*3.5. Determination of Encapsulation Efficiency and Loading Capacity*

To quantify the CBLO content in CS@CBLO NPs, the TGA/ DTG (derivative thermal gravimetric) techniques were employed for evaluating loading capacity (LC) and encapsulation efficiency (EE). Freeze dried CS NPs and CS@CBLO NPs were, respectively, analyzed by a TGA furnace at 25–600 ◦C with a heating rate of 10 ◦C/min under a nitrogen atmosphere. The percentage of weight loss of each composition during thermal decomposition obtained from TGA was used to determine the content of CBLO in CS@CBLO NPs. The equations for the calculation of the loading capacity (Equation (1)) and encapsulation efficiency (Equation (2)) are listed below:

$$\text{LC} = \frac{\text{Weight of loaded CBLO}}{\text{Weight of sample}} \times 100\tag{1}$$

$$EE = \frac{\text{Weight of loaded CBLO}}{\text{Weight of initial CBLO}} \times 100\tag{2}$$

## *3.6. Characterization using FTIR, TGA and XRD*

The FTIR spectra of CBLO, CS NPs, and CS@CBLO NPs were measured by a Horiba FT-730 spectrometer (Kyoto, Japan). A sample of about 6 mg was mixed with 100 mg of KBr and then pressed in a standard device using a pressure of 6000 W to produce 13 mmdiameter pellets. For the CBLO spectral measurement, the liquid sample (~2 μL) was deposited onto a KBr disk. The spectra (4000–400 cm<sup>−</sup>1) were recorded with a resolution of 4 cm−<sup>1</sup> and 64 scans were performed per sample.

Thermogravimetric analysis (TGA) was performed with a TA2000/2960 thermogravimetric analyzer. Each freeze-dried sample (~5 mg) was placed into the TGA furnace and measurements were carried out under a nitrogen atmosphere with a heating rate of 10 ◦C/min from 25 to 600 ◦C.

X-ray diffraction (XRD) patterns of samples were recorded in the scanning mode on a Bruker D8ADVANCE diffractometer operated at a voltage of 40 kV and a current of 30 mA with Cu Kα radiation (λ = 1.5405 Å). The scanning angle (2θ) travelled from 5◦ to 50◦ with a scanning speed of 3◦ min<sup>−</sup>1.
