International Journal of Molecular Sciences
Article Surfactin as a Green Agent Controlling the Growth of Porous Calcite Microstructures
Anna Bastrzyk 1,*, Marta Fiedot-Toboła 2,* , Halina Maniak 3 , Izabela Polowczyk 1 and Gra˙zynaPłaza 4 1 Department of Process Engineering and Technology of Polymer and Carbon Materials, Faculty of Chemistry, Wrocław University of Science and Technology, Wybrze˙zeWyspia´nskiego27, 50-370 Wrocław, Poland; [email protected] 2 Łukasiewicz Research Network-PORT Polish Center for Technology Development, Stabłowicka 147, 54-066 Wrocław, Poland 3 Department of Micro, Nano and Bioprocess Engineering, Faculty of Chemistry, Wrocław University of Science and Technology, Wybrze˙zeWyspia´nskiego27, 50-370 Wrocław, Poland; [email protected] 4 Department of Environmental Microbiology, Institute for Ecology of Industrial Areas, Kossutha 6, 40-844 Katowice, Poland; [email protected] * Correspondence: [email protected] (A.B.); marta.fi[email protected] (M.F.-T.); Tel.: +48-71-320-32-39 (A.B.); +48-71-734-71-54 (M.F.-T.) Received: 8 July 2020; Accepted: 31 July 2020; Published: 1 August 2020
Abstract: This study presents a new, simple way to obtain mesoporous calcite structures via a green method using an eco-friendly surface-active compound, surfactin, as a controlling agent. The effects of synthesis time and surfactin concentration were investigated. The obtained structures were characterized by X-ray powder diffraction (XRD), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC) coupled with gas mass spectrometry (QMS) analysis. The experimental data showed that surfactin molecules significantly changed the morphology of the calcite crystals, roughening and deforming the surface and creating a greater specific surface area, even at low biosurfactant concentrations (10 ppm). The size of the crystals was reduced, and the zeta potential value of calcium carbonate was more negative when more biosurfactant was added. The XRD data revealed that the biomolecules were incorporated into the crystals and slowed the transformation of vaterite into calcite. It has been shown that as long as vaterite is present in the medium, the calcite surface will be less deformed. The strong influence of surfactin molecules on the crystal growth of calcium carbonate was due to the interaction of surfactin molecules with free calcium ions in the solution as well as the biomolecules adsorption at the formed crystal surface. The role of micelles in crystal growth was examined, and the mechanism of mesoporous calcium carbonate formation was presented.
Keywords: biomineralization; calcite; vaterite; biosurfactant; Turbiscan; TGA; XRD
1. Introduction
Calcium carbonate (CaCO3) is a common biomaterial produced by living organisms for building shells and exoskeletons such as seashells, avian eggshells, and bones to support and protect their bodies [1]. This biomaterial possesses unique properties (high surface area, high porosity, high mechanical strength, and non-toxicity) that can offer wide potential applications in industry as filler materials for paints, pigments, coatings, paper and plastics, carriers of active compounds and nanoparticles for drug delivery, and matrices for polymeric capsules [2–7]. CaCO3 occurs in different crystalline polymorphs: anhydrous phases of aragonite, vaterite and calcite, and hydrated phases of calcium carbonate monohydrate, calcium carbonate hexahydrate and amorphous calcium carbonate [8]. Each of these crystalline forms is characterized by various morphologies and physicochemical properties. For instance, spherical hexagonal
Int. J. Mol. Sci. 2020, 21, 5526; doi:10.3390/ijms21155526 www.mdpi.com/journal/ijms Int. J. Mol. Sci. 2020, 21, 5526 2 of 15 vaterite exhibits higher porosity and can decompose rapidly under relatively mild conditions compared with rhombohedral calcite [2]. Aragonite, on the other hand, forms thin needles and has a strong tendency to form spherulitic clusters with high porosity and density [9]. In biology, biomineralization can occur at specific sites on the cell wall, in spaces between closely packed cells, inside enclosed compartments within a cell or in spaces outside a cell. The specific mineralization sites can include phospholipid vesicles, protein vesicles, cellular assemblies, and molecular frameworks [10]. The formation of crystalline polymorphs with desired properties is controlled by different and occasionally synergic processes, such as the regulation of respective ion concentrations, the formation of initial amorphous and other metastable precursor phases, and the presence of specific biomolecular additives [11]. Extensive research has shown that living organisms require biomolecular additives such as proteins and polysaccharides with carboxyl, phosphate and sulfate groups to control the hierarchical structure and the mechanical properties of calcium carbonate [10,12]. These biomolecules can act as templates and control the polymorphs, orientation and morphology of calcium carbonate. Thus, research has been focused for many years on learning and mimicking the biosynthesis of calcium carbonate. The ability to control the particle shape, size and morphology is fundamental from the viewpoint of technical applications [13]. Therefore, finding a way to obtain calcium carbonate with specific properties remains a challenge, especially with the use of “green additives”. These “green compounds” include biosurfactants, the surface-active molecules produced by living cells. Compared to synthetic surfactants, biosurfactants are non-toxic, easily biodegradable and can be synthesized from renewable waste with an appropriate ratio of carbohydrates and lipids to support optimal bacterial growth [14]. Surfactin (C53H93N7O13) is an anionic lipopeptide biosurfactant with two carboxyl groups that has been widely used in the “green synthesis” of silver and gold nanoparticles [15]. Our previous research has shown that surfactin can also be used effectively as a calcium carbonate growth controlling agent. CaCO3 structures obtained in the presence of surfactin were characterized by large irregularities (terraces and depressions) on the surface of the crystals during a short ageing period [16]. To better understand the mechanism of crystal growth, this study investigated how surfactin concentration can affect the morphology, polymorphs and properties of calcium carbonate precipitated in an aqueous solution after prolonged ageing. The kinetics of crystal growth in the presence of surfactin molecules were also examined. As a result, we identified a new role of surfactin in the biosynthesis of porous calcite. The porous calcite formed and modified by biosurfactants can offer many new possibilities in biomedical and environmental applications.
2. Results and Discussion The calcium carbonate particles were synthesized in a spontaneous, rapid reaction in which calcium chloride and sodium carbonate were used as sources of calcium and carbonate ions, respectively. In the control test (without biosurfactant), turbidity appeared immediately after the Na2CO3 solution (pH 11.3) was poured into the CaCl2 solution without biosurfactant (pH 8.0), indicating the precipitation of CaCO3 particles. After 5 min of reaction, the sample vial was inserted into the measuring cell of the Turbiscan apparatus, and changes in the value of light passing through and reflected from the sample were recorded. Over time, the light transmittance value (T) increased significantly from 20 to approximately 75, reaching a plateau after approximately 15 min (Figure1A). A high value of T (%) indicates very unstable systems [17], in this case due to the formation of larger calcium carbonate particles that sediment. Similar results were observed where Ca2+ ions were in contact with biosurfactant particles for 24 h. However, in the samples with surfactin, the T (%) value increased much more slowly, reaching a value of approximately 70 after 1 h (see Figure1A). This is the result of the interaction of biomolecules during the process (nucleation and further growth of particles). To better understand the process, the ageing time was extended to 24 h, different surfactin concentrations were applied, and the polymorphic form of the crystals and morphology of the structures were analysed. Physicochemical and thermal properties of the biomineral were also measured. Int. J. Mol. Sci. 2020, 21, 5526 3 of 15 Int. J. Mol. Sci. 2020, 21, x FOR PEER REVIEW 3 of 15
Figure 1. (A) T (%) value changes during the time of reaction. (B) Zeta potential of calcium carbonate Figure 1. (A) T (%) value changes during the time of reaction. (B) Zeta potential of calcium carbonate precipitated at various surfactin concentrations after 24 h of ageing (NaCl concentration was 1 mM, precipitated at various surfactin concentrations after 24 h of ageing (NaCl concentration was 1 mM, standard deviation for zeta potential values was lower than 1 mV). standard deviation for zeta potential values was lower than 1 mV). 2.1. Particle Size Distribution and Zeta Potential of Calcium Carbonate after 24 H of Ageing To better understand the process, the ageing time was extended to 24 h, different surfactin The particle size distributions and the diameters of CaCO3 particles obtained in the control system concentrations were applied, and the polymorphic form of the crystals and morphology of the and with surfactin after 24 h of ageing are shown in Figure S1 and Table1, respectively. The addition of structuressurfactin towere the reactionanalysed. system Physicochemical resulted in a significantand thermal particle properties size reduction. of the Thebiomineral median diameterwere also measured. (d50) of CaCO3 formed without the biosurfactant was 24.4 µm. Increasing the concentration of surfactin molecules up to 20 ppm decreased the value of d50 to 10.6 µm, and more particles with diameters lower 2.1.than Particle 5 µm Size were Distribution formed. Comparing and Zeta Potent the dataial ofobtained Calcium Carbonate with 10 and after 20 24 ppm H of surfactin, Ageing one can see
thatThe the particle difference size in thedistributi size ofons the resultingand the particlesdiameters was of notCaCO significant.3 particles This obtained means that in surfactinthe control systemeffectively and with blocked surfactin the growth after of24 CaCO h of ageing3 crystals, are even shown at 10 in ppm. Figure S1 and Table 1, respectively. The addition of surfactin to the reaction system resulted in a significant particle size reduction. The Table 1. Median diameter (d50), lower (d10) and upper (d90) deciles of calcium carbonate particles median diameter (d50) of CaCO3 formed without the biosurfactant was 24.4 μm. Increasing the obtained in the presence of biosurfactant after 24 h of ageing. (Standard deviation for d values was concentrationlower than of 1surfactinµ). molecules up to 20 ppm decreased the value of d50 to 10.6 μm, and more particles with diameters lower than 5 μm were formed. Comparing the data obtained with 10 and 20 ppm surfactin, one Surfactincan see that Concentration the difference [ppm] in thed10 size[µm] of thed resulting50 [µm] particlesd90 [µm] was not significant. This means that surfactin effectively0 blocked the growth 12.5 of CaCO 24.43 crystals, 44.6even at 10 ppm. 5 8.4 15.4 26.0 10 6.3 12.4 21.6 Table 1. Median diameter (d50), lower (d10) and upper (d90) deciles of calcium carbonate particles 20 5.0 10.6 19.6 obtained in the presence of biosurfactant after 24 h of ageing. (Standard deviation for d values was lower than 1 μ). The presence of surfactin molecules in the reaction system changes not only the size but also the Surfactin Concentration [ppm] d10 [µm] d50 [µm] d90 [µm] zeta potential value of CaCO3 particles. Based on the data presented in Figure1B, it can be seen that the zeta potential value of particles0 in the control sample12.5 changed from24.4 8.1 to 44.621.1 mV in a pH range − − from 7 to 9.8. The addition of surfactin5 at concentrations of8.4 10 and 2015.4 ppm increased26.0 the negative value of the zeta potential to 30 and 36.110 mV, respectively, at6.3 a pH of approximately12.4 21.610 0.3. The changes − − ± in zeta potential value were also20 observed by other researchers5.0 using10.6 polypeptides,19.6 fulvic acid or stearicThe acidpresence [18,19 of]. Fromsurfactin previous molecules data in in the the literature, reaction system it is known changes that biomolecules not only the cansize adsorb but also on the zetasolid potential faces formed value asof aCaCO result3 particles. of the crystallization Based on the process data presented and change in the Figure interfacial 1B, it energycan be [seen20,21 that]. theThis zeta may potential explain value the decrease of particles in particle in the size control in the sample presence changed of surfactin, from as − shown8.1 to in−21.1 Figure mV S1 in and a pH rangeTable from1. The 7 highto 9.8. negative The addition value of of zeta surfactin potential at ofconcentrations crystals obtained of 10 in theand presence 20 ppm ofincreased surfactin the negativeproves thatvalue biosurfactant of the zeta potential particles are to − present30 and on −36.1 the surfacemV, respectively, of particles. at a pH of approximately 10 ± 0.3. The changes in zeta potential value were also observed by other researchers using polypeptides, fulvic acid or stearic acid [18,19]. From previous data in the literature, it is known that biomolecules can adsorb on solid faces formed as a result of the crystallization process and change the interfacial energy [20,21]. This may explain the decrease in particle size in the presence of surfactin, as shown in Figure S1 and Table 1. The high negative value of zeta potential of crystals obtained in the presence of surfactin proves that biosurfactant particles are present on the surface of particles.
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2.2. XRD Analysis of Calcium Carbonate after 24 H of Ageing 2.2. XRD Analysis of Calcium Carbonate after 24 H of Ageing CrystallineCrystalline calcium calcium carbonate carbonate polymorphspolymorphs obtained obtained after after 24 h of24 ageingh of ageing were determined were determined by XRD by XRDanalysis. analysis. From From Figure Figure2, it can2, it be can seen be that seen in thethat control in the sample control (without sample surfactin) (without and surfactin) in the presence and in the presenceof 5 ppm of surfactin,5 ppm onlysurfactin, calcite only crystals calcite were obtained.crystals Thiswere means obtained. the complete This means transformation the complete of transformationthe metastable of phase, the metastable vaterite, into phase, a thermodynamically vaterite, into stablea thermodynamically phase, calcite, under stable these conditions.phase, calcite, underA furtherthese increaseconditions. in surfactin A further concentration increase upin tosurf 20actin ppm resultedconcentration in a mixture up to of calcite20 ppm and resulted vaterite in a mixtureparticles, of calcite which and was vaterite revealed particles, by the characteristic which was peaks revealed in Figure by the2. characteristic peaks in Figure 2.
FigureFigure 2. XRD 2. XRD patterns patterns of ofcalcium calcium carbonate carbonate precipitated precipitated at at di differentfferent surfactin surfactin concentrations concentrations after after 24 24
h of hageing. of ageing. The The concentration concentration of of CaCl CaCl22 andand Na Na2COCO3 was 55 mMmM (c-calcite, (c-calcite, v-vaterite). v-vaterite).
For 10 and 20 ppm surfactin in the reaction system, the vaterite content was found to be 1.4% and For 10 and 20 ppm surfactin in the reaction system, the vaterite content was found to be 1.4% and 14.9%, respectively (Table2). This means that the higher concentrations of surfactin molecules can 14.9%,effectively respectively delay the(Table transformation 2). This means of vaterite that tothe calcite. higher In concentrations all investigated samples,of surfactin a trace molecules amount can effectively delay the transformation of vaterite to calcite. In all investigated samples, a trace amount of Na2CO3 (0.1%, at 2Ө= 28◦) was found. On the basis of the calculated unit cell parameters of the of Napolymorphs2CO3 (0.1%, (Table at 22Ө), = a 28 possible˚) was incorporation found. On the of surfactinbasis of intothe calculated the crystal structureunit cell ofparameters calcite was of the polymorphsfound. For (Table that purpose, 2), a possible the relative incorporation changes (∆ l,of∆ Vsurfactin, %) in the into lengths the ofcrystal a and structure c edges and of volume calcite was found.in the For elemental that purpose, cell for the pure relative calcite changes without surfactin(Δl, ΔV, (%)l0, Vin0 )the and lengths for systems of a and with c varied edges amounts and volume of surfactin (ls, Vs) were calculated: ∆l = (ls l )/l 100. In all cases studied, a slight elongation of in the elemental cell for pure calcite without surfactin− 0 0· (l0, V0) and for systems with varied amounts of surfactin0.046–0.054% (ls, Vs) andwere 0–0.04% calculated: for the Δ al and= (l cs–l edges0)/l0·100 of the. In calcite all cases elemental studied, cell, respectively,a slight elongation was observed. of 0.046– 0.054%Nevertheless, and 0–0.04% the increasefor the a in and the edgec edges dimensions of the ca didlcite not elemental correspond cell, directly respectively, to an increment was observed. of surfactin concentration added to the reaction medium and seemed to become rather independent of its Nevertheless, the increase in the edge dimensions did not correspond directly to an increment of higher values (10 and 20 ppm). This small elongation of the edges, however, influences the volume of surfactin concentration added to the reaction medium and seemed to become rather independent of the calcite elemental cell (Table2). For the subsequent concentrations of 5, 10 and 20 ppm surfactin, its higherthe calculated values relative(10 and changes 20 ppm). in the This volume small of elongation calcite were of 0.085, the 0.142 edges, and however, 0.144%, respectively. influences the volumeSimilar of the results calcite were elemental observed cell for biogenic(Table 2). calcite For the and subsequent for calcite obtained concentrations in the presence of 5, 10 of and amino 20 ppm surfactin,acids and the proteins calculated [22,23 relative]. The changes changes in the in elementalthe volume cell parametersof calcite seemwere to 0.085, indicate 0.142 that surfactinand 0.144%, respectively.may induce Similar anisotropic results positive were lattice observed distortion for duebiogenic to the possiblecalcite and incorporation for calcite of biomoleculesobtained in the presenceinto the of crystalamino lattice. acids and This proteins phenomenon [22,23]. may The influence changes the crystalin the structureelemental of ce calcitell parameters in all studied seem to indicate that surfactin may induce anisotropic positive lattice distortion due to the possible incorporation of biomolecules into the crystal lattice. This phenomenon may influence the crystal structure of calcite in all studied systems. The effect of surfactin on the changes in the edge length and volume of the vaterite elemental cell is difficult to determine, as Table 2 shows, the increase in the values of particular parameters is in the range of standard deviation.
Int. J. Mol. Sci. 2020, 21, 5526 5 of 15 systems. The effect of surfactin on the changes in the edge length and volume of the vaterite elemental cell is difficult to determine, as Table2 shows, the increase in the values of particular parameters is in the range of standard deviation.
Table 2. Phase composition and the elemental cell parameters of the precipitated calcium carbonate calculated from the diffraction patterns. (Standard deviation for calcite or vaterite content was lower than 2%. Standard deviation for calcite cell parameters a, c and V were 0.003–0.008%, 0.001 0.006%, − 0.006 0.013%, respectively. Standard deviation for vaterite cell parameters a, c and V were 0.011–0.013%, − 0.025–0.029%, 0.23–0.34%, respectively).
Surfactin Concentration Calcite Vaterite [ppm] Content [%] Structural Parameters Content [%] Structural Parameters a = b = 4.9900(2) Å c = 17.0550(7) Å 0 100 -- α = β = 90◦, γ = 120◦ V = 367.776 (Å)3 a = b = 4.9923(3) Å c = 17.0537(14) Å 5 100 -- α = β = 90◦, γ = 120◦ V = 368.087 (Å)3 a = b = 4.9927(1) Å a = b = 4.1268(4) Å c = 17.0608(6) Å c = 8.4791(20) Å 10 98.6 1.4 α = β = 90◦, γ = 120◦ α = β = 90◦, γ = 120◦ V = 368.299 (Å)3 V = 125.057 (Å)3 a = b = 4.9926(2) Å a = b = 4.1299(5) Å c = 17.0619(9) Å c = 8.4699(25) Å 20 85.1 14.9 α = β = 90◦, γ = 120◦ α = β = 90◦, γ = 120◦ V = 368.308 (Å)3 V = 125.109 (Å)3
2.3. Morphology of Calcium Carbonate Crystals after 24 H of Ageing The morphology of the obtained calcium carbonate crystals was analysed using scanning electron microscopy,and the images are presented in Figure3. In Figure3A, it can be seen that without biomolecules, rhombohedral structures with a smooth surface and sharp edges characteristic of calcite were formed. The morphology changed when the biosurfactant was added to the reaction medium (Figure3B–F). The surface of the crystal formed in the presence of 5 ppm surfactin was deformed as some irregularities appeared (Figure3B). By analysing the images in Figure3, with higher biosurfactant concentrations, the presence of spherical structures characteristic of vaterite was observed, which was also confirmed by XRD analysis (Figure2). However, the most interesting morphology was observed for calcite. Increased surfactin concentration resulted in a greater surface roughness of the calcite crystals, with spherical and oval cavities on the surface. Moreover, the edges of the crystals were not so sharp and were significantly deformed (Figure3C–F). The deformation of calcite crystal microstructures after adding compounds of biological origin has been widely described in the literature [24–28]. Biomolecules, especially anionic molecules, can bind calcium ions in solution as well as on the surface of created crystals, changing the direction of growth or blocking the growth of the crystal surface [2]. Surfactin is a surfactant that reduces the surface tension of water to 28 mNm 1 at a critical micellization concentration (CMC) in − the range of 10–20 ppm, depending on pH and ionic strength [29]. Our previous studies showed that in a 5 mM aqueous solution of CaCl2 at pH 8, the CMC value for surfactin was 4.6 ppm [16]. Thus, the porosity of the calcite surface was a result of the formation of biosurfactant micelles in the reaction medium at surfactin concentrations above the CMC value. Similar results were obtained in the presence of nanosized spherules and worms of polymers and casein micelles [27,30]. Int. J. Mol. Sci. 2020, 21, 5526 6 of 15 Int. J. Mol. Sci. 2020, 21, x FOR PEER REVIEW 6 of 15
FigureFigure 3. SEM 3. SEM images images of of calcium calcium carbonate carbonate precipitation atat various various surfactin surfactin concentrations concentrations after after 24 h24 h ofof ageing.ageing. ( A()A control) control sample, sample, (B) 5 ppm,(B) 5 ( Cppm,,D) 10 ppm,(C,D) ( E10,F) 20ppm, ppm ( ofE,F surfactin.) 20 ppm The of concentration surfactin. The of
concentrationCaCl2 and Na of2 CaClCO3 2was and 5mM. Na2CO3 was 5mM.
In Inour our studies, studies, it was it was noted noted that that more more micelles micelles present present in the systemsystem leadlead to to more more porous porous calcite calcite crystals.crystals. Comparing Comparing these these results results with with previous previous results results [16], [16], it was observedobserved thatthat by by extending extending the the reactionreaction time time to to 24 h,h, porous porous crystals crystals are formedare form ated twice at thetwice biosurfactant the biosurfactant concentration. concentration. These structures These may have great potential for use as inorganic matrices or carriers of active compounds or nanoparticles. structures may have great potential for use as inorganic matrices or carriers of active compounds or It can be assumed that micelle concentration and ageing time play a crucial role in the formation of nanoparticles. It can be assumed that micelle concentration and ageing time play a crucial role in the porous calcite structures. formation of porous calcite structures. 2.4. BET Analysis of Calcium Carbonate after 24 H of Ageing 2.4. BET Analysis of Calcium Carbonate after 24 H of Ageing One of the most important parameters describing inorganic matrices is the value of specific surface areaOne (SSA) of the as well most as important the size and parameters volume of pores. describing The hysteresis inorganic loops matrices observed is inthe Figure value S2 of indicate specific surfacethe mesoporous area (SSA) structureas well as of the CaCO size3. Itand was volume observed of thatpores. particles The hysteresis obtained without loops observed biomolecules in Figure had 2 1 S2 aindicate small SSA the value mesoporous of 0.18 m structureg− (Table 3of). CaCO This is3. the It resultwas observed of rhombohedral that particles calcite formationobtained withwithout a biomoleculessmooth surface, had a as small shown SSA in Figurevalue3 ofA. 0.18 The m increase2g−1 (Table in surfactin 3). This concentration is the result of to 10rhombohedral ppm and 20 ppmcalcite 2 1 formationsignificantly with increased a smooth the surface, value of as SSA shown to 1.67 in andFigure 4.87 3A. m gThe− , respectively. increase in Asurfactin higher valueconcentration of the SSA to 10 arisesppm and from 20 the ppm roughness significantly of the calciteincreased surface the formedvalue of in SSA the presenceto 1.67 and of biosurfactant 4.87 m2g−1, respectively. (Figure3E,F). A higherA larger value specific of the surface SSA arises area was from obtained the roughness when a higher of the concentration calcite surface of biosurfactant formed in the was presence added to of biosurfactantthe reaction (Figure mixture. 3E–F). From A the larger data inspecific Table3 surf, it canace be area seen was that obtained the average when pore a higher diameter concentration increased of biosurfactantfrom 8.7 to 11.7 was nm added when surfactinto the reaction at higher mixture. concentrations From the was data added. in Table Moreover, 3, it can the be average seen that pore the averagevolume pore doubled diameter with increasingincreased biosurfactantfrom 8.7 to 11.7 concentration. nm when This surfactin may be at due higher to the concentrations formation of more was added.micelles Moreover, in the solution the average as well aspore the volume fact that atdoubled a 20 ppm with concentration increasing of biosurfactant biosurfactant, moreconcentration. vaterite Thiswas may formed be due (Table to the2). formation It is known of from more the micelles literature in that the vateritesolution has as awell higher as the SSA fact than that calcite at a [2031 ].ppm concentration of biosurfactant, more vaterite was formed (Table 2). It is known from the literature Table 3. The characteristic surface parameters of CaCO3 precipitate. that vaterite has a higher SSA than calcite [31]. Surfactin Concentration SSA Average Pores Volume Average Pores Diameter [ppm] 2 1 3 1 [nm] Table 3. The characteristic[m g− ] surface parameters[cm g− ] of CaCO3 precipitate. 0 0.18 0.05 - - ± Surfactin10 1.67 0.1 0.00513Average0.0002 Pores Average8.7 Pores1.0 SSA± ± ± Concentration20 4.87 0.09 0.0123Volume0.001 Diameter11.7 1.2 [m± 2g−1] ± ± [ppm] [cm3g−1] [nm] Comparing these0 data with our 0.18 previous± 0.05 results [16 -], one can say that - after 24 h of ageing, 0.00513 ± 8.7 ± 1.0 2 1 the specific surface area10 and average 1.67 pore ± volume0.1 slightly decreased from 7.11 to 4.87 m g− and from 0.0002 20 4.87 ± 0.09 0.0123 ± 0.001 11.7 ± 1.2 Comparing these data with our previous results [16], one can say that after 24 h of ageing, the specific surface area and average pore volume slightly decreased from 7.11 to 4.87 m2g−1 and from
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0.0202 to 0.0123 cm3g−1, respectively. This was related to the reduction of vaterite content and the 3 1 0.0202incorporation to 0.0123 of cm moreg− biosurfactant, respectively. molecules This was into related calcite to thecrystals. reduction of vaterite content and the incorporation of more biosurfactant molecules into calcite crystals. 2.5. Thermogravimetric Characteristics of Calcium Carbonate Structures 2.5. Thermogravimetric Characteristics of Calcium Carbonate Structures −1 Thermogravimetric analysis of calcium carbonate samples with 5 Kmin1 heating rate was also Thermogravimetric analysis of calcium carbonate samples with 5 Kmin− heating rate was also performed. To show the effect of surfactin on the thermal behaviour of CaCO3, samples with the performed. To show the effect of surfactin on the thermal behaviour of CaCO , samples with the most most differing surfactin content were selected, namely, without and with 203 ppm surfactin. It was differing surfactin content were selected, namely, without and with 20 ppm surfactin. It was observed observed that in both samples, with and without biosurfactant, the main degradation process started that in both samples, with and without biosurfactant, the main degradation process started above 690 C, above 690°C, which was related to the conversion of calcite into calcium oxide (Figure 4A). Despite◦ which was related to the conversion of calcite into calcium oxide (Figure4A). Despite the fact that two the fact that two additional weight losses were detected at lower temperatures (Figure 4A). The first additional weight losses were detected at lower temperatures (Figure4A). The first loss had no specified loss had no specified maximum on the DTG curve. The second loss was observed only for the maximum on the DTG curve. The second loss was observed only for the sample with surfactin and sample with surfactin and had a broad DTG peak with maximum about 510°C (Figure 4B). From the hadliterature, a broad it DTG is known peak that with the maximum first weight about loss 510 is ◦relatedC (Figure to 4theB). evaporation From the literature, of moisture it is contained known that in the first weight loss is related to the evaporation of moisture contained in the samples, and the second the samples, and the second weight loss is related to the evaporation of water bound in the CaCO3 weightstructure loss and/or is related transformation to the evaporation of vaterite of water to calc boundite [32,33]. in the To CaCO compare3 structure the chemical and/or transformationcomposition of ofthe vaterite sample to with calcite and [32 ,without33]. To comparesurfactin, the the chemical weight compositionloss observed of during the sample this withtwo recesses and without was surfactin,determined. the weightThe temperature loss observed ranges during was thisselected two recessesaccording was to determined.DTG results. TheIt was temperature shown that ranges in the wasfirst selected range (25–400 according °C), to the DTG weight results. ofIt the was materi shownal thatwith in surfactin the first rangedecreased (25–400 by ◦0.95%.C), the Continued weight of theheating material resulted with surfactinin a further decreased decrease by in 0.95%. material Continued weight to heating the same resulted value. in For a further a sample decrease without in materialsurfactin, weight it was to not the samepossible value. to divide For a sample this te withoutmperature surfactin, region itfor was two not processes, possible toas divide any DTG this temperaturemaximum regionin this forarea two was processes, observed. as any For DTG this maximum reason, the in thisweight area wasloss observed.of this material For this reason,in one thetemperature weight loss range of this from material 25 to in 566 one °C temperature was measur rangeed. Its from value 25 was to 566 approximately◦C was measured. 0.71%. Its The value higher was approximatelyweight loss value 0.71%. in The this higher temperature weight loss range value (25– in566 this °C) temperature for the sample range (25–566 with surfactin◦C) for the could sample be withconnected surfactin with could better be connected moisture with adsorption better moisture and surfactin adsorption molecule and surfactin degradation. molecule degradation.Also, these Also,differences these differences were visible were in the visible total inweight the total loss weightof the samples, loss of the which samples, was higher which for was the higher sample for with the samplesurfactin with by surfactin approximately by approximately 1%. The maximum 1%. The maximumDTG associated DTG associatedwith calcite with conversion calcite conversion was also washigher also for higher this sample for this samplewith surfactin, with surfactin, which could which demonstrate could demonstrate obstruction obstruction of this process of this processby the use by theof usebiosurfactant of biosurfactant (Figure (Figure 4 and 4Table and Table4). 4).
FigureFigure 4. 4.TG TG and and DTG DTG curves curves of of the the calcium calcium carbonate carbonate structures structures obtained obtained without without and and with with 20 20 ppm ppm of biosurfactant.of biosurfactant. Time Time of reaction of reaction was was 24 h. 24 The h. concentrationThe concentration of CaCl of CaCl2 and2 Naand2 CONa32COwas3 was 5 mM. 5 mM. (A) TG;(A) (BTG;) DTG. (B) DTG.
Table 4. Thermogravimetry results of the samples (∆m–weight loss, T–maximum on the DTG curve, Table 4. Thermogravimetry results of the samples (Δm–weight loss, T–maximum on the DTG curve, onset-temperature- of main degradation start). onset-temperature- of main degradation start). Surfactin Concentration Surfactin Concentration∆m 1 (25–400 ◦C) [%] ∆m2 (400–566 ◦C) [%] T1 [◦C] TT21[ ◦C] T2 ∆ mtotal∆mtotal[%] [ppm] ∆m1 (25–400 °C) [%] ∆m2 (400–566 °C) [%] [ppm] [°C] [°C] [%] 0 0.71 - 717 43.69 200 0.950.71 0.95 510- 724 717 44.7143.69 20 0.95 0.95 510 724 44.71
Int. J. Mol. Sci. 2020, 21, x FOR PEER REVIEW 8 of 15
In addition, model-free kinetics were used to determine activation energy values as a function of material conversion [34]. To achieve this, both samples were analysed by the thermogravimetric method with four different heating rates (5, 10, 15, and 20 Kmin−1) and presented as a conversion curve (Figure S3). The activation energy value was calculated as follows (1):