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A de ned stoichiometry is generally not relevant to describe macromolecular complexes, even in dilute regime (ie, with only isolated chains and no polymer polymer aggregates) In practice, the number of bound nanoparticles per polymer chain and the fraction of bound azobenzene are the two primary structural parameters that can be measured (Fig 75b) First, we focus on the binding of the azo chromophores Proton NMR shifts (May et al, 2005), red-shift on the UV spectra (Labarthet et al, 2000), variation of circular dichroism (Shimomura et al, 2002; Suzuki et al, 1991) are among the experimental signals that can be used to probe the solvatation of the chromophore Accordingly, it is possible to determine the fraction of free azobenzene groups and whether the association takes place gradually or not.

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Our measurements show that the fraction of azobenzene side group that are bound into micelles is most typically signi cantly smaller than 100% Fig 76a gives representative UV-visible spectra of azo-modi ed polymers in water, as a function of the concentration of surfactant The absorbance obviously increases and the main peak is red-shifted with increasing the surfactant concentration, two features that re ect the transfer of azobenzene from water into a less polar environment The presence of isobestic points betrays that the azo groups are either in contact with water or tightly bound to the surfactant and no intermediate solvatation state is detected (not shown) The absorbance at a xed wavelength indicates accordingly the degree of binding of the azobenzene groups In Fig 76b, the absorbance abruptly increases above the critical micelle concentration (Khoukh et al, 2007).

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In the case of other nanoparticles, such as proteins or polymers of CDs, the absorbance increases immediately upon addition of the particles (Pouliquen et al, 2007; Pouliquen and Tribet, 2006) At incipient binding, the association was considered very tight and quantitative, and it has not been possible to estimate a binding constant (not shown) The association was presumably strengthened because of multipoint attachments (see later) A saturation plateau can be reached at high concentrations of the nanoparticles Once normalized by the concentration of azobenzene and a reference shift of extinction coef cient (eg, as measured with azobenzene in water and dodecane), the plateau value should indicate the fraction of bound chromophore In practice, the normalized plateau value depends on the integration level of the azobenzene and intrachain repulsion (as modulated by ionic strength in the case of our azo-modi ed polyelectrolyte).

In other words, variability of the molar absorption in the saturated state clearly indicates that a variable fraction of the chromophore remains unbound, even in presence of large excess of surfactant With neutral micelles of Triton X-100 and C12E4, the saturation could correspond to less than 50 mol% association (Fig 76b, right axis), which means formation of long polymer loops rather than short ones upon binding Incomplete association of the chromophores is presumably because of.

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