Salting-out extraction of allicin from garlic (Allium sativum L.) based on ethanol/ammonium sulfate in laboratory and pilot scale
Introduction
Garlic (Allium sativum L.) is a well-known edible and medicinal plant since ancient China. Allicin (diallylthiosulfinate) is an organosulfur compound, and it is one major biological active substance in garlic (Tyagi, Pradhan, Srivastava, & Mehrotra, 2014). Actually, allicin is converted from alliin after crushing of the garlic clove under the action of alliinase (Amagase et al., 2001, Ankri and Mirelman, 1999). Allicin was first studied for its antibacterial properties in the middle of 20th century (Cavallito & Bailey, 1944), then its other pharmacological actions of anti-oxidant, antifungal, antihypertensive, anti-inflammatory, and inhibition of tumor were also found (El-Kashef et al., 2015, Hirsch et al., 2000, Liu et al., 2015). Up to now, the solvent extraction using water or ethanol aqueous solution (Arzanlou and Bohlooli, 2010, Bocchini et al., 2001, Wang et al., 2014) and supercritical fluid extraction (SFE) (del Valle et al., 2012, Liang et al., 2012, Rybak et al., 2004) are the most widely used methods for the extraction of allicin from garlic in laboratory, pilot or large scale. However, the solvent extraction can obtain the crude extract and the samples need further purification; SFE requires sophisticated instrument and high cost.
Aqueous two-phase extraction (ATPE) was first introduced by Albersson, and the most commonly used two aqueous two-phase systems (ATPSs) were PEG/salt and PEG/dextran (Albertsson, 1986). ATPS based on low molecular organic solvents (e.g. methanol, ethanol, acetone, and n-propanol) and inorganic salts had been developed in recent years, which can also be called salting-out extraction (SOE) system (Dong et al., 2016). Compared with polymer ATPS, SOE has the advantages of lower cost, lower viscosity, quicker phase separation time, relatively lower environmental toxicity and easier to scale up (Amid et al., 2012, Fu et al., 2015, Liu et al., 2014, Ooi et al., 2009, Wang et al., 2010). SOE systems had been used to extract various bioactive compounds from different plants resources, such as anthocyanins from grape juice (Wu et al., 2014), alkaloids from Sophora flavescens Ait. (Zhang et al., 2015), phenolic compounds from Ficus carica L. (Feng et al., 2015), rutin from acerola waste (Reis et al., 2014), lignans from Zanthoxylum armatum (Guo, Su, Huang, Wang, & Li, 2015), and polysaccharides from Semen Cassiae (Chen et al., 2016).
The objective of this study is to use SOE for the extraction and preliminary purification of allicin from garlic powder. The allicin is extracted into alcohol-rich phase, while partial impurities are extracted into the salt-rich phase. The extraction conditions were optimized at laboratory scale, SOE was scaled up to the pilot scale under the optimized conditions. The phase-forming components of ethanol and ammonium sulfate were recycled and reused.
Section snippets
Materials and reagents
The garlic samples were originated in Shandong province and bought from the Vanguard supermarket in Changsha City, Hunan Province. The allicin standard was purchased from National Institutes for Food and Drug Control (Beijing, China) with HPLC purity larger than 98%. HPLC grade acetonitrile was purchased from TEDIA Company, Inc. (Fairfield, OH, USA). The analytical reagents of different organic solvents (ethanol, n-proanol, isopropanol, acetone and acetonitrile) and salts (ammonium sulfate,
Selection of the optimum SOE system
To screen the optimal SOE system for the extraction and isolation of allicin, various organic solvents (ethanol, n-propanol, isopropanol, acetone and acetonitrile) and salts (ammonium sulfate, sodium dihydrogen phosphate, sodium sulfate, and potassium phosphate) were considered as the phase-forming components. The binodal curves were shown in Fig. 1, which imply that the closer the binodal curves to the coordinates, the less salt (organic solvent) required for forming two phases under the same
Conclusions
In this work, ethanol/ammonium sulfate ATPS was successfully applied for the SOE of allicin from garlic in laboratory and pilot scale. The factors influencing the SOE were investigated in detail. The obtained optimum results were as follows: 24% (w/w) ethanol concentration, 23% (w/w) ammonium sulfate concentration, 40 g/L loaded sample at room temperature with pH being not adjusted. The major three influencing factors were further optimized by RSM. The optimized conditions were 22.57% (w/w)
Acknowledgement
This work was financially supported by the National Natural Science Foundation of China (No. 21406262).
References (39)
- et al.
Intake of garlic and its bioactive components
The Journal of Nutrition
(2001) - et al.
Purification of serine protease from mango (Mangifera Indica Cv. Chokanan) peel using an alcohol/salt aqueous two phase system
Food Chemistry
(2012) - et al.
Antimicrobial properties of allicin from garlic
Microbes and Infection
(1999) - et al.
Introducing of green garlic plant as a new source of allicin
Food Chemistry
(2010) - et al.
Antiproliferative and antioxidant properties of an enzymatic hydrolysate from brown alga, Ecklonia cava
Food and Chemical Toxicology
(2006) - et al.
Determination of diallyl thiosulfinate (allicin) in garlic (Allium sativum L.) by high-performance liquid chromatography with a post-column photochemical reactor
Analytica Chimica Acta
(2001) - et al.
Extraction and characterization of polysaccharides from Semen Cassiae by microwave-assisted aqueous two-phase extraction coupled with spectroscopy and HPLC
Carbohydrate Polymers
(2016) - et al.
Supercritical CO2 extraction of allicin from garlic flakes: Screening and kinetic studies
Food Research International
(2012) - et al.
Extraction and purification of IgG by hydrophilic organic solvent salting-out extraction
Journal of Chromatography B
(2016) - et al.
Protective effect of allicin against gentamicin-induced nephrotoxicity in rats
International Immunopharmacology
(2015)
Phase separation in a salting-out extraction system of ethanol-ammonium sulfate
Separation and Purification Technology
Observations on the use of statistical methods in food science and technology
Food Research International
Combination of aqueous two-phase extraction and cation-exchange chromatography: New strategies for separation and purification of alliin from garlic powder
Journal of Chromatography B
Aqueous two-phase extraction of 2,3-butanediol from fermentation broths using an ethanol/ammonium sulfate system
Process Biochemistry
Hydrophilic organic/salt-containing aqueous two-phase solvent system for counter-current chromatography: A novel technique for separation of polar compounds
Journal of Chromatography A
Allicin protects spinal cord neurons from glutamate-induced oxidative stress through regulating the heat shock protein 70/inducible nitric oxide synthase pathway
Food & Function
Optimisation of aqueous two-phase extraction of anthocyanins from purple sweet potatoes by response surface methodology
Food Chemistry
Thermodynamic study of aqueous two phase systems for some aliphatic alcohols plus sodium thiosulfate plus water
Fluid Phase Equilibria
Purification of lipase derived from Burkholderia pseudomallei with alcohol/salt-based aqueous two-phase systems
Process Biochemistry
Cited by (55)
Allicin: A review of its important pharmacological activities
2023, Pharmacological Research - Modern Chinese MedicineA review of recent developments in sugars and polyol based soluting out separation processes
2023, Separation and Purification TechnologyOrganosulfur in food samples: Recent updates on sampling, pretreatment and determination technologies
2023, Journal of Chromatography AProcess intensification on the enhancement of allicin yield from Allium sativum through ultrasound attenuated nonionic micellar extraction
2021, Chemical Engineering and Processing - Process Intensification