Phytochemical Diversity Research Articles

Phytochemical variability during vegetation of Chamerion angustifolium (L.) Holub genotypes derived from in vitro cultures

The purpose of the study was to evaluate Chamerion angustifolium (L.) Holub genotypes for preliminary selection and further breeding programs aimed at obtaining a suitable industrial form for the pharmaceutical applications. Clonally propagated plants representing 10 genotypes of Ch. angustifolium were regenerated under in vitro conditions, hardened and planted in the field. Studies included an evaluation of shoot proliferation, phytochemical assessment of in vitro and ex vitro plants as well as investigations of intraspecies variability regarding four phenological stages: vegetative, beginning of blooming, full blooming, and green fruit phases. Quantitative and qualitative analyses of bioactive compounds were performed using high-performance liquid chromatography coupled with diode array detector and tandem mass spectrometer (HPLC–DAD–MS/MS) and high-performance liquid chromatography (HPLC) methods. The efficiency of shoot multiplication varied between genotypes from 8.12 to 21.48 shoots per explant. A high reproduction rate (> 20 shoots per explant) was recorded for four lines (PL_45, PL_44, PL_58, DE_2). Plants grown in vitro synthesized oenothein B (11.2–22.3 mg g−1 DW) and caffeic acid derivatives. Plants harvested from field contained the full spectrum of polyphenols characteristic for this species, and oenothein B and quercetin 3-O-glucuronide were the most abundant. The maximal content of oenothein B was determined in the vegetative phase of fireweed, while some flavonoids were found in the highest amount in full blooming phase. The results of analysis of variance indicated significant differences among genotypes in oenothein B, 3-O-caffeoylquinic acid and flavonoids accumulation in four phenological phases. PL_44 plants were characterized by high content of oenothein B and quercetin 3-O-glucuronide as well as a relatively high level of other flavonoids. Based on our phytochemical and micropropagation studies, PL_44 genotype was the best candidate for early selection and further breeding programs.

Metabolomics by UHPLC-Q-TOF Reveals Host Tree-Dependent Phytochemical Variation in Viscum album L.

Viscum album L., commonly known as European mistletoe, is a hemi-parasitic plant of the Santalaceae family. The in vitro and in vivo effects of V. album differ, according to its host tree. However, little is known about the host-dependent phytochemical diversity in V. album. In this study, the metabolic profiles of V. album ssp. album from Malus domestica Bork., Quercus robur L., and Ulmus carpinifolia Gled were compared. Leaves, stems, and berries were collected in Switzerland, by the same procedure, in September 2016 and 2017. The methanolic extracts were analyzed by ultra-performance liquid chromatography, coupled to electrospray quadrupole time-of-flight mass spectrometry in positive ionization mode. The data were submitted to partial-least square discriminant analysis (PLS-DA) and the results showed that the V. album ssp. album samples were clustered into three groups, according to the three distinct host trees. Seven compounds, with high VIP scores (variable importance in projection), were responsible for this differentiation. The following four compounds were detected in both the harvest years: arginine, pipecolic acid or lysine, dimethoxycoumarin, and sinapyl alcohol, suggesting their use as host specific V. album biomarkers. The present work highlights the importance of standardized harvest and analytical procedures for the reproducibility of the chemical results of herbal materials.

Morphological Diversity, Genetic Characterization, and Phytochemical Assessment of the Cypriot Tomato Germplasm.

Tomato (Solanum lycopersicum L.) is considered one of the most valuable and versatile vegetable crops globally and also serves as a significant model species for fruit developmental biology. Despite its significance, a severe genetic bottleneck and intense selection of genotypes with specific qualitative traits have resulted in the prevalence of a restricted number of (geno)types, also causing a lack of diversity across widespread cultivated types. As a result, the re-emergence of landraces as well as traditional and heirloom varieties is largely acknowledged as a countermeasure to restore phenotypic, phytochemical and genetic diversity while enriching the aroma/taste tomato palette. On those grounds, the Cypriot tomato germplasm was assessed and characterized. Ten landrace accessions were evaluated under greenhouse conditions and data were collected for 24 IPGRI discrete phenotypic traits. Grouping of accessions largely reflected the fruit shape and size; four different fruit types were recorded across accessions (flattened, heart-shaped, rounded and highly rounded). Moreover, a single run panel consisting of ten SSRs was developed and applied in order to genetically characterize 190 Cypriot genotypes and foreign heirloom varieties. Based on genetic indexes it was established that tomato landraces have a rather low level of heterogeneity and genetic variation. Finally, mineral and phytochemical analyses were conducted in order to estimate biochemical attributes (total phenolics, ascorbic acid, lycopene, β-carotene, total soluble content, titratable acidity) across genotypes; thus, ascertaining that the Cypriot panel has a high nutritional value. Due to the thermo-drought adaptation and tolerance of these genotypes, the current study serves as a roadmap for future breeding efforts in order to incorporate desirable traits or develop novel tomato lines combining resilience and alimentary value.

Intraspecific Genetic Diversity of Cistus creticus L. and Evolutionary Relationships to Cistus albidus L. (Cistaceae): Meeting of the Generations?

Cistus (Cistaceae) comprises a number of white- and purple-flowering shrub species widely distributed in the Mediterranean basin. Within genus Cistus, many taxa are subject to various taxonomic uncertainties. Cistus creticus, a prominent member of the purple-flowered clade, is a prime case of the current taxonomic troubles. Floras and databases approve different species names and utilise different or additional/fewer synonyms. Various intraspecific classification systems based on subspecies or varieties are in use. The inconsistent determination of plant material makes it difficult to compare literature regarding the phytochemical diversity and biological activities of plant material and impedes a systematic utilization of the manifold medicinal properties of C. creticus. In the present investigation, we used DNA sequence data from one nuclear region (ITS) and two chloroplast regions (trnL-trnF, rpl32-trnL) to test the intraspecific genetic diversity of C. creticus and its evolutionary relationships to the closely related C. albidus. The combined DNA data confirmed C. creticus as a rather heterogeneous species that integrates two major evolutionary lineages with clearly different genetic characteristics. The ‘Eastern Mediterranean clade’ seems to represent old and ancestral characteristics. This lineage exhibits a close relationship to the geographically distant C. albidus, expressed by very closely related ribotypes and an interspecifically shared chlorotype. The ‘Western Mediterranean clade’ is characterized by a distinctive ITS polymorphism (co-occurring paralogous ribotypes) and more distantly related chlorotypes. The formation of the genetically complex ‘Western Mediterranean clade’ seems to have involved hybridization and recurrent formation or migration movements.

Standardization, Quality Control, and Bio- Enhancement of Botanical Insecticides: a Review

Botanicals are substances extracted from plants for use in various applications, such as the production of insecticides. Botanical insecticides (BIs) have recently attracted awareness in pest management owing to their potential to substitute synthetic pesticides. BIs are eco-friendly and more sustainable due to their ability to breakdown after use without generating toxic residues and diverse approach actions on targeted pests. Nevertheless, BIs are still not readily accepted, because the supporting proofs are very traditional, raising doubts about their quality. Additionally, the phytochemical variations of plants yield uneven and sometimes unfamiliar pesticide activity. This paper discusses challenges to overcome and presents the most noteworthy knowledge on BIs, their standardization, quality control, and bio- enhancement to be useful in agriculture and to improve human health.

Morphological, phytochemical and genetic diversity of threatened Polygonatum verticillatum (L.) All. populations of different altitudes and habitat types in Himalayan region.

The online version contains supplementary material available at 10.1007/s12298-021-01044-9.

Growth response of maize seed-borne fungi to cereal phenolic acid mixtures

Phenolic acids (PAs) are potential antifungal agents of stored grain cereals. Previous studies have mostly focused on the action of single compounds, consequently providing limited information on their effect as mixtures. We tested the hypothesis that higher phytochemical diversity and concentration of phenolic acids provide better defense against pathogenic fungi of cereals. Five maize seed-borne fungi with different degrees of specialization as cereal pathogens were exposed in vitro to five phenolic acids found in cereal grains either alone or in combinations. We detected that the inhibitory effect of PAs depended on their concentration and the fungus specialization, but the tested hypothesis was not functional for any plant-fungal species relationship. Moreover, various PAs or PAs-mixtures were almost equally inhibitory against the fungal species. We also found that phytochemical diversity had a differential effect on the phytopathogens according to their degree of specialization. Thus, a high phytochemical diversity in high concentration would protect a plant against a fungal community, but not necessarily against a single-fungus species.

Implications of the foliar phytochemical diversity of the avocado crop Persea americana cv. Hass in its susceptibility to pests and pathogens.

Phytochemical diversity (PD) can be considered as a defensive trait; it can operate through single plant secondary metabolites or usually as complex mixtures of them. We tested the more diversity-better defense hypothesis correlating the leaf plant secondary metabolites (PSMs) with the incidence of plant enemies on Hass avocado trees. We expected a negative correlation between the occurrence of plant enemies and PD metrics. Also, as intraspecific PSMs polymorphisms in plant populations are common, we studied the incidence of plant enemies on Hass avocado trees representing chemical variants (chemotypes). We expected a differential incidence of plant enemies among trees grouped by their mono and sesquiterpene + phenylpropanoid chemotypes. We analyzed foliar hexane extracts from 236 trees in 17 orchards by gas chromatography and for the incidence of red mite, thrips, whitefly, avocado branch borer, fruit rot, scab, and peduncle collar blight. The predicted negative correlation between the plant enemies’ incidence and the phytochemical metrics did not occur. To determine the relationship between enemy incidence and chemotypes we grouped the trees by cluster analysis using a matrix of PSMs in each tree. Most trees were grouped under four out of 23 chemotypes. Branch borers attacked trees of low-frequency chemotypes more frequently than trees with common chemotypes. The incidence of five plant enemies was different among the predominant chemotypes. The hypothesis of more diversity-better defense was not supported by the correlations between the phytochemical diversity and the incidence of pests and pathogens in Hass avocado orchards. Based on our results, we hypothesize that phytochemical diversity function as a defensive trait relies more on differentiation among individuals in a population than on the sole increase of chemical diversity. Also, the differential incidence of pests and pathogens on trees classified by their foliar chemotypes implies that these susceptibility or resistance markers represent potential useful tools for Hass avocado orchard pest management.

Chemical characterization and therapeutics of Dalbergia latifolia Roxb: A review

Dalbergia latifolia Roxb. is recognized as a medicinally important tree of Indian forest flora providing diverse source of phytochemicals having huge potential in Ayurveda and other materia medica. D. latifolia is rich in alkaloids, carbohydrates, glycosides, flavonoids, alkaloids, phenolic compounds and tannin. Biomolecules such as dalbergin, latifolin, (R)-dalbergione, dalbinol, dalbin, latinone and dalcriodain were reported profoundly from D. latifolia wood. It is a rich source of triterpenoids, benzofuran, neoflavonoids and rotenoids. Extracts from D. latifolia heartwood, leaves, seeds, bark and roots exhibited antimicrobial, antioxidant, anthelmintic, anticancer, antimutagenic, anti-termite, anti-obesity, cerebroprotective etc. activities. Current article provides an inclusive account on phytochemical diversity and the range of pharmacological activities of D. latifolia.

Morphological and phytochemical variations in Leonurus cardiaca L. under water deficit stress

Morphological and phytochemical variations in <i>Leonurus cardiaca</i> L. under water deficit stress

Fruits, frugivores, and the evolution of phytochemical diversity

Plants produce an enormous diversity of secondary metabolites, but the evolutionary mechanisms that maintain this diversity are still unclear. The interaction diversity hypothesis suggests that complex chemical phenotypes are maintained because different metabolites benefit plants in different pairwise interactions with a diversity of other organisms. In this synthesis, we extend the interaction diversity hypothesis to consider that fruits, as potential hotspots of interactions with both antagonists and mutualists, are likely important incubators of phytochemical diversity. We provide a case study focused on the Neotropical shrub Piper reticulatum that demonstrates: 1) secondary metabolites in fruits have complex and cascading effects for shaping the outcome of both mutualistic and antagonistic fruit–frugivore interactions, and; 2) fruits can harbor substantially higher levels of phytochemical diversity than leaves, even though leaves have been the primary focus of plant chemical ecology research for decades. We then suggest a number of research priorities for integrating chemical ecology with fruit–frugivore interaction research and make specific, testable predictions for patterns that should emerge if fruit interaction diversity has helped shape phytochemical diversity. Testing these predictions in a range of systems will provide new insight into the mechanisms driving frugivory and seed dispersal and shape an improved, whole‐plant perspective on plant chemical trait evolution.

Structural and compositional dimensions of phytochemical diversity in the genus Piper reflect distinct ecological modes of action

Abstract An increasing number of ecological studies have used chemical diversity as a functionally relevant, scalable measure of phytochemical mixtures, demanding more rigorous attention to how chemical diversity is estimated. Most studies have focused on the composition of phytochemical mixtures and have largely ignored structural concerns, which may have greater importance for ecological function. Here, we explore the development of structural complexity and compositional diversity resulting from different biotic and abiotic interactions in Piper kelleyi Tepe (Piperaceae). We also describe how variation in structural complexity and compositional diversity differs between two congeners, P. kelleyi and P. reticulatum. To better interpret these results, we have developed a framework for interpreting these dimensions of chemical diversity in phytochemical mixtures. We used the tropical shrub, P. kelleyi, as a model system to examine interactions between ecological factors and dimensions of phytochemical diversity. We also compared compositional diversity and metabolic complexity in P. kelleyi and P. reticulatum using liquid chromatography and 1H NMR, respectively, to examine trade‐offs between compositional diversity and structural complexity. A framework is provided to generate meaningful estimates of the structural complexity of phytochemical mixtures as measured by 1H NMR. Piper is an abundant plant genus that supports diverse insect communities throughout the tropics. Subtle changes in understorey forest light were associated with increases in herbivory that directly increased compositional diversity and indirectly decreased structural complexity in P. kelleyi. This was attributed to the production of oxidation products resulting from herbivory‐driven decomposition of structurally complex defence compounds. This type of complex result would remain undetected using standard chemical ecology approaches and accounts for the detailed molecular changes that are likely to affect species interactions. Synthesis. Our quantitative framework provides a method for considering trade‐offs between structural complexity and compositional diversity and the interpretation of analytical approaches for each. This methodology will provide new theoretical insights and a more sophisticated model for examining the ecology and evolution of chemically mediated interactions.

Veterinary Phytomedicine in India : A Review

Nature is a unique source of structures of high phytochemical diversity, many of them possessing interesting biological activities and medicinal properties. Phytochemicals ensure safe, eco-friendly and most effective drug for animal use. Veterinarians use these phytochemicals with the knowledge from traditional system and modern pharmacology. High cost of allopathic medicine is one of the major contributing factors in loss of productive livestock and animal productivity. Many plants have been identified which show promising abilities in disease prevention and cure. Phytochemicals can be used either by feeding whole plant parts or by the process of extraction of these chemicals and delivering in concentrated forms. The pharmacological activities of plants are attributed to certain active principles, namely alkaloids, glycosides, fats, oils, tannins, saponins etc. The use of cheap and easily available phytochemicals can thus be made by taking a leaf out of the ethno-veterinary practices. Although the effects of natural remedies may seem slower, the results are sometimes better on the long run especially with respect to chronic diseases. The horizon of phytochemicals has to be explored and exploited to create a “green treatment” protocol to develop an ecofriendly and toxin free production system that is beneficial for existence of both human and animal population in the near future.

Assessment of phytochemical and genetic diversity analysis of Plumbago zeylanica L. accessions

Assessment of phytochemical and genetic diversity analysis of Plumbago zeylanica L. accessions

Short Communication: Phytochemical diversity and bioactivity of Malaysian Premna cordifolia (Lamiaceae)

Abstract. Daud D, Dewa MSAM, Mahbob ENM, Razak WRWA. 2021. Short Communication: Phytochemical diversity and bioactivity of Malaysian Premna cordifolia (Lamiaceae). Biodiversitas 22: 3245-3248. The aims of the present study are to determine the chelation potential and protective effect of Premna cordifolia leaves ethanolic extract in lead intoxicated mice. Twenty-four male mice were divided into four groups with six animals each. Group 1 was healthy mice and received 2 ml/kg bwt distilled water. Group 2 was healthy mice and supplemented with 200 mg/kg bwt P. cordifolia leaves ethanolic extract. Group 3 was lead intoxicated mice and received 2 ml/kg bwt distilled water. Group 4 was lead intoxicated mice and supplemented with 200 mg/kg bwt P. cordifolia leaves ethanolic extract. All treatments were given for 14 days. Chelation activity was determined by measuring the concentrations of lead in the liver, kidney and testis by using atomic absorption spectroscopy. Sperm quality was tested according to World Health Organization manual and testosterone was estimated by using ELISA kit. Spectroscopic data showed that P. cordifolia leaves ethanolic extract was able to lower lead concentrations in liver, kidney and testis of lead intoxicated mice. Following a decrease in lead concentrations, sperm quality and testosterone levels were restored in lead intoxicated mice supplemented with P. cordifolia leaves ethanolic extract. In conclusion, P. cordifolia leaves ethanolic extract has the potential to be developed as an alternative remedy for treating lead poisoning effects.

Exploring the phytochemical variation of non-volatile metabolites within three South African Salvia species using UPLC-MS fingerprinting and chemometric analysis

The South African Salvia species, Salvia africana-lutea, S. lanceolata and S. chamelaeagnea, are widely used to treat fever and inflammation associated with skin and lung infections. The aim of this study was to explore the non-volatile secondary metabolites and the phytochemical variation within these lesser known species, to support product development and commercialisation. Chemical profiles of the methanol extracts of 81 wild-harvested samples were obtained using ultra performance-quadrupole-Time-of-Flight-mass spectrometry (UPLC-qToF-MS). Forty-one compounds, including caffeic acid, rosmarinic acid, carnosol, carnosic acid and ursolic acid, were detected and confirmed across the three species. Nineteen compounds were tentatively identified of which 14 have not been reported in these species. Principal component analysis revealed distinct clusters corresponding to the three species, confirming chemical differences. Marker compounds for each species were revealed using orthogonal projection to latent structures-discriminant analysis. Further chemometric analysis reflected a degree of intraspecies variation, although the chemistry within populations was mostly conserved. Potential chemotypes for each species were identified through unique compounds associated with each group. The concentrations of medicinally important metabolites, namely, rosmarinic acid, carnosol, carnosic acid and ursolic acid, were determined, using validated UPLC-PDA methods. Ursolic acid was present at levels up to 38.2 mg/g, confirming that these species are a rich source of this compound. No similar studies combining liquid chromatography with chemometric analysis, and utilising a large sample size from various habitats, have been reported for these three Salvia species. The results will guide selection of cultivars with the best attributes for the intended therapeutic application, thereby protecting wild populations from over-exploitation.

Variation in volatile organic compounds in fruits of Iranian Capparis spinosa L. accessions.

In the present study, essential oil (EO) content and phytochemical variation were evaluated in the fruits of 10 Iranian Capparis spinosa accessions. The accessions were collected from their natural habitats of Iran and grown together in field conditions. The EOs content ranged from 0.55 to 1.46%. The correlation analysis revealed a significant negative correlation between the EO content and altitude of the sampling locations (P < 0.01; r = −0.84). GC–MS analysis revealed 31 compounds in the EOs of the accessions, mainly being isopropyl isothiocyanate (5.5–13.7%), methyl sulfonyl heptyl isothiocyanate (4.6–15.6%), butyl isothiocyanate (3.6–10.6%), γ-terpinene (4.4–9.2%), and thymol (22.9–37.1%). According to the cluster analysis, the accessions were classified into three groups. Principal component analysis (PCA) demonstrated that the first and second PCs1 confirmed more than 76% of the total variation in the phytochemical components among the C. spinosa samples. Our results revealed that the sampling altitude was the most effective factor in explaining this variation.

Genetic diversity and population structure of Seriphidium Sub-genus of Artemisia from different terrains of Balochistan, Pakistan

Abstract. Rehman A, Saeed S, Ahmed A. 2021. Genetic diversity and population structure of Seriphidium Sub-genus of Artemisia from different terrains of Balochistan, Pakistan. Biodiversitas 22: 2658-2664. Seriphidium is one of the largest sub-genus of Artemisia growing in different geographical populations and terrains of Balochistan Pakistan. This genus is economically important and used as folk medicine. The genus is highly variable, besides morphological characterization, molecular authentication is also needed for novel drug discovery. Three significant species of the genus viz., S. quettense, S. oliverianum, and S. maritimum were selected for morphological and molecular characterization and phytochemical investigation. Genetic variability assessment was carried out by polymorphic bands analysis by using molecular markers in 18 accessions collected from two sites, namely, Ziarat and Hazarganji-Chiltan National Park, Balochistan, Pakistan. A combined molecular marker system by using 17 Randomly Amplified Polymorphic DNA (RAPD) and 9 Inter-Simple Sequence Repeats (ISSR) amplified 231 loci, of which 111 were polymorphic (48% polymorphism). Phenolic contents and Flavonoids were estimated for phytochemical assessment. Results revealed that the highest phenolic compounds were in S. quettense when compared with other investigated taxa. In conclusion, molecular marker profiling together with phytochemical variation of all three species exhibited genetic diversity and chemical variation. The findings of the current investigation can further be used as a baseline study for the implementation of conservation strategies. In addition, phytochemical assessment can be utilized for modern drug discovery

Analysis of Peganum harmala, Melia azedarach and Morus alba extracts against six lethal human cancer cells and oxidative stress along with chemical characterization through advance Fourier Transform and Nuclear Magnetic Resonance spectroscopic methods towards green chemotherapeutic agents

Traditional medicines implicate consumption of plant crude extracts, which may consist of extensive phytochemical diversity. Overall, the most biologically active extract of Peganum harmala (seeds) exhibited significant cytotoxic activity on Artemia salina with LC50 value of 61.547 µg/mL, while P. harmala (roots) [LC50 = 124.229 µg/mL] and M. azedarach (fruits) [LC50 = 147.813 µg/mL] showed moderate cytotoxic potential. P. harmala (seeds) extract also showed the maximum antitumor potential with 52.278 µg/mL LC50. Branches of P. harmala and Morus alba were not active in both bioassays. These outcomes were further reinforced by the levels of phenolics and flavonoids checked against gallic acid and quercetin equivalents, respectively, by standard curves. Current study aims to isolate, structurally characterize and analyze the bioactive compound from plant extracts by using chromatographic and spectrophotometric techniques. Bioactivity guided isolation of extracts led to the isolation of PH-HM-16 from ethyl acetate fraction P. harmala seeds. Chemical structure of PH-HM-16 was elucidated by ESI-MS, 1H NMR, 13C NMR, HSQC and IR spectrum. The results demonstrated significant positive anticancer activities against six human cancer cell lines assessed through MTT cancer cell growth inhibition assay. PH-HM-16 was most effective against prostate cancer cell lines [IC50 = 17.63 µg/mL] followed by breast cancer cell line MCF7 [IC50 value of 41.81 µg/mL]. IC50 value of PH-HM-16 against human myeloid leukemia cell line HL-60 and human colorectal tumor cells HCT-116 was observed as 68.77 µg/mL and 71.54 µg/mL respectively. The IC 50 value of PH-HM-16 compound was not significant against human gastric cancer SGC-7901 (111.89 µg/mL) and human lung adenocarcinoma epithelial cell line A549 (176.04 µg/mL). Isolated bioactive metabolite PH-HM-16 possesses significant antitumor potential so this could be the first step to develop an effective anticancer agent. Hence, this compound represents a promising potential to be chemically standardized or developed into pharmaceuticals for the chemoprevention and/or the treatment of certain types of cancer, especially as adjuvant phytotherapeutics in conventional chemotherapy.

Custard Apple (Annona squamosa L.) Leaves: Nutritional Composition, Phytochemical Profile, and Health-Promoting Biological Activities.

Annona squamosa L. (custard apple) belongs to the family Annonaceae and is an important tropical fruit cultivated in the West Indies, South and Central America, Ecuador, Peru, Brazil, India, Mexico, the Bahamas, Bermuda, and Egypt. Leaves of custard apple plants have been studied for their health benefits, which are attributed to a considerable diversity of phytochemicals. These compounds include phenol-based compounds, e.g., proanthocyanidins, comprising 18 different phenolic compounds, mainly alkaloids and flavonoids. Extracts from Annona squamosa leaves (ASLs) have been studied for their biological activities, including anticancer, antidiabetic, antioxidant, antimicrobial, antiobesity, lipid-lowering, and hepatoprotective functions. In the current article, we discussed the nutritional and phytochemical diversity of ASLs. Additionally, ASL extracts were discussed with respect to their biological activities, which were established by in vivo and in vitro experiments. A survey of the literature based on the phytochemical profile and health-promoting effects of ASLs showed that they can be used as potential ingredients for the development of pharmaceutical drugs and functional foods. Although there are sufficient findings available from in vitro and in vivo investigations, clinical trials are still needed to determine the exact effects of ASL extracts on human health.