Optimization of elemental selenium (Se(0)) determination in yeasts by anion-exchange HPLC-ICP-MS.

An analytical method was developed for the speciation of elemental selenium (Se(0)) in selenized yeasts by anion-exchange HPLC-ICP-MS after its chemical transformation into SeSO32- by reaction with sodium sulfite. The presence of Se(0) in the yeasts was further confirmed by single-particle ICP-MS. Indeed, Se nanoparticles, if present, are expected to be, at least partly, Se(0). X-ray photoelectron spectroscopy, a well-recognized technique for chemical element speciation in the solid state, was also used with this objective. Both methods were able to confirm the presence of Se(0) in the selenized yeasts but failed to provide reliable quantitative results. Analytical performances of the HPLC-ICP-MS method were then evaluated for Se(0) determination. Quantification limits of 1 mg/kg were reached. The recovery levels from an added quantity comprised between 93 and 101%. Within-run and between-run precisions were both below 8%. The procedure developed was finally applied to quantify Se(0) content in a series of seven yeast batches from different suppliers. Se(0) was found to be present in all the studied yeasts and represented on average 10-15% of the total Se.

Technological challenges in the preclinical development of an HIV nanovaccine candidate.

Despite a very active research in the field of nanomedicine, only a few nano-based drug delivery systems have reached the market. The "death valley" between research and commercialization has been partially attributed to the limited characterization and reproducibility of the nanoformulations. Our group has previously reported the potential of a peptide-based nanovaccine candidate for the prevention of SIV infection in macaques. This vaccine candidate is composed of chitosan/dextran sulfate nanoparticles containing twelve SIV peptide antigens. The aim of this work was to rigorously characterize one of these nanoformulations containing a specific peptide, following a quality-by-design approach. The evaluation of the different quality attributes was performed by several complementary techniques, such as dynamic light scattering, nanoparticle tracking analysis, and electron microscopy for particle size characterization. The inter-batch reproducibility was validated by three independent laboratories. Finally, the long-term stability and scalability of the manufacturing technique were assessed. Overall, these data, together with the in vivo efficacy results obtained in macaques, underline the promise this new vaccine holds with regard to its translation to clinical trials. Graphical abstract.

Advanced nanomedicine characterization by DLS and AF4-UV-MALS: Application to a HIV nanovaccine.

Nanoformulations are complex systems where physicochemical properties determine their therapeutic efficacy and safety. In the case of nanovaccines, particle size and shape play a crucial role on the immune response generated. Furthermore, the antigen's integrity is also a key aspect to control when producing a nanovaccine. The determination of all those physicochemical properties is still an analytical challenge and the lack of well-established methods hinders the access of new therapeutics to the market. In this work, robust methods for the characterization of a novel HIV nanoparticle-based vaccine produced in good manufacturing practice (GMPs)-like environment were developed. With slightly polydisperse particles (< 0.2) close to 180 nm of size, batch-mode Dynamic Light Scattering (DLS) was validated to be used as a quality control technique in the pilot production plant. In addition, a high size resolution method using Asymmetrical Flow Field Flow Fractionation (AF4) demonstrated its ability to determine not only size and size distribution but also shape modification across the size and accurate quantification of the free active ingredient. Results showed a monomodal distribution of particles from 60 to 700 nm, most of them (> 90%) with size lower than 250 nm, consistent with more traditional techniques, and revealed a slight change in the structure of the particles induced by the presence of the antigen. Finally, a batch to batch variability lower than 20% was obtained by both DLS and AF4 methods indicating that preparation method was highly reproducible.

Study of the presence of micro- and nanoparticles in drinks and foods by multiple analytical techniques.

A variety of food and drink samples (n = 21) were analyzed to evaluate the presence of (nano-) particles in their composition. After assessment of the sample pre-treatment step, a fast screening analysis was performed for drinks by Dynamic Light Scattering showing particles from 10 to 300 nm that could correspond to organic or metallic NPs. Metallic NPs were identified in foods by Single-Particle mode Inductively Coupled Plasma Mass Spectrometry and Asymmetrical Flow Field-Flow Fractionation coupled to Multiangle Laser Light Scattering and Inductively-Coupled Plasma Mass Spectrometry. The determination of Ti, Si and Ag concentration in the initial food suspensions, after filtration and centrifugal ultrafiltration enabled to estimate the ionic and nanoparticles content. Si-containing particles can be present in cappuccino powder as large aggregates and Si- and Al-containing particles in hot chocolate. Ti-containing NPs (80-200 nm) were found in chewing gum and Ag NPs in silver pearls (50-150 nm) used for decoration pastry.

Screening of TiO2 and Au nanoparticles in cosmetics and determination of elemental impurities by multiple techniques (DLS, SP-ICP-MS, ICP-MS and ICP-OES).

Cosmetics are part of the daily life of most of the people. Thus, a complete characterization of the products we applied in our skin is necessary. In this work, an analytical investigation of a wide variety of cosmetics from the point of view of total element content and metallic nanoparticles (NPs) has been performed. Firstly, we analyzed the total element content by ICP-MS and ICP-OES after acid digestion as an assessment of the presence of metal impurities. Prohibited elements in cosmetics, according to the European Commission regulation No 1223/2009, were not detected, and only elements mentioned in the label were found (e.g. Al, Fe, Ti and Si). Secondly, a screening of the presence of NPs has been performed by Dynamic Light Scattering (DLS) and Single Particle Inductively-Coupled Plasma Mass Spectrometry (SP-ICP-MS). Two sample preparation procedures were applied. The first protocol consisted in the preparation of suspensions in 0.1% w/v SDS and the second based on defatting with hexane followed by resuspension in water. DLS was employed as a routine method for a fast analysis of NPs, but this technique showed limitations due to the lack of specificity. SP-ICP-MS analyses were then performed, first as a screening technique to evaluate the presence of TiO2 and Au NPs in cosmetics suspensions prepared in SDS; and second, when a positive answer was obtained about the presence of NPs from the screening, SP-ICP-MS was used for particle size determination. Results showed that only TiO2 NPs were present in two sunscreens, one anti-wrinkle day cream, one lip balm protector labeled as 'nano' and in one brand of toothpaste not labeled as 'nano'. Sizes obtained for both sample preparations were compared and ranged from 30 to 120nm in most of the samples.