Journal of Molecular Structure (Theochem) 419 (1997) 139–154

Vitamin C at 120 K: experimental and theoretical study of the charge density1
M. Milanesio a, R. Bianchi b, P. Ugliengo a, C. Roetti a, D. Viterbo a,*

Dipartimento di Chimica IFM, Universita’ di Torino, Via P. Giuria 7, I-10125 Torino, Italy ` Chimica, CNR, Via C. Golgi 19, 20133 Milano, Italy Centro di Studio per le Relazioni tra Struttura e Reattivita Received 1 November 1996; accepted 12 February 1997


Abstract Vitamin C is an extremely interesting molecule, not only for its important biological properties, but also for its crystal structure in which the formation of several hydrogen bonds plays an important role. We have collected X-ray diffraction data at ˚ −1. The crystals do not undergo any phase transformation and the space group remains P2 1 with − 153ЊC up to sinv/l = 1.17 A two independent molecules in the asymmetric unit. In the final difference Fourier map non-spherical electron density in the bonding regions was clearly indicated. A study by means of a multipole expansion of the electron density has been performed and the experimental electrostatic potential maps obtained. We have also computed the electron density by means of the crystalorbital approach as coded in the program crystal95 [R. Dovesi et al., crystal95 User’s Manual, University of Torino]. A complete ab initio conformational analysis of the isolated molecule has also been carried out at HF-SCF and density functional level, with reasonably large basis sets. The results of this analysis are: (i) vitamin C molecules are tightly bound in the crystal by a network of eight intermolecular H-bonds per molecule. The relative strength of these bonds is well accounted for by the experimental electrostatic potential. No relevant intramolecular interactions are present; (ii) crystal95 calculations give a value of 44 kcal mol −1 as the sublimation energy per mole of molecules, computed at correlated level with 6-21G(d,p) and Perdew 91 potential [J.P. Perdew and Y. Wang, Phys. Rev. B, 45 (1992) 13244; M. Causa ` and A. Zupan, Chem. Phys. Lett., 220 (1994) 145]; (iii) the crystal95 electron density maps are in good agreement with the experimental ones; (iv) the calculations on the isolated molecule reveal that the most stable conformers are different from the X-ray structure and are characterized by intramolecular H-bonds; the maximum energy difference among the 11 conformers taken into account is less than 6 kcal mol −1; (v) the NMR analysis in water reveals that the least populated side-chain conformation around bond C5–C6 is that found in our computed gas-phase conformers with the most folded-up side chain, which are indeed less prone to be solvated. ᭧ 1997 Elsevier Science B.V.

1. Introduction Vitamin C was first isolated by the Hungarian scientist Albert Szent-Gyo ¨ rgyi from the adrenal cortex of cattle, and later from orange juice and cabbage water.
* Corresponding author. E-mail: 1 Dedicated to Linus Pauling, a giant in chemistry and in life.

The molecular formula C 6H 8O 6 was also established and the new molecule was named huxuronic acid [1].

0166-1280/97/$17.00 ᭧ 1997 Elsevier Science B.V. All rights reserved. PII S 0 16 6- 1 28 0 (9 7 )0 0 25 2 -2

17184 A transparent prismatic crystal.2 Bond distances and angles Tables of observed and calculated structure factors of the anisotropic thermal displacement parameters and of the other parameters used in the multipole expansion may be obtained from the authors. 3 (ethanol). this property being much less evident in non-aqueous media. insoluble in chloroform.4. increase of circulating levels of lipoprotein (HDL). fats and oils 1. molar mass pK a Specific rotation Solubility (g/100 cm 3 at 20ЊC) Density (g cm −3) Vitamin C is an acid in aqueous solution. 2.14 1st (C(3)–OH) 4. The preferred conformation in the solid state is one in which the side chain is in an extended conformation. Finally. thereby preventing the arterial deposition of cholesterol.140 M. 1. l-xilo-ascorbic acid. several uses of vitamin C as a drug may be recalled: • • • treatment after surgery. which play an important role in cell degeneration. using gaussian94 [6]. because the ionization of the enolic OH linked to C-3 (pK a = 4. using the multipole expansion method. as illustrated in Fig. Results and discussion 2. The final atomic coordinates are given in Table 3. using a diffraction data up to sinv/l = 1. petroleum ether. thus reducing the risk of coronary heart disease. 13 C NMR studies [4] have shown that the same conformation found in the crystal is the dominant one also in water solution. treatment of anaemia in order to improve iron absorption. Indeed. detailed analysis. cevitamic acid. Among its various functions. We have collected accurate low-temperature single crystal diffraction data in order to obtain the experimental electron density distribution of vitamin C. 2.and intra-molecular hydrogen bonds in dictating the most stable conformations. in order to assess the importance of inter. The structure of the two independent molecules (a and b) is very close to that determined by Hvoslef [3] and is shown in Fig.5.25 2nd (C(2)–OH) 11. in which the pattern of hydrogen bonds is also indicated. 1 (glycerol). l-ascorbic acid. we have performed a complete ab initio conformational analysis of the isolated molecule. 120 K crystal structure analysis We have collected low temperature (−153ЊC) X-ray ˚ −1. 1.6-pentahydroxyhex-2-enoicacid-4-lactone 190–192 (with decomposition) 176. It is also a powerful reducing agent in aqueous solution. In the present work we have carried out a more . benzene. The most relevant physical data for vitamin C are listed in Table 1. 3. 2 • The first definite X-ray and neutron studies on vitamin C were carried out by Hvoslef [3]. The experimental electron density maps where then compared with the charge density maps calculated using the periodic Hartree–Fock (HF) method as implemented in crystal95 [5].25) gives a delocalized ascorbate anion.3. The most important biochemical properties of vitamin C [2] are due to its reducing power. More-detailed data collection information is available in Table 2. redoxon. physical trauma and ulcer in order to enhance the body’s immune response in fighting infections. Milanesio et al.79 [a]20 D = + 23Њ in water [a]28 D = + 49Њ in methanol 33 (water). vitamin C neutralizes free radicals. 1. The first stage of its oxidation is readily reversible and yields l-dehydroascorbic acid. which can be divided into three parts.65 Melting point (ЊC) Rel./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 Table 1 Most relevant physical data of vitamin C Nomenclature Hexuronic acid.1. prevention of cataract and other eye diseases. l-threo-2.

was used. The final difference Fourier map shows significant residual maxima. 2. together with positions.48r) respectively. therefore.48r) and r exp(−2. The multipole expansion included a monopole and a dipole with radial functions given by exp(−2. finally. the electron density of vitamin C was obtained by two different approaches: first the experimental electron density was derived by the multipole expansion method using accurate diffraction data. the network of intermolecular hydrogen bonds (eight hydrogen bonds per molecule) is retained. The multipole expansion method The aspherical-atom formalism developed by Stewart [7] and implemented in the valray set of programs [8]. Fig. 2. The electron population and isotropic thermal parameters were varied.3 for dipole. also. Structure of the two independent molecules of vitamin C in the unit cell at 120 K. 1. this is shown in Fig. Non-spherical bonding electron density is. with the highest maxima located on the double bonds. The two monopoles of each atom were derived from the canonical SCF s. describing the contraction/expansion of the spherical component of the valence electron density [10]. Milanesio et al. The hydrogen atoms were fixed in the position obtained by a multipole refinement in which they are polarized in the direction of their bonded atom. 2. anisotropic thermal factors. midway between all bonds between non-hydrogen atoms. The adopted multipole model was as follows.2. then the electron density was computed using the crystal95 [5] ab initio method. which displays a section of the map through the five-member ring of molecule a. quadrupole and octopole respectively were used. clearly indicated.M./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 141 Fig. between non-hydrogen atoms do not show relevant variations with respect to the room temperature structure. The multipole terms were considered up to the octopole level. Two radial exponents (a values) one for carbons and the other for oxygens were refined.2. Vitamin C electron density studies As already mentioned. . the results of the two approaches were compared. k parameters and electron population coefficients. H atoms.and p-orbitals [9]. C and O atoms.1. 2.2. Slater-type radial functions r n exp( − ar) with n = 2. The outer monopole was shaped with a variable scaling parameter k. Section of the final difference Fourier map through the fivemember ring of molecule a.

etc.82% 2.99023 0.08722 1. phase transitions at high pressures.44172 0.92249 0.72837 1. b = 6.20090 1.75774 0./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 Crystal appearance and dimensions Space group Temperature Cell dimensions ¯) Collected reflections (hkl and hkl ¯ not merged) Independent reflections (hkl and hkl Reflection with I Ͼ 2j(I) Final R Prismatic and transparent 0.44667 0.22542 0.39265 0.42994 0. on the whole.59573 0.82478 0.99681 1. The program can compute a number of crystalline bulk properties: electron density and Mulliken analysis.49476 0.62332 0.47239 0.53693 1.24170 0.57766 0.90760 0.68836 0.08191 1.2.42427 0.46091 1.88825 0. analogous to the procedure for molecular calculations. These are in turn combinations of Gaussiantype functions (GTFs) whose exponents and coefficients are defined in the input stream (s.61993 0. lattice energy.06551 0.34834 1.39871 0.24488 0.45492 0.15990 1.38740 Z 0. ‘‘LCAO’’ in the present case means that each crystalline orbital is a linear combination of Bloch functions defined in terms of local functions.43633 0.28019 0.45082 0.44825 0. band structure.35447 1.18561 1.80239 0. Electron correlation has been Table 3 Vitamin C atomic coordinates Atom Molecule a X O1 O2 O3 O4 O5 O6 C1 C2 C3 C4 C5 C6 H2O H3O H4 H5 H5O H6A H6B H6O 1.390(1).51523 0.39301 0.40156 0.26579 0. c = 17. II) at increasing pressure were investigated [14].37119 0.42707 0. the structures of the crystalline phases of ice (ice VIII.55148 Y 0.36230 1.44892 0.40737 1.74626 1.84077 0.42156 0.59841 0.36(2)Њ 13618 10817 (5043 after merging) 8957 3. Milanesio et al.56998 0.79001 1.09348 0.127(4) A b = 99.82658 0.16250 1.80327 1.83058 0.44218 0.26616 1.262(1).80708 0.41183 0.60191 1.85091 0.09838 1.32658 1. X-ray structure factors.15442 1.15714 1.79137 0.54546 0.44051 0.36727 1.61722 1.05734 1. p and d shells of GTFs can be used).11612 0.18215 0.95352 0.43013 1.43617 0.42491 0. IX.66932 incorporated using the density functional approach.42843 0.72914 0.33260 0.19028 0. as implemented in the crystal95 ab initio program [5] is.37259 Z 1. So far.13170 Y 0.60979 0.72422 0.96837 0.35773 0.31791 1.63492 1.42698 0. Molecule b X 1.01116 1.32648 0.09700 0.43878 0.56096 0.13527 .41528 1.44311 0.38244 1.44 × 0.36398 1.76293 0. recently.43835 0. the correlation energy was calculated a posteriori with the Perdew 91 potential [12] from the HF-SCF density.29865 0.68485 0.85332 0.52 mm 3 P2 1 −153ЊC ˚ a = 6.60765 0.44762 0.2.47208 0.12788 1.90467 1. crystal95 ab initio method The periodic HF crystal-orbital as linear combination of atomic orbitals (HF-CO-LCAO) method [11].27078 0.01242 1.42 × 0. molecular crystals have not been studied much by periodic calculations [13].142 Table 2 Data collection parameters M.

In Fig./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 143 Fig. Two sections through the five-member rings of molecules a and b of the difference Fourier map r obs − r iam obtained by the multipole expansion method. The manifestation of the electron lone pairs of the O atoms is evident in all maps of Figs. 4 and 5. For comparison. 4. and they reveal essentially the same bond features. 4(a) and Fig. 3. 5(a) and Fig. The final residual density map is shown in Fig. . r obs − r iam (iam: independent atom model). Fig. Deformation electron density at 120 K At the end of the multipole expansion refinement.3. Milanesio et al. This indicates that the multipole model reproduces correctly the aspherical bonding density contained in the experimental X-ray data. are shown.27%.M. Final residual density map.2. the same two sections of the r cal − r iam (r cal: calculated with the multipole model) deformation density map are shown in Fig. the final agreement factor between observed and calculated structure factors was R = 2. 3 and its appearance is an indication of the lack of significant systematic errors in the experimental X-ray data. 4(b) the two sections through the rings of the two independent molecules a and b of the difference map. 5(b). 2.

which is the same used for the study of crystalline urea [13]. In Fig.p) basis set. In order to asses the influence of the intermolecular interactions. 2. 37 of Ref. the values computed with the reduced basis set are smaller than those evaluated by the 621G(d. as shown in Table 4 by the stabilization of about 2 kcal mol −1/H-bond. r(r) bulk. 5./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 Fig. [5]). Deformation density maps r cal − r iam through the five-member rings of molecules a and b. this does not agree with the results of molecular calculations. In the table E bulk is the crystal95 total energy and E molecule is the total energy of an isolated molecule. in which the polarization of the molecules in the crystal field is a relevant stabilizing factor. Milanesio et al. is compared with the density r(r) split calculated as r(r)split = r(r)1 + r(r)2 + … + r(r)K (non-interacting molecules) where the summation is over all K molecules in the cell and the intermolecular interactions were cancelled by an artificial expansion of the lattice (see p. A similar trend was also found for the urea crystal.4. which is quite sizable in all cases. together with the values per mole of molecules and per mole of hydrogen bonds (in the rather crude approximation that all the binding is due to four equivalent hydrogen bonds per molecule). It may be inferred that this trend is due to the periodic nature of the crystals. The binding energy of a mole of unit cells (four molecules per cell) has been evaluated at different levels of approximation for both basis sets and the values are given in Table 4. 6 a difference map r(r) bulk − r(r) split is shown on the plane of the fivemember ring. initially performed with a minimal basis set MINI-1. The pattern of the residual density is in agreement with the expected effects of hydrogen bond formation: electron depletion from the intermolecular region resulting in a denuded hydrogen atom and in a more polar O–H bond.p) basis set. crystal95 calculations on the molecular crystal of vitamin C The calculations.2.144 M. together with . Electron correlation estimated with the HF-SCF density plus the Perdew 91 [12] correlation functional is also relevant. were then carried out with the wmore extended 6-21G(d. where it is found that too small a basis set tends to overestimate the binding energies. As may be seen. DE has also been corrected for BSSE error. the bulk electron density. which is not accounted for by the MINI-1 basis set.

p) a delocalized effect due to rearrangements of the molecular periphery [15].8 26. may be rather difficult to perform in direct space and it becomes much easier when it is carried out in reciprocal space. DE H-Bond = DE Mol/4 Basis set Binding energy per DE (kcal mol −1) No correction MINI-1 Cell Molecule H-Bond Cell Molecule H-Bond 103. the overall discrepancy factor between the two sets of data is very low: R bs = ∑ lFbulk − Fsplit l=[ ∑ Fbulk ] = 0:40 It is important to note that both structure factors refer to atoms at rest and are purely theoretical results free of any experimental error. shows that the standard deviations are minimal in the intermolecular regions.b).2. 2.9 142. the agreement between these two sets of data is also very good: R bc = ∑ lFbulk − me Fcalc l= ∑ Fbulk = 2:37 The small value of R bs indicates the very small effect of the intermolecular interactions. Considering that the latter values are affected by the experimental errors. and it might seem surprising that the multipole expansion maps not only clearly show all intermolecular hydrogen bonds.0 11. whereas experimental errors are random. the corresponding structure factors F bulk and F split may be computed and compared. calculated with the 6-21G(d. the small value of % % . Enlarged views of the r(r) bulk and r(r) bulk − r(r) split maps in the region of the shortest and the longest hydrogen bonds are shown in Fig.7 12.7 15. Indeed.5 35.9 BSSE and Correlation — — — 176. Difference map r(r) bulk − r(r) split on the plane of the fivemember ring. Comparison of the computed structure factors obtained by crystal95 and by the X-ray analysis The comparison of similar electron density maps Fig. DEMol = DECell /4.5.0 145 6-21G(d. Furthermore. Two reasons can be given to explain this fact: (i) the intermolecular interactions give a small systematic signal.p) basis set. 6. Milanesio et al. which are the same for both sets of data. F bulk may also be compared with the structure factors calculated from the multipole expansion density and back-corrected for the thermal displacement effects (me Fcalc ).6 8.6 50. They are only affected by the choice of the basis set and by the approximations made in crystal95. 10(a. (ii) as we shall see later./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 Table 4 Binding energies calculated by crystal95 : DECell = Ebulk − 4Emolecule . and the effects of a strong and a weak interaction may be compared. 8(a. the error analysis of the electron density maps obtained by the multipole expansion approach.0 44.b) and Fig. from the electron densities. Their effect on the residual is much smaller than that of experimental errors.b).4 3.7 BSSE 61. but can also discriminate their strength. The maps may also be compared with those obtained by the multipole analysis of the diffraction data and shown in Fig. 7(a.5 202.M.0 6.

35%) when it is computed with F bulk values calculated using the MINI-1 minimal basis set. 9(b). . 7.6. 8. removed from the crystal. It is worth pointing out that the agreement is not as good (R bc = 3. 2./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 Fig. They were computed assuming the generalized scattering factors as obtained by the multipole Fig. Milanesio et al.2. R bc shows that the model given by crystal95 is in very good agreement with the experimental data. are shown in Fig. Effects of the strongest hydrogen bond on the charge density of vitamin C: (a) r(r) bulk map and (b) r(r) bulk − r(r) split map. Effects of the weakest hydrogen bond on the charge density of vitamin C: (a) r(r) bulk map and (b) r(r) bulk − r(r) split map.146 M. 9(a) and Fig. Electrostatic potential and electron density maps from 120 K X-ray data Electrostatic potential maps relative to molecules a and b.

627 2.93 0.786 2.707 2. H…O (A 1.76 2.749 2.78 1. .666 2.08 ˚) Dist.82 1. as discussed later) is shown by the less pronounced minimum.87 1.75 1. the two maps are similar and clearly indicate the most attractive zones for protons in hydrogen-bond formation.88 0.M.692 2. Milanesio et al.769 2.96 0.93 1.95 0. O–H (A 0. whereas the low acceptor ability of O3 (corresponding to the most acidic OH group.90 1./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 147 Fig. On the whole.907 2.935 Angle O–H…O (deg) 169 a 169 b 171 171 158 165 157 162 168 167 164 161 162 156 149 149 Present analysis.98 0.95 ˚) Dist.649 2.08 0.72 1.85 0. the deepest minima are close to O1 and O2 (the features around the side-chain oxygen atoms cannot be seen on the maps). Neutron diffraction.77 1.05 0.86 1. refinement and the figures illustrate the sections through the five-member rings.645 2. 9.590 2.658 2.768 2.63 1.79 1.656 2.65 1.88 0. O…O (A 2.88 0. This is in agreement with the pattern of Table 5 Most relevant geometrical features of the eight independent hydrogen bonds in vitamin C Bond (1) O2b–H…O6a (2) O2a–H…O5aЈ (3) O3b–H…O1bЈ (4) O3a–H…O1aЈ (5) O5a–H…O6bЈ (6) O6a–H…O5bЈ (7) O5b–H…O2bЈ (8) O6b–H…O2aЈ a b ˚) Dist.10 2.94 0.612 2.85 1.96 0.90 0. Electrostatic potential maps relative to molecules a and b.73 1.85 1.

d) similar maps relative to the two hydrogen bonds (3 and 4)./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 Fig.786 A ˚ ) link. 10. 1.b) the electron density maps on the planes of the shortest (1) and longest (8) hydrogen bonds are shown. as determined by the present study.148 M. 10(a. In Fig. 10(c. (b) the longest hydrogen bond. The most significant difference between the two maps is in the value of the minima close to O2. . are compared with those obtained by Hvoslef [3] from neutron diffraction data at room temperature. are illustrated. in which O(3)–H acts as donor. which is stronger than the O6bЈ –O2a (2. and in Fig. (c) the O(3)–H…O(1bЈ) and (d) the O(3)– H…O(1aЈ) interactions. In Table 5 the most relevant geometrical features of the eight independent hydrogen bonds. it is worth noting that the deeper minimum near O2b cor˚ ) hydrogen bond responds to the O5bЈ –O2b (2.935 A The electrostatic potential maps around all hydrogen bond regions have also been examined for a more detailed analysis of the hydrogen bonding patterns found in the crystal. hydrogen bonds found in the crystal and shown in Fig. Electron density maps of (a) the shortest hydrogen bond. Milanesio et al.

p) is then used throughout the paper. The main results of this topological study are in good agreement with the description of the chemical bonds of vitamin C in terms of deformation electron density.p) basis sets.56 3-21G(d. No geometrical constraints were imposed during the optimization.3. 4 and 5. since it adds an extra diffuse set to the standard 6-31G(d. The standard notation. HF basis set effect The HF basis set dependence of the relative stabilities of a subset of six conformers is shown in Table 7.35 −0. has been used in single point (SP) energy calculations carried out on the HF/6-31G(d. The 6-31+G(d. C. D.240 j(r) 0. F.49 2.91 149 6-31G(d.410 0.36 0.p) level. B.36 3. 2. while the conformer resulting from the optimization of X s is named X o. A quick evaluation of the relative stabilities of each conformer was carried out by optimizing the geometry at the HF level with both MINI-1 and 3-21G(d.3. 2.p) geometries. In Table 6 we give the values of the electron density at the bond critical point (a point where ٌr(r) vanishes) only for the hydrogen bonds shown in Fig.024 r/j(r) 81 75 83 10 A B C D E Xo Table 7 Basis set effect on the relative stabilities at HF level Conformer DE (kcal mol −1) MINI-1 2.0 4.p) level with respect to the absolute minimum G are shown in Fig.16 1.1. The recently proposed hybrid density functional method B3-LYP. Electron correlation effects on the geometrical features were evaluated for five selected conformers by geometry optimization carried out at the B3LYP/6-31+G(d. The experimental X-ray conformation is labelled X s.200 0.86 2. Milanesio et al.M. Recently.090 0. different from that found in the X-ray structure.p) basis set is the best compromise between accuracy and efficiency at the correlated level.88 0. B3-LYP/6311+G(2d.34 0. 1. 10.0 1.08 4.0024 0. To increase flexibility in the basis set. Theoretical calculations on the isolated molecule Molecular manipulation by means of the moldraw [18] program was used to generate all relevant conformers stabilized by intra-molecular hydrogen bonds. The main geometrical features of each conformer.2p) set has been adopted.61 3. which consists of the Becke [19] three-parameters gradient-corrected exchange term plus the gradientcorrected correlation component due to Lee et al. The selected conformers are the only available at . This result appears to be of similar quality to that obtained for l-alanine [17] from X-ray data at 23 K.37 2. 11. were finally selected and named in the following as A. Starting from the optimum HF/3-21G(d. 3. Even though at these points the electron density is very low./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 Table 6 Electron density and its standard deviation at the critical points of same selected H-bonds H-bond O2b–H…O6a O6b–H…O2a O3b–H…O4b O3a–H…O4a r(r) 0.p) 3.0012 0. E. a full optimization with the 6-31G(d. it has also been shown that this basis set is very reliable in treating hydrogen bonds [21].19 0. Nine conformers.43 The Bader [16] analysis of the experimental electron density distribution was also carried out. The following computational strategy was adopted. and they are not reported here. as shown in Figs.p) approach ensures good geometric and reasonable energetic results. the double polarized 6-311+G(2d. 2. Electron correlation effects on the relative stability of each conformer were taken into account by the density functional method. G. [20]. H and I. From experience accumulated over the past years it has been shown that the HF/6-31G(d.66 0.p) geometries.2p)//HF/6-31G(d.p) used at the HF level.p) basis set was carried out.005 0. 4.p) 1. its evaluation by means of the multipole model employed to fit our data is highly significant. as well as the relative stabilities (RS) at the HF/631G(d.

Milanesio et al./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 Fig.p) basis sets overestimate intra-molecular hydrogen bond interactions. whereas the HF/6-31G(d. 11.3. while the HF/6-31G(d.1 kcal mol −1 above that of conformer E. Electron correlation effects As previously mentioned. Conformer C is a good example of the basis set effect. and the relative stabilities of the conformers are changed. G.2. so that conformers F.p) method gives a more realistic structure with an energy 2.150 M. all energies are given relative to the most stable conformer E. H.p) method gives a more realistic picture of these interactions. I are not included in Table 7. critical for a proper evaluation of the effects of intra-molecular hydrogen bond interactions. Within the selected set. The choice of a sufficiently extended basis set is. 2. Geometrical features of the 11 conformers of vitamin C taken into account. Indeed HF/MINI-1 gives such short hydrogen bonds that this conformer becomes the absolute minimum.p) levels. X s. Both MINI-1 and 3-21G(d. two different procedures were considered in order to take into account . MINI-1 and 3-21G(d. minimal basis sets are inadequate to describe these weak interactions. therefore.

carried out on the X o.p) level. The relative stabilities of the various conformers calculated with the different methods are shown in Table 8. the effects of the geometry relaxation induced by electron correlation. Hydrogen bond features When the molecule is isolated. Milanesio et al.3. have been assessed by full optimization (OPT) at the B3-LYP/631+G(d.M./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 151 Fig. as indicated in the . and the stabilities of some of the conformers are changed with respect to those obtained at the HF/6-31G(d. On the one hand. as resulting from HF/6-31G(d. E. All conformers present at least two hydrogen bonds. the formation of intra-molecular hydrogen bonds may become a stabilizing factor. but only some interactions have stabilizing effects. G and I conformers. These changes cannot be ascribed to the lack of geometry optimization because B3-LYP(SP) and B3-LYP(OPT) data are almost identical. has been accounted for by SP calculations at the B3LYP/6-311G+G(2d.p) calculations. Electron correlation plays a significant role.p) level. correction to the relative stability of each conformer. F.2p)//HF/6-31G(d. (continued ) electron correlation effects. 2.p) level. On the other hand. As a consequence. we shall only discuss the data obtained at B3LYP(SP).3. 11.

G C. which is not involved in any Hbond in the crystal. 11. G.5 +2. E. (continued ) following table: Interaction O(2)–H…O(1) O(2)–H…O(3) O(3)–H…O(6) O(3)–H…O(2) O(5)–H…O(4) Conformer E.0 −2. C X o. being the one responsible for the low value of the pK a. D E. Conformer E shows a medium strength hydrogen bond between O3–H and the oxygen atom O6: it is . Milanesio et al. 11): in both conformers O3–H acts only as a proton donor. O3–H is the most acidic group of vitamin C. behaves as a good proton acceptor.5 −2. I C. is a very poor proton acceptor. The minimum energy gas-phase conformations G and E are particularly stable because of the presence of favourable intra-molecular hydrogen bonds (Fig. F. at the same time. In the crystal the O3–H group is the only one which does not behave as hydrogen acceptor from the neighbouring hydrogen donor groups. whereas O4.0 −2. G Effect Stabilizing Destabilizing Stabilizing Stabilizing Stabilizing Energy gain (kcal mol −1) −2.152 M. B. Indeed. H. It is. then. because no relevant steric hindrance is present around the O3 atom.0 It is worth noting that the O3–H group prefers to act as proton donor. This is due to electronic factors./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 Fig. no surprise that it acts as the strongest hydrogen donor and.

Newmann projections along bond C5–C6. the largest energy difference (between D and G) is less than 6 kcal mol −1.2 3./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 Table 8 Electron correlation effect on relative stabilities Conformer DE (kcal mol −1) HF/6-31G(d.2p)//HF/6-31G(d.p) A B C D E F G I H Xs Xo 2.1 1. E and H have a significant population. The alternative hydrogen bond formed by the O2–H group with the sp 3 oxygen O3. B3-LYP(OPT) = B3-LYP/6-31+G(d.0 0. Nevertheless.1 3.0 3.1 1.p) basis sets give energetic results in contrast with the chemical properties of vitamin C.0 1. Fig.5 1.4 B3-LYP(SP) = B3-LYP/6-311+G(2d. .9 1.17 2. with respect to conformers E and F.1 3.0 3. A further stabilizing interaction. Conclusions The results of the B3-LYP/6-311+G(2d. and it must be expected that the formation of intermolecular hydrogen bonds with polar solvent molecules or with neighbouring molecules in the crystal can easily alter the relative stabilities of the different conformers.6 3.9 2. Indeed.p) level Conformer G E H I C F Xo B A Xs D DE B3-LYP(SP) (kcal mol −1) 0. is unfavourable because of the poor H-acceptor behaviour of the O3–H group.2 2.2p)//HF/6-31G(d.0 0.2p)//HF/631G(d. 11).M. This is further evidence that the MINI-1 and 321G(d.p) or more extended basis sets are adopted.p).61 B3-LYP (SP) 3.5 1. which are only accounted for by the more accurate calculations.1 B3-LYP (OPT) — — — — −0.9 3.4 0.7 3.3 4. This is certainly the case of the X s conformation found in the crystal.7 7. will be used in the present discussion of the gas-phase conformations. O1. E and F. as found in conformers C and D (Fig.0 0.0 1.3 3. shown in Table 9. Indeed the formation of an extensive network of inter-molecular hydrogen bonds in the crystal favours a conformation with the worth noting that this interaction is particularly strong because of the resonance-assisted hydrogen bonding mechanism [22]. 3.p)//B3-LYP/6-31+G(d.p).2 3.3 5.5 153 Table 9 Relative stabilities at B3-LYP/6-311+G(2d.9 5.p) SP calculations. a net decrease in stabilization is computed for these two latter conformers. when 631G(d.2 3. which is only present in conformers G.1 4.0 1. it can be concluded that for the isolated molecule of vitamin C only the three conformers G.0 5. 12. From a simple Boltzmann analysis of the values of the relative energies listed in Table 9. Milanesio et al.9 — — 3.9 0. is the intra-molecular hydrogen bond between O2–H and the sp 2 oxygen atom.

Lee. Third Electronic Computational Chemistry Conf. P. Acta Crystallogr. J. side chain stretching out of the five-member ring to maximize the possible interactions with the surrounding molecules. R.). adopt the +syn disposition with a lower population. Dovesi. C and E. R. (b) C. Causa ` . J. 12) shows that all three possible dispositions (anti.. Berlin. Italy. crystal95 User’s Manual. F. Merati. conformers G. USA. D. R. adopt the least populated −syn disposition. Wang. B: 24 (1968) 23. Chem. Saunders. Electrostatic properties of molecules from diffraction data. M. Roetti. Royal Society of Chemistry. Acta Crystallogr. Ugliengo. R. Carnegie-Mellon University. pp. L. Orlando. J. 1996. A. 63 and references cited therein. Lett.A. V. Bianchi. November 1–30. Phys. Roetti. 45 (1992) 13244. Bertolasi. when the molecule is isolated. Szent-Gyo ¨ rgyi. 98 (1993) 5648. Gilli. Pisani. 1991. 207 (1993) 9–23. V. Reid. Lett. Dovesi.. an analysis of the Newman projections along bond C5–C6 (Fig. R. J. 1988. 1995. P.R. H and I. Biochem. in: A. Harrison.U. Z. J. Stewart. J. W. with the preferred anti disposition. A. the side chain stretches most out of the five-member ring to allow the formation of inter-molecular hydrogen bonds with polar solvent molecules. Perdew. F. D. B: 49 (1993) 564–576. p. Data Nucl. Chem.R.unito. Dovesi.B. Parr. Hartree–Fock Ab Initio Treatment of Crystalline Systems. 48. are represented among the considered conformers: • in X s. Clementi. Zupan. R. the formation of favourable intra-molecular hydrogen bonds becomes the main stabilizing factor. This work was supported by the Italian MURST and CNR. 1996. Chem. Phys. Piniella (Eds. P. USA. At. Dovesi. R. C. P. 1983. Pittsburgh. van der Wal. Lecture Notes in Chemistry. Destro. Phys. Gatti. M. J. Sect. Sect. Garrone. 100 (1994) 2128. Orlando.R. P. T. V. with the side chain curled up to form a hydrogen bond between O6 and O3. On the other hand. B. Partridge. Luck. University of Torino. Oxford University Press. (b) M. C. Gaussian Inc. 1990. Atoms in Molecules: A Quantum Theory. Roetti. Rev. Gatti. Phys. Y. N. As far as the side chain conformation is concerned. Phys. M. K. Plenum. R. Pittsburgh. C. J. E. Becke. 220 (1994) 145. Roetti. Apra ` . Viterbo. E. Phys. R. Zeegers-Huyskens (Eds. J. gaussian94. V.S. available at http://www. Educ. Heidelberg.G. Revision C. Paper 22. Jeffrey. Huyskens. Davies.154 M. Chiari.F. Ferretti. G.. Saunders. R. Hermansson. Chem. Gilli. with a less stretchedout side chain. [15] [16] [17] [18] [19] [20] [21] References [1] A. Springer. 101 (1994) 10686. M.P. conformers B. Kristallogr. Chem.M. (a) R.J. B 37 (1988) 785. Milanesio et al. W. Yang. Springer. FreyriaFava and Dr. R. +syn and −syn). C. Sect. considered by Reid [4] in his NMR study of vitamin C in water solution. Saunders. Chem. Phys.W. Causa ` .D. Intermolecular Forces. [2] M. V. valray User’s Manual. (a) J. A. Torino./Journal of Molecular Structure (Theochem) 419 (1997) 139–154 Chemistry and Biochemistry. Hvoslef.3. Ugliengo. Chem. Causa ` . Frisch et al. D and F. Rev. Data Tables 14 (1974) 177–478. Roetti. also acknowledges the allowance of free computing time by the CINECA Supercomputing Centre. Stewart. C.html. 22 (1928) 1387. Vitamin C: Its [22] . Acta Crystallogr. Austin. C. Spackman.J. Saunders.F. Stewart. 1991. The Application of Charge Density Research to Chemistry and Drug Design. Department of Chemistry.. G.A.. C. Orlando for producing electron density maps with crystal95. 41–43. Ojamae. Bader. R. C. Roetti.). 66 (1989) 344–345. R.R. B. A: 40 (1984) 587–593..A. 1991. C.F.F.F. 186 (1991) 47.P. E. Phys. 92 (1990) 7402. V. [3] [4] [5] [6] [7] [8] [9] [10] [11] • • [12] [13] [14] Acknowledgements We wish to express our thanks to paper22/frame. R. Civalleri. vol. PA.