Lc 16

In order to obtain sunscreen-loaded microcapsules with good sunscreen effect, it was necessary to study the OMC microcapsules by using the different wall materials with the good ultraviolet absorption. Several kinds of wall materials, such as modified starch (C001), sodium carboxymethyl cellulose (CMC), arabic gum (GA), sodium alginate (ALG), chitosan (CS), sodium caseinate (SC), maltodextrin (MD), and β-cyclodextrin (β-CD), were measured by UV spectrophotometer (LC-16, Shimadzu, Kyoto, Japan) during 200–400 nm.

AMFs with clean and intact surfaces were immersed in a certain amount of physiological medium. AMFs were taken out repeatedly and rinsed with deionized water at each time point, the residual liquid was collected to quickly freeze in a −80 °C refrigerator and it was treated with vacuum freeze-drying for 24 h. The freeze-dried powder was dissolved in 400 μL methanol and transferred to a centrifuge tube through a 0.22 μm membrane filter. The content of antimicrobial peptide Ac.X2 was analyzed by HPLC (LC-16, SHIMADZU). Note: # Indicates that the film was a disc with a diameter of 0.6 cm. ## The concentration of exudative Ac.X2 and 100 μL culture medium after homogenization was calculated for comparison of cell viability.

The concentrations of encapsulated DHA and CQ were determined by HPLC (LC-16, Shimadzu, Suzhou, China) with a UV detector (SPD-16, Shimadzu, Suzhou, China). LNPs were dissolved in acetonitrile and sonicated for 20 min to release the drugs. The mobile phase used were water:acetonitrile (40:60, v/v) for DHA and 7.2 mM sodium hexanesulfonate solution (containing 10 ml acetic acid and adjusted to pH 3.5 using triethylamine):acetonitrile (70:30, v/v) for CQ. Chromatographic separation was performed on a Hedera ODS-C18 column (250 × 4.6 mm, 5 ⎧m, HanBon, Jiangsu, China) at a flow rate of 1 ml/min; DHA and CQ were detected at 210 and 343 nm, respectively. The LC and LR of DHA and CQ were calculated according to Eqs. 1, 2 below. $LC(\%)=\frac{W_{drug loaded}}{W_{total liposome} }\times 100$
$LR(\%)=\frac{W_{drug loaded}}{W_{drug added}}\times 100$

About 5 × 10⁶ cells were resuspended in 500 μl methanol (80%, −80 °C precold) and incubated for 20 min at −80 °C, centrifuged for about 40 min, 12000g to remove the sediment, then subjected to detect the UDP-GlcNAc level by HPLC (LC-16, Shimadzu). In brief, samples and a series of standard UDP-GlcNAc (Sigma) solutions were filtered at 0.22 μm. Ten microliter aliquots of samples were directly injected into HPLC C18 column (250 mm × 4.6 cm, 5 μm, 100 Å, Kromasil), which is suitable to separate the nucleotide sugars (45 (link)), while a pump system supplied moving phase A (0.1% trifluoroacetic acid [TFA] in distilled water) and moving phase B (0.1% TFA in acetonitrile [ACN]). Absorbance measurements were made at 214 nm on a UV-detector (SPD-10A, Shimadzu). The profile of the gradient moving phase was as follows: 0∼3 min, 90% A, 10% B, and the flow rate was 1 ml/min. The level of UDP-GlcNAc was calculated based on the peak area referenced to standards.

TC, a typical antibiotic found at high levels (above 920 mg/L) in pharmaceutical wastewater^{12 (link)}, was employed to assess the adsorption performance of the synthesised erdite nanorods. A stock solution of 2000 mg/L TC was prepared, and its pH was adjusted to 5 by adding 1.44 mol/L HCl and 1.32 mol/L NaOH. A series of dilute solutions was prepared from the stock by addition of deionised water. Each dilution (20 mL) was mixed with 0.01 g of the erdite particles in an Erlenmeyer flask. The flasks were sealed with parafilm and shaken at 200 rpm in an incubator (THZ-98A, Yiheng, Shanghai, China) at room temperature. After 24 h, the flasks were taken from the incubator, and the erdite particles were separated by agitation at 5500 rpm for 5 min. The TC concentration in the supernatant was determined by high-performance liquid chromatography (LC-16, Shimadzu, Japan) using a mobile phase composed of 0.1% phosphoric acid and methanol at a ratio of 60:40 (v/v). The absorbance at 268 nm was read by using a UV detector, and the retention time was approximately 5 min. The adsorption capacity (q_e, mg/g) of erdite was calculated by using the following equation: $$q_e=\frac{(C_0-C_e)\times V}{m},$$ where C₀ and C_e are the initial and equilibrium concentrations of TC (mg/L), respectively, V is the solution volume (L), and m is the particle dosage (g).