This study aimed to optimize subcritical water extraction process, characterize chemical composition and investigate the biological activities of Nitraria sibirica Pall. (NSP) anthocyanin. Overall, the optimization was achieved under following conditions: extraction temperature 140 °C, extraction time 45 min and flow rate 7 mL/min with the extraction yield of 1.075 mg/g. 3 cyanidin, 3 petunidin, 1 delphinidin and 1 pelargonidin compounds were identified in the anthocyanic extract from NSP via UPLC-Triple-TOF-MS/MS. NSP anthocyanin exhibited better DPPH free-radical scavenging activity than ascorbic acid. It displayed superior α-glucosidase inhibition activity, which was ∼14 times higher than that of acarbose. Moreover, enzyme kinetics results indicated that NSP anthocyanin behaved as a reversible, mixed-type inhibitor. Molecular docking and molecular dynamics simulation results revealed that NSP anthocyanin interacted with α-glucosidase mainly via van der Waals forces, hydrogen bond and possessed fairly stable configuration. Therefore, NSP anthocyanin is a promising α-glucosidase inhibitor for diabetes mellitus.

Pectin was extracted from red dragon fruit (Hylocereus polyrhizus) peel using two different extraction methods: subcritical water extraction (SCWE) and conventional acid extraction (AE), from two different types of peels, fresh peel puree and dried peel powder. SCWE method on fresh peel puree showed an ∼18.88 % increase in pectin yield compared to AE. Extracted pectin is classified as low methoxyl pectin (DE: 8.51-50.64 %), with an average molecular weight ranging from 115.23 kDa to 577.84 kDa and a Gal-A content of 44.09 % - 53.90 %. The potential of pectin from fresh peel puree to be applied as a biodegradable film was further explored. Different pectin concentrations (3-5 % w/v) were used to prepare the films. Regarding the film performance, PF-S5, which was produced from SCWE with 5 % of pectin concentration, exhibits better thermal stability (Tdmax 250 °C, residue of 28.69 %) and higher moisture barrier (WVP 5.59 × 10-11 g.cm-1.s-1.Pa-1). In comparison, PF-A showed lower water solubility (45.14-69.15 %), higher water contact angle (33.01° - 44.35°), and better mechanical properties (TS: 2.12-4.11 MPa, EB: 48.72-61.39 %). Higher molecular weight accompanied by higher DE and Gal-A content contributes to better pectin film properties.

Subcritical water extraction (SWE) is an efficient and eco-friendly technology that rapidly extracts valuable compounds from natural materials. In this study, response surface methodology (RSM) was utilized to determine the optimal extraction conditions for Gastrodiae Rhizoma using SWE (GRP-S). The optimum conditions were found to be 161 °C extraction temperature, 41 min extraction time, and a solid-to-liquid ratio of 1.55 mg/mL. Under these optimal conditions, the experimental yield of GRP-S was 66.32% ± 0.10% (n = 3), demonstrating a significant increase compared to hot water reflux extraction (HWE) in the extraction yield of polysaccharides. Characterization studies employing SEM, FT-IR, and HPAEC-PAD confirmed the differences between GRP-S and GRP-H (GRP obtained by HWE). Furthermore, both GRP-S and GRP-H exhibited a significant ability to protect HepG2 cells from ethanol-induced damage, with GRP-S showcasing a superior effect. The widespread adoption of SWE technology can lead to high GRP content in extracts and promote the green and sustainable development of natural products extraction processes.

Subcritical water extraction was used to extract bioactive phenolic compounds from Vaccinium dunalianum Wight leaves. The optimal extraction conditions were determined as an extraction temperature of 150 °C, an extraction time of 40 min, and a liquid-solid ratio of 35 : 1 mL/g. The total phenolic content reached 21.35 mg gallic acid /g, which was 16 % higher than that by hot water extraction. The subcritical water extraction extract exhibited strong scavenging activity of DPPH free radical and ABTS+ free radical, as well as significant tyrosinase inhibitory activity. The study suggests that subcritical water extraction can alter the composition of the extracts, leading to the production of various phenolic compounds, effective antioxidants, and tyrosinase inhibitors from Vaccinium dulciana Wight leaves. These findings confirm the potential of Vaccinium dunalianum Wight as a natural antioxidant molecule source for the medicine and food industries, and for the therapy of skin pigmentation disorders.

Hemerocallis citrina Borani as a low-cost vegetable, has various health benefits. However, the industry of H. citrina Borani is in the state of primary processing, with poor economic benefits. This study aimed to investigate the physicochemical properties, and the antioxidant activity of H. citrina Borani polysaccharide (HCBP) using subcritical water extraction (SWE) at different temperatures, to expand the value of H. citrina Borani. HCBP mainly composed of nine monosaccharides (glucose, galactose, rhamnose, fucose, mannose, arabinose, xylose, galacturonic acid, and glucuronic acid), among which the content of neutral sugar was higher and uronic acid was lower. HCBP contained glycosidic bond of β-configurations and trace quantities protein. The molecular weight of HCBP decreased with increasing temperature. Shear thinning occurred in HCBP with the increase of shear rate (0.01-1 s-1), and the apparent viscosity of HCBP decreased at higher temperature (150-160°C) with the increase continuously of shear rate (1-10 s-1), but almost remained constant at lower temperature (130-140°C). Scanning electron microscope showed that HCBP had rough surface, loose structure, obvious particle gap, and irregular shape. In addition, HCBP extracted at 160°C had strong FRAP activity, and HCBP extracted at 130 and 140°C had better ABTS radical scavenging activity. This study suggests that HCBP extracted by SWE could provide a cheap raw material as food thickening agent and natural antioxidants.

Subcritical water refers to high-temperature and high-pressure water. A unique and useful characteristic of subcritical water is that its polarity can be dramatically decreased with increasing temperature. Therefore, subcritical water can behave similar to methanol or ethanol. This makes subcritical water a green extraction fluid used for a variety of organic species. This review focuses on the subcritical water extraction (SBWE) of natural products. The extracted materials include medicinal and seasoning herbs, vegetables, fruits, food by-products, algae, shrubs, tea leaves, grains, and seeds. A wide range of natural products such as alkaloids, carbohydrates, essential oil, flavonoids, glycosides, lignans, organic acids, polyphenolics, quinones, steroids, and terpenes have been extracted using subcritical water. Various SBWE systems and their advantages and drawbacks have also been discussed in this review. In addition, we have reviewed co-solvents including ethanol, methanol, salts, and ionic liquids used to assist SBWE. Other extraction techniques such as microwave and sonication combined with SBWE are also covered in this review. It is very clear that temperature has the most significant effect on SBWE efficiency, and thus, it can be optimized. The optimal temperature ranges from 130 to 240 °C for extracting the natural products mentioned above. This review can help readers learn more about the SBWE technology, especially for readers with an interest in the field of green extraction of natural products. The major advantage of SBWE of natural products is that water is nontoxic, and therefore, it is more suitable for the extraction of herbs, vegetables, and fruits. Another advantage is that no liquid waste disposal is required after SBWE. Compared with organic solvents, subcritical water not only has advantages in ecology, economy, and safety, but also its density, ion product, and dielectric constant can be adjusted by temperature. These tunable properties allow subcritical water to carry out class selective extractions such as extracting polar compounds at lower temperatures and less polar ingredients at higher temperatures. SBWE can mimic the traditional herbal decoction for preparing herbal medication and with higher extraction efficiency. Since SBWE employs high-temperature and high-pressure, great caution is needed for safe operation. Another challenge for application of SBWE is potential organic degradation under high temperature conditions. We highly recommend conducting analyte stability checks when carrying out SBWE. For analytes with poor SBWE efficiency, a small number of organic modifiers such as ethanol, surfactants, or ionic liquids may be added.

This study investigated the effects of ultrasound assisted-subcritical water (U-SW), subcritical water (SW), ultrasound (U) and hot water (HW) pretreatments and acid hydrolysis (AH) and alkaline hydrolysis (AlkH) on the phenolic composition, antioxidant potential and cytotoxicity of Tartary buckwheat hull extracts. The Folin Ciocalteu assay and HPLC-MS were used to characterize and quantify phenolics of the extracts. The ABTS, FRAP and TEAC assays were used to measure antioxidant activity and the MTT assay was used to measure cytotoxicity of the extracts in HepG2 human liver cancer cells. Results showed that U-SW gave the best AH yield of phenolics (128.45), followed by SW (85.82) and U (64.70), compared to the control, HW (35.82 mgg-1). The same trend was observed for phenols extracted using AlkH. U-SW had the highest antioxidant activity, followed by SW and U regardless of hydrolytic method used. Cytotoxicity followed a similar trend with U-SW and SW being the most cytotoxic to liver cancer cells, followed by U, with the least being HW. The findings suggested that plant materials such as Tartary buckwheat hulls can be pretreated with U-SW, SW and U prior to hydrolytic recovery of bound polyphenols. Also, AH was more efficient than AlkH for phenol extraction, and gave extracts with higher antioxidant activity and cytotoxicity in HepG2 liver carcinoma cells. This application allows for beneficial usage of agricultural biomass and help diversify income sources and products for industry.

The effects of ultrasound-assisted (UAE), subcritical water (SWE) and ultrasound assisted-subcritical water (UA-SWE) treatments on tartary buckwheat polyphenol yield, composition, antioxidant activity and cytotoxicity in human liver carcinoma cells were studied. Folin Ciocalteu assay was used to measure total free phenol content (TFPC), and ABTS, DPPH, FRAP and TEAC assays were used to measure antioxidant activity (AA). Polyphenol characterization was done by LC-MS and cell antioxidant activity (CAA) and cytotoxicity were done using the 2,2\'-Azobis-(2-amidinopropane) dihydrochloride [ABAP] and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide [MTT] assays respectively. The highest polyphenol yield was obtained by SWE (53.3 mg g-1), followed by UA-SWE (31.8 mg g-1), UAE (19.3 mg g-1) and HWE (4.2 mg g-1). Also, SWE had the highest TFPC (7.9 mgGAE/gdw). UAE and UA-SWE showed no differences with TFPC being 6.6 and 6.8 mgGAE/gdw, respectively. The control method (HWE) had the largest number of phenolic compounds identified (25), followed by UAE, SWE and UA-SWE which had 20, 13 and 11 phenolics respectively. Beside phenolic acids, all treatments extracted a number of flavonoids such as flavan-3-ols (catechin-7-O-glucoside, epigallocatechin-3-gallate, epigallocatechin, epicatechin), flavonols (kaempferol-3-O-glucoside, kaempferol, kaempferol-3-rutinoside, rutin, quercetin, quercetin-3-O-glucuronide hyperin), flavones (vitexin, isovitexin, orientin, isoorientin) and anthocyanins (cyanidin-3-O-rutinoside, Cyanidin 3-O-galactoside, cyanidin-3-O-glucoside). SWE gave the highest AA for all tests. However, the AA of those obtained by UAE and UA-SWE did not vary (P < 0.05), but were higher than HWE. Different extracts had best AA at different concentrations (HWE, 300; UAE, 250; SWE, 150; UA-SWE, 200 μg/mL). The IC50 of AA were 270.8 ± 21.3, 198.1 ± 16.0, 97.9 ± 13.5, and 150.4 ± 12.8 μg/mL, respectively for HWE, UAE, SWE and UA-SWE. Generally, SWE and UA-SWE showed the highest cytotoxic activities, followed by UAE, with HWE being the lowest. IC50 of cytotoxicity were 76.1 ± 3.3, 79.5 ± 7.0 and 92.6 ± 4.9 µg/mL for SWE, UA-SWE and UAE, respectively. SWE is a promising method for polyphenol extraction and its combination with ultrasound should be optimized for high yield and conservation of bioactivity.

To investigate and compare the preliminary structural characteristics and biological activity in vitro of polysaccharides from Sagittaria sagittifolia L. (SSs) by different extration methods, three polysaccharides (SSW, SSU, and SSP) were obtained with hot water, ultrasound-assisted, and subcritical water extraction. Their structural features were elucidated using High Performance Liquid Chromatography (HPLC), Gas Chromatography (GC), Scanning Electron Microscopy (SEM), Infrared Spectroscopy (IR), Atomic Force Microscopy (AFM), Zeta Potential and Congo red methods. Furthermore, the antioxidant activity and immunostimulatory effects were investigated in vitro. Molecular weight and monosaccharide composition analysis exhibited that SSW (2275.0 kDa), SSU (148.7 kDa), and SSP (1984.0 kDa) were heteropolysaccharide with dramatically different monosaccharide species and mole ratios. In addition, SSP exhibited stronger antioxidant activity in vitro and more potent immunomodulatory activity than SSW and SSU. SSP has greater potential to be explored as biologicalagents for use in complementary medicine or functional foods.

Wheat germ protein (WGP) was extracted with subcritical water and then hydrolyzed with Alcalase 2.4 L to obtain antioxidant hydrolysates. Wheat germ peptides (WG-P, Mw < 1 kDa) were purified by using Sephadex G-15 column chromatography. The results showed that WG-P-4 possessed the strongest DPPH radical scavenging activity in comparison with other peptides fractions. In addition, free amino acids and LC-MS/MS analysis showed that Gly-Pro-Phe, Gly-Pro-Glu, and Phe-Gly-Glu were the major peptides of WG-P-4. Interestingly, the WG-P-4 fractions had good absorption characteristic. Moreover, the ratio of Papp both sides of apical compartment (AP) and basolateral compartment (BL) were between 0.5 and 1.0 on Caco-2 cell model, which indicated that transmembrane transportation was mainly passive transport. Therefore, WG-P could exert an effective antioxidant action by across the intestinal epithelium.