Phytochemical Profile and In Vitro Bioactivities of Wild Asparagus stipularis.

In this study, Asparagus stipularis was characterized concerning its phytochemical composition, antioxidant potential, cytotoxicity, and pancreatic lipase inhibitory activities. Twenty-seven compounds were identified and quantified by HPLC-DAD-MS in the leaf, stem, pericarp, and rhizome of ethanolic extracts. Seven steroidal saponins were detected, and the highest content was quantified in rhizome and pericap. A. stipularis also contained significant amounts of flavonoids in the aerial part. Isorhamnetin tetra-glycoside, quercetin-3-glucosyl-rutinoside, and rutin were the main flavonoid derivatives in leaf, stem, and pericarp extracts, respectively. In addition, eleven phenolic acids were also detected; among them, caffeic acid, protocatechuic acid, p-hydroxybenzoic acid, and ferulic acid were the predominant phenolics, with these having the highest amounts quantified in the rhizome extracts. All the tested extracts possessed antioxidant capacities, with pericarp and rhizome extracts exhibiting the highest activity in DPPH, ABTS, and FRAP assays. The extracts from pericarp and rhizome were revealed to also be the strongest inhibitors of pancreatic lipase. The rhizome extracts exhibited potent cytotoxic activity against HCT-116 and HepG2 with IC50 values of 30 and 54 µg/mL after 48 h of treatment. The present study demonstrated that A. stipularis can be used as a new source of natural antioxidants and potential anticancer and antiobesity compounds.

Date Seeds (Phoenix dactylifera): Antioxidant Potential and Profile of Free and Bound Polyphenols from Different Cultivars.

In this study, the antioxidant activities and detailed phenolic profiling of extracts from seven cultivars of date seeds were investigated. Significant differences were detected among cultivars. Total phenolic content (TP) ranged between 135.9±12.1 and 284.86±21.9 mg GAE/g DM. The total flavonoid value varied between 34.20±0.34 and 94.46±1.04 mg RE/g DM. The condensed tannin ranged from 24.17±1.13 to 201.60±9.95 mg CTE/g DM. Phloroglucinolysis was used to depolymerize the bound polyphenols. Results show the presence of phenolic acids: Hydroxybenzoic acid, hydroxycinnamic acid, and a high amount of flavan-3ols (monomers, dimers, and trimers). Before depolymerization, the highest amount of total polyphenols was identified in Kenta (8.48 g/kg) and the lowest was detected in Hessa (4.74 g/kg). After depolymerization, the flavan-3-ols increased significantly, ranging between 46.91g/kg in Hessa and 72.38 g/kg in Deglet Nour, with a high degree of polymerization (DP) in all cultivars. It can be concluded that date seeds represent a good source of bioactive compounds.

A Sustainable Approach for the Valorization of Underutilized Date Fruits.

Secondary varieties of date fruits are often discarded because they do not have commercial value. However, their phytochemicals are very similar to those of the primary ones and therefore, they can be valorized as a source of compounds of interest, mainly phenols and dietary fiber. Their chemical composition changes with ripening, so their characterization throughout this process is of great significance. Date fruit samples were harvested at Khalal, Rutab, and Tamer stages, and a mixture of fruits from ornamental date trees was also analyzed. Aqueous and ethanolic extracts were studied for their phenolic composition. In aqueous extracts, phenols decreased with ripening, while in the ethanolic ones having higher phenolic content. Chelidonic acid, a γ-pyrone, was the major compound found in all extracts, but in the ethanolic ones, flavonoids were also present in similar amounts. After purification by adsorption chromatography, all extracts were assayed for their antimicrobial activity. Those from the Tamer stage showed the highest activity, especially against Gram-positive bacteria. The fibrous residues after aqueous and ethanolic extractions were also characterized. Their chemical composition suggested that they can be considered as a good source of prebiotic arabinoxylans and antioxidant fiber, whose antiradical activity correlated with their phenolic content. Date fruits from secondary varieties are promising as a worthwhile starting point for obtaining new value-added products.

Phytochemical Characterization and Bioactivity of Asparagus acutifolius: A Focus on Antioxidant, Cytotoxic, Lipase Inhibitory and Antimicrobial Activities.

The phytochemical composition of leaves, stems, pericarps and rhizomes ethanolic extracts of Asparagus acutifolius were characterized by HPLC-DAD-MS. A. acutifolius samples contain at least eleven simple phenolics, one flavonon, two flavonols and six steroidal saponins. The stem extracts showed the highest total phenolic acid and flavonoid contents, where cafeic acid and rutin were the main compounds. No flavonoids were detected in the leaf, pericarp or rhizome while caffeic acid and ferulic acid were the predominant. Steroidal saponins were detected in the different plant parts of A. acutifolius, and the highest contents were found in the rhizome extracts. The stem extracts exhibited the highest antioxidant activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) and the highest 2,2-azino-bis (3 ethylbenzothiazoline-6-sulphonic acid) (ABTS) scavenging activity was found in the pericarp extracts. The rhizome and leaf extracts showed a potent cytotoxic activity against HCT-116 and HepG2 cell lines. Moreover, the pericarp and rhizome extracts revealed a moderate lipase inhibitory activity. The leaf and rhizome extracts were screened for their antimicrobial activity against human pathogenic isolates. The leaf extract exhibited a powerful inhibitory activity against all the bacteria and fungi tested.

In Vitro Toxicity of Asparagus Saponins in Distinct Multidrug-Resistant Colon Cancer Cells.

Colorectal cancer is the third most common cancer in the world. Many efforts have focused on finding natural molecules with potential chemo-preventive activity due to their low toxicity compared to synthetic drugs. However, comprehensive information on the bioactive fractions and components is still missing. In this study, we developed a method for the quantitative separation and isolation of saponins from asparagus genotypes consisting of an adsorption chromatography and subsequent liquid chromatographic separation on a reversed-phase column. The saponins isolated were tested for their cytotoxic activity against human colon cancer cell lines, which could develop cross-resistance to a wide variety of chemotherapeutic drugs. Our results showed that Huétor-Tájar asparagus saponins (HTSAP), mainly protodioscin and HTSAP-10 have higher cytotoxic activity than HTSAP-1, HTSAP-6, and HTSAP-8. This study links the potential anticancer effect of asparagus to specific saponins and unveils the triguero Huétor-Tájar asparagus as a nutraceutical particularly in colon cancer therapies.

Applications of Saponin Extract from Asparagus Roots as Functional Ingredient.

When replanting an asparagus field, the roots of the previous crop are crushed and incorporated into the soil, creating problems of autotoxicity and fungal infections. Asparagus roots can be considered as a valuable byproduct, since they are very rich in saponins (3-6%), compounds currently considered as bio-emulsifiers. The objective is to evaluate the emulsifying and foaming capacity of a saponin extract from asparagus roots (ARS) and compare it with other commercial extracts. ARS was obtained using a process patented by our research group. The results have shown that ARS has activity similar to Quillaja extract. Its critical micellar concentration falls between that of Quillaja and Tribulus extracts (0.064, 0.043, and 0.094 g/100 mL, respectively). Both emulsifying and foaming activities are affected by pH, salt, and sucrose to a similar extent as the other extracts. Additionally, it has demonstrated an inhibitory effect on pancreatic lipase, which is even better than the other two studied extracts, as indicated by its IC50 value (0.7887, 1.6366, and 2.0107 mg/mL for asparagus, Quillaja, and Tribulus, respectively). These results suggest that ARS could serve as a natural emulsifying/foaming agent for healthier and safer food products and as a potential aid in treatments for obesity and hyperlipidemia.

Sustainable valorization of co-products from asparagus cultivation by obtaining bioactive compounds.

Asparagus cultivation generates every year a significant amount of by-products that consist of root and frond. Leaving these residues on the fields after harvesting negatively affects the following asparagus crops, since they release autotoxic (allelopathic) substances into the soil, whose accumulation causes that asparagus yields gradually decrease over the years, becoming an unprofitable crop in a period of about 10 to 15 years. This phenomenon is known as decay and forces the entire asparagus plantation to be lifted (abandoned). On the other hand, once a certain plantation has been lifted, it is not profitable to immediately re-plant new asparagus plants, since the yields that are achieved are never more than half of normal ones. It is necessary to wait an average of 4 or 5 years before replanting asparagus in these lands. This phenomenon is known as the replanting problem, and causes the need to continually search for new land for growing asparagus. Another added problem for farmers is that the elimination of those plant residues from asparagus cultivation entails significant economic costs. For all these reasons, it is essential to seek alternatives for the management of that waste that improve the sustainability of the crop within the scope of the circular economy. In this context, this work proposes the valorization of asparagus by-products by obtaining bioactive compounds. Main objectives of the present work include: i) phytochemical analyses of asparagus fronds and roots; ii) obtaining bioactive extracts, with distinct technological and nutritional functionalities, by using an environmentally sustainable extraction process, easy to implement in the practice of a food industry and with methods compatible with food use. Characterization of asparagus by-products shown that fronds had an average flavonoid content of 2.637 ± 0.014 g/Kg fresh weight, which is up to 5-6 times higher than that of the spears; and roots contained up to 10 times more saponins (2.25 g/Kg fresh weight), which were accompanied by lower quantities of phenolic acids (368 mg/Kg fresh weight). Statistical analysis revealed that those phytochemical contents were mainly determined by location and phase of the vegetative cycle, whereas genetic factors did not significantly influence them. Based on the results of the present work, the proposal for the recovery and valorization of asparagus by-products is based on obtaining two bioactive extracts, the first being an antioxidant extract enriched in flavonoids, with an average yield of 10.7 g/Kg fresh frond and a flavonoid richness of 17%; and the second, a saponins extract with an average yield of 10.3 g/Kg fresh root and a richness of 51%. These natural extracts have great techno-functional potential in the agri-food industry and some of them are already being tested as additives in the preparation of soups, breads and meat products.

Nutritional composition of green asparagus (Asparagus officinalis L.), edible part and by-products, and assessment of their effect on the growth of human gut-associated bacteria.

Asparagus is considered a healthy food with a high content of bioactive compounds. In this study, the proximate and mineral composition, non-digestible carbohydrates and bioactive compounds of edible spear, spear by-product and root have been evaluated. Their activity on the growth of human gut-associated bacteria has been studied. The results support the high nutritional and functional value of the asparagus, including its by-products, highlighting the potential of the non-edible parts to be used as prebiotics. A remarkable content in xylose, inulin, flavonoids and saponins has been found. It has been shown that the spear by-product can be selectively used to promote the growth of commensal or probiotic lactobacilli and bifidobacteria strains. It has been confirmed that any part of the asparagus has a potential future as a healthy food or as health-promoting ingredients, however more work is required to identify the compounds able to modulate the human gut microbiota.

Asparagus Roots: From an Agricultural By-Product to a Valuable Source of Fructans.

Asparagus roots are by-products from asparagus cultivation and they could be considered one of the best sources of fructans. These polymers are interesting food ingredients for their prebiotic and immuno-stimulating characteristics. The aim of this work is to characterize the fructan profile from the roots of several asparagus varieties grown at different locations and pickled at three vegetative statuses in order to valorize these by-products as fructan source. Fructans were extracted with hot water and fractionated into three pools according to their molecular weight (MW). Their average MW was studied by HPSEC and their degree of polymerization by HPAEC. The fructan content was up to 12.5% on fresh weight basis, depending on variety and sampling date. The relative abundance of the three pools also depended on the picking moment as after the spear harvest period their total content and MW increased. The average MW of the three fractions was similar among varieties with 4.8, 8.4 and 9 sugar units, although fructans up to 30 units were identified by HPAEC. These characteristics make them similar to the commercialized Orafti®-GR inulin, a common additive to food products. Therefore, the concept of asparagus roots as cultivation waste must be changed to a new feedstock for sustainable agriculture and industry.

Asparagus Fructans as Emerging Prebiotics.

Commercial fructans (inulin and oligofructose) are generally obtained from crops such as chicory, Jerusalem artichoke or agave. However, there are agricultural by-products, namely asparagus roots, which could be considered potential sources of fructans. In this work, the fructans extracted from asparagus roots and three commercial ones from chicory and agave were studied in order to compare their composition, physicochemical characteristics, and potential health effects. Asparagus fructans had similar chemical composition to the others, especially in moisture, simple sugars and total fructan contents. However, its contents of ash, protein and phenolic compounds were higher. FTIR analysis confirmed these differences in composition. Orafti®GR showed the highest degree of polymerization (DP) of up to 40, with asparagus fructans (up to 25) falling between Orafti®GR and the others (DP 10-11). Although asparagus fructan powder had a lower fructan content and lower DP than Orafti®GR, its viscosity was higher, probably due to the presence of proteins. The existence of phenolic compounds lent antioxidant activity to asparagus fructans. The prebiotic activity in vitro of the four samples was similar and, in preliminary assays, asparagus fructan extract presented health effects related to infertility and diabetes diseases. All these characteristics confer a great potential for asparagus fructans to be included in the prebiotics market.

Date Seeds: A Promising Source of Oil with Functional Properties.

The cultivation of the date palm (Phoenix dactylifera L.) is the main activity and source of livelihood for people from arid and semiarid regions of the world. Date production is increasing every year. In addition, pitted date exportation is rising and great amounts of date seeds are produced. This biomass represents a problem for manufacturing companies. At the moment, date seeds are normally discarded or used as animal feed ingredients. However, this co-product can be used for many other applications due to its valuable chemical composition. Oil is one of the most interesting components of the date seed. In fact, date seeds contain 5-13% oil. Date seed oil contains saturated and unsaturated fatty acids with lauric and oleic as the main ones, respectively. Tocopherols, tocotrienols, phytosterols, and phenolic compounds are also present in significant amounts. These phytochemicals confer added value to date seed oil, which could be used for many applications, such as food product formulations, cosmetics, and pharmaceuticals. This review provides up-to-date data on the different extraction techniques and the chemical composition of date seed oils. The applications of date seed oil have also been reviewed.

Identification of flavonoid diglycosides in several genotypes of asparagus from the Huétor-Tájar population variety.

The qualitative and quantitative composition of flavonoids from the Huétor-Tájar population variety of asparagus (commonly known as " triguero") was investigated. Flavonoids were analyzed by reversed-phase high-performance liquid chromatography-diode array detection (HPLC-DAD). Liquid chromatography-mass spectrometry (LC-MS) under identical HPLC conditions was used to verify the identities of the flavonoid glycosides from triguero asparagus. The quantities of asparagus flavonoids were calculated according to concentration curves constructed with authentic standards. Total flavonoid contents, calculated as the sum of individual compounds, were determined and ranged from 400 to 700 mg/kg fresh weight. The most abundant was rutin, which represented 55-98% of the total flavonoid complement. Triguero asparagus were revealed to be an important source of not only quercetin derivatives but also kaempferol and isorhamnetin glycosides. Significant differences (p < 0.05) in the content and relative composition of flavonoids were found among the spears of the distinct asparagus genotypes from the Huétor-Tájar population variety.

Saponin Profile of Wild Asparagus Species.

The aim of this work was to study the saponin profiles from spears of different wild asparagus species in the context of its genetic diversity aside from geographical seed origin. They included Asparagus pseudoscaber Grecescu, Asparagus maritimus (L.) Mill., Asparagus brachiphyllus Turcz., Asparagus prostrates Dumort., and Asparagus officinalis L. The saponin analysis by LC-MS has shown that saponin profile from wild asparagus is similar to that previously described for triguero asparagus from Huétor-Tájar landrace (triguero HT), which had not ever been reported in the edible part of asparagus. All the samples, except A. officinalis, were characterized for having saponins distinct to protodioscin and the total saponin contents were 10-fold higher than those described for commercial hybrids of green asparagus. In particular, A. maritimus from different origins were rich in saponins previously found in triguero HT. These findings supported previous suggestion, based on genetic analysis, about A. maritimus being the origin of triguero HT. Multivariate statistics including principal component analysis and hierarchical clustering analysis were used to define both similarities and differences among samples. The results showed that the greatest variance of the tested wild asparagus could be attributed to differences in the concentration of particular saponins and this knowledge could be a tool for identifying similar species.

The phytochemical and bioactivity profiles of wild Asparagus albus L. plant.

The ethanolic extracts from the leaves, pericarps and rhizomes of Asparagus albus L. were investigated for their phytochemical composition, antioxidant (DPPH and FRAP assays), anti-microbial against human pathogenic isolates and cytotoxic (human colon carcinoma HCT-116 cells) activities. The highest flavonoid content was obtained in the leaf extract followed by the pericarp but there were no flavonoids detected in the rhizome. However, the rhizome had a high concentration of saponins. Flavonoid and saponin profiles were similar to those previously described for the triguero Huetor Tajar asparagus landrace. It was found that the pericarp ethanolic extract exhibited higher antioxidant activity than rhizome and leaf extracts. Moreover, the rhizome possessed more evident cytotoxic activity against HCT-116 cells in comparison to leaf and pericarp. All extracts showed varying degrees of antimicrobial activity against most of the human pathogenic isolates. In addition, the leaves showed more powerful inhibitory activities against the maximum number of bacteria and all the fungai isolated and the highest activity was in the pericarp extract against multidrug resistant Pseudomonas aeruginosa (MDR) and Erythromycin resistant Streptococcus agalactiae (ER) with an inhibition zone of 21mm and 19mm, respectively. The results show that A. albus could be a new crop with pharmaceutical interest because its richness in bioactive compounds provides considerable benefits for human health.

Effect of storage on wall-bound phenolics in green asparagus.

The cell walls of green asparagus spears have been analyzed for their phenolic and carbohydrate composition as modified by postharvest storage. Esterified phenolic components were released by sequential alkaline hydrolysis and identified and quantified by diode array HPLC. Significant quantities of ferulic acid (FA) and its derivatives were found to increase at least 3-fold during storage, particularly in walls from the lower parts of the stem, where accompanying changes in sugar composition were also observed. In fresh asparagus, >60% of the total FA was in the form of diferulic acid, and this increased to approximately 70% after 3 days of storage. The main FA dehydrodimers were 8-8-, 8-O-4-, and 8-5-diferulates. These have been detected in other monocotyledonous and dicotyledonous plants, but as a smaller proportion of the total FA. The possible roles phenolic esters might have in relation to the mechanical, textural, and wound-response properties of asparagus spears are discussed.

Ferulic acid crosslinks in asparagus cell walls in relation to texture.

Post-harvest toughening of asparagus spears is associated with a large increase in monomeric and diferulic acids in the cell walls of stem tissues. The purpose of this study has been to investigate the distribution of these phenolic components among cell wall polymers and the role they play in the formation of associated pectic-xylan-phenolic complexes in relation to post-harvest toughening. The phenolic esters are found in all the extractable polysaccharide fractions, particularly the 0.5 M KOH fraction, as well as the insoluble cellulose-rich residue. The storage-related increase occurs in all fractions but is most prominent in the 0.5 M KOH-soluble components. Degradation of 0.5 M KOH subfractions with pure polysaccharide degrading enzymes has confirmed the occurrence of pectic-xylan-phenolic complexes in which ferulic acid and its dehydrodimers are attached to the xylan component but not to the pectic component. Studies on cell separation show that the maturation- and storage-related increase in thermal stability of cell adhesion (and therefore texture) is probably due to an increase in phenolic cross linking of xylans mainly in the parenchyma tissues. This overcomes the thermal lability of the pectic polysaccharides that are responsible for cell adhesion in immature tissues. The storage-induced appearance of some of the diferulic acid moieties in a number of wall polymer fractions supports the hypothesis that the storage affect is a wound-induced response rather than a continuation of maturation-related activity.

Cell wall bound anionic peroxidases from asparagus byproducts.

Asparagus byproducts are a good source of cationic soluble peroxidases (CAP) useful for the bioremediation of phenol-contaminated wastewaters. In this study, cell wall bound peroxidases (POD) from the same byproducts have been purified and characterized. The covalent forms of POD represent >90% of the total cell wall bound POD. Isoelectric focusing showed that whereas the covalent fraction is constituted primarily by anionic isoenzymes, the ionic fraction is a mixture of anionic, neutral, and cationic isoenzymes. Covalently bound peroxidases were purified by means of ion exchange chromatography and affinity chromatography. In vitro detoxification studies showed that although CAP are more effective for the removal of 4-CP and 2,4-DCP, anionic asparagus peroxidase (AAP) is a better option for the removal of hydroxytyrosol (HT), the main phenol present in olive mill wastewaters.

Asparagus byproducts as a new source of peroxidases.

Soluble peroxidase (POD) from asparagus byproducts was purified by ion exchange chromatographies, and its kinetic and catalytic properties were studied. The isoelectric point of the purified isoperoxidases was 9.1, and the optimum pH and temperature values were 4.0 and 25 °C, respectively. The cationic asparagus POD (CAP) midpoint inactivation temperature was 57 °C, which favors its use in industrial processes. The Km values of cationic asparagus POD for H2O2 and ABTS were 0.318 and 0.634 mM, respectively. The purified CAP is economically obtained from raw materials using a simple protocol and possesses features that make it advantageous for the potential use of this enzyme in a large number of processes with demonstrated requirements of thermostable POD. The results indicate that CAP can be used as a potential candidate for removing phenolic contaminants.

Saponin profile of green asparagus genotypes.

The main goal of this study was to determine the saponin profiles of different "triguero" asparagus genotypes and to compare them to green asparagus commercial hybrids. The samples consisted of 31 commercial hybrids and 58 genotypes from the Huétor-Tájar (HT) population variety ("triguero"). The saponin analysis by high-performance liquid chromatography-mass spectrometry allowed for the determination of 12 saponins derived from a furostan-type steroidal genin, 4 of which had never been described in the edible part of asparagus. The saponin profile of "triguero" asparagus was a combination of these new saponins and protodioscin. Although protodioscin was the major saponin found in commercial hybrids, some of these 12 saponins were detected as major components in some of the commercial hybrids. The total contents of saponins described in some of these HT genotypes reach values as high as 10-100 times higher than those found in commercial hybrids.

Optimization of a method for the profiling and quantification of saponins in different green asparagus genotypes.

The main goal of this study was the optimization of a HPLC-MS method for the qualitative and quantitative analysis of asparagus saponins. The method includes extraction with aqueous ethanol, cleanup by solid phase extraction, separation by reverse phase chromatography, electrospray ionization, and detection in a single quadrupole mass analyzer. The method was used for the comparison of selected genotypes of Huétor-Tájar asparagus landrace and selected varieties of commercial diploid hybrids of green asparagus. The results showed that while protodioscin was almost the only saponin detected in the commercial hybrids, eight different saponins were detected in the Huétor-Tájar asparagus genotypes. The mass spectra indicated that HT saponins are derived from a furostan type steroidal genin having a single bond between carbons 5 and 6 of the B ring. The total concentration of saponins was found to be higher in triguero asparagus than in commercial hybrids.