Michigan). Dry ingredients were mixed for20 s at speed 4, then liquid ingredients were slowly andcontinuously added while the mixing action was continued.All ingredients were then mixed for 15 min at speed 4. Thedough was then covered with a cloth and allowed to rest for5 min at 25 °C and was successively passed among groovedrollers eight times using a conventional dough sheeter(Unika Storci, Italy) to obtain pasta sheet 1.5±0.02 mmthick. Three productions of each fresh pasta type werecarried out in different days. Physicochemical characteriza-tion of fresh pasta sheets was carried out 1 h afterproduction.

Macroscopic Quality Attributes

Colour L* (Brightness), a* (redness), b* (yellowness) andthe overall colour difference (ΔE, from STD) of each pastatype were obtained (CIE 1978) using a colorimeter (CM

Food Bioprocess Technol (2012) 5:1642–1652 1643

2600d, Minolta Co., Osaka, Japan) equipped with astandard illuminate D65 using a 2° position of the standardobserver. Ten punctual colour determinations were takenfor each pasta type at each pasta production. The followingequation was used to calculate the ΔE (CIE 1978; Kumaret al. 2006; Patras et al. 2010) that is the overall colourdifference (of the L*, a* and b* colour parameters) fromSTD.

ΔE ¼ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi

L»sample � L

»STD

� �

þ a»sample � a

»STD

� �

þ b»sample � b

»STD

� �2r

Texture Force at rupture (maximum force required to shearthe sample, N) and extensibility (deformation at rupture,mm) were obtained with a two-dimensional extensibilitytest (Bejosano et al. 2005) using a TA.XT2 TextureAnalyzer (Stable Micro Systems, Goldalming, U.K.). ATA-108 Texture fixture and an acrylic sphere (2.54 cmdiameter at edges) were used; the test was conducted incompression mode at a constant speed of 3 mm/s. Texturalproperties of ten samples of each pasta type for each pastaproduction were analysed.

Cooking Loss Cooking loss (the amount of solid substancelost to cooking water) was determined according to the AACCMethod (1999). The analysis was performed in triplicate foreach fresh pasta type for each pasta production.

Water Status

Water Activity Water activity (aw) of fresh pasta sampleswas measured at 25 °C using a Decagon Aqualab meterTE8255 (Pullman, WA). Fresh pasta sheets were brokeninto small pieces immediately before water activity mea-surement. Water activity of two samples of each pasta typewas analysed in duplicate for each pasta production.

Moisture Content Moisture content (MC, g water/100 gproduct) of fresh pasta was determined from weight loss bydrying in a forced-air oven at 105 °C until constant weight.

Moisture content of two samples of each fresh pasta typewas analysed in duplicate for each pasta production.

Thermal Properties of Ice Melting Peak Thermal propertieswere measured using a differential scanning calorimeter(DSC Q 100 TA Instruments, New Castle, DE, USA),calibrated with n-dodecane. Pasta samples (5–8 mg) wereplaced into aluminium pans (Perkin Elmer, Somerset, NJ,USA), equilibrated at −50 °C and heated to 40 °C with aheating rate of 5 °C/min. The Universal Analysis Software,Version 3.9A (TA Instruments, New Castle, DE) was usedto analyse the thermograms. The thermal event observed(endothermic peak around 0 °C) was characterised forenthalpy (ΔH, J/g), onset (Ton), and offset (Toff) temper-atures of the transition.

“Frozen” water (FW) content was calculated using thefollowing equation (Baik and Chinachoti 2001; Vittadiniet al. 2004):

FW ¼ Enthalpy ice fusion� 1

latent heat ice fusion

� �

� 1

MC

� �

� 100

where:

FW = frozen water [%, g frozen water/g water]Enthalpy ice fusion [J/g product]Latent heat of ice fusion = 334 J/g iceUnfrozen water (UFW) was calculated as the difference

between sample moisture content and frozen water amount,as previously reported (Vittadini and Vodovotz 2003).

Three DSC scans of two samples of each fresh pastatype for each pasta production were analysed.

Proton Nuclear Magnetic Resonance Mobility (1H NMRMobility) 1H NMR mobility was measured by low resolu-tion (20 MHz) 1H NMR spectrometer (the miniSpec,Bruker Biospin, Milano, Italy) connected with a thermostatoperating at 25.0±0.1 °C. Approximately 3 g of samplewere placed into a 10 mm NMR tube that was then sealed

Ingredient (%) STD SF SM S-M CF CJ C-J SF-CJ CF-SM

Semolina (durum) 75.0 41.5 72.3 39.4 48.6 71.9 48.1 44.2 44.5

Whole soy flour – 24.6 – 25.2 – – – 22.1 –

Carrot flour – – – – 20.8 – 20.6 – 22.3

Wheat gluten – 3.1 – 3.2 2.8 – 2.7 3.0 3.0

Water 25.0 30.8 – – 27.8 – – – –

Defatted soy milk – – 27.7 32.2 – – – – 30.2

Carrot juice – – – – – 28.1 28.6 30.7 –

Table 1 Fresh pasta samplesformulations: control (STD),soy flour enriched (SF); soymilk enriched (SM); soy ingre-dients enriched (S-M); carrotflour enriched (CF); carrot juiceenriched (CJ); carrot ingredientsenriched (C-J); soy flour andcarrot juice enriched (SF-CJ)and carrot flour and soy milkenriched (CF-SM)

1644 Food Bioprocess Technol (2012) 5:1642–1652

with parafilm to prevent moisture loss during the NMRexperiment.

Free induction decay (FID) T2 (spin–spin relaxation timeusing a CPMG pulse sequence) and T1 (spin–latticerelaxation time, using an inversion recovery pulse se-quence) experiments were carried out. FIDs were acquiredusing a single 90° pulse, followed by dwell time of 7 μsand a recycle delay of 0.8 s. T2 was obtained with a recycledelay of 1.5 s (that must be at least ≥5T1) and interpulsespacing of 0.04 ms. T1 experiment was acquired with arecycle delay of 1.5 s, an inter pulse spacing ranging from0.1 to 1,500 ms, depending on sample’s relaxation time. T2and T1 curves were analysed as quasi-continuous distribu-tions of relaxation times using a UPEN software (Borgiaet al. 1998; Borgia et al. 2000). Duplicated analyses on twopasta samples for each fresh pasta type for each pastaproduction were carried out for a total of 4 NMR analysesfor each NMR experiment.

Statistical Analysis

Means and standard deviations (SD) were calculated withSPSS statistical software (Version 13.0, SPSS Inc., Chicago,IL, USA). SPSS was used to verify significant differences tothe parameters among fresh pasta samples produced withdifferent formulation by one-way-analysis of variance fol-lowed by least significant difference test (LSD) at p<0.05.

Results and Discussion

Soy and/or carrot products were added to a standard freshpasta formulation in the maximum amount that still allowedobtainment of an acceptable product as determined by atrained cook. Soy and carrot were added as dry (soy andcarrot flour) and liquid (soy milk and carrot juice)ingredients with the intent to understand how differentamounts of functional ingredients as well as in hydrated ordry form may alter water dynamics and physicochemicalproperties of fresh pasta. Fresh pasta formulations (Table 1)were, therefore, obtained varying the relative proportions ofdry and liquid ingredients and resulted in products withdifferent moisture contents (Table 3).

The effect of addition of soy ingredients increasedsignificantly the product protein content of the fresh pasta(i.e., from 9.0% of STD to 9.5% in SM and ≥16% in SF, S-M, and SF-CJ; calculated from product compositionreported on the ingredients label) and the effect of soy onwater state and physicochemical properties was, therefore,mainly attributed to soy proteins. On the contrary, theaddition of carrot flour and juice primarily affected thesugar and fibre content of the final product (i.e. sugars:

from 0.5% of STD to 7.8% in CF, 8.8 in C-J, and 9.3% inCF-SM; fibre: from 0.1% of STD to 4.7% in CF, 4.9 in C-J,and 5.0% in CF-SM), and, therefore, the effect of carrotingredients on water state and physicochemical propertieswas mainly attributed to these constituents.

Colour, Texture and Cooking Loss

The colour parameters of the fresh pasta samples producedfor this study are reported in Table 2 and the observabledifferences are ascribable only to the different ingredientsused in the formulations since the processing steps (mixingand lamination) were consistent among all products. All thecolour parameters measured (L*, a* and b*, Table 2) weresignificantly affected by the addition of different ingre-dients to the standard formulation. STD had L*=∼73,a*=∼0.5 and b*=∼23. Colour of the soy-enriched freshpasta samples (SF, SM and S-M samples) decreased in L*and increased in a* and b*. Addition of carrot-basedingredients (CF, CJ and C-J samples) decreased L* anddrastically increased a* and b* because of colouredpigments in those samples. The overall differences incolour, ΔE, indicated that all fresh pasta samples had adifferent colour when compared to STD sample.

Textural properties were found to be formulation depen-dent as shown in Fig. 1a. Force at rupture was 5.58±0.26 Nfor STD and significantly lower for all modified fresh pastaformulations, indicating a weaker macroscopic matrix inthese products. The presence of whole soy flour in theformulation resulted in the lowest forces at rupture (SF andS-M samples) attributable, possibly, to the higher moisturecontent of these formulations and/or to an altered/not propergluten network development. Soy proteins are known tohave a great affinity for water and to affect proper glutendevelopment (Knorr and Betschart 1978; Doxastakis et al.2002; Traynham et al. 2007). The presence of SM slightlydecreased the force at rupture of the product as compared tothe STD. SM remained significantly harder than the othersoy flour-based formulations (SF and S-M) due, possibly, tothe lower amount of soy proteins expected in the product.

When carrot ingredients were used (CF, CJ and C-J), theforce at rupture decreased, probably because of an alteredgluten matrix development due to the presence of sugarsand fibre in the recipes. Both sugars and fibre are known tohave high affinity for water (Chinachoti 1993; Wang et al.2002) that was, possibly, only partially available for glutennetwork development. Forces at rupture of CF and C-J weresignificantly higher than SF and S-M probably due to thesignificantly lower moisture content and/or lower amountof semolina substituted with carrot than with soy flour. Inthe SF-CJ and CF-SM samples the forces at rupturereflected the behaviour observed when soy flour or carrotflour were added.

Food Bioprocess Technol (2012) 5:1642–1652 1645

Extensibility (Fig. 1b) of STD was 28.7±0.8 mm,comparable to fresh pasta containing SM, while it wassignificantly increased in the presence of CJ (31.5±1.3 mm). The presence of whole soy and carrot floursdecreased the extensibility of the respective products(Fig. 1b). Similar results were reported also by Scazzinaet al. (2008) that investigated the textural properties of

enriched tortillas with whole soy flour and carrot juice incomparison with a standard tortilla.

The amount of solids residue in the cooking water hasbeen drastically affected by the changes in formulation.STD lost 3.32±0.01 g solids/100 g sample and all freshpasta types developed (with exception of SM) showed asignificant higher solids release during cooking (Fig. 2)suggesting the development of a weaker gluten network insamples containing whole soy flour and dehydrated carrot.In particular, solids loss during cooking was drasticallyhigher in the presence of carrot flour (CF, C-J and CF-SMsamples, Fig. 2).

Water Status

Water macroscopic status of fresh pasta samples wasstudied by means of water activity and moisture content.The necessity to obtain good quality products implied thevariation of the amount of water in the recipes and,therefore, the moisture content differences of pasta samples(Table 3) had to be primarily attributed to the differentamount of water added to the formulation.

STD had water activity equal to 0.975±0.001 and moisturecontent equal to 31.9±0.3% (g water/100 g sample). The

g gf

e d

b

a

c

b

STD SF SM S-M CF CJ C-J SF-CJCF-SM0

2

4

6

8

10

12cooking loss

g gf

e d

b

a

c

b

(g s

olid

s/10

0 g

sam

ple)

Fig. 2 Cooking loss for fresh pasta samples. Different letters abovethe bars indicate significant difference among fresh pasta samples(LSD test, p≤0.05)

a

f

b

f

c

d

c

e

c

b

cd

b

f

cd

a

e

cd

d

STD SF SM S-M CF CJ C-J SF-CJ CF-SM

STD SF SM S-M CF CJ C-J SF-CJ CF-SM

N

0

1

2

3

4

5

6

mm

10

14

18

22

26

30

34

force at rupture

extensibility

a

f

b

f

c

d

c

e

c

b

cd

b

f

cd

a

e

cd

d

a

b

Fig. 1 Force at rupture (a) and extensibility (b) properties for freshpasta samples. Different letters above the bars indicate significantdifference among fresh uncooked pasta samples (LSD test, p≤0.05)

Table 2 Brightness (L*, mean±standard deviation), redness (a*, mean ± standard deviation), yellowness (b*, mean ± standard deviation) and ΔE(mean ± standard deviation) of fresh pasta samples

STD SF SM S-M CF CJ C-J SF-CJ CF-SM

L* 73.3±0.6 63.2±1.2 71.4±0.6 64.3±1.0 63.2±1.7 67.3±0.8 62.6±1.4 60.5±1.7 62.3±1.1

a* 0.5±0.1 4.3±0.6 0.7±0.1 4.4±0.5 15.0±1.4 13.6±0.5 18.5±0.8 10.7±0.5 11.2±0.8

b* 23.0±0.7 24.0±1.2 24.2±1.0 23.5±1.3 32.0±1.7 42.2±1.8 32.6±1.4 33.8±2.5 31.1±1.2

ΔE Ref 11.0±1.4 2.5±0.8 9.9±1.2 20.0±2.0 24.3±1.5 23.1±1.6 19.8±2.1 17.5±1.2

1646 Food Bioprocess Technol (2012) 5:1642–1652

substitution of part of semolina with whole soy flour increasedthe amount of water needed in the formulations due to thegreat capacity to soy proteins to “bind” water molecules, aspreviously reported (Doxastakis et al. 2002; Traynham et al.2007). This fact was reflected on products’ moisture contentsthat was found to be ∼37, ∼36 and ∼33% g water/100 gsample for SF, S-M and SF-CJ, respectively. Although waterwas present in larger amounts, water activity of SF, SM andS-M was significantly lower than in the STD sample(Table 3) possibly because of “stronger” water–soy proteinsinteractions detectable by a water activity macroscopicmeasurement. The presence of sugars and fibre originatedfrom dehydrated carrot and carrot juice did not change themoisture content of samples (Table 3) with the exception ofC-J pasta that had a significantly lower moisture content(30.7±0.9% g water/100 g sample). However, carrot-basedingredients significantly reduced the water activity in CF, CJ,C-J and CF-SM samples (Table 3), possibly due to thehigher sugar content of these products.

The interaction of solids and water molecules at macromo-lecular level was studied by means of the thermal properties ofthe icemelting peak obtained byDSC. DSC thermograms of allsamples exhibited one major endothermic transition (i.e. icemelting) as the samples were heated from −50 °C to 40 °C(Fig. 3a). STD ice melting peak shape sample was comparablethat of to SF, SM and S-M samples. The formulationscontaining dehydrated carrot and carrot juice presented, on thecontrary, a broader peak indicating the presence of moreheterogeneous solids-water interactions. Transition temper-atures (Ton and Toff, Fig. 3b) of peaks were formulationdependent with Ton and Toff of STD equal to approximately−9 and 2 °C, respectively. The presence of whole soy flour inthe formulation (SF and SM) slightly shifted the transitiontoward lower temperatures and increased the temperaturemelting range indicating the presence of more heterogeneousand stronger water–solids interactions. Soy milk did not affectthe temperature transitions if compared to STD. A slightdecrease of the temperature onset in soy-containing bread waspreviously reported also by Vittadini and Vodovotz 2003.Carrot ingredients significantly shifted the ice melting peaktowards lower temperatures possibly because of the freezingpoint depression induced by sugars (Ross 1995). The frozenand unfrozen water content (FW and UFW, at the selectedDSC experimental conditions) of fresh pasta samples (Fig. 3c)were found to be formulation dependent. STD sample had aFW=38.0±0.6 g frozen water/100 g water; SOY addedproducts had FW either significantly higher (SF), lower(SM) or comparable (S-M) with STD. Carrot-based ingre-dients significantly decreased FW (∼21, ∼29 and ∼23 g frozenwater/100 g water for CF, CJ and C-J, respectively) ascompared to STD possibly due to the presence of solubilisedsugars that increased the viscosity of the hydrophilic phaseresulting in a decrease in motion of the water molecules thatT

able

3Moisturecontent(g

water/100

gsample,

mean±standard

deviation)

andwater

activ

ity(m

ean±standard

deviation)

offreshpastasamples

STD

SF

SM

S-M

CF

CJ

C-J

SF-CJ

CF-SM

MC

31.9±0.3c

37.1±0.5a

31.7±0.5c

35.8±0.5b

33.1±0.9c

32.3±0.3c

30.7±0.9d

32.5±1.4c

32.4±0.5c

a w0.97

9±0.00

1a0.97

5±0.00

5b0.97

5±0.00

5b0.97

2±0.00

1c0.93

2±0.00

2f0.97

0±0.00

0d0.92

2±0.00

1g

0.96

8±0.00

1e0.93

4±0.00

1f

Different

letters

follo

wingthestandard

deviations

indicate

sign

ificantdifference

amon

gfreshpastasamples

(LSD

test,p≤0

.05)

Food Bioprocess Technol (2012) 5:1642–1652 1647

could form ice crystals detectable by DSC (Chinachoti 1993;Vittadini et al. 2004).

Molecular mobility of fresh pasta was studied by lowresolution 1H NMR, an analytical technique that has beenproven to provide an important insight about the propertiesand dynamics of food materials (Bertram et al. 2002; Wanget al. 2004; Mariette and Lucas 2005; Vittadini and Vodovotz2003; Sereno et al. 2007; Serventi et al. 2009). 1H NNRmobility of fresh pasta was investigated with multiple lowresolution 1H NMR experiments in an attempt to covermolecular mobility ranging from ∼10 μm to ∼500 ms. Itmust be emphasised that the 1H NMR analysis is not specificfor water as the signal detected may arise from any protonpresent in the sample relaxing in the time frame character-istic of the experiment (Halle and Wennerstroem 1981;Schmidt and Lai 1991; Colquhoun and Goodfellow 1994;Ruan and Chen 2001). 1H NMR analysis allows, therefore,to study proton dynamics in the matrix. Mobility of the leastmobile 1H fractions of fresh pasta was analysed with a FIDexperiment while the more mobile protons fractions were

characterised in terms of 1H T2 and T1 relaxation timesdistributions.

Characteristic representative 1H FID curves werereported in the 7–100 μs range to ensure field homogeneityand are shown in Fig. 4a. STD, SM and CJ samples showeda comparable decay (Fig. 4a and b) indicating the presenceof a similar fast relaxing solid-like 1H population. On thecontrary, the other samples (SF, S-M, CF, C-J, SF-CJ, CF-SM) were found to have a more mobile rigid protonsfraction (comparable among them) as indicated by the lesssharp decay of the representative curves, if compared withSTD sample. The fast relaxing 1H population observed withthe FID might arise from protons in solid-like components,such as starch and proteins and water molecules tightlyassociated with those solids (Kim and Cornillon 2001). Thepresence of soy and carrot flours increased the mobility ofthe solid-like protons because of a possibly altered water–solid interaction and/or a water molecules displacementtowards different domains resulting in different molecularmobility.

0

20

40

60

80

100

-40 -30 -20 -10 0 10 20-5

-4

-3

-2

-1

0g

“fro

zen

wat

er”/

100

g w

ater

UFW

FW

-40

-30

-20

-10

0

10

20

Ton

Toff

temperature transition (˚C)

temperature (˚C)

heat

flo

w (

W/g

)

SF SM S-M CF CJ C-J SF-CJ CF-SMSTD

SF SM S-M CF CJ C-J SF-CJ CF-SMSTD

ba

c

Fig. 3 Thermal properties of ice melting peak from differentialscanning calorimetry, DSC. a Characteristic thermograms of STDsample in the −40–20 °C temperature range; b temperatures transition

of ice melting peak for fresh pastas; c frozen water content (FW) andunfrozen water content (UFW) for fresh pastas

1648 Food Bioprocess Technol (2012) 5:1642–1652

1H T2 and T1 experimental curves were analysed as quasi-continuous distributions of relaxation times and the results arereported in Fig. 5. Characteristic 1H T2 distribution of STDindicated the presence of two well–resolved 1H populations(Fig. 5a) relaxing in the ∼0.1–∼1.5 ms range (peak centredat ∼0.17 ms; ∼20% of the total detectable protons) and inthe ∼1.6–∼87 ms range (peak at 6.4 ms; ∼80% of the totaldetectable protons). These two 1H populations were tenta-tively attributed to 1H of the gluten domain and starchdomain, respectively, based on the findings of 1H NMR lowresolution reports carried out on bread, tortillas and modelsystems containing starch and gluten (Doona and Baik 2007;Wang et al. 2004; Engelsen et al. 2001). Similar results werepreviously reported in fresh pasta (Carini et al. 2009a, b).

SM and CJ 1H T2 distributions were similar with theSTD (Fig. 5a) but the shape of the slower relaxing 1H

population (centred at ∼6.4 ms) was broader than in STD.These differences indicated the presence of protons relaxingover a large range of relaxation times suggesting a highermobility heterogeneity of the protons belonging to thispopulation. The 1H molecules dynamics of SM and CJwere overall comparable to STD. The presence of wholesoy flour in the formulations (SF, S-M and SF-CJ samples)resulted in more complex 1H T2 relaxation line-shapessuggesting the presence of three peaks originating fromthree not completely resolved and strongly exchanging 1HT2 populations (Fig. 5). The prevalent population wasshifted towards slower relaxation times (∼10 ms) andoverlapped with another minor 1H population that may beassociated to soy proteins contribution (Serventi et al.2009). Fresh pasta enriched with dehydrated carrot (CF andC-J) presented a 1H T2 complex line-shape originating fromat least two overlapping 1H populations exchanging withinthe NMR T2 time frame.

As previously reported, the fastest 1H population wasattributed by some authors to protons belonging to thegluten domain (Engelsen et al. 2001). It might bespeculated, therefore, that the presence of a not resolvedpeak in the soy- and carrot-flour-containing pastas couldbe related to the lack of a proper gluten networkdevelopment and it could bear out the attribution of theseprotons to those associated to the gluten phase. Further

a

SFS-MSSTDSMno

rmal

ized

inte

nsity

b

CFC-J

CJ norm

aliz

ed in

tens

ity

STD

c

CF-SM

STD

SF-CJ

norm

aliz

ed in

tens

ity

Fig. 4 1H FID for fresh pasta samples. a STD vs. soy ingredients-based samples; b STD vs. carrot ingredients-based samples; c STD vs.SF-CJ and CF-SM

1 1a bHT2 HT1

CF-SM

SF CJSF-CJ

C JC-J

CJCJ

CFCF

S-M

SM

SF

STD

T2 (ms) T2 (ms)

Fig. 5 1H T2 distribution (a) and 1H T1 distribution (b) for fresh pastasamples

Food Bioprocess Technol (2012) 5:1642–1652 1649

experimental evidence must be created for a definiteattribution of the fastest relaxing 1H population to gluten,but it is interesting to notice that this signal becomes lessdefined and less resolved when the product formulationreduces the possibility of the formation of a good glutennetwork (e.g. increase in non gluten forming ingredients inthe formulation).

The 1H T1 distribution was unimodal in all fresh samples(Fig. 5b) indicating that protons were in “fast exchange”regime in the NMR experimental time window. STD hadthe more mobile 1H T1 population peaked at ∼86 ms. Thepresence of soy ingredients added to the standard formu-lation decreased the mobility of T1 population (∼74, ∼79and ∼72 ms for SF, SM and S-M, respectively) and,especially, broadened the peak (less markedly in SM,Fig. 5b) indicating a more heterogeneous relaxation processin this time frame. Also the presence of carrot ingredientsdecreased 1H T1 (∼64, ∼73 and ∼51 ms for CF, CJ and C-J,respectively), while peak width remained relatively narrow(with the exception of C-J). CF-SM and SF-CJ had 1H T1equal to ∼70 and ∼55 ms, respectively. Inclusion ofingredients with great affinity for water (soy proteins,sugars and fibre) modified the water dynamics (as alsoshowed with other water status parameters) and probablydecreased the intermolecular spacing as the lower 1H T1measured when the standard formulation was modified. Thepresence of soy flour increased the peak size increasing theheterogeneity of the separations between nuclei and themolecular structure suggesting that soy proteins, besidescausing higher 1H T2 mobility, could be able to developinteraction with water at different molecular mobilityranges.

Conclusions

Fresh pastas samples enriched with ingredients of welldocumented nutritional functionality (soy and carrot) weredeveloped and the effect of formulation on physicochemicalproperties and water status were studied. Different for-mulations significantly affected macroscopic propertiessuch as colour, texture and cooking loss.

The pasta formulations in which semolina was partiallysubstituted with soy and carrot (both in solid or liquid form)decreased the force at rupture and the extensibility andincreased the solids released during cooking, if comparedwith STD. These macroscopic phenomena may be due tothe formation of a weaker and less elastic gluten structure.The presence of components such as soy proteins; carrotsugars and fibre may have either sterically hindered thegluten proteins interaction or altered water distribution anddynamics in the pasta dough leading to improper glutenhydration.

The macroscopic properties of fresh pasta measured inthis study can also affect the texture of cooked pasta thatmay result in different quality than a cooked pasta obtainedwith a standard formulation and, therefore, make it moresuitable for particular applications or to satisfy the specificrequest of different consumers.

The water status was also significantly affected by theformulation either at macroscopic (water activity), macro-molecular (frozen water content) or molecular (1H NMRmobility) level. In particular, soy and carrot significantlydecreased water activity of fresh pasta regardless of theproduct moisture content. Macromolecular and molecularwater status was, on the contrary, significantly differentwhen soy and carrot were added. Moreover, a dependenceupon the physical status of the functional ingredient addedwas found. In particular, soy flour increased both the frozenwater content and the overall 1H NMR mobility of freshpasta while the macromolecular and molecular water statusof SM pasta did not significantly differ from STD. Thepresence of both carrot flour and carrot juice caused asignificant decrease in the frozen water content of freshpasta but, at a molecular level, carrot flour altered theproton molecular mobility (e.g. slower 1H FID decay andlack of 1H T2 peak at 0.17 ms), while pasta containingcarrot juice did not differed from the STD.

Soy and carrot ingredients were found to significantlyalter the water dynamics at different space-time levels offresh pasta, underlying that also small changes in formula-tion may significantly affect properties of food materials.The role of each water status parameter in defying not onlythe physicochemical but also the microbial and nutritionalattributes and stability of fresh pasta should be object offurther investigation in fresh pasta as well as in a largervariety of food products.

Acknowledgement The authors would like to thank Sandro Salardifor his precious help in optimising pasta formulations and for carryingout part of the experiments.

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