Porous starch as an effective carrier for sustained release and bioactivity protection of cannabidiol full spectrum oil.

Abstract

Full-spectrum cannabidiol oil (CFSO) has attracted growing attention across industries for its multiple health benefits, but its poor water solubility and low bioavailability remain unresolved challenges. In this study, porous starch (PS) was used as a wall material to encapsulate CFSO, and the physicochemical properties, water solubility, bioavailability, and antioxidant activity of the encapsulated product were evaluated. Results confirmed the successful preparation of PS-CFSO. Using Box-Behnken design response surface methodology, the maximum encapsulation efficiency (80.69 ± 0.71 %) was achieved under optimal parameters: CFSO concentration of 75 mg/mL, PS-to-CFSO mass ratio of 7.6:1 (g/g), and loading time of 41 min. CFSO was encapsulated in PS in an amorphous state, and PS increased CFSO's thermal decomposition temperature. Compared with free CFSO, PS-CFSO showed a 17.92-fold increase in water solubility, a 4.74-fold higher release rate in simulated gastric fluid, and a 4.13-fold higher release rate in simulated intestinal fluid. PS-CFSO release followed pseudo-Fickian diffusion and first-order kinetics. Additionally, its antioxidant activity was enhanced by over 20 %. Overall, This study expands the application of starch-based carriers and provides critical guidance for developing CFSO delivery systems with high encapsulation efficiency, stability, and bioavailability, thereby broadening CFSO's prospects in the food and pharmaceutical industries.

Untitled section

Keywords: Cannabidiol full-spectrum oil, Porous starch, Solubility, Bioavailability, Delivery systems

Graphical abstract

Highlights

•Cannabidiol full-spectrum oil (CFSO) delivery system was prepared by porous starch.
•PS-CFSO binds through non-covalent interactions.
•The solubility and stability of CFSO were enhanced.
•The bioavailability and antioxidant activity of CFSO were improved.

Article notes

Untitled section

Received 2025 Aug 28; Revised 2025 Nov 25; Accepted 2025 Nov 26; Collection date 2025.

Author notes

1.Introduction

Industrial hemp (Cannabis sativa L.) is an economic crop with tetrahydrocannabinol content below 0.3 %, widely utilized in industrial, pharmaceutical, and food sectors (Baidoo et al., 2025). Cannabidiol (CBD) is the most abundant non-psychoactive cannabinoid in industrial hemp (Vardanega et al., 2024). It exhibits multiple beneficial effects, including hypoglycemic activity (Ghasemi-Gojani et al., 2022), anticancer properties (Yan et al., 2023), pain relief (Charles et al., 2024), and anxiolytic characteristics (Vardanega et al., 2024). Its applications in the food industry continue to expand, with the EU CBD market projected to reach 1.6 billion euros by 2020 (Lachenmeier et al., 2023). However, purifying CBD to high purity is complex and costly. In contrast, full-spectrum cannabidiol oil (CFSO) is a crude extract rich in CBD, with small amounts of polyphenols, terpenes, and other components (Li et al., 2023a), and it is more widely applicable in practice. CFSO is easier to prepare and exhibits synergistic effects between its components (Li et al., 2022). For example, it has shown greater efficacy than purified CBD in treating refractory epilepsy (Pamplona et al., 2018). However, CFSO has extremely poor water solubility, which limits its oral bioavailability. Additionally, it is prone to degradation under high temperatures and during prolonged storage (Li et al., 2022). To our knowledge, only one study has reported the encapsulation of CFSO using 2,6-di-O-methyl-β-cyclodextrin, which effectively improved its thermal stability and bioavailability (Li et al., 2022). However, 2,6-di-O-methyl-β-cyclodextrin (DM-β-CD) is typically synthesized from β-cyclodextrin through a methylation reaction, involving a complex and costly synthesis process. Therefore, identifying additional natural, low-cost green carriers that can effectively enhance the water solubility and stability of CFSO holds significant practical importance for its broader application across multiple fields (Yang et al., 2025b).

Porous starch (PS) is a modified starch featuring a honeycomb-like porous structure, with its core advantages lying in high pore volume and large specific surface area (Zhu et al., 2024). It also exhibits excellent biocompatibility and biodegradability (Yang et al., 2025c), making it one of the most commonly used encapsulation materials in the food and pharmaceutical industries (Lei et al., 2018). PS raw materials are widely available, with natural starches from grains (corn, rice), legumes, tubers (potato, tapioca), palm, fruits, and stems all suitable for PS modification (Chen et al., 2020). Rice porous starch significantly enhances insulin bioavailability (Du et al., 2025), while lotus seed porous starch improves proanthocyanidin thermal stability and UV radiation tolerance (Wang et al., 2025). PS preparation methods include physical, chemical, enzymatic, and synergistic modification approaches. Among these, enzymatic modification is currently the most widely applied due to its mild reaction conditions, high catalytic efficiency, and strong substrate specificity (Kavya et al., 2025). Corn PS treated with 33 units of α-amyloglucosidase for 2 h forms the largest pore size (1.42 μm), making it more suitable for loading lipophilic active ingredients like curcumin and resveratrol (Wahab and Janaswamy, 2024). The synergistic process combining high-shear processing with dual-enzyme hydrolysis (α-amylase + saccharifying enzyme) further increases the specific surface area of PS, achieving a camellia oil encapsulation efficiency (EE) of 93.42 ± 0.04 % (Wu et al., 2025). Furthermore, citric acid crosslinking modification can regulate PS water solubility, reducing it by approximately 40 % in anthocyanin encapsulation systems (Quilez-Molina et al., 2024). Encapsulation process conditions also play a role, such as ultrasonic treatment at 500 W for 30 min which facilitates naringin dispersion (Cao et al., 2025, Cao and Lu, 2025). These studies confirm that PS's loading performance is regulated by its structural properties, modification degree, and encapsulation process conditions. Additionally, compared to the DM-β-CD mentioned above, PS offers a more environmentally friendly preparation process and is significantly less expensive (only 2.6–20 % of the cost). However, how PS parameters affect CFSO encapsulation remains unclear, emphasizing the need for targeted optimization to address this scientific gap.

Therefore, this study prepared microspheres using CFSO as the core material and PS as the shell material. Through response surface optimization experiments, the optimal preparation process for PS-CFSO microspheres was ultimately determined. Their structure and thermal properties were characterized using scanning electron microscopy (SEM), fourier transform infrared spectroscopy (FT-IR), and thermogravimetric analysis (TGA). Further investigations explored their bioavailability in simulated gastrointestinal conditions and in vitro antioxidant activity. This study provides robust data support for developing porous starch-based delivery systems for CFSO. It not only offers new insights for enhancing CFSO's bioavailability and stability but also paves new pathways for innovative applications of CFSO in the health food sector.

2.Materials and methods

2.1.Materials

CFSO (with an 80 % CBD content) was jointly provided by Heilongjiang Zhanqi Biotechnology Co., Ltd. (Harbin, China) and the Heilongjiang Provincial Key Laboratory of Hemp Industry Technology. PS is prepared by treating corn starch with α-amylase and glucoamylase in weak acid. It has a purity >99 %, amylose content of 26.32 %, and molecular weight (M_W) of 1.044 × 10⁴ kDa, and was procured from Liaoning Lida Biotechnology Co., Ltd. (Liaoning, China). Other chemical reagents, including artificial gastric juice and artificial intestinal juice, were purchased from Shanghai Chemical Reagent Co., Ltd. (Shanghai, China).

2.2.High performance liquid chromatography (HPLC)

High-performance liquid chromatography (HPLC, Thermo Fisher Scientific, Germany) detection conditions for CFSO: A reversed-phase C18 column (Agilent Technologies, Inc.) was used, with specifications of 250 mm × 4.6 mm and 5 μm, and the column temperature was set at 30 °C. The mobile phase consisted of acetonitrile and 0.1 % glacial acetic acid aqueous solution at a volume ratio of 75:25, with a flow rate of 0.8 mL/min and a detection wavelength of 220 nm. CFSO standard solutions (0.1, 0.4, 0.6, 0.7, 0.8 mg/10 mL, and 0.1 mg/mL) were prepared using the mobile phase. The injection volume was 20 μL. The CFSO HPLC chromatogram and standard curve are provided as supporting materials (Fig. S1).

2.3.Preparation of PS-CFSO

PS-CFSO was prepared via magnetic stirring. A specific amount of CFSO was added to 10 mL of methanol solvent, then processed for 3 min using an ultrasonic cell disruptor (JY92-II, Ningbo Xinzhi Biotechnology Co., Ltd.) to promote its dissolution and dispersion. Subsequently, PS is added according to the predetermined ratio. A rotor with a specification of 5 cm was placed in the solution, which was then stirred at 100 rpm for a specific duration in a thermostatic magnetic water bath at 37 °C. The specific research factors were determined according to the design of single-factor experiments. After stirring, the solution was centrifuged at 10,000 rpm for 20 min. The supernatant was collected, and the resulting precipitate was dried to obtain PS-CFSO.

2.4.Analysis and calculation of PS-CFSO

After collecting the aforementioned supernatant, the mass of CFSO loaded in PS-CFSO can be calculated by quantifying the mass of CFSO in the supernatant (Li et al., 2022). Specifically, high-performance liquid chromatography (HPLC) is used to measure the peak area of CFSO. The mass of CFSO in the supernatant is then calculated using the standard curve, and the mass of CFSO in PS-CFSO is derived from this value. Subsequently, the encapsulation efficiency and drug-loading rate of CFSO in PS-CFSO are calculated. The formulas for encapsulation efficiency and drug-loading rate are presented in Equations (1) and (2):

(1)

E = \frac{M_{0}}{M_{P S - C F S O}} \times 100 %

(2)

D = \frac{M_{0} - M_{i}}{M_{0}} \times 100 %

Where E represents the encapsulation efficiency; M_PS-CFSO is the total mass of the loaded substance; D represents the loading rate; M₀ is the total mass of CFSO input; M_i is the mass of CFSO in the supernatant.

2.5.Optimization of the preparation process of PS-CFSO

The effects of cannabidiol full-spectrum oil (CFSO) concentration (10–100 mg/mL), porous starch (PS)-to-CFSO mass ratio (1:1–8:1, g/g), and adsorption time (10–60 min) on the encapsulation efficiency of PS-CFSO were investigated separately. For single-factor experiments: (1) To evaluate CFSO concentration, the PS-to-CFSO mass ratio was fixed at 6:1 (g/g) and adsorption time at 30 min; (2) To evaluate the PS-to-CFSO mass ratio, CFSO concentration was fixed at 50 mg/mL and adsorption time at 30 min; (3) To evaluate adsorption time, CFSO concentration was fixed at 50 mg/mL and the PS-to-CFSO mass ratio at 6:1 (g/g). Based on the single-factor experimental results, a three-factor, three-level response surface methodology (RSM) optimization experiment was designed to determine the optimal preparation process for PS-CFSO. The factors and their respective levels were: CFSO concentration (60, 70, 80 mg/mL), PS-to-CFSO mass ratio (6:1, 7:1, 8:1, g/g), and adsorption time (35, 40, 45 min).

2.6.Physicochemical properties characterization of PS-CFSO

2.6.1.Scanning electron microscopy (SEM)

CFSO, PS, and PS-CFSO were attached to conductive adhesive. After gold sputtering, their micro-surface morphologies were observed under 30 kV Li et al., 2024b.

2.6.2.Particle size and zeta potential analysis

The particle size and zeta potential were determined using a dynamic light scattering (DLS) instrument (model LT2200E, Zhuhai Zhenshi Optical Instruments Co., Ltd., China). Samples were dispersed in water, diluted, and then measured (Lu et al., 2025).

2.6.3.Fourier transform infrared spectroscopy (FT-IR)

An infrared spectrophotometer (Bruker Alpha II, Bruker Corporation, USA) was used with the potassium bromide pellet method. The test temperature was 20 °C, and the scanning wavelength range was 400–4000 cm⁻¹ (Wang et al., 2025b; Li et al., 2023b ).

2.6.4.X-ray diffraction (XRD)

An X-ray diffractometer (D8 ADVANCE, Bruker Corporation, USA) was employed. The detection range was 3° ≤ 2θ ≤ 60° under the conditions of 30 kV and 35 mA.

2.7.Thermogravimetric analysis (TGA)

TGA was performed using a Simultaneous TG-DSC Thermal Analyzer (STA 449 F3 Jupiter®, NETZSCH, Germany). The temperature program was set from 50 to 600 °C at a heating rate of 10 °C/min under argon flow (20 mL/min).

2.8.Determination of aqueous solubility

2.8.1.Hydrophilicity

CFSO and PS-CFSO were mechanically compressed into tablets, placed on a clean glass slide, and 20 μL of double-distilled water was dropped onto the drug tablets for hydrophilicity testing.

2.8.2.Water solubility

CFSO and PS-CFSO samples were mixed with deionized water, and the solutions were magnetically stirred at 37 °C for 48 h. The mixtures were then centrifuged for 10 min, and the resulting solutions were collected and detected by HPLC at 220 nm.

2.9.Cumulative release rate of simulated digestion in vitro

First, CFSO and PS-CFSO were respectively placed in 12 mL of simulated gastric fluid or simulated intestinal fluid for 3 h. Further, after the CFSO and PS-CFSO samples were digested in simulated gastric fluid at 37 °C for 2 h, the digested simulated gastric fluid was mixed with an equal volume of simulated intestinal fluid. The pH of the resulting mixture was adjusted to 7.0, followed by digestion at 37 °C for 3 h. Samples were taken every 0.5 h to determine the cumulative release rate. The release kinetics were fitted using the zero-order equation (3), first-order equation (4), and Higuchi equation (5), while the Peppas equation (6) was employed to investigate the release mechanism of PS-CFSO (Gu et al., 2021):

(3)

y = a x

(4)

y = 1 - e^{- a x}

(5)

y = a x^{1 / 2}

(6)

y = a x^{b}

where y represents the cumulative release ratio of CFSO at x min, a is the structural and geometrical characteristic constants of PS-CFSO, and b is the release index.

2.10.Determination of antioxidant activity

According to our previous studies, the DPPH and ABTS free radical scavenging activity assays were performed (Li et al., 2023a; Lu et al., 2025). Briefly, the sample was mixed with an equal volume of DPPH solution. After a 30-min reaction in the dark, the absorbance was measured at 517 nm. The calculation formula is as follows: DPPH free radical scavenging activity (%) = [1 - (A₁ - A₂)/A₀] × 100. Where A₁ represents the absorbance of the DPPH-sample mixture, A₂ is the absorbance of the ethanol-sample mixture, and A₀ is the absorbance of the blank control group without sample. In addition, the sample was mixed with ABTS solution at a volume ratio of 1:3. After a 6-min reaction at room temperature in the dark, the absorbance was determined at 734 nm. The calculation formula is: ABTS free radical scavenging activity (%) = [1 - (A₁ - A₂)/A₀] × 100. Where A₁ is the absorbance of the ABTS-sample mixture, A₂ is the absorbance of the solvent-sample mixture, and A₀ is the absorbance of the blank control group.

2.11.Statistical analysis

Each measurement was performed at least three times. The data are presented as mean ± standard deviation (SD). Statistical significance of differences among the mean values was evaluated at the P < 0.05 level using SPSS 25.0 software (IBM Corp., Armonk, NY, USA), via one-way analysis of variance (ANOVA) followed by Duncan's multiple range test for pairwise comparisons.

3.Results and discussion

3.1.RSM optimization of PS-CFSO

The experimental values for Box-Behnken design (BBD) was shown in Table 1 for the factors of CFSO concentration (A), PS-CFSO mass ratio (B), adsorption time (C). Multiple regression was performed on the data utilizing Design-Expert 13. A quadratic polynomial equation between the Encapsulation efficiency and the parameter:

(7)

Y = 81.56 + 4.06 A + 1.98 B + 0.6569 C - 1.29 A B + 2.28 A C + 1.15 B C - 3.06 A^{2} - 3.08 B^{2} - 2.35 C^{2}

Table 1Experimental design and the results.

Code	A CFSO concentration	B Adsorption time	C PS-CFSO ratio	E	D
Code	(mg/mL)	(min)	(g/g)	(%)	(%)
1	60	35	7	67.93	8.49
2	80	35	7	78.62	9.83
3	60	45	7	74.22	9.28
4	80	45	7	79.77	9.97
5	60	40	6	74.31	10.61
6	80	40	6	77.87	11.12
7	70	40	8	76.29	8.48
8	70	40	8	81.07	9.01
9	70	35	6	73.93	10.56
10	70	45	6	75.84	10.83
11	70	35	8	74.73	8.30
12	70	45	8	81.23	9.03
13	70	40	7	81.19	10.15
14	70	40	7	81.02	10.13
15	70	40	7	82.99	10.37
16	70	40	7	83.84	10.48
17	70	40	7	79.97	9.99

As shown in Table 2, the significance test of the analysis of variance (ANOVA) for the regression model was conducted using F-values and P-values. The model is statistically significant (F = 33.29, P < 0.0001), but its lack of fit is not significant (F = 1.09, P > 0.05). This indicates that the model has good applicability in predicting and analyzing the PS-CFSO loading process. These data demonstrate the reliability of the experimental results (Wang et al., 2025a).

Table 2Coefficients from standard regression models.

Source	Sum of squares	df	Square	F	p-value
Model	12.08	9	1.34	33.29	<0.0001
A-CFSO concentration	1.18	1	1.18	29.21	0.0010
B-Adsorption time	0.46	1	0.46	11.50	0.0116
C-PS-CFSO ratio	8.66	1	8.66	214.78	<0.0001
AB	0.10	1	0.10	2.55	0.1543
AC	1.28	1	1.28	3.17	0.9567
BC	0.05	1	0.05	1.25	0.3003
A²	0.53	1	0.53	13.09	0.0085
B²	0.97	1	0.97	23.97	0.0018
C²	0.02	1	0.02	0.44	0.5266
Residual	0.28	7	0.04
Lack of Fit	0.13	3	0.04	1.09	0.4490
Pure Error	0.16	4	0.039
Cor Total	12.36	16

Factors A, B, and C all have significant effects on the loading process (P < 0.05), with the order of influence being C > A > B. Furthermore, the interaction effects of any two factors on the PS-CFSO loading process were investigated (Fig. 1). With the introduction of the three factors, the loading capacity first increased and then decreased (or tended to stabilize). The contour lines in each interaction plot are elliptical, indicating that there are interaction effects between any two factors, and steeper contour lines indicate more significant interaction effects (Li et al., 2025b;Zhou et al., 2022).

Fig. 1Three-dimensional surface plots and contour plots for the interaction between any two variables. A₁-A_2: CFSO concentration (A) & Adsorption time (B), B₁-B₂: CFSO concentration (A) & PS-CFSO ratio (C), C₁-C₂: Adsorption time (B) & PS-CFSO ratio (C). The color gradient represents the magnitude of the response values, increasing gradually from blue (lowest value) to red (highest value).

The model predicted optimal parameters: CFSO concentration of 75.109 mg/mL, PS-to-CFSO mass ratio of 7.601:1 (g/g), loading time of 41.325 min, with a predicted encapsulation efficiency of 83.27 % and drug-loading capacity of 9.70 %. For practical operability (Wang et al., 2025), parameters were adjusted to 75 mg/mL, 7.6:1 (g/g), and 41 min. Under these conditions, the actual encapsulation efficiency was 80.69 ± 0.71 % and drug-loading capacity was 9.13 ± 0.09 %, with a theoretical deviation of only 0.97 %. This small deviation confirms the reliability of the interaction analysis and the model's practical applicability (Ye et al., 2025; Lu et al., 2025).

3.2.Physicochemical properties of PS-CFSO

3.2.1.Morphological observations

The micro-morphologies of PS, CFSO, and PS-CFSO (with a loading rate of 80.69 %) were observed under different magnifications (2000 × and 4700 × ) (Fig. 2). Fig. 2A₁-2A₂ show CFSO, which is lipid-soluble and exists as a solid at room temperature. It has a particle size range of approximately 30–200 nm, presents an irregular aggregated state, and is poorly soluble in water (Li et al., 2022). As can be seen from Fig. 2B₁-2B₂, PS features a smooth surface with a porous interconnected structure, having a particle size range of 5–20 μm and a pore size range of 1–2 μm. PS can be gradually dissolved after being degraded by gastrointestinal enzymes (Cao and Lu, 2025). Fig. 2C₁-2C₂ display PS-CFSO, whose surface is rough with visible particulate substances. This suggests that CFSO was likely successfully loaded onto PS (Yu et al., 2024). Additionally, compared with the original morphologies of CFSO and PS, PS-CFSO shows a more uniform distribution and a more consistent shape. This is consistent with previous studies (Cao and Lu, 2025; Wahab and Janaswamy, 2024; Wu et al., 2025).

3.2.2.Particle size and zeta potential analysis

The particle sizes and zeta potentials of PS, CFSO, and PS-CFSO microparticles were determined via dynamic light scattering (DLS), with results presented in Fig. 3A. The particle size of PS was 17.63 ± 1.8 μm, that of CFSO was 164.18 ± 1.6 nm, and that of PS-CFSO microparticles was 20.76 ± 0.9 μm. It can be observed that the adsorption of CFSO on the surface and within the pores of PS alters the surface hydrophobicity of the resulting PS-CFSO. Additionally, due to PS's high hygroscopicity, it may absorb water and swell during the experiment. This leads to a slightly larger average particle size of PS-CFSO compared to PS (Piloni et al., 2022).

The zeta potentials of PS, CFSO, and PS-CFSO microparticles are shown in Fig. 3B. The zeta potential values of PS, CFSO, and PS-CFSO are −34.33 ± 1.97 mV, −31.27 ± 0.31 mV, and −33.13 ± 0.78 mV, respectively. All of these values exhibit a certain degree of electronegativity. The absolute value of PS-CFSO's zeta potential is significantly higher than that of CFSO, indicating that solutions containing PS-CFSO particles are more stable and thus easier to store (Wu et al., 2025). Notably, the slight reduction in surface negativity of PS after CFSO loading (rather than a dramatic charge shift or reversal) serves as direct evidence of successful CFSO loading. This phenomenon reflects the physical shielding effect of CFSO molecules on the negative charge sites of PS, verifying that CFSO has been effectively immobilized on/into PS (rather than remaining free in the system). Further insights into this interaction will be analyzed in conjunction with the FT-IR data presented in Section 3.2.3.

3.2.3.Infrared spectroscopy analysis

The results are presented in Fig. 4A. CFSO shows typical hydroxyl (-OH) stretching vibration peaks as broad peaks at 3518 and 3411 cm⁻¹. These peaks originate from C-H stretching vibrations in the benzene ring (Li et al., 2022; Yang et al., 2025a). Peaks at 1629, 1585, 1518, and 1444 cm⁻¹ correspond to skeletal stretching vibrations of the benzene ring. Peaks at 1377 and 1216 cm⁻¹ are attributed to -CH₃ bending vibration and C-O stretching vibration, respectively (Li et al., 2022). For PS, the characteristic broad peak near 3400 cm⁻¹ corresponds to hydrogen bond stretching vibrations. The characteristic broad peak at 2928 cm⁻¹ is assigned to C-H stretching vibration of methylene groups in glucose units (Quilez-Molina et al., 2024). The peak near 1641 cm⁻¹ is attributed to O-H stretching vibration in amorphous regions and asymmetric stretching vibration of carboxylate groups (RCOO⁻) in PS. This indicates the presence of amylopectin or amylose molecular chains in PS (Wahab and Janaswamy, 2024). The characteristic broad peak near 1415 cm⁻¹ corresponds to C-H stretching vibration in -CH₂-. Peaks near 1156 cm⁻¹ and 1047 cm⁻¹ are assigned to C-C stretching and C-O stretching in the crystalline region, respectively (Han et al., 2023). The characteristic broad peak near 928 cm⁻¹ corresponds to the skeletal mode vibration of α-1,4 glycosidic bonds. PS-CFSO exhibits a characteristic broad peak near 3400 cm⁻¹ corresponding to hydrogen bond stretching vibrations, which is similar to that of PS. The characteristic peaks of porous starch are more prominent in the FTIR spectrum of PS-CFSO. Due to the binding between PS and CFSO, a similar characteristic absorption peak is observed at 1634 cm⁻¹. This peak is slightly shifted from the 1629 cm⁻¹ peak of CFSO. Some peaks of CFSO are masked because of its low content in PS-CFSO, and interactions occur between the two components. As a result, the characteristic hydroxyl (-OH) stretching vibration peak of CFSO at approximately 3518 cm⁻¹ disappears in PS-CFSO. Weak peaks related to CFSO are detected at around 1159 and 928 cm⁻¹, and this indirectly confirms that CFSO was likely successfully loaded onto PS (Wahab and Janaswamy, 2024). The spectrum of PS-CFSO shows that the intensity at 1044 cm⁻¹ is significantly higher than that of the 1047 cm⁻¹ characteristic peak in the PS spectrum. A peak at 1048 cm⁻¹ is also observed in the CFSO spectrum, and this may be attributed to the binding of CFSO to PS. As shown by the SEM images in Fig. 2, a small amount of CFSO is distributed on the surface of PS. Thus, CFSO can be detected in the FTIR analysis of PS-CFSO. The above results indicate that no chemical reaction occurred between PS and CFSO, and no new groups were generated (Wu et al., 2025).

Overall, combining zeta potential data with FT-IR analysis indicated that the interaction between PS and CFSO is physical (non-covalent). This phenomenon primarily stems from the synergistic effects of both hydrophobic interactions and hydrogen bonding (Lu et al., 2025). Firstly, hydrophobic interactions serve as the primary driving force. As a lipophilic oil, CFSO spontaneously adsorbs and embeds into the hydrophobic channels of phosphatidylserine (PS), thereby minimizing contact with the aqueous phase. This thermodynamic advantage promotes stable binding between the two. Secondly, weak hydrogen bonding further enhances the stability of this association. The abundant hydroxyl (-OH) groups on the PS surface form weak hydrogen bonds with the phenolic hydroxyl/ester groups of CFSO. This maintains the nature of non-covalent interactions while reinforcing the anchoring effect of CFSO on or within the PS surface.

3.2.4.Crystal structure

XRD is an effective method for analyzing microcrystalline or powder states, and Fig. 4B shows the crystal structures of the samples. CFSO exhibits characteristic diffraction peaks in the 10–30° range, which indicates that CBD is present in a crystalline form. This aligns with the crystal structure of CBD extracts reported in previous studies (Li et al., 2022; Vardanega et al., 2024). PS displays distinct diffraction peaks at 14.3°, 17.55°, and 23.87°, which are characteristic of the type A crystalline structure of cereal starch (Cao and Lu, 2025; Niu et al., 2025). PS-CFSO has an almost identical diffraction pattern to PS. This can be attributed to two factors: first, CFSO is loaded onto starch in a molecular or amorphous form; second, starch acts as a carrier and exerts a dispersing effect that inhibits CFSO crystallization (Yu et al., 2024). This result confirms that PS effectively loads CFSO. It is consistent with previous findings on PS-based loading of naringin (Cao and Lu, 2025) and nanoliposome-based delivery of CFSO (Vardanega et al., 2024).

3.2.5.Thermal property

As shown in Fig. 4C, all samples gradually lose mass as temperature increases. CFSO begins to undergo significant weight loss at approximately 170 °C. This aligns with trends reported in previous studies, where CBD starts to lose weight significantly at 200 °C (Lv et al., 2019). Both PS and PS-CFSO start to show rapid mass loss at approximately 270 °C. This temperature is much higher than CFSO's thermal decomposition temperature of 170 °C. This indicates that complexation between PS and CFSO effectively enhances CFSO's thermal stability. A similar phenomenon was observed when CBD was encapsulated by cyclodextrins (Lv et al., 2019).

Derivative thermogravimetric (DTG) curves were obtained by taking the first derivative of thermogravimetric (TG) weight loss curves (Fig. 4D). The peak temperatures are 252.84 °C for CFSO, 318.12 °C for PS, and 311.24 °C for PS-CFSO. These results further confirm that the PS-CFSO complex has better stability than free CFSO. This stability is beneficial for the product's further processing and application (Cao and Lu, 2025; Li et al., 2025a; Wu et al., 2025).

3.3.Aqueous solubility

Contact angle measurements can characterize the hydrophilicity and hydrophobicity of samples, with a larger contact angle indicating stronger hydrophobicity (Li et al., 2023a, Li et al., 2024a; Li et al., 2024b ). As shown in Fig. 5A, the contact angle of CFSO is 91.2°, while that of PS-CFSO is 60.6°. The contact angle of PS-CFSO is significantly smaller than that of CFSO. This can be explained by two factors: CFSO has a hydrophobic skeleton and undergoes hydrophobic interactions, whereas PS contains abundant hydrophilic groups such as hydroxyl groups (Chen et al., 2021). During magnetic stirring, the two components combine to form PS-CFSO. This combination weakens the hydrophobicity of CFSO, leading to a reduced contact angle.

The solubility of CFSO and PS-CFSO in water was further determined (Fig. 5B). The results show that the saturated solubility of CFSO is 0.38 ± 0.04 μg/mL, while that of PS-CFSO is 6.81 ± 2.04 μg/mL. In comparison, the water solubility of PS-CFSO is 17.92 times higher than that of CFSO, confirming that the adsorption by PS effectively improves the solubility of CFSO. In previous report, PS also increased the water solubility of curcumin by 17.81 times, which is similar to the results of this study (Niu et al., 2025). This similarity arises from two physical mechanisms of PS. First, amorphous solid dispersion is critical: XRD (Section 3.2.4, Fig. 4B) showed that crystalline CFSO, originally with characteristic peaks at 10–30°, converts to an amorphous state in PS. This transition eliminates intermolecular aggregation, a major barrier to water solubility, consistent with Niu et al. (2025), who observed amorphous dispersion of curcumin (another crystalline lipophilic compound) in PS. Second, hydrophilic matrix wrapping, stabilized by non-covalent hydrogen bonds, enhances solubility. FT-IR results (Section 3.2.3, Fig. 4A) revealed that abundant hydroxyl groups (-OH) in PS (broad peak at ∼3400 cm⁻¹) form hydrogen bonds with electronegative oxygen atoms in CFSO (e.g., benzene rings) and curcumin (e.g., phenolic groups). This weak interaction anchors lipophilic compounds in PS's porous structure, while PS's hydrophilic matrix forms a “water-friendly shell” that reduces interfacial tension with water. No new peaks in FT-IR spectra confirm no chemical reactions occurred, further validating the physical nature of this process. This consistency observed across structurally distinct compounds (CFSO and curcumin) highlights the advantage of PS as a universal carrier: its solubilizing effect relies on physical mechanisms applicable to various lipophilic substances. Existing report have shown that naringin encapsulated by PS also achieves a similar enhancement effect (11.63 times), which further verifies this universality (Wang et al., 2024). Such versatility reduces the need for customized carriers and lowers the costs of research and industrial applications.

The dissolution status of CFSO and PS-CFSO in water can be visually observed in Fig. 5C. The original CFSO exhibits obvious hydrophobic characteristics, evidenced by the phenomena of floating on the surface and adhering to the wall, indicating its extremely low water solubility. In contrast, the PS-CFSO complex shows superior dispersion properties. A consistent phenomenon is also observed in studies on the encapsulation of CFSO by cyclodextrins (Li et al., 2022).

3.4.Release in simulated gastrointestinal fluid

The controlled release of CFSO in the human gastrointestinal tract is highly significant, as it influences the bioaccessibility and bioavailability of CFSO (Wang et al., 2022). The in vitro release profiles of CFSO in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) are presented in Fig. 6A and B. After 3 h of digestion in SGF and SIF, only 14.35 ± 0.66 % and 20.11 ± 1.23 % of CBD were detected from free CFSO, respectively. This indicates that free CFSO has low bioaccessibility. The cumulative release rates of PS-CFSO were 68.65 ± 1.29 % (in SGF) and 83.50 ± 2.32 % (in SIF), which are 4.78 and 4.16 times higher than those of free CFSO, respectively. This observation can be explained by two factors. First, the particle size of CBD loaded in the PS-CFSO complex is smaller, as observed via SEM. Second, PS may increase the solubility of CFSO (Xiang et al., 2021). In addition, the release of free CFSO tended to stabilize after approximately 1 h. By contrast, the release of PS-CFSO stabilized at around 2.5 h in SGF and continued even after 3 h in SIF. This demonstrates that PS-CFSO has a superior sustained-release effect. This effect can be attributed to PS absorbing water and swelling, which causes pore shrinkage. This shrinkage requires more time for CFSO to escape from the complex. Furthermore, the amphiphilic groups in PS-CFSO may form a structure with a hydrophilic exterior and hydrophobic interior, thereby exerting a shielding effect on CFSO (Cao and Lu, 2025). Similar enhanced bioavailability has been observed when porous starch was used to deliver curcumin and naringin (Cao and Lu, 2025; Niu et al., 2025).

Fig. 6Release in simulated gastrointestinal fluid and antioxidant activity. (A) Simulated gastric fluid, (B) Simulated intestinal fluid, (C) DPPH, (D) ABTS. Lowercase letters indicate statistical significance among different groups at p < 0.05.

The release curves were fitted (Table S1), and the R² and adjusted R² (R_adj²) values of each model were comparable, indicating good consistency in the simulation of actual release processes. From the perspective of R² values, the first-order kinetics model yielded the highest R² values, which were 0.9636 (in simulated gastric fluid) and 0.9388 (in simulated intestinal fluid), with the corresponding fitting curves shown in Fig. 6A and B. The in vitro release of curcumin encapsulated in PS also conformed to a first-order kinetic model (Han et al., 2023). The release mechanism of PS-CFSO in simulated gastrointestinal fluids was more consistent with the Peppas model, which gave release exponents (n) of 0.3524 and 0.3199. When n < 0.45, it indicates that the release follows a pseudo-Fickian diffusion mechanism, where the release curve is similar to that of Fickian diffusion but requires a longer time to reach equilibrium (Gu et al., 2021).

3.5.Antioxidant activity

CFSO acts as an antioxidant and can effectively scavenge free radicals. However, complexation of active substances may alter their antioxidant capacity. This effect depends on two factors: the orientation of hydroxyl groups in the PS cavity (which are responsible for free radical scavenging) and the strength of host guest molecular interactions (Li et al., 2022). For PS-CFSO, two properties are crucial: maintaining antioxidant capacity and retaining the ability to release CFSO effectively for free radical scavenging. As shown in Fig. 6C and D, both CFSO and PS-CFSO exhibit strong DPPH and ABTS free radical scavenging activities. The DPPH IC₅₀ of CFSO is 168.14 ± 3.57 μg/mL, and its ABTS IC₅₀ is 36.25 ± 0.88 μg/mL. These values are consistent with those reported in previous studies (Erukainure et al., 2023; Li et al., 2022). Compared with free CFSO, PS-CFSO shows a 20.24 % increase in DPPH free radical scavenging activity and a 22.22 % increase in ABTS free radical scavenging activity. This indicates that PS encapsulation effectively enhances CFSO's antioxidant capacity. This enhancement may be explained by the behavior of CFSO molecules when entrapped in the PS cavity: the hydroxyl groups of CFSO form intramolecular and intermolecular hydrogen bonds. These bonds increase the degree of hydroxylation of CFSO and improve the stability of the guest molecule (CFSO), thereby promoting its reaction with ABTS and DPPH free radicals (Olga et al., 2015). Notably, PS encapsulation has also been shown to enhance the antioxidant activity of curcumin (Han et al., 2023; Li et al., 2021).

4.Conclusion

In this study, a process was pioneered for preparing CFSO-PS as the wall material. The physicochemical properties, bioavailability, and antioxidant activity of the microspheres were systematically evaluated. It was found that CFSO was successfully loaded into PS in an amorphous form, exhibiting significantly enhanced thermal stability, water solubility, and antioxidant activity compared to its free state. In simulated gastrointestinal fluids, PS-CFSO exhibited sustained-release behavior, substantially improving CFSO's bioavailability. Overall, this work promotes the advancement of starch-based delivery technologies and provides a viable solution for CFSO applications in functional foods (e.g., fortified beverages), pharmaceuticals (e.g., oral formulations), and cosmetics (e.g., antioxidant skincare products). It also offers a reference for delivering other liposoluble bioactive compounds. However, the current assessment of CFSO's bioavailability and antioxidant activity relies on in vitro experiments, which may not fully reflect actual physiological responses and metabolic processes in vivo. Therefore, future validation can be further pursued through in vivo animal studies and clinical trials. Additionally, deeper exploration of PS-CFSO's behavior in complex food or drug matrices (including interactions with other ingredients and stability during processing and storage) is warranted to contribute to its practical implementation.

Author contribution

Jiangfei Li: Investigation, Writing–original draft, Visualization, Writing–review & editing. Naicheng Xin, Guanjun Ye: Investigation, Conceptualization. Zhilong Zhou, Junwen Wang, Ting Ju, Heqi Gao, Yang Gao, Xianghui Meng: Methodology. Alejandro Cifuentes: Resources. Weihong Lu & Yingchun Zhang: Resources, Writing–review & editing, Funding acquisition.

Data availability declaration

Data will be made available on request.

Formatting of funding sources

Funding for this study was provided by the Key Research and Development Project Innovation Base of Heilongjiang Province (JD2023SJ22) and the Fundamental Research Funds for the Central Universities (HIT-XTCX-5).

Declaration of competing interest

The authors declare that they have no conflicts of interest.

Untitled section

Handling Editor: Dr. Xing Chen

Footnotes

Footnote Group

Contributor Information

Weihong Lu, Email: lwh@hit.edu.cn.

Yingchun Zhang, Email: zhangyingchun@hit.edu.cn.

Appendix A.Supplementary data

The following is the Supplementary data to this article.

References

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Associated Data

Supplementary Materials

Data Availability Statement

Data will be made available on request.

Studies

Conditions

Cannabinoids and active compounds

Related research

Abstract

Untitled section

Graphical abstract

Highlights

Article notes

Untitled section

Author notes

1.Introduction

2.Materials and methods

2.1.Materials

2.2.High performance liquid chromatography (HPLC)

2.3.Preparation of PS-CFSO

2.4.Analysis and calculation of PS-CFSO

2.5.Optimization of the preparation process of PS-CFSO

2.6.Physicochemical properties characterization of PS-CFSO

2.6.1.Scanning electron microscopy (SEM)

2.6.2.Particle size and zeta potential analysis

2.6.3.Fourier transform infrared spectroscopy (FT-IR)

2.6.4.X-ray diffraction (XRD)

2.7.Thermogravimetric analysis (TGA)

2.8.Determination of aqueous solubility

2.8.1.Hydrophilicity

2.8.2.Water solubility

2.9.Cumulative release rate of simulated digestion in vitro

2.10.Determination of antioxidant activity

2.11.Statistical analysis

3.Results and discussion

3.1.RSM optimization of PS-CFSO

3.2.Physicochemical properties of PS-CFSO

3.2.1.Morphological observations

3.2.2.Particle size and zeta potential analysis

3.2.3.Infrared spectroscopy analysis

3.2.4.Crystal structure

3.2.5.Thermal property

3.3.Aqueous solubility

3.4.Release in simulated gastrointestinal fluid

3.5.Antioxidant activity

4.Conclusion

Author contribution

Data availability declaration

Formatting of funding sources

Declaration of competing interest

Untitled section

Footnotes

Footnote Group

Contributor Information

Appendix A.Supplementary data

References

Untitled section

References

Associated Data

Supplementary Materials

Data Availability Statement

Additional materials