J Am Oil Chem Soc DOI 10.

1007/s11746-011-1952-3

LETTER TO THE EDITOR

Triacylglycerol Polymorphism: What Can We Learn from Space Groups and Crystalline Tendency?
R. John Craven Robert W. Lencki

Received: 20 July 2011 / Revised: 16 September 2011 / Accepted: 6 October 2011 AOCS 2011

Polymorphism strongly affects the functionality of fat and fat-containing foods. For instance, margarine in the desirable b0 form is smooth and creamy, whereas, the more stable b polymorph is associated with a grainy texture. Similarly, cocoa butter in form V produces chocolate that is glossy, snaps nicely, and melts in your mouth, whereas, the more stable form VI is a dull white or grey lm that does not melt as readily [1]. Many organic compounds are polymorphic and this polymorphism occurs via numerous mechanismssome of which are ill-dened [2]. Similarly, the current understanding of triacylglycerol (TAG) polymorphism is, in many ways, incomplete. A number of factors are thought to contribute to TAG polymorphism. In the solid phase, TAG molecules adopt one of several possible chain-length (double or triple chain length aka 2L or 3L) and glycerol conformation (chair or tuning fork) structures. These structures are determined to a large part by the TAGs substituent fatty acids and their relative afnity for each other, with acyl chains congregating due to similarities in length and degree of saturation [3, 4]. In addition, variations in acyl-chain tilt as well as acyl-chainand methyl-end-packing are also thought to contribute to, or be indicative of, polymorphic behavior [5, 6]. While this descriptive mechanism has been useful, particularly for molecules with minimal stereochemistry (i.e. n-parafns and waxes) [7] it is incapable of predicting or explaining some common polymorphic behavior of TAG. For example the current model cannot explain why, while simple (monoacid)
R. J. Craven R. W. Lencki (&) Department of Food Science, University of Guelph, Guelph, ON N1G 2W1, Canada e-mail: rlencki@uoguelph.ca R. J. Craven e-mail: craven@uoguelph.ca

TAG are b-tending, enantiopure mixed (di- and triacid) TAG (e.g. milk fat, sn-10:0-10:0-16:0 and sn-16:0-16:0-14:0) are b0 -stable [810]. Moreover, it does not explain why enantiopure TAG (sn-10:0-10:0-16:0 and sn-16:0-16:0-14:0) are b0 -stable while the corresponding racemic mixtures (rac-10:0-10:0-16:0 and rac-16:0-16:0-14:0) are b-stable [9, 10]. Perhaps the current descriptive mechanism for TAG polymorphism could be improved by including some consideration for the stereochemical conformation of constituent molecules.

Crystalline Tendency The relative stereochemistry of molecules within the unit cell can be determined by spectroscopic means (single-crystaland powder diffraction X-ray) or from the phase behavior of enantiomeric mixtures (crystalline tendency) [1113]. In our lab we employed the latter technique to understand the relationship between polymorphism and stereochemistry for a chiral TAG system (sn-10:0-10:0-16:0 and sn-16:0-10:0-10:0). Samples of enantiopure 1,2-bisdecanoyl-3-palmitoyl-sn-glycerol (sn-10:0-10:0-16:0; E-TAG) and racemic bisdecanoyl-1(3)-palmitoyl-rac-glycerol (rac-10:0-10:0-16:0 : 50% sn-10:0-10:0-16:0 ? 50% sn-16:0-10:0-10:0; R-TAG) were prepared in [99% purity (by GC) [9]. While their melting points were similar (sn-10:0-10:0-16:0: Tp = 32.9 C; rac-10:0-10:0-16:0: Tp = 33.38 C) their polymorphism (b0 -tending E-TAG vs b-tending R-TAG) and consequently their crystallization and melting behavior was remarkably different [9]. A similar polymorphic tendency has been reported for sn-16:0-16:0-14:0 (b0 -tending) and rac-16:0-16:0-14:0 (b-tending) [10]. Liquidus data was used to construct a phase diagram for enantiomeric mixtures of sn-10:0-10:0-16:0 and

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sn-16:0-10:0-10:0 (cf. Fig. 12 in Reference 9). One half of the phase diagram was derived with the understanding that the other half of the diagram is an identical mirror image. Analysis of this data revealed that blends of sn-10:0-10:0-16:0 and sn-16:0-10:0-10:0 form a meta-stable eutectic (i.e. conglomerate in the b0 polymorph) and a stable 1:1 molecular compound (i.e. racemic compound in the b polymorph). In addition, the most thermodynamically-stable polymorph for enantiopure sn-10:0-10:0-16:0 and by analogy sn-16:0-10:0-10:0 is b0 . Thus, for the subject compounds, molecules of opposite enantiomers occupy separate unit cells in the b0 -form and are matched in the unit cell of the b-form [9]. These results are summarized under the heading crystalline tendency in Tables 1, 2.

Table 2 Unit cell stereochemistry for b0 -form triacylglycerols One stereoisomer in the unit cell Crystalline tendency [9] Pure enantiomer (sn-10:0-10:0-16:0) is b-tending Racemic mixture (sn-10:0-10:0-16:0 and sn-16:0-10:0-10:0) forms a metastable conglomerate (eutectic) Single-crystal X-ray 12:0-14:0-12:0 C2 [28], sn-16:0-16:0-14:0 C2 [10 ] X-ray powder diffraction 12:0-14:0-12:0, 14:0-16:0-14:0 (at 250 K) I2 [29] 16:0-18:1t-16:0, 16:0-18:0-16:0, 16:0-16:0-18:1t, 16:0-16:018:0 I2 [6] Both stereoisomers in the unit cell Single-crystal X-ray

Stereochemistry of the Unit Cell for b- and b0 -Form Triacylglycerols As mentioned above, the relative stereochemistry of molecules within the unit cell can also be determined by X-ray spectroscopy (single-crystal and powder diffraction). This information is expressed by the crystallographic space group determined for the crystal structure [11, 12]. Crystallographic space groups have been assigned for many b- and b0 -form TAG and as a result the relative stereochemistry of their unit cells can be determined (Tables 1, 2).

11:0-11:0-11:0 P21/c [30]twinning observed but not taken into account X-ray powder diffraction 10:0-12:0-10:0, 14:0-16:0-14:0 (at 298 K), 16:0-18:0-16:0 Iba2 [29], 16:0-18:0-18:0 C2/c [6]difculties reported but twinning not considered

Table 1 Unit cell stereochemistry for b-form triacylglycerols Both stereoisomers in the unit cell Crystalline tendency [9] sn-10:0-10:0-16:0 and sn-16:0-10:0-10:0 (racemic mixture) form a stable 1:1 molecular compound (racemic compound) Single-crystal X-ray " 10:0-10:0-10:0 P1 [18], 12:0-12:0-12:0 P" [19], 18:0-18:0-18:0 1 [20], 10:0-C11Br:0-10:0 P" [21], 16:0-16:0-16:0 P" [22], 1 1 18:1t-18:1t-18:1t P" [23] 1 X-ray powder diffraction 14:0-14:0-14:0, 18:0-18:0-18:0 P" [24], 13:0-13:0-13:0, 1 15:0-15:0-15:0, 17:0-17:0-17:0, 19:0-19:0-19:0 P" [25], 1 18:0-18:1-18:0 P" [1], 14:0-18:1-14:0, 16:0-18:1-16:0, 1 18:0-18:1-18:0, 16:0-18:1-18:0, 18:0-18:1-20:0 P21/n [26], 16:0-18:1-16:0, 18:0-18:1-18:0, 16:0-18:1-18:0, 18:0-18:1-20:0 Cc [27], 18:0-18:0-18:1t, 16:0-18:0-18:0, 16:0-16:0-18:0, 16:0-16:0-18:1t, 14:0-14:0-16:0, 12:0-14:0-14:0, 12:0-12:0-14:0, 18:0-18:1t-18:0, 16:0-18:1t-16:0, 16:0-18:0-16:0 P" [5] 1 One stereoisomer in the unit cell Single-crystal X-ray sn-16:0-16:0-2:0 P21 [14]exceptional polymorphic behavior common for acetoyl acylglycerols

Based on crystalline tendency and crystallographic space group assignments (P" for the most part) the unit cell 1 of b-form TAG generally contains both stereoisomers (Table 1). Not surprisingly, 1,2-dipalmitoyl-3-acetoyl-snglycerol (sn-16:0-16:0-2:0) [14] is an exception as are many acetoyl acylglycerols [15]. While the stereochemical composition of the unit cell for b-form TAG is clear-cut, the situation appears more complex for b0 -form TAG (Table 2). However, if the role of crystal twinning in TAG b0 forms is taken into account the matter is simplied. For b0 forms of TAG, the determination of crystal structure by X-ray (single-crystal and powder) is frequently confounded by the growth of crystal twins [4]; thus, variations in crystal growth conditions coupled with crystal twinning effects are a probable cause for the multitude of b0 forms reported for numerous TAG including tristearin (18:0-18:0-18:0) and tripalmitin (16:0-16:0-16:0). [Crystal twinning is] the growth of two or more differently oriented domains of a single structure into a twinned crystal twinning can be described in terms of a symmetry element, the twin-element, which, unlike normal symmetry elements, does not occur in every unit cell but relatively few timesor even only onceon a macroscopic scale [16]. When results from X-ray studies where crystal twinning was negligible or was properly accounted for are included with the crystalline tendency results the unit cell for b0 form TAG contains only one stereoisomer (Table 2). It appears that the unit cell of all b-form TAG contains both stereoisomers (Table 1)the exceptional polymorphism of acetoyl acylglycerols notwithstanding (viz.

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J Am Oil Chem Soc Fig. 1 Schematic for a b and b b0 unit cells of achiral TAG and racemic mixtures of chiral TAG

sn-16:0-16:0-2:0). Pure enantiomers must therefore assume the b0 form because the unit cell for these compounds can only contain one stereoisomer. Moreover, when the effects of twinning are taken into account the current data indicates that the unit cell for b0 forms of chiral and achiral TAG (e.g. the CnCn?2Cn series) contain only one stereoisomer (Table 2). This preliminary analysis demonstrates the key role unit cell stereochemistry plays in TAG polymorphism (Fig. 1). At present, crystallographic space group assignments, drawn from X-ray data, are available for many b- and b0 -form TAG (Tables 1, 2). However, only one study to determine the crystalline tendency of chiral TAG via the binary phase behavior of enantiomeric mixtures has been conducted (Tables 1, 2) [9]. Further work in this area will indicate whether the trends noted here are system-specic or whether they are the general rule. Given the role of crystalline tendency in the polymorphism of chiral systems [13] and new insights regarding conformational polymorphism [17] we hypothesize that these observations will apply to the general case.

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