Preparation and physicochemical stability of hemp seed oil liposomes

https://doi.org/10.1016/j.indcrop.2021.113283Get rights and content

Highlights

  • Hemp seed oil liposomes were prepared by ethanol injection-ultrasonic dispersion (EIUD).

  • The encapsulation efficiency and oil loading ratio were used as the response indicators.

  • The average particle size, polydispersity index and zeta potential were assessed.

  • TEM and cryo-SEM showed round and regular HSOL surface.

  • In HSO, TPC, DPPH inhibition and Δ9-THC stability were enhanced by HSOL.

Abstract

Hemp seed oil (HSO) liposomes (HSOLs) were prepared by ethanol injection-ultrasonic dispersion (EIUD). The encapsulation efficiency (EE) and oil loading ratio (OLR) were used as the response indicators to optimize the preparation conditions of the HSOLs. The average particle size (Z-Ave) of the prepared liposomes was 122 ± 1.64 nm, the polydispersity index (PDI) was 0.21 ± 0.06, the zeta potential (ZP) was -36.5 ± 0.32 mV, the encapsulation efficiency (EE) was 81.3 ± 1.62 % and the oil loading rate (LOR) was 6.13 ± 0.07 %. Transmission electron microscopy (TEM) and cryo-scanning electron microscopy (cryo-SEM) showed that the morphology of the HSOLs was round, and the cut surface was regular. The storage and oxidative stability tests of the HSOLs showed that the Z-Ave was 152 ± 1.87 nm and the polydispersity index (PDI) was 0.72 ± 0.03 after storage for 70 days under natural light at 25 °C. The zeta potential change showed no trend. The polyphenolic content in the HSOLs was 2.25 mg GAE/100 g, the DPPH clearance rate was 60.7 %, and the Δ9-tetrahydrocannabinol (Δ9-THC) content was 85.43 mg/kg.

Introduction

Hemp seeds contain a large amount of unsaturated fatty acids and proteins but do not contain cholesterol. Therefore, hemp seeds have attracted increasing attention as health products (Yang et al., 2017). The edible derivative of hemp seed is mainly hemp seed oil (HSO), which is an excellent edible oil source for humans (Smeriglio et al., 2016). HSO contains tetrahydrocannabinol and a large amount of phenolic compounds with high antioxidant properties (Pellegrini et al., 2005). Certain refining and storage methods can destroy the phenolic compounds in HSO because the oil is easily oxidized, which reduces the oil content and the antioxidant effects of the product. The content of unsaturated fatty acids in the HSO approximately 80 % (Dunford, 2015; Kostić et al., 2013), and the oxidative stability index (OSI) value is 8.15 h. The oil is easily oxidized, leading to rancidness and a decline in quality (Simopoulos, 2002). Therefore, HSO has a high risk of oxidation in direct applications.

Liposomes are an emerging encapsulating vector. According to the literature, many components that are sensitive to oxygen, temperature and light are enhanced by liposome encapsulation to extend their shelf life (Aditya et al., 2015; Ni et al., 2015). Liposomes encapsulated licorice extract and retained high biological activities over time (Castangia et al., 2015). Curcumin-loaded vesicles prepared from phospholipids (P90 G) and immobilized with sodium hyaluronate showed high stability (Manca et al., 2019). Liposomes encapsulated mangiferin and improved its effectiveness against oxidative stress in fibroblasts (Pleguezuelos-Villa et al., 2020). Liposomes encapsulated grape seed extract, enhanced the activity of grape polyphenols, and improved its protective and nursing effects on the human intestines (Manca et al., 2020). The use of liposomes to encapsulate fish oil (Ghorbanzade et al., 2017), tea polyphenol oil (Lu et al., 2011), and artemisinin oil (Sinico et al., 2005) achieved significant results. Many methods have been used to prepare liposomes, including the film dispersion method, freeze-thaw method, reverse evaporation method (Wu et al., 2019), and ethanol injection method.

Conventional liposome experiments use chloroform or diethyl ether to embed and dissolve liposome wall materials. These reagents cannot be removed by thin film dispersion. Therefore, ethanol injection-ultrasonic dispersion (EIUD) is used to prepare low risk materials in a safe and non-toxic manner. The application of liposome encapsulation and release technology in pharmaceutical domain has great potential. To date, no studies have evaluated liposome encapsulated HSO. This study investigated liposome encapsulation to prevent the destruction of polyphenols inside the hemp seed oil liposomes (HSOLs) to decrease the risk of HSO oxidation after refining. The microscopic biological morphology and physical stability of these liposomes were studied, and the effects of liposome encapsulation on the antioxidant activity of the polyphenol compounds in HSO were evaluated. This study is expected to enable HSOLs to achieve the safe delivery and controlled release of bioactive molecules in target products, improve its bioavailability, and provide more benefits for achieving human health.

Section snippets

Preparation of liposomes

HSOLs were prepared by the EIUD described by Charcosset et al. (2015). In short, according to the optimal combination, an appropriate amount of the sample was weighed, dissolved in 100 mL of ethanol, and gently heated at 40 ± 1 °C to increase the miscibility. The ethanol solution was injected into 200 mL of PBS containing 5 % trehalose at 55 ± 2 °C through an injection needle at a constant injection rate of 1.5 mL/min to improve the micro-mixed phase in the aqueous phase. After the injection,

Optimization and verification of the preparation conditions

The results of the orthogonal L9 (34) test are shown in Supplementary Table 2. The differences in ΔR values show that with EE as the index, the primary and secondary order of the effects of each factor were: B > A>C > D, and the optimal combination was B1A3C1D2. Taking OLR as the index, the order of influence of each factor was B > A>C > D, and the optimal combination was B1A3C2D2. The main factors affecting the encapsulation efficiency (EE) and oil loading rate (OLR) were B: HSO / (EPC + Chol)

Conclusion

HSO was encapsulated in liposomes composed of egg yolk lecithin (EPC) and cholesterol (Chol). The size distribution of the HSOLs was uniform, and the shape was spherical and full. When stored under natural light at 25 °C, after 70 days the average particle size of the HSOLs increased by only 19.7 %, the polydispersity index (PDI) increased by 70.8 %, the retention rate of polyphenols increased by 28.0 %, and the ability to remove DPPH increased by 26.2 %. The retention rate of

CRediT authorship contribution statement

Yanguo Shi: Project administration, Supervision, Methodology. Wen Wang: Writing - original draft, Conceptualization, Methodology. Xiuqing Zhu: Visualization, Investigation, Formal analysis. Bing Wang: Formal analysis, Validation, Visualization. Yue Hao: Writing - review & editing, Funding acquisition. Liqi Wang: Data curation, Formal analysis. Dianyu Yu: Funding acquisition, Investigation, Project administration, Resources, Supervision, Writing - review & editing. Walid Elfalleh:

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgements

This work was supported by a grant from the National Natural Science Foundation of China (NSFC): Construction of a three-dimensional enzyme electrode and its correlation with phospholipid content in oil (No: 32072259).

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