Intramolecular stabilization of a catalytic [FeFe]- hydrogenase mimic investigated by experiment and theory†

The mono-substituted complex [Fe2(CO)5(μ-naphthalene-2-thiolate)2(P(PhOMe-p)3)] was prepared taking after the structural principles from both [NiFe] and [FeFe]-hydrogenase enzymes. Crystal structures are reported for this complex and the all carbonyl analogue. The bridging naphthalene thiolates resemble μ-bridging cysteine amino acids. One of the naphthyl moieties forms π–π stacking interactions with the terminal bulky phosphine ligand in the crystal structure and in calculations. This interaction stabilizes the reduced and protonated forms during electrocatalytic proton reduction in the presence of acetic acid and hinders the rotation of the phosphine ligand. The intramolecular π–π stabilization, the electrochemistry and the mechanism of the hydrogen evolution reaction were investigated using computational approaches.


Introduction
Complexes of the type μ-S 2 Fe 2 (CO) 6 were first reported in 1965 1-3 and early chemistry for this class of complexes was developed by Seyferth and co-workers. [4][5][6] It was only after the report of the first crystal structure of the [FeFe]-hydrogenase active site that this class of complexes was thoroughly investigated. 7 The H-cluster has a binuclear active site with an azadithiolate bridging ligand and carbonyl and cyanide ligands to iron atoms. This [2Fe-2S] cluster unit in the active site catalyses the reversible reduction of protons to hydrogen (see Fig. 1). 8,9 Based on the structural similarity of this class of complexes to the active site a large number of organometallic [2Fe-2S] clusters have been reported as electrocatalysts for proton reduction. [10][11][12][13][14] Whereas the Fe atoms of the [FeFe]-hydrogenase are connected via a bridging carbonyl and an azadithiolate ligand, the Ni and Fe atoms of the [NiFe]-hydrogenase enzyme are covalently bound to two bridging cysteinate amino acid residues. We here present a catalytically active homobimetallic Fe model complex mimicking the [FeFe]-enzyme with an asymmetric ligand environment and two bridging monothiolate ligands resembling the [NiFe]-enzyme (see Fig. 1).
A large number of model complexes have been reported either as structural or functional mimics of the active site. [10][11][12][13][14][15] Most of the mimics use dithiolate ligands with or without an amine group in the metal linker, see for example ref. 16-22. Only a few examples of complexes are known with monothiolate types of ligands, in which there is no covalent linkage between the two sulphur atoms. [23][24][25] The consequences of removing the linker between the sulphur atoms have been investigated by Lichtenberger et al.  2 (P(PhOMe-p) 3 )] complex (below) takes up design principles from both catalytic centres.
for the alkyl thiolato complex, [Fe 2 (CO) 6 (µ-SMe) 2 ] A. 23 Since π-electron rich polyaromatic thiolate ligands are known to influence the stability and electrochemical properties of the [Fe 2 (CO) 6 ] core, the precursor complex, [Fe 2 (CO) 6 (µ-naphthalene-2-thiolate) 2 ] 1 and electron rich tris (4-methoxyphenyl) phosphine (L = P(PhOMe-p) 3 ) were used to synthesize the diiron carbonyl complex [Fe 2 (CO) 5 (µ-naphthalene-2-thiolate) 2 (P(PhOMe-p) 3 )] 2. The monodentate phosphine ligand was used to provide an electron rich environment around the iron center and study its influence on the stability and electrochemical properties of 2. The steric demand of the phosphine influences does not only affect the electronic structure of the iron atoms but also hinders the phosphine ligand rotation by forming π-π stacking interactions between the bridging monothiolate and the terminal phosphine ligand.

Preparation and structural characterization
The starting complex [Fe 2 (CO) 6 (µ-naphthalene-2-thiolato) 2 ] 1 was synthesized as reported in the literature. 25 The synthesis of the target complex [Fe 2 (CO) 5 (µ-naphthalene-2-thiolate) 2 (P(PhOMe-p) 3 )] 2 is presented in Scheme 1. As shown in the scheme, complex 2 was prepared by the substitution of one of the CO ligands on 1 with the electron-donating phosphine ligand (L = P(PhOMe-p) 3 ). A dichloromethane solution of 1 was added to an acetonitrile solution of Me 3 NO·2H 2 O and stirred for 10 min under an argon atmosphere at room temperature. This solution was then stirred at room temperature for 6 h under an argon atmosphere upon addition of the phosphine ligand. The complex was obtained as a blood-red solid after the removal of the solvent followed by column chromatography on a silica gel column using hexane-dichloromethane solution (3 : 2). Blood-red coloured crystals were obtained for 2 from hexane-dichloromethane solution at low temperature. The molecular structures were determined by X-ray diffraction analysis. The X-ray crystal structure of 1 not reported earlier was also obtained and is given in the ESI. † The crystal structure of 2 is shown in Fig. 2. The crystallographic parameters are given in the ESI. † New crystallographic and structural parameters of complex 1 were available before but were determined here and are also given in the ESI. † The Fe-Fe bond lengths for 1 and 2 are 2.49 and 2.52 Å, respectively which are close to that reported for the active site of the hydrogenase enzyme (2.6 Å). 7 The slightly elongated Fe-Fe bond is due to the stronger σ-donor properties of the phos-phine ligand in 2 compared to the carbonyl group. The phosphine ligand occupies the apical position in the diiron complex 2 which is similar to that seen in the reported phosphine substituted complexes. 10,12,[26][27][28][29] Both complexes display a butterfly structure ( Fig. 2 and ESI †) with the two naphthyl groups in an equatorial conformation in 1. In 2 one of the p-PhOMe moieties of the terminal phosphine ligand leads to a spatial reorientation of one of the naphthyl rings perpendicular to the second to form π-interactions. The two naphthalene ligands thus occupy one equatorial and one rotated perpendicular axial orientation.
Complex 2 was characterized by FTIR, NMR and UV-Vis spectroscopies. The FTIR spectrum of complex 2 is shown in Fig. 3. The FTIR spectra of the mono-substituted complex 2 show several strong absorption bands in the region of 2040-1930 cm −1 in dichloromethane, that are assigned to terminal carbonyl groups.
The ν CO bands for 2 are slightly shifted towards lower wavenumbers (by ∼60 cm −1 ) compared to 1 (2070-2000 cm −1 , see Table 1) but similar to those observed for analogous phosphine substituted complexes. 12,25,30 The shift towards lower wavenumbers is due to the attachment of a basic phosphine ligand to one of the iron centres. The shift is slightly more pronounced than that observed for the reported triphenyl-Scheme 1 Synthesis of complex 2 from its precursor 1.  phosphine substituted complex [Fe 2 (CO) 5 (µ-naphthalene-2thiolate) 2 (PPh 3 )]. 25 This indicates that P(PhOMe-p) 3 with the methoxy groups in the para-position of the phenyl rings is a stronger electron donating ligand than PPh 3 . The calculated spectra for complexes 1 and 2 are given in the ESI † and allow an assignment of the FTIR bands to symmetric and asymmetric molecular stretching modes of the CvO ligands.
The 1 H NMR spectrum of 2 in CDCl 3 (see the ESI †) displayed peaks in the aromatic region (8.0-7.0 ppm) for the phenyl and naphthyl ring protons, in addition to a singlet at 3.76 ppm for the methyl protons of OMe. A singlet with a chemical shift value of 52.56 ppm was observed in 31 P { 1 H} NMR spectra which confirms the presence of the phosphine ligand. A fast rotation of the phosphine ligand around the P-Fe bond is not possible since one of the p-PhOMe ligands forms π-stacking interactions with one of the μ-bridging naphthylthiolates (see Fig. 2 and below) unlike the fast rotation of terminal cis-1,2-C 2 H 2 (PPh 2 ) 2 ligands in a dithiolate complex. 31 The presence of a phosphine ligand in the mono-substituted complex 2 leads to weak metal-centered d-d transitions in the visible region and ligand-centered π-π* transitions in the UV-Vis spectra in acetonitrile and dichloromethane (see the ESI †). The appearance of the bands at a higher wavelength for 2 as opposed to those at 249 and 346 nm for the all carbonyl complex 1 is because of greater electron density at one of the iron centers due to the attachment of a phosphine ligand in the apical position.
The cyclic voltammograms (CVs) for 2 were measured in acetonitrile under an argon atmosphere. The CVs for 2 displayed two one-electron irreversible reduction waves (E pc = −1.66 and −2.37 V versus Fc/Fc + ) that can be assigned Fe I Fe I → Fe I Fe 0 and Fe I Fe 0 → Fe 0 Fe 0 (see Fig. 4). The calculated redox potentials of −1.62 V and −2.19 V support this assignment. Upon one-reduction the calculated Fe-Fe bond distance increases from 2.57 Å in 2 (2.52 Å in the crystal structure) to 2.88 Å in 2 − . This can be rationalized when inspecting the LUMO of 2 which is an anti-bonding combination of the iron d z 2 orbitals (Fig. 5). The coordination sphere of the iron atoms remains unchanged (compared to 1 − , see below). Upon oneelectron reduction of complex 1, one of the Fe-S thiolate bonds is broken (Fe⋯S distance 3.96 Å) and the excess electron is localized on the four-coordinate Fe only (unpaired spin density 0.95). The second reduction to yield 1 2− and 2 2− is accompanied by an increase in iron-iron bond distances to 2.75 Å and 3.54 Å, respectively, whereas the coordination sphere of 2 2− is unchanged, in 1 2− the Fe-S bond is fully dissociated (with a Fe⋯S distance of 4.49 Å; see the ESI † for details).
In the crystal structure of 2, one of the aromatic phenyl rings forms π-stacking interactions with one of the bridging S-naphthalates (see Fig. 2 and 5).
The calculated unpaired spin density ( Fig. 6) shows that in the mono-reduced Fe I Fe 0 species 2 − the electron is non-uniformly distributed over both iron atoms with atomic spin densities for Fe(CO) 3 of 0.25 and 0.63 for Fe(CO) 2 (P(PhOMe-p) 3 ). This is in agreement with the shifted reduction potentials of 2/ 2 − due to the strong electron donating phosphine ligand (see below).   In the two-fold reduced state, an additional π-stacking interaction is formed between a different aromatic ring of the terminal PhOMe-p substituent and the second bridging thiolate (see Fig. 7). This is accomplished by the rotation of this naphthyl ring from an equatorial into an axial orientation. This extra intramolecular interaction helps to stabilize 2 2− and retain the coordination environment.
Two one-electron irreversible oxidations were also observed at E pa = 0.29 and 1.00 V versus Fc/Fc + which might be ascribed as Fe I Fe I → Fe I Fe II and Fe I Fe II → Fe II Fe II , respectively (see the ESI †). The assignment of one-electron oxidation events is supported by calculations in which BP86 performs superior to B3LYP (see Table 2). The reduction of compound 2 occurs at a more negative potential than 1 and its oxidation at a less positive potential than complex 1 ( Table 2). 25 This is due to the substitution of CO for P(PhOMe-p) 3 making the [Fe-Fe] center of 2 more electron-rich than 1. The electrochemical data are consistent with the results of similar model complexes reported in the literature. 26 Furthermore, the reduction potentials for 2 are slightly more negative in comparison with that of [Fe 2 (CO) 5 (µ-naphthalene-2-thiolate) 2 (PPh 3 )] B thus, indicating better electron donating properties of the phosphine ligand in 2 ( Table 2).
Upon oxidation of 2 to 2 + and 2 2+ , the Fe I Fe II and Fe II Fe II states preserve their structural integrity. All covalent bonds remain intact. Complex 2 was examined for its ability to act as an electrocatalyst for the reduction of protons to molecular hydrogen in the presence of acetic acid (Fig. 8). The complex was found to be unstable in the presence of stronger acids ( perchloric acid). CVs of 2 in the presence of acetic acid show a new peak at −2.02 V versus Fc/Fc + which shifts cathodically with an increase in the amount of acid. The increase of current at this reduction potential with an increase in the amount of acid can be attributed to the reduction of protons to molecular hydrogen. 26 The first reduction peak at −1.66 V observed in the absence of an acid is shifted anodically to −1.60 V upon addition of acid without much increase in current with increasing amounts of acid.
The diiron-based HER catalysts operate via an intermediate mixed-valence hydride species considered to be a rare class of complex 10,32 and difficult to isolate. 12 After reduction to 2 − (see Fig. 5), such a hydride species is obtained from the first protonation in an ECEC mechanism. One-electron reduction of mono- 33,34 and diiron-complexes 35 with bridging dithiolates usually leads to an elongation of one Fe-S bond to afford initial protonation of the μ 1 -thiolate. In the presence of a second coordination shell basic site 36,37 an initial ligand protonation might be kinetically driven but eventually a thermodynamically favoured metal-hydride species is subsequently formed. 37 Both BP86 and B3LYP calculations were shown to yield proton affinities within 1-2 kcal mol −1 with respect to CCSD(T) calculations and experiments. 38 BP86 slightly overestimates and B3LYP slightly under-estimates proton affinities. In the absence of a ligand protonation site (such as nitrogen), 2 − can be protonated to yield either a terminal Fe-hydride ( proton affinity of 8 kcal mol −1 ) or a μ-bridging hydride ( proton affinity of 17 kcal mol −1 ) (complex 2H, see Table 3). A terminal ironhydride can be thermodynamically unstable and convert to the bridging hydride by an intramolecular isomerization even at   low temperature. 39 In the absence of a vacant terminal protonation site, a 'rotated' structure with a semi-bridging CO ligand is obtained for 2H terminal , which is also observed for related dithiolate complexes. 40 The equilibrium between 'rotated' and 'unrotated' structures is mediated by subtle steric effects of bidentate dithiolate ligands (ethanedithiolate versus propanedithiolate). 42 Protonation of a bridging naphthalenethiolate is less favoured with a calculated proton affinity of ∼3 kcal mol −1 which is also found in ref. 41. The protonated thiolate might also re-arrange to a terminal iron hydride. Protonation of one of the μ 2 -naphthalenethiolates might be the reason for the catalyst instability and degradation in strong acids ( perchloric acid).
In the plot of i cat /i p versus acid concentration for complex 2, the current first increases and then levels off at a maximum of 62.65 mM of acid in solution (see the ESI †). The initial linearity indicates that the reaction is second order with respect to acid concentration (see eqn (1)) for a scan rate of 0.1 V s −1 (see the ESI †).
Also, the ratio i cat /i p for two different acetic acid concentrations (20.8 and 62 mM) varies linearly with the square root of the scan rate (ν −1/2 ) confirming the validity of eqn (1) (see the ESI †). Scan rate independence is achieved only at a scan rate of 0.4 V s −1 and greater (see the ESI †). The plot of the amount of acetic acid versus k obs (turnover frequency (TOF) from eqn (2) and (3)) for complex 2 calculated from the i cat /i p ratio is shown in Fig. 9.
A value of i cat /i p of 21.43 was obtained for 2 (1.62 mM) for the highest acid concentration investigated (62.65 mM), corres-ponding to a turnover frequency of 88 s −1 . 42 The TOF of 2 was determined by taking the successive cyclic voltammograms of the reaction mixtures for which the concentration of acetic acid was systematically increased until the k obs (TOF) remained constant. For comparison, the data generated for complexes 1, B and [Fe 2 (CO) 6 (µ-SPh) 2 ] C have also been included (see the ESI †). 24,25 Furthermore, the catalytic efficiency was examined by determining the overpotential in MeCN. The overpotential for 2, calculated as described by Evans et al., was found to be −0.56 V ( Table 2). 26 Complex 2 was found to be a better catalyst in comparison with complexes 1, B and C. Also, complex 2 catalyzes the reduction of protons at a slightly more negative potential than 1 and B and at a less negative potential than C.
The background current due to the direct reduction of protons at the glassy carbon electrode without the catalyst in the presence of acetic acid was found to be negligible for potentials in the range of −1.4 to −2.1 V. Hence, the acidinduced currents in the presence of catalysts can be attributed to catalytic turnover. Similar background measurements for different acids in the absence of a catalyst have also been recently looked into by McCarthy et al. 43 The second one electron reduction from 2H to 2H − was calculated to occur at redox potentials of −1.67 V/−1.98 V (BP86/ B3LYP) for 2H bridging compared to −2.19 V for 2 − /2 2− reduction of the sole catalyst in the absence of an acid. The final chemical (C) protonation step from 2H − to 2HH is unlikely to occur at an iron atom since a dihydride species in a formal Fe I Fe I complex would be formed. The metal oxidation state can be preserved when protonation occurs at a sulfur atom of the bridging μ-thionaphthalate. Such a protonation is thermodynamically feasible with a calculated proton affinity of 19 kcal mol −1 (BP86) and 25 kcal mol −1 (B3LYP). This ECEC̲ step, however, leads to a cul-de-sac since the doubly reduced species displays two π-π stacking interactions between the bridging μ-thiolates and the terminal P(PhOMe-p) ligands (see Fig. 7 and Fig. 10). This interaction hinders the intramolecular rotational isomerization of the sterically demanding μ-naphthalene around the S-C carbon which would be necessary to bring the protonated sulfur and the bridging hydride in close spatial vicinity.
The monothiolate bridging ligands, unlike bridging dithiolates, enable a direct protonation of the μ-hydride species to form a Fe 0 -H 2 -Fe 0 complex. Such a protonation retains the Table 3 Computed differences in proton affinities for 2 − to afford 2H. Gibbs free energy differences between the terminal 2H terminal and bridging 2H bridging hydrides are given in kcal mol −1 . Systematic errors are thus expected to cancel out. A positive value indicates a preference for a μ-hydride metal oxidation states to 2HH from which molecular hydrogen is easily released and complex 2 is recovered (see Scheme 2). During the path of the electrocatalytic proton reduction, the Fe⋯Fe distances only change by less than 0.8 Å (see the ESI †). The suggested mechanism resembles the reverse reaction of a [NiFe]-hydrogenase enzyme when oxidizing H 2 . Molecular hydrogen is activated between the Ni and Fe atoms of the active site and a μ-hydride is obtained as a stable intermediate Ni-C.

Conclusions
A new diiron carbonyl complex [Fe 2 (CO) 5 (µ-naphthalene-2thiolate) 2 (P(PhOMe-p) 3 )] 2 was prepared from [Fe 2 (CO) 6 (µnaphthalene-2-thiolate) 2 ] 1 using the monodentate phosphine ligand, P(PhOMe-p) 3 and characterized by different spectroscopic techniques. The crystal structures for both complexes 1 and 2 are reported. The electrochemical investigations of complex 2 were performed in acetonitrile in the presence of acetic acid. Complex 2 was found to successfully electro-catalyze the reduction of acetic acid to H 2 with an overpotential of −0.56 V. This value of overpotential is towards the lower side in comparison with similar other reported complexes. 25,26,44,45 The bridging monothiolates in 2 resemble the bridging cysteine motif in [NiFe]-hydrogenase enzymes (see Fig. 1). Their rotational flexibility is limited due to the interactions with the protein matrix. In the model complex, this is accomplished by intramolecular stabilization. The monothiolates in the enzyme and in the model complex are sufficiently small to allow a direct protonation of the metal-metal bond to form a μ-hydride species. 46,47 This is not possible in the case of[FeFe]hydrogenase enzymes. In [FeFe] model complexes with bidentate thiolate ligands, a terminal hydride is formed initially which requires an intramolecular isomerization to form the bridging hydride. Such a re-orientation is not feasible in the [FeFe] enzyme since the terminal inorganic ligands form strong hydrogen bonds with the surrounding amino acid residue. 48 The monothiolate-bridged [FeFe] complex 2 is remarkably stable and can be reduced and oxidized two times without any structural distortion or degradation. This is accomplished by a combination of sufficient molecular flexibility from monothiolate bridging ligands and an electron rich and sterically demanding terminal P(PhOMe-p) 3 ligand which forms hydrophobic interactions with naphthalene-thiolates.
Previous attempts to stabilize hydrogen evolving catalysts have mainly focused on the covalent fixation of small molecules on electrode surfaces 49,50 incorporating into metalorganic frameworks 51 or hydrogel films 52 to improve the number of catalytic turnovers and the long-term catalyst stability. The concept of introducing stabilizing hydrophobic π-π interactions by incorporating both aromatic μ-bridging monothiolate ligands with a sufficient degree of rotational flexibility and terminal ligands with aromatic substituents has not been explored before. These apparently weak π-π interactions are sufficient to afford a compromise between structural flexibility of the catalyst and the integrity and stability of the coordination environment. Due to the hydrophobic nature of the π-interactions these are also expected to hold in aqueous media.

Methods and materials
All the experiments were carried out under an inert atmosphere using Schlenk techniques unless otherwise specified. Complex [Fe 2 (CO) 6 (µ-naphthalene-2-thiolate) 2 ] 1 was prepared according to the reported procedure. 25 All the anhydrous solvents (dichloromethane, acetonitrile and toluene) and starting materials were obtained from Sigma-Aldrich and used without further purification. The deuterated solvents were also obtained from Sigma-Aldrich. The 1 H and 31 P NMR spectra were recorded at room temperature in CDCl 3 solution with a JEOL 400 MHz NMR Spectrometer. FTIR spectra were recorded from dichloromethane solutions of the samples over the range of 400-4000 cm −1 on a Perkin Elmer FTIR Spectrometer. The UV-Vis spectra for complex 1 were recorded on a PerkinElmer Fig. 10 The 2HH species with (i) a μ-bridging hydride and (ii) a protonated naphthalene-thiolate after two one electron reductions. The two protons added are circled in red. π-π stacking interactions between bridging naphthalene ligands and terminal phenyl rings are indicated as dashed lines. This stabilization hinders an intramolecular isomerization required to release molecular hydrogen.
Scheme 2 Suggested mechanism for the electrocatalytic proton reduction of complex 2 (see the text for details).
Lambda-25 spectrophotometer and for complex 2 on an Avantes Avaspec-2048-USB2. The elemental analyses were carried out with a Vario Micro Cube elemental analyzer.

)]
A dichloromethane solution of 1 (200 mg, 0.334 mmol, 15 mL) was added to an acetonitrile solution of trimethylamine N-oxide dihydrate (Me 3 NO·2H 2 O) (37.1 mg, 0.334 mmol, 15 mL) and stirred for 10 min under an argon atmosphere. This was followed by the addition of tris(4-methoxyphenyl) phosphine (118 mg, 0.334 mmol) dissolved in 5 mL of CH 2 Cl 2 . The solution was stirred at room temperature for 6 h and the solvent was removed by using a rotary evaporator. The residue was chromatographed on a silica gel column. Elution with a mixture of hexane/dichloromethane (3 : 2) afforded a red coloured solution. Complex 2 was obtained as an air-stable blood-red powder after the removal of solvent. The complex was then recrystallized from hexane/CH 2 Cl 2 by a layering method at −10°C. Yield

X-ray crystallography
Single crystals of 1 and 2 were grown by slow evaporation of hexane/dichloromethane solutions at low temperature. X-Ray data of 1 and 2 were collected on an Oxford X-Calibur-S single crystal X-ray diffractometer using Mo-K α radiation. Significant crystallographic parameters and refinement details are listed in the ESI. † The structures were solved and refined by fullmatrix least-squares techniques on F 2 using the SHELX-97 (SHELXTL program package). 53 CCDC reference numbers 1054472 (2) and 1054473 (1) † contain the supplementary crystallographic data for this paper.

Electrochemistry
Electrochemical measurements were conducted in acetonitrile with 0.1 M tetrabutylammoniumhexafluorophosphate (Sigma-Aldrich, electrochemical grade) as the supporting electrolyte that was dried in a vacuum at 383 K. Cyclic voltammetry was carried out using an Autolab potentiostat with a GPES electrochemical interface (Eco Chemie). The working electrode was a glassy carbon disc (diameter 3 mm, freshly polished) for cyclic voltammetry. Platinum was used as the counter electrode. The reference electrode was a non-aqueous Ag/Ag + electrode (CH Instruments, 0.010 M AgNO 3 in acetonitrile). All the potentials are quoted against the ferrocene-ferrocenium couple (Fc/Fc + ); ferrocene was added as an internal standard at the end of the experiments. All solutions were prepared from dry acetonitrile (Sigma-Aldrich, spectroscopic grade, dried with molecular sieves 3 Å).
Computational details ADF2016 54,55 was used with the BP86 56,57 and B3LYP 58,59 exchange-correlation functionals, Grimme's dispersion correction with Becke-Johnson damping 60 and a TZP Slater-type basis set for all atoms. The solvent (acetonitrile) was used in a COSMO solvation model 61,62 using the Klamt atomic radii and surface definition with corrections for outlying charges. Thermodynamic corrections were obtained using statistical thermodynamics from BP86 calculations under standard conditions. 63 Redox potentials were calculated following ref. 64 and are given relative to a Fc/Fc + reference electrode in acetonitrile (calculated absolute potentials are 4.94 V for BP86 and 5.18 V for B3LYP). The BP86 value agrees well with the experiment (4.98 V (ref. 65)).

Conflicts of interest
There are no conflicts to declare.