Abstract
In the present work, four green processes have been compared to evaluate their potential to obtain rosemary extracts with in vitro anti-proliferative activity against two colon cancer cell lines (HT-29 and HCT116). The processes, carried out under optimal conditions, were: (1) pressurized liquid extraction (PLE, using an hydroalcoholic mixture as solvent) at lab-scale; (2) Single-step supercritical fluid extraction (SFE) at pilot scale; (3) Intensified two-step sequential SFE at pilot scale; (4) Integrated PLE plus supercritical antisolvent fractionation (SAF) at pilot scale. Although higher extraction yields were achieved by using PLE (38.46% dry weight), this extract provided the lowest anti-proliferative activity with no observed cytotoxic effects at the assayed concentrations. On the other hand, extracts obtained using the PLE + SAF process provided the most active rosemary extracts against both colon cancer cell lines, with LC50 ranging from 11.2 to 12.4 µg/mL and from 21.8 to 31.9 µg/mL for HCT116 and HT-29, respectively. In general, active rosemary extracts were characterized by containing carnosic acid (CA) and carnosol (CS) at concentrations above 263.7 and 33.9 mg/g extract, respectively. Some distinct compounds have been identified in the SAF extracts (rosmaridiphenol and safficinolide), suggesting their possible role as additional contributors to the observed strong anti-proliferative activity of CA and CS in SAF extracts.
Highlights
Nowadays, supercritical fluid extraction (SFE) employing CO2 is an established industrial process for the production of high added-value products
Four green processes were selected based on previous works focused on the enrichment of rosemary extracts on carnosic acid (CA) and CS [41,42], to investigate their potential for the production of rosemary leaves’ extracts and fractions with potent in vitro anti-proliferative activity against colon cancer cells
Pressurized Liquid Extraction (PLE) process was chosen since it is suitable for providing a rosemary extract at lab scale that can be used to obtain the starting material required for other processes
Summary
Supercritical fluid extraction (SFE) employing CO2 is an established industrial process for the production of high added-value products. Process integration includes prior unit operation (fermentation, extraction, enzyme pre-treatment, physical fractionation or size reduction) followed by sub-or supercritical extraction or fractionation processes (supercritical chromatography, enzymatic conversion, precipitation and coating of solutes, among others) [45] Regarding this approach, recent reports suggest the combined use of extraction processes, such as supercritical CO2 followed by pressurized liquid extraction (PLE, employing ethanol and water) for improving the recovery of compounds with different polarities and bio-functionalities [44,48,49,50]. Once the processes have been optimized at lab-scale, it is
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