Food Engineering, Materials Science, & Nanotechnology Green Formulation of Triglyceride/Phospholipid- Based Nanocarriers as a Novel Vehicle for Oral Coenzyme Q10 Delivery Sonia Alavi, Sarah Akhlaghi, Simin Dadashzadeh, and Azadeh Haeri Abstract: This study was aimed to develop a novel nanocarrier for coenzyme Q10 (CoQ10) by a green process that prevented the use of surfactants and organic solvents. Triglyceride/phospholipid-based nanocarriers were developed through high-pressure homogenization (an industrial feasible process), and a 2 5-1 fractional factorial design was adopted to assess the influences of formulation variables on the considered responses, including vesicle size, entrapment efficiency, loading capacity, and solubility of the vehicles in simulated gastrointestinal fluids. The optimized formulation was further in-depth characterized in terms of morphology, release behavior, biocompatibility (Caco-2 cell cytotoxicity and histological examination), thermal behavior, and Fourier transform infrared analysis. Optimal nanocarriers were found to have mean particle size of 75 nm, narrow particle distribution, and CoQ10 entrapment of 95%. The optimized formulation was stable upon incubation in simulated gastrointestinal fluids without considerable leakage of cargo, which was in agreement with their sustained release behavior. Microscopic observations also confirmed nanosized nature of the vesicles and revealed their spherical shape. Moreover, toxicity evaluations at the cellular and tissue levels revealed their nontoxic nature. In conclusion, triglyceride/phospholipid-based nanocarriers proved to be a green safe vehicle for delivery of CoQ10 with industrial-scale production capability and could provide a new horizon for delivery of hydrophobic nutraceuticals. Keywords: Coenzyme Q10, lipid-based nanocarrier, green formulation, factorial design, triglyceride, phospholipid Practical application: Green nanostructure formulation approaches have recently gained tremendous attraction for their safe profile especially when it comes to supplements, which are generally recommended for daily use. However, their sufficient association with cargoes and industrial-scale production have remained considerable challenges. This study focuses on the development of lipid-based nanocarriers for CoQ10 by an industrial feasible process that prevents the use of any surfactants or organic solvents. Introduction Coenzyme Q10 (CoQ10, 2,3-dimethoxy-5-methyl-6- decaprenyl-1,4-benzoquinone) is a lipophilic vitamin analog, present in all cells of human origin (Alcazar-Fabra, Navas, & Brea-Calvo, 2016). It scavenges free radicals, restrains lipid peroxidation and functions as a coenzyme in various metabolic pathways (Potgieter, Pretorius, & Pepper, 2013). However, the level of this antioxidant declines with aging and its deficiency can interrupt cellular mitochondrial functions, triggering an oxidative cycle through destabilization of lysosomal membrane (Bentinger, Brismar, & Dallner, 2007; Sohal & Forster, 2007). Thus, biological functions of CoQ10 highlight the rationale for its growing use as a nutritional supplement as well as therapeutic adjuvant in the management of cardiovascular, neurodegenerative and neuromuscular disorders, periodontal disease, cancers, and diabetes (Garrido-Maraver et al., 2014; Rybalka, Timpani, Stathis, Hayes, & Cooke, 2015). JFDS-2019-0652 Submitted 5/1/2019, Accepted 7/18/2019. Authors Alavi, Akhlaghi, Dadashzadeh, and Haeri are with Dept. of Pharmaceutics, School of Phar- macy, Shahid Beheshti Univ. of Medical Sciences, Tehran, Iran. Author Haeri is also with Pharmaceutical Sciences Research Center, Shahid Beheshti Univ. of Med- ical Sciences, Tehran, Iran. Direct inquiries to author: Dr. Azadeh Haeri (E-mail: a_haeri@sbmu.ac.ir) However, regarding its large molecular weight (863 Da), low aqueous solubility (log P > 10), and thermolability, the oral bioavailability of CoQ10 is extremely low and has limited its clinical use (Borekova, Hojerova, Koprda, & Bauerova, 2008; Jankowski, Korzeniowska, Cieslewicz, & Jablecka, 2016). To over- come these obstacles, various formulations have been proposed, including complexation with cyclodextrins (Fir, Smidovnik, Milivojevic, Zmitek, & Prosek, 2009), complexation with albumin hydrolysate (Matsushita, Oshima, Takahashi, & Baba, 2013), solid dispersions (Bhandari et al., 2007), and nanoformulations (Cheuk et al., 2015; Kaci et al., 2018; Lee, Suh, Kim, & Lee, 2017; Qin et al., 2017). As most of these approaches led to low-to-moderate success (Hatanaka, Kimura, Lai-Fu, Onoue, & Yamada, 2008; Piao et al., 2011; Zhou et al., 2013), new strategies, especially green ones (Chemat et al., 2017), are in great demand, aiming not only to increase the solubility of CoQ10, but also to eliminate the use of any surfactant or organic solvent. These chemical additives are frequently employed to deal with hydrophobic compounds; however, most of which are not inert from the perspective of pharmaceutical product quality and lead to a variety of vehicle- related side effects (Abrantes, Duarte, & Reis, 2016). Hence, green approaches are of undeniable value, especially when it comes to supplements that are generally recommended for daily use. Among the recent efforts in this regard are triglyceride/ phospholipid-based nanocarriers, which have been proposed to C  2019 Institute of Food Technologists R  doi: 10.1111/1750-3841.14763 Vol. 0, Iss. 0, 2019  Journal of Food Science 1 Further reproduction without permission is prohibited