Functional and quality profile evaluation of butters, spreadable fats, and shortenings available from Czech market

Ci a ion: Lapˇcíko á, B.; Lapˇcík, L.; Valen a, T.; Kuˇce o á, T. Func ional and Quali y P o ile E alua ion o Bu e s, Sp eadable Fa s, and Sho enings A ailable om Czech Ma ke . Foods 2022,11, 3437. h ps://doi.o g/10.3390/ oods11213437 Academic Edi o : C is ina Alamp ese Recei ed: 5 Oc obe 2022 Accep ed: 26 Oc obe 2022 Published: 29 Oc obe 2022 Publishe ’s No e: MDPI s ays neu al wi h ega d o ju isdic ional claims in published maps and ins i u ional a il- ia ions. Copy igh : © 2022 by he au ho s. Licensee MDPI, Basel, Swi ze land. This a icle is an open access a icle dis ibu ed unde he e ms and condi ions o he C ea i e Commons A ibu ion (CC BY) license (h ps:// c ea i ecommons.o g/licenses/by/ 4.0/). oods A icle Func ional and Quali y P o ile E alua ion o Bu e s, Sp eadable Fa s, and Sho enings A ailable om Czech Ma ke Ba bo a Lapˇcíko á, Lubomí Lapˇcík * , Tomáš Valen a and Te eza Kuˇce o á Facul y o Technology, Depa men o Food Technology, Tomas Ba a Uni e si y in Zlín, Nám. T. G. Masa yka 5555, 760 01 Zlin, Czech Republic *Co espondence: [email p o ec ed]; Tel.: +420-576-035-115 Abs ac : The aim o his s udy was o assess he unc ional p ope ies o bu e s, sp eadable a s, and sho enings, collec ed om he Czech ma ke , in co ela ion wi h hei nu i ional alues decla ed by he p oduce s. Va ious me hods we e applied o de e mine ele an pa ame e s o he p oduc s. Using pene a ion es s, samples we e cha ac e ized by speci ic ex u al a ibu es acco ding o hei com- posi ion and p ocessing ype, pa icula ly o he p esence o milk/ ege able a s. Using di e en ial scanning calo ime y (DSC), he mal peaks co esponding o medium- and high-mel ing iacylglyc- e ol ac ions we e de ec ed in he anges 15–16 ◦ C and 31.5–34.5 ◦ C, espec i ely. Rheological analysis e ealed ha he iscoelas ici y o samples was ela ed o equency beha io o he a s uc u e, cha ac e ized by he dominance o elas ic modulus (G 0 ) o e iscous modulus (G 00 ) up o he equency o 10 Hz. This indica ed good emulsion s abili y o he p oduc s in he egion o linea iscoelas ici y. Fo sp eadable a s, he s uc u e was esis an o phase sepa a ion in he whole equency ange unde s udy (0.1–100 Hz). The esul s showed ha he applied echniques can be success ully used o cha ac e ize he p ocessing and composi ional quali y o bu e s and ege able a s. Keywo ds: bu e s; sp eadable a s; sho enings; ex u e p o ile analysis; ee a y acids; di e en ial scanning calo ime y; heology; luo escence spec ome y 1. In oduc ion Bu e s and dai y al e na i es, such as sp eadable a s and sho enings, a e in ol ed in many ood applica ions. Milk a is a main componen o bu e p oduc s. O e a wide ange o empe a u es, milk a is a mix u e o solid c ys als connec ed in a ne wo k and embedded in a liquid phase. S uc u al and composi ional cha ac e is ics o hese c ys als e ol e as unc ion o empe a u e and ime. C ys alliza ion p ope ies and polymo phism o milk a in bu e and ege able a s in al e na i e p oduc s can be a ec ed by many ac o s, e.g., sample dispe sion s a e, cooling a e, shea applied du ing he p ocessing, p esence o mino lipid compounds ( ee a y acids, phospholipids, mono- and diacylglyce ols), and iacylglyce ols (TAG) composi ion [1]. Milk (c eam) om di e en sou ces can be u ilized o he o ma ion o bu e s and ela ed p oduc s. As a low-cos and e icien al e na i e o bu e s, sp eadable a s and sho enings a e p oduced. They a e cha ac e ized by speci ic mix u es o ege able a s/oils and hei p ocessing. Cu en ly, sp eadable a s a e p oduced by anses e i ica ion o iacylglyce ols. Sho enings a e p epa ed by con olled c ys alliza ion o a subs ance, whe eas he dispe sion o TAG c ys als is o med (ideally in he o m o spa ial ne wo k), and he dispe sion medium is c ea ed by liquid iacylglyce ols [2]. C ys alliza ion o milk a globules du ing aging o c eam plays an impo an ole in bu e ’s unc ional p ope ies. The numbe and size o c ys als in c eam can be adjus ed by con olling he empe a u e and ime o he aging p ocess and by empe a u e luc ua ions be o e bu e chu ning. Rapid cooling can cause a la ge quan i y o small c ys als in he c eam, which can p oduce a i m bu e , whe eas slow cooling o a long ime may yield a so bu e [3–5]. Foods 2022,11, 3437. h ps://doi.o g/10.3390/ oods11213437 h ps://www.mdpi.com/jou nal/ oods Foods 2022,11, 3437 2 o 15 A con enien and as echnique, which enables de e mining he composi ion and c ys alliza ion p o ile o bu e s and ege able a s, is di e en ial scanning calo ime y (DSC), acili a ing he analysis o a ious ood componen s. DSC is nei he expensi e no labo -in ensi e, and i can be applied in many kinds o ood quali y e alua ions, including assessmen o milk a adul e a ion by ege able oils o milk a ac ion cha ac e iza ion. DSC s udies o milk and ege able a mel ing and c ys alliza ion in ol e he measu ing o eleased o abso bed hea (no malized in J/g) a ele an hea ing/cooling a es. The p inciple o analysis lies in he ac ha e e y a has a unique composi ion o a y acids and iacylglyce ols, which a e mani es ed in cha ac e is ic mel ing and c ys alliza ion cu es [6–8]. The he mal beha io o milk a , like all a s, is a di ec e lec ion o he s o age condi- ions unde which he a has been c ys allized. I is ela i ely easy o se up a me as able c ys alline s a e ( β0 modi ica ion) in milk a , which pe sis s un il he empe a u e is aised o he mel ing poin o he me as able o m. Repea ed empe a u e cycling o bu e p od- uc s om e ige a ed o oom condi ions allows he con e sion o a me as able s a e o a mo e s able o m ( β ), being ad e sely a ec ed in he p ocess o sp eadabili y [ 9 ]. Fo sp eadable a s, small β0 c ys als a e a o ed since hey enable he o ma ion o a good c ys al ne wo k h oughou he con inuous lipid phase which is ela ed o desi able senso y and ex u al p ope ies [10]. Tex u e is one o he signi ican quali y a ibu es o edible a s. I is based on he a ’s TAG p o ile and a y-acid con en [ 11 ]. Tex u al pa ame e s a e associa ed wi h he sp eadabili y, appea ance, and consume accep abili y o he p oduc s. Bu e ex u e can be a ec ed by di e en p ocessing me hods and ea men s. I is a ibu ed o he h ee- dimensional ne wo k o a c ys als in a con inuous oil phase. The ex en o c ys alliza ion and he a io o solid o liquid a a e he p ima y de e minan s o bu e consis ency and o he ex u al pa ame e s [4,12]. In his s udy, he e ec o p ocessing ype and composi ion on unc ional p ope - ies o bu e s and ege able a s was examined. In pa icula , we ocused on ex u al, heological, c ys alliza ion, and mel ing p ope ies, in es iga ed using a combina ion o sui able echniques. The aim o his s udy was o access he p ope ies and quali y o ma ke bu e s, sp eadable a s, and sho enings om a ious unc ional aspec s, and o p esen alida ion and p oduc cha ac e iza ion. The applied me hods we e simul aneously u ilized o e alua e he ele an pa ame e s o bu e s and bu e al e na i es o de e mine he co ela ion be ween he da a decla ed by he p oduce s and he unc ional p ope ies. On he basis o he esul s, cha ac e is ic pa ame e s o he samples could be de i ed and used o gene alize he unc ional p o ile o in es iga ed p oduc s. 2. Ma e ials and Me hods 2.1. Ma e ials Di e en comme cial b ands o bu e s, sp eadable a s, and sho enings, collec ed om he Czech ma ke in he yea 2018, we e in es iga ed in ou s udy. The p oduc s we e selec ed on a andom basis o p o ide he highes a iabili y o p ocessing ype and composi ion as possible. Samples we e p oduced in Czech Republic, excep o Roki nianka bu e om Poland. They included i e ypes o bu e s (No. 1, No. 2, No. 3, No. 4, and No. 5), h ee ypes o sp eadable a s (No. 6, No. 7, and No. 8), and wo ypes o sho enings (No. 9 and No. 10). Th ee ba ches o each sample we e es ed. Be o e he analyses, samples we e s o ed a e ige a ed empe a u es s anda dly used in he dis ibu ion chain (4 ±1◦C) o 1 week. In Table 1, he p oduc s a e cha ac e ized, including hei p oduce s, ene gy in ake, and nu i ional alues. All chemicals used in his s udy we e pu chased om ce i ied companies and we e o analy ical g ade. Cyclohexane ( ≥ 99.8%) was deli e ed by Sigma-Ald ich (USA), die hyl e he ( ≥ 99.8%) was deli e ed by Ch omse is s. .o. (Czech Republic), and e hanol absolu e ( ≥ 99.8%) was deli e ed by Pen a Chemicals Unlimi ed (Czech Republic). Po assium hy- d oxide and phenolph halein indica o we e ob ained om Lachne , s. .o. (Czech Republic). Foods 2022,11, 3437 3 o 15 Table 1. Nu i ional cha ac e is ics o bu e s, sp eadable a s, and sho enings pe 100 g o he p oduc s. Nu i ional Values/Ene gy In ake pe 100 g P oduc s Name, P oduce Roki nianka Bu e (No. 1), Mlecza ska Roki nianka Olomoucké Bu e (No. 2), OLMA, a.s. Ta a Fa mᡠské Bu e (No. 3), Mléká na Hlinsko, a.s. ˇ CeskáChu ’ Bu e (No. 4), OLMA, a.s. D . Halíˇ Bu e (No. 5), Mléká na ˇ Ceje iˇcky, spol. s .o. Ene gy in ake (kJ/kcal) 3060/744 3095/739 3134/762 3095/739 3056/743 To al a (g) a (o which sa u a ed a y acids) 82.1 (57.4) 83.0 (51.0) 84.0 (55.0) 83.0 (51.0) 82.0 (52.0) Saccha ides (g) (o which monosaccha ides) 0.6 (0.6) 0.8 (0.8) 0.8 (0.8) 0.8 (0.8) 0.7 (0.7) P o eins (g) 0.7 0.6 0.7 0.6 0.6 Sal (g) 0.02 0.02 0.10 0.02 0.02 Vi amin A (µg) - - - - - Vi amin D (µg) - - - - - Vi amin E (mg) - - - - - Pe la (No. 6), UNILEVER ˇ CR, spol. s .o. Rama Classic (No. 7), UNILEVER ˇ CR, spol. s .o. Flo a (No. 8), UNILEVER ˇ CR, spol. s .o. He a (No. 9), UNILEVER ˇ CR, spol. s .o. Zla áHaná (No. 10), OLMA, a.s. Ene gy in ake (kJ/kcal) 1450/346 2241/535 1673/405 2666/637 2762/659 To al a (g) a (o which sa u a ed a y acids) 39.0 (10.0) 60.0 (24.0) 45.0 b (10.0) 72.0 (34.0) 74.0 c (35.0) Saccha ides (g) (o which monosaccha ides) 0 (0) 0.5 (0.5) <0.5 (<0.5) <0.5 (<0.5) 0.8 (0.8) P o eins (g) 0 <0.5 <0.5 <0.5 0.6 Sal (g) 0.32 0.23 0.52 0.21 0.02 Vi amin A (µg) 800 (100%) d800 (100%) d800 (100%) d800 (100%) d- Vi amin D (µg) 7.5 (150%) d7.5 (150%) d7.5 (150%) d7.5 (150%) d- Vi amin E (mg) 18 (150%) d7.5 (60%) d14 (120%) d- - a The alues o o al a a e ela ed o milk a in bu e s and ege able a s in sp eadable a s and sho enings. b Fo No. 8, he p oduce decla ed he p esence o polyunsa u a ed a y acids (PUFAs) in o al a , i.e., omega-3 and omega-6 essen ial a y acids. c Fo No. 10, he p oduce decla ed 58% ege able a s and 15% milk a pe 100 g o he p oduc . d The pe cen ages o i amins a e ela ed o he e e ence daily ene gy in ake o an a e age weigh adul pe son (8400 kJ/2000 kcal). A hyphen indica es ha no alue was decla ed by he p oduce . 2.2. Tex u e P o ile Analysis Tex u e p o ile analysis was pe o med using a TA.XT. plus Tex u e Analyze (S able Mic o Sys ems, Uni ed Kingdom). Wi h he help o a cu ing cylinde , samples o s anda d size 15 × 35 mm ( hickness × diame e ) we e p epa ed, as can be seen in Figu e 1. An aluminum cylind ical p obe SMS P/50 (o 50 mm diame e ) was used o double com- p ession o samples equilib a ed a empe a u e 15 ± 0.5 ◦ C. This empe a u e was used wi h espec o he a iacylglyce ol mel ing p ope ies [ 13 ] o achie e op imal sample pene a ion. Two measu emen modes (p obe se ings) we e applied. In he i s mode, samples we e squeezed wice o 5 mm dis ance (sample pene a ion) using a igge o ce o 10 × g. In he second mode, samples we e comp essed o 10% o hei ini ial heigh (10% s ain) using a igge o ce o 5 × g. In bo h modes, he p obe es speed was 0.5 mm/s, wi h p e- es and pos - es speeds o 1 and 10 mm/s, espec i ely. The o ce and displacemen o he p obe we e eco ded as a unc ion o ime. The samples’ ex u al pa ame e s we e de e mined. Ha dness ( i mness) (N) ep esen ed he maximum o ce ha occu ed du ing a comp ession cycle, and he Young elas ic modulus (N/s) was he modulus o sample de o mabili y calcula ed as a g adien . Sp inginess (elas ici y) (mm) was he heigh which sample eco e ed a e i s de o ma ion be ween Foods 2022,11, 3437 4 o 15 he end o he i s and beginning o he second comp ession. Adhesi eness (N) was he maximum nega i e o ce equi ed o pull he p obe away om sample, i.e., o o e come a ac i e o ces be ween he p obe and sample su ace. Cohesi eness (-) was ela ed o he s eng h o sample in e nal bonds, and i was calcula ed as he a io o a ea unde o ce– ime cu e a e he second comp ession o he a ea a e he i s comp ession. In addi ion o hese p ima y ex u al cha ac e is ics, seconda y pa ame e s we e de e mined. S inginess (mm) was he dis ance o which he sample was ex ended du ing decomp ession be o e sepa a ing om he p obe. Gumminess (N) ep esen ed he ene gy needed o disin eg a e a semisolid sample o a s a e eady o be swallowed. Gumminess was calcula ed using he ollowing equa ion: Gumminess =A2 A1 ×F1, (1) whe e A 2 is he a ea unde he o ce– ime cu e a e he second comp ession (n.s), A 1 is he a ea unde he o ce– ime cu e a e he i s comp ession (n.s), and F 1 is he peak o ce du ing he i s comp ession (n). Foods 2022, 11, x FOR PEER REVIEW 7 o 17 Figu e 1. Tex u e p o ile analysis o bu e s (A), sp eadable a s, and sho enings (B) using 5 mm dis ance pene a ion es (ba cha wi h 5% e o ba s). Tex u al pa ame e s a e desc ibed in he legend wi h he ele an uni s in he b acke s; adhesi eness is exp essed in absolu e alues. Di e - en supe sc ip le e s o he same ex u al pa ame e indica e s a is ically signi ican di e ences be ween he samples (α ≤ 0.05, Tukey es ). Figu e 1. Tex u e p o ile analysis o bu e s ( A ), sp eadable a s, and sho enings ( B ) using 5 mm dis ance pene a ion es (ba cha wi h 5% e o ba s). Tex u al pa ame e s a e desc ibed in he legend wi h he ele an uni s in he b acke s; adhesi eness is exp essed in absolu e alues. Di e en supe sc ip le e s o he same ex u al pa ame e indica e s a is ically signi ican di e ences be ween he samples (α≤0.05, Tukey es ). Chewiness (N) was conside ed as ene gy equi ed o chew a solid sample o a s a e eady o be swallowed. This pa ame e was de e mined using he ollowing equa ion: Foods 2022,11, 3437 5 o 15 Chewiness =Gumminess ×L2 L1, (2) whe e L 2 is he dis ance om he beginning o he second comp ession o he peak o ce o he second comp ession (L 2 is equal o he sp inginess) (mm), and L 1 is he dis ance om he beginning o he i s comp ession o he peak o ce o his comp ession (mm) [14–16]. 2.3. F ee Fa y Acids and Acid Numbe The ee a y acid index and acid numbe o bu e s, sp eadable a s, and sho enings we e de e mined using he i a ion me hod. B ie ly, 5 ± 0.5 g o a sample was mel ed a 30–40 ◦ C, and hen mixed wi h an app op ia e amoun (10 mL) o p e-neu alized o ganic sol en (composed o 99.8% die hyl e he and absolu e e hanol in he a io 1:1). Unde a con inuous mild hea ing in a wa e ba h, he solu ion was i a ed using a po assium hyd oxide alcoholic solu ion o s anda dized concen a ion 0.1 mol/L. Phenolph halein alcoholic solu ion (1 w .%) was used as an indica o . The i an olume measu ed du ing sample i a ion was sub ac ed om he i a ion olume o he blank sample (wi hou he a ). F ee a y acid con en (w .%) was calcula ed using he ollowing equa ion: F ee a y acids (FFA)index =V×c×M 10 ×m, (3) whe e Vis he i an olume du ing sample i a ion educed by i an olume du ing blank sample i a ion (ml), cis he concen a ion o po assium hyd oxide solu ion (mol/L), Mis he mola weigh o oleic acid (282.47 g/mol) used as a ep esen a i e a y acid, and mis he weigh o sample (g). Acid numbe is he measu e o ee a y acids amoun in he a and is exp essed as he weigh o po assium hyd oxide (mg) needed o neu alize he acids ( ep esen ed by oleic acid) con ained in 1 g o a . The ollowing o mula was used o calcula e he acid numbe (mg/g): Acid numbe =56.11 ×V×c m, (4) whe e Vis he i an olume du ing sample i a ion educed by i an olume du ing blank sample i a ion (mL), cis he concen a ion o po assium hyd oxide solu ion (mol/L), and m he weigh o sample (g). 2.4. DSC Analysis Di e en ial scanning calo ime y (DSC) was pe o med on a DSC 1 S a Sys em (Me le -Toledo, G ei ensee, Swi ze land). The de ice was calib a ed using an indium s anda d o empe a u e co ec ion. App oxima ely 15 ± 1 mg o sample was inse ed in o an aluminum pan (ME-27331, 40 µ L pan wi h pin) and sealed he me ically. An emp y sealed pan was used as a e e ence. All samples we e measu ed unde he ollowing empe a u e egime: dynamic hea ing om +25 ◦ C o +70 ◦ C a a hea ing a e o 10 ◦ C/min, and holding a his empe a u e o 5 min o ensu e ha all nuclei o c ys als we e elimina ed. Subsequen ly, cooling was applied om +70 ◦ C o − 40 ◦ C a a cooling a e o 5 ◦ C/min. Then, iso he mal cooling was held a − 40 ◦ C o 3 min, ollowed by inal hea ing o +70 ◦ C (a hea ing a e 5 ◦ C/min). Expe imen s we e ealized unde ni ogen a mosphe e a a low a e o 50 mL/min. The mally induced s uc u al ansi ions ( he mal e en s) we e cha ac e ized by T o (onse ), T p (peak), and T e (endse ) empe a u es on DSC cu es. Mel ing and c ys alliza ion peak empe a u es ( ◦ C) we e e alua ed, and co esponding en halpies (J/g) we e de e mined acco ding o he a ea unde DSC cu e [1,17–21]. 2.5. Rheology Rheological analysis was pe o med on a heome e HAAKE RheoS ess 1 (The mo Fishe Scien i ic, Wal ham, MA, USA). Pla e–pla e geome y (pla e P35TiL, measu ing pla e Foods 2022,11, 3437 6 o 15 co e MPC35) was applied wi h a measu ing gap (i.e., dis ance be ween he pla es) o 2 mm. The ollowing pa ame e s we e obse ed: elas ic s o age modulus G 0 (Pa), loss iscous modulus G 00 (Pa), complex iscosi y η * (Pa · s), and phase loss angle ( an δ= G00/G0) . Measu emen s we e ca ied ou a a equency o 0.1–10 Hz, p essu e o 20 Pa, and em- pe a u e o 15 ±0.5 ◦C. Tempe a u e condi ions we e ensu ed by a ci cula ing wa e ba h The mos a HAAKE AC 200 (The mo Fishe Scien i ic, Wal ham, MA, USA). Using HAAKE RheoWin so wa e (The mo Fishe Scien i ic, Wal ham, MA, USA), sample dependencies o moduli and iscosi y on equency we e e alua ed. 2.6. Fluo escence Spec ome y Fluo escence spec ome y o bu e s and ege able a s was ealized using a Shi- madzu RF-1501 spec opho ome e (Shimadzu, Kyo o, Japan). The exci a ion wa eleng h was 220–360 nm , and he emission wa eleng h was 300–800 nm. Fluo escence in ensi y was ead au oma ically in a bi a y uni s (AU) a speci ic wa eleng hs (nm) o exci a- ion/emission spec a. Qua z cu e es 6030-UV He asil (Hellma Analy ics GmbH & Co. KG, Müllheim, Ge many) o 10 mm op ical leng h adjus ed o he UV/Vis egion we e used o he analyses. Samples o bu e s, sp eadable a s, and sho enings we e mel ed a a empe a u e be ween 30 and 40 ◦ C. Liquid samples we e homogenized by addi ion o nonpola sol en (99.9% cyclohexane). Concen a ions o solu ions we e adjus ed o de ec op imal luo es- cence in ensi y dependen on he wa eleng h: o bu e s, 1% and 2% (w/ ) solu ions, i.e., 1 g/100 mL and 2 g/100 mL, espec i ely; o sp eadable a s and sho enings, 10% and 50% (w/ ) solu ions. 2.7. S a is ical Analysis Da a we e analyzed using one-way analysis o a iance (ANOVA me hod). Di e ences in he mean alues among s a is ical g oups we e es ed a a signi icance le el o α≤ 0.05. The Tukey es was applied o mul iple compa isons (s a is ical anking) o he mean esponses o ea men g oups ( α≤ 0.05) o e alua e i hey a e g ea e han would be expec ed by chance. S a is ical so wa e SigmaS a e sion 2.03 (Sys a So wa e, Inc., Chicago, IL, USA) was used o da a es ing. All expe imen s we e pe o med in a leas h ee eplica es. 3. Resul s 3.1. Tex u e P o ile Analysis As p esen ed in Figu es 1and 2, simila ex u al beha io was de e mined o samples o he same p oduc ype (bu e s, sp eadable a s, and sho enings) applying di e en me hods (5 mm dis ance, 10% s ain). Acco dingly, he same gene aliza ions can be made o bu e s, as well as sp eadable a s. In es iga ed bu e s showed ela i ely high alues o ha dness, elas ic modulus, gumminess, and chewiness, in compa ison o sp eadable a s. Rela i ely la ge di e ences wi hin he g oup o bu e s we e ound o he Young elas ic moduli. These indings a e consis en wi h he obse a ions o Espe e al. [ 22 ], who cha ac e ized bu e s o highe ha dness compa ed o sp eadable a s bu lowe compa ed o anhyd ous milk a s. Lowe sp eadabili y and adhesi eness o bu e p oduc s, p epa ed om o dina y cow milk, can be associa ed wi h hei a y-acid composi ion, cha ac e ized by highe long-chain sa u a ed a y-acid con en [23]. Rema kable indings we e obse ed o sho enings No. 9 and No. 10 which co e- sponded o bu e s wi h se e al ex u al a ibu es ( ela i ely high alues o chewiness, gumminess, Young elas ic modulus, and ha dness). This ac can be ela ed o he nu- i ional cha ac e is ics o he sho enings, as p esen ed in Table 1; he sho enings had ela i ely high o al a con en , compa ed o sp eadable a p oduc s. In pa icula , he ela i ely high con en o sa u a ed a y acids in he sho enings could lead o a mo e igid s uc u e. Mo eo e , sho ening No. 10 con ained 15% milk a , and sho ening No. 9 in ol ed d ied whey powde in he ecipe, which may ha e in luenced hei ex u e Foods 2022,11, 3437 7 o 15 p o iles. In he case o sho ening No. 10, ela i ely lowe alues o ha dness compa ed o bu e s and ano he sho ening could be explained by he p esence o sp ing milk a , which ends o be so e han summe milk a [ 20 ]. This ac can also be con i med by he DSC analysis esul s, which e ealed he p esence o low-mel ing TAG ac ions in sample No. 10, ypical o sp ing milk a , as discussed below (see Sec ion 3.3). In con as o bu e s and sho enings, samples No. 6, No. 7, and No. 8 beha ed as ypical sp eadable a s wi h ela i ely low ha dness, gumminess, and chewiness. This may be a ibu ed o ela i ely low o al a con en and/o emulsi ie p esen in he a s, a ec ing hei c ys al ne wo k s uc u e and emulsion s abili y [22,24]. Foods 2022, 11, x FOR PEER REVIEW 8 o 17 Figu e 2. Tex u e p o ile analysis o bu e s (A), sp eadable a s and sho enings (B) using 10 % s ain pene a ion es (ba cha wi h 5% e o ba s). Tex u al pa ame e s a e desc ibed in he leg- end wi h he ele an uni s in he b acke s; adhesi eness is exp essed in absolu e alues. Di e en supe sc ip le e s o he same ex u al pa ame e indica e s a is ically signi ican di e ences be- ween he samples (α ≤ 0.05, Tukey es ). 3.2. F ee Fa y Acids and Acid Numbe The ee a y acid (FFA) index (w .%) is an essen ial pa ame e used o con ol he quali y o dai y p oduc s. We applied his index o assess he quali y o bu e s and com- pa e i wi h ege able a s. The FFA index ep esen s he pe cen age o ee a y acids, such as oleic, my is ic, palmi ic, and o he acids in ol ed in he p oduc . Values o he FFA index (w .%) a e p esen ed in Figu e 3. The ela ions among he samples co espond o he acid numbe exp essed in mg o po assium hyd oxide pe 1 g o a . Using one-way analysis o a iance (ANOVA), he samples we e conside ed as one s a is ical se which di e ed signi ican ly (a signi icance le el α ≤ 0.05). As e iden om he g aph (Figu e 3), he ee a y-acid con en o all samples was below he limi o 1 w .% FFA (2 mg/g). A op imal condi ions, FFA should be close o ze o. The ela i ely high FFA con en in sho ening No. 10 (0.77 w .%, 1.53 mg/g) could be explained by he a ious s o age condi ions wi hin he supply chain [25]. In addi ion o 85% ege able a s, sho ening No. 10 con ained 15% milk a , as s a ed in Table 1. On he o he hand, sp eadable a No. 6 exhibi ed he lowes FFA con en (0.215 mg/g) among all samples. This can be explained by he ela i ely low o al a con en (39 w .%) in he p oduc . Howe e , no gene al ela ionship was ound be ween he amoun o ee a y acids and o al a con en in he examined g oup o ma ke p oduc s. Figu e 2. Tex u e p o ile analysis o bu e s ( A ), sp eadable a s and sho enings ( B ) using 10% s ain pene a ion es (ba cha wi h 5% e o ba s). Tex u al pa ame e s a e desc ibed in he legend wi h he ele an uni s in he b acke s; adhesi eness is exp essed in absolu e alues. Di e en supe sc ip le e s o he same ex u al pa ame e indica e s a is ically signi ican di e ences be ween he samples (α≤0.05, Tukey es ). 3.2. F ee Fa y Acids and Acid Numbe The ee a y acid (FFA) index (w .%) is an essen ial pa ame e used o con ol he quali y o dai y p oduc s. We applied his index o assess he quali y o bu e s and compa e i wi h ege able a s. The FFA index ep esen s he pe cen age o ee a y acids, such as oleic, my is ic, palmi ic, and o he acids in ol ed in he p oduc . Values o he FFA index (w .%) a e p esen ed in Figu e 3. The ela ions among he samples co espond o he acid numbe exp essed in mg o po assium hyd oxide pe 1 g o a . Using one-way analysis o a iance (ANOVA), he samples we e conside ed as one s a is ical se which di e ed signi ican ly (a signi icance le el α≤0.05). As e iden om he g aph (Figu e 3), he ee a y-acid con en o all samples was below he limi o 1 w .% FFA (2 mg/g). A op imal condi ions, FFA should be close o Foods 2022,11, 3437 8 o 15 ze o. The ela i ely high FFA con en in sho ening No. 10 (0.77 w .%, 1.53 mg/g) could be explained by he a ious s o age condi ions wi hin he supply chain [ 25 ]. In addi ion o 85% ege able a s, sho ening No. 10 con ained 15% milk a , as s a ed in Table 1. On he o he hand, sp eadable a No. 6 exhibi ed he lowes FFA con en (0.215 mg/g) among all samples. This can be explained by he ela i ely low o al a con en (39 w .%) in he p oduc . Howe e , no gene al ela ionship was ound be ween he amoun o ee a y acids and o al a con en in he examined g oup o ma ke p oduc s. Foods 2022, 11, x FOR PEER REVIEW 9 o 17 Figu e 3. F ee a y acids (w .%) in bu e s, sp eadable a s, and sho enings (ba cha wi h 5% e o ba s). Di e en supe sc ip le e s abo e he ba s indica e s a is ically signi ican di e ences be ween he samples (α ≤ 0.05, Tukey es ). 3.3. DSC Analysis On he basis o DSC cu es, a c ys alliza ion and mel ing peaks we e analyzed, as summa ized in Table 2. The mog ams a e p esen ed in Figu es S1 o S10 o Supplemen- a y Ma e ials. Fo cooling cycle o bu e s, he i s c ys alliza ion peak ela ed o medium mel ing ac ions was obse ed in a empe a u e ange o 15.0 ± 1.5 °C. The second c ys- alliza ion peak associa ed wi h low mel ing ac ions was de e mined a 9.0 ± 0.7 °C, de- penden on he sample examined [17] ( he peaks a e no s a ed in he able). Wi hin he c ys alliza ion p ocess o wa e , a sha p exo he mic peak was de ec ed in he empe a u e ange om −9 o −20 °C due o he wa e supe cooling phenomenon o ming s able nuclei o c ys al size [9]. De ec ed alues we e in ela ion o he con en and cha ac e o wa e p esen in he emulsion sys em and a ied om one sample o ano he , because nuclea- ion is a s ochas ic e en [26]. Du ing ehea ing, he i s endo he mic peak ela ed o ice mel ing was obse ed a ound he empe a u e o 0 °C, depending on he sal con en in he p oduc s. Fo bu e s, he ice mel ing peak luc ua ed in a na ow ange ( om 0.02 o 0.37 °C), due o a low amoun o sal (Table 1). The wo endo he mic peaks we e associa ed wi h mel ing o milk a polymo phs. The i s peak was de ec ed a 15.1 ± 0.5 °C o Czech bu e p oduc s, and a 16.1 °C o No. 1 bu e om Poland, as p esen ed in Table 2. This peak can be ela ed o iacylglyce ol medium-mel ing ac ions, when a polymo phic ansi ion om he α- o me as able β′- o m o milk a occu s [17]. The second endo he mic peak a 31.5–34.5 °C can be a ibu ed o high-mel ing ac ions, cha ac e ized by longe hyd oca bon chains [7], i.e., o he mel ing p ocess o he mos s able milk a c ys alliza ion o m (β) [9]. Abo e his empe a u e, he liquid phase was o med. As p esen ed in Table 2, ele an he mog ams we e also de ec ed o sp eadable a s (No. 6, No. 7, and No. 8). Fo sp eadable a s and sho enings, he a ia ions in ice mel ing peak empe a u e we e highe ( om −0.12 o +0.83 °C), compa ed o bu e s. The p ocess o ice mel ing was p obably in luenced by he emulsi ie s p esen in he ege able a s and he p ocessing echnology. In compa ison o bu e s, wa e in sp eadable a s c ys allized in a ela i ely na ow empe a u e ange o app oxima ely −14 o −17 °C, which could Figu e 3. F ee a y acids (w .%) in bu e s, sp eadable a s, and sho enings (ba cha wi h 5% e o ba s). Di e en supe sc ip le e s abo e he ba s indica e s a is ically signi ican di e ences be ween he samples (α≤0.05, Tukey es ). 3.3. DSC Analysis On he basis o DSC cu es, a c ys alliza ion and mel ing peaks we e analyzed, as summa ized in Table 2. The mog ams a e p esen ed in Figu es S1–S10 o Supplemen a y Ma e ials. Fo cooling cycle o bu e s, he i s c ys alliza ion peak ela ed o medium mel ing ac ions was obse ed in a empe a u e ange o 15.0 ± 1.5 ◦ C. The second c ys alliza ion peak associa ed wi h low mel ing ac ions was de e mined a 9.0 ± 0.7 ◦ C, dependen on he sample examined [ 17 ] ( he peaks a e no s a ed in he able). Wi hin he c ys alliza ion p ocess o wa e , a sha p exo he mic peak was de ec ed in he empe a u e ange om − 9 o − 20 ◦ C due o he wa e supe cooling phenomenon o ming s able nuclei o c ys al size [ 9 ]. De ec ed alues we e in ela ion o he con en and cha ac e o wa e p esen in he emulsion sys em and a ied om one sample o ano he , because nuclea ion is a s ochas ic e en [26]. Du ing ehea ing, he i s endo he mic peak ela ed o ice mel ing was obse ed a ound he empe a u e o 0 ◦ C, depending on he sal con en in he p oduc s. Fo bu e s, he ice mel ing peak luc ua ed in a na ow ange ( om 0.02 o 0.37 ◦ C), due o a low amoun o sal (Table 1). The wo endo he mic peaks we e associa ed wi h mel ing o milk a polymo phs. The i s peak was de ec ed a 15.1 ± 0.5 ◦ C o Czech bu e p oduc s, and a 16.1 ◦ C o No. 1 bu e om Poland, as p esen ed in Table 2. This peak can be ela ed o iacylglyce ol medium-mel ing ac ions, when a polymo phic ansi ion om he α - o me as able β0 - o m o milk a occu s [ 17 ]. The second endo he mic peak a 31.5–34.5 ◦ C can be a ibu ed o high-mel ing ac ions, cha ac e ized by longe hyd oca bon chains [ 7 ], i.e., o he mel ing p ocess o he mos s able milk a c ys alliza ion o m ( β ) [ 9 ]. Abo e his empe a u e, he liquid phase was o med. Foods 2022,11, 3437 9 o 15 As p esen ed in Table 2, ele an he mog ams we e also de ec ed o sp eadable a s (No. 6, No. 7, and No. 8). Fo sp eadable a s and sho enings, he a ia ions in ice mel ing peak empe a u e we e highe ( om − 0.12 o +0.83 ◦ C), compa ed o bu e s. The p ocess o ice mel ing was p obably in luenced by he emulsi ie s p esen in he ege able a s and he p ocessing echnology. In compa ison o bu e s, wa e in sp eadable a s c ys allized in a ela i ely na ow empe a u e ange o app oxima ely − 14 o − 17 ◦ C, which could indica e a good dispe sion o wa e phase in he oil phase (i.e., s able sys ems wi h educed phase sepa a ion). I can be expec ed ha he a io be ween ege able a s and oils in hese p oduc s (such as palm oil, sun lowe oil, coconu a , Shea bu e , e c.) a ec s hei c ys alliza ion p ope ies. Beyond he wa e c ys alliza ion peak, small exo he mic peaks in he empe a u e ange o − 20.5 o − 30.5 ◦ C we e de ec ed (no shown in he able). These peaks o mino en halpy change can be ela ed o he c ys alliza ion o ege able oils, and hei pa ame e s we e dependen on he unsa u a ed a y-acid con en in in es iga ed p oduc s. Fo he sho enings (No. 9, No. 10), small c ys alliza ion peaks in he same empe a u e ange we e obse ed. In he ehea ing cycle, se e al mel ing ac ions o TAGs we e indica ed o sp ead- able a s. De ec ed endo he mic peaks co esponded o he occu ence o sa u a ed and unsa u a ed a y acids p esen in anses e i ied a s o he p oduc s. Peaks ecoded a ela i ely high empe a u es (be ween app oxima ely 44 and 51 ◦ C) could be explained by he con en o TAGs highe mel ing ac ions [20]. In he case o sho enings, a mel ing peak a 8.5 ◦ C was eco ded o sample No. 10. This peak can be a ibu ed o he p esence o low-mel ing milk a ac ions; he e was 15% milk a decla ed by he p oduce , which migh ha e come om sp ing milk, con aining mo e low-mel ing TAG ac ions [ 20 ]. Fo bo h sho enings (No. 9 and No. 10), an endo he mic peak a empe a u e a ound 37 ◦ C was obse ed, associa ed wi h highe mel ing ac ions o TAGs. Table 2. DSC esul s o bu e s, sp eadable a s and sho enings applying cooling p ocess om +70 ◦ C o − 40 ◦ C (a cooling a e 5 ◦ C/min), and hea ing p ocess om − 40 ◦ C o +70 ◦ C (a hea ing a e 5◦C/min). Sample C ys alliza ion Peak (Cooling) 1s Mel ing Peak (Hea ing) 2nd/3 d Mel ing Peak (Hea ing) Tc(◦C) ∆H (J/g) Tp(◦C) ∆H (J/g) Tp(◦C) ∆H (J/g) No. 1 −14.09 ±0.14 a32.22 ±0.47 a16.15 ±0.12 a18.13 ±0.24 a34.20 ±0.21 a192.41 ±0.48 a No. 2 −19.66 ±0.18 b34.21 ±0.20 b15.10 ±0.13 b30.60 ±0.35 b31.55 ±0.25 b66.79 ±0.29 b No. 3 −16.92 ±0.22 c30.61 ±0.28 c15.09 ±0.10 b34.35 ±0.26 c31.24 ±0.12 b40.59 ±0.37 c No. 4 −13.32 ±0.19 d38.87 ±0.30 d15.34 ±0.11 b14.21 ±0.15 d31.84 ±0.27 b,c 38.54 ±0.31 d No. 5 −9.16 ±0.15 e35.20 ±0.28 e15.49 ±0.22 b14.54 ±0.17 d34.38 ±0.19 a148.99 ±0.52 e No. 6 −15.83 ±0.10 42.61 ±0.31 - - - 44.57 ±0.23 d- 335.97 ±0.40 No. 7 −16.91 ±0.18 c74.34 ±0.29 g- - 32.30 ±0.19 c 51.11 ±0.17 e417.88 ±0.55 g 8.50 ±0.12 h No. 8 −14.46 ±0.09 a42.90 ±0.15 24.57 ±0.16 c83.69 ±0.35 e- 50.83 ±0.14 e- 57.29 ±0.20 i No. 9 −16.71 ±0.12 c58.72 ±0.26 h- - 37.57 ±0.28 50.01 ±0.19 g316.11 ±0.48 j 26.56 ±0.11 k No. 10 −21.60 ±0.28 g39.01 ±0.15 d8.48 ±0.20 d22.18 ±0.14 34.12 ±0.10 a43.35 ±0.19 l T c —c ys alliza ion peak; ∆ H—en halpy change; T p —mel ing peak. Resul s o he mal pa ame e s a e s a ed as he a i hme ic mean ± s anda d de ia ion o h ee means o sample ba ch iplica e measu emen s. The hyphen means ha no peak in he co esponding empe a u e ange was de ec ed. Di e en supe sc ip le e s in he same column indica e s a is ically signi ican di e ences be ween he samples (α≤0.05, Tukey es ).