Data of characterization and related assays of lipid-core nanocapsule formulations and their hydrolysis mechanism

The data presented here are related to the research paper entitled “Chemical stability, mass loss and hydrolysis mechanism of sterile and non-sterile lipid-core nanocapsules: the influence of the molar mass of the polymer wall,” [1]. Experimental details of the nanoemulsion and nanosphere preparation. Sterilization methodology and their efficacy by microbiological analyses (turbidimetry and fungi and bacteria detection). Characterization data of formulations, LNC 1, LNC 2 and LNC 3, analyzed by laser diffraction and DLS analysis, as well as, characterization data of degradation by SEC, including all statistics analyses.


Data format
Raw, analyzed Experimental factors SEC analyses were performed after extraction procedure of all constituents of LNC and separation of precipitate (polymer) and supernatant (lower molar mass constituents). Microscopy images, turbidimetry and diameter analyses of LNC were obtained without pre-treatment of samples.

Experimental features
Details of experimental methodologies used in this study such as preparation of nanoemulsion and nanosphere formulations. Sterilization process of LNC formulations and the efficacy of this technique by microbiological analyses. Characterization of LNC formulations after storage by SEC to identify the predominant hydrolysis mechanism. Particle size analyses to characterize physicochemical stability.

Data source location
Porto Alegre, Brazil Data accessibility Data is provided with this article Related research article [1] S. Calgaroto, L. E. Fauri, L. Frank, K. Paese, S. S. Guterres, A. R. Pohlmann, Chemical stability, mass loss and hydrolysis mechanism of sterile and non-sterile lipid-core nanocapsules: the influence of the molar mass of the polymer wall, Reactive and Functional Polymers, 2018 Value of the data SEC molar mass profiles of all the material constituents of polymeric nanocapsules is innovative for the scientific community since for most of the investigations just the polymer wall is evaluated for complex colloidal systems.
Microbiological analyses of sterile and non-sterile LNC formulations provided information on the efficiency of the sterilization process and were are useful for their evaluation.
Laser diffraction and dynamic light scattering (DLS) were compared with data from other works when analyzed the storage of similar delivery system and prove that physical parameters does not suffer alteration during the storage.

Data
The data presented in Section 1.1 is the initial physicochemical characterization of LNC formulations, prior and after sterilization process. The size distribution profiles of formulations are showed in Fig. 1 and the DLS profile in Fig. 2. Section 1.2 involves the determination of each LNC constituent material by size exclusion chromatography (SEC) (Fig. 3). The data presented in Section 1.3 includes the determination of crystallinity degree for the LNC constituents and LNC formulations, prior and after sterilization process (Table 1). Section 1.4 brings data referent to sterilization process and microbiological analyses that prove their efficacy (  Section 1.5 is related to the SEC profiles and molecular weight changes for the LNC constituents, prior and after sterilization ( Fig. 6 and Table 3). Section 1.6 show data referent to physicochemical characterization for nanocapsules formulations, non-sterile (LNC) and sterile (LNCS), storage at 5°C (60 days) by laser diffraction (Fig. 7) and DLS analyses (Table 4). Section 1.7 presented the changes on molar mass of nanocapsules formulations (precipitate - Fig. 8 and supernatant -Fig. 9) storage at 5°C (60 days). The statistical analyses applied in all SEC results are presented in the same section (Tables 5, 6 and 7).     Table 1.

Experimental design, materials and methods
The methodologies to obtain the data exposed here are described in Calgaroto et al. [1].

Preparation of Nanoemulsion (NE) and Nanosphere (NS) formulations
To prepare NE formulation, an organic phase containing 0.038 g of sorbitan monostearate, 0.160 g of oil (capric/caprylic triglyceride) and 27 mL acetone was injected into an aqueous phase containing 0.080 g polysorbate 80 and 53 mL ultrapure water, under magnetic stirring at 40°C. To prepare NS, the organic phase was composed by 0.100 g poly(ε-caprolactone), 0.038 g of sorbitan monostearate solubilized in 27 mL acetone. This phase was injected into an aqueous phase containing 0.080 g polysorbate 80 and 53 mL ultrapure water, under magnetic stirring at 40°C. For both formulations, after 10 min, the organic solvent was removed under reduced pressure at 40°C using a rotary evaporator (Büchi, Switzerland), having their volume reduced to 10 mL. The formulations prepared in triplicate batches (n ¼ 3).

Sterilization process
Sterilization was performed in Horizontal Autoclave (Phoenix AB42, São Paulo, Brazil) at 134°C for 10 min and 2.10 bar as previously described [2]. Initially, 5 mL of each formulation was packed in

Particle size analyses
The nanocapsule formulations were evaluated (particle size distribution) by laser diffraction (LD) using a Malvern Mastersizer s 2000 instrument (Malvern Instruments, UK). The sample (n ¼3) was placed in the equipment using a micropipette (#Z646598) (BRAND s Transferpette s S pipette, single channel) acquired from Sigma-Aldrich (Steinheim, Germany) device wet unit (Hydro 2000SM -AWM2002 -Malvern, UK) in an amount sufficient to obtain more than 2% obscuration. Mie theory of light scattering was used to calculate the particle size distribution. Mean diameter was expressed as volume-weighted mean diameter (d4,3), and polydispersity (Span) was calculated using Eq. (1), where d0.9, d0.1, and d0.5 are respectively the diameters at percentiles 90, 10, and 50 of the cumulative size distribution curve. The median diameter by number of particles (d0.5)n was also determined for each sample using the distribution curve based on the number of particles.
Dynamic light scattering (DLS) was carried out to determine the mean hydrodynamic diameter and the polydispersity of the submicrometric particle populations in a Nanoseries s ZetaSizer ZS (Malvern, UK) equipment. LNC formulations (20 μL) were diluted in MilliQ s water (10 mL) previously filtered (0.45 μm, hydrophilic membrane (#HVLP) (Durapore s , Merck, Germany). Each sample was poured into a quartz flow cell (#ZEN0023, Malvern, UK). The scattered light was detected at an angle of 173°. The correlograms were fit using the method of Cumulants to calculate the z-average diameters. Experiments were conducted with three batches for each sample. Table 4 z-Average diameters and polydispersity (PDI) determined by dynamic light scattering (DLS) for formulations before and after the storage time (60 days) (semi-dilute regimen).

Turbidimetry test
The microbiological contamination was evaluated by a turbidimetry method to identify the presence of microorganisms in the formulation prior (LNC) and after sterilization process (LNCS). The presence of contaminants in the sample is related with the increase in absorbance [3]. The absorbance of formulations without incubation (LNC and LNCS) was determined as the controls. The formulations (LNC and LNCS) were incubated at 37 7 1°C during 48 h using three different concentrations by adding 1, 5 or 10 μL of each sample into 1 mL of Luria Bertani (LB) medium. The experiments were performed in triplicate, and the absorbance values were measured by spectrometry (Spectramax M2e -SoftMax Pro Software Interface 5) at 370 nm.

Fungi and bacteria detection
The detection of fungi and bacteria in the formulations was performed by inoculating 20 μL of each formulation (LNC and LNCS) for 48 h at 37 7 1 and 35 7 1°C, respectively, a blood agar plate for the bacterial growing and in a Sabouraud plate for the fungal growing. Table 5 Statistical analysis of the weight loss [ΔM w (%)] and dispersity (Đ ¼ M w /M n ) for the non-sterile and sterile nanocapsule formulations (precipitate) under storage, at 5°C, by SEC. Peaks (1), (2) and (3)