Simultaneous Determination of Two Isomers of Asarone in Piper sarmentosum Roxburgh (Piperaceae) Extracts using Different Chromatographic Columns

Purpose: To develop a rapid and reliable reverse phase high performance liquid chromatography (RPHPLC) method to quantify the two isomers of asarones in P. sarmentosum extracts using two different columns under similar analytical conditions.Methods: Two isomers, α- and β-asarone, were analyzed using two types of C 18 columns with 0.1 % orthophosphoric acid: acetonitrile: methanol (50: 40: 10) as mobile phase. The developed method was applied to determine the contents of α- and β-asarone in extracts of different parts of P. sarmentosum.Results: Column A retention times for the elution of α- and β-asarone were 11.890 ± 0.008 and 10.80 ± 0.004 min, respectively, and were significantly shorter than those of column B (15.110 ± 0.024 and 13.290 ± 0.018, respectively, p < 0.001). Column B showed better resolution (1.82 ± 0.025 of the isomers than column A (1.10 ± 0.01, p < 0.001). Both columns showed comparable sensitivity, precision and selectivity of the compounds investigated. α-Asarone level was in the range 0.36 - 5.14 % in ethanol and 50 % ethanol extracts, but absent in all water extracts. β-Asarone occurred in the range of 0.01 - 0.15 % in ethanol and 50 % ethanol extracts but was absent in all water extracts of P. sarmentosum.Conclusion: The results indicate that the developed method is a suitable quality assurance method for determining α- and β-asarone isomers in herbal extracts and food preparations.Keywords: α- Asarone, Isomers, Piper sarmentosum, Herbal extracts, Retention tim

Neuroprotective Effects and Mechanism of β

Emerging evidence suggests that activated astrocytes play important roles in AD, and β-asarone, a major component of Acorus tatarinowii Schott, was shown to be a potential therapeutic candidate for AD. While our previous study found that β-asarone could improve the cognitive function of rats hippocampally injected with Aβ, the effects of β-asarone on astrocytes remain unclear, and this study aimed to investigate these effects. A rat model of Aβ1–42 (10 μg) was established, and the rats were intragastrically treated with β-asarone at doses of 10, 20, and 30 mg/kg or donepezil at a dose of 0.75 mg/kg. The sham and model groups were intragastrically injected with an equal volume of saline. Animals were sacrificed on the 28th day after administration of the drugs. In addition, a cellular model of Aβ1–42 (1.1 μM, 6 h) was established, and cells were treated with β-asarone at doses of 0, 2.06, 6.17, 18.5, 55.6, and 166.7 μg/mL. β-Asarone improved cognitive impairment, alleviated Aβ deposition and hippocampal damage, and inhibited GFAP, AQP4, IL-1β, and TNF-α expression. These results suggested that β-asarone could alleviate the symptoms of AD by protecting astrocytes, possibly by inhibiting TNF-α and IL-1β secretion and then downregulating AQP4 expression

Insecticidal Activities of Compounds from Sweet Flag (Acorus Calamus) against Red Imported Fire Ants Solenopsis invicta (Hymenoptera: Formicidae)

Due to public health and environmental issues, alternatives of synthetic pesticides have been researched for a long time. We also evaluated the toxicity and repellency of the sweet flag (Acorus calamus) powder and two bioactive compounds (α-asarone and β-asarone) against workers of the red imported fire ant (RIFA), Solenopsis invicta under laboratory conditions. Sweet flag powder applied at 1 mg/cm2 or more provided 100% ant mortality within 18 hours, and repelled almost 97% of ants within one hour. Β-asarone was the faster acting compound against RIFA compared to α-asarone and sweet flag powders. The LT50 values inclined exponentially with the increase in the application rate of the test items. On the other hand, repellency did not increase with the increase in the application rate of the test items, but did with the increase in exposure time. Based upon the results of this study, α-asarone and β-asarone, as well as sweet flag powders could be another alternative tool to control the RIFA

HPLC Profiling of B Asarone Content and Cytogenetic Studies of Medicinally Important Indian Acorus Calamus l., Accessions

Acorus calamus L. is a well- known herb for its traditional medicinal and pharmaceutical potentials. The rhizome of the plant is mainly used for medicinal purposes because it yields an essential oil known as “Calamus Oil”. The main component of the essential oil is â-asarone. In our present study, 20 different species of Acorus calamus were collected from different parts of South India and North East India. An attempt has been made to characterize all the accessions on the basis of their ploidy level by cytogenetic studies, and determining â- asarone content by HPLC analysis. All the accessions were screened for their ploidy level by staining the root chromosomes at metaphase stage. The plants observed were either diploid or triploid ruling out any tetraploids. HPLC analysis of powdered rhizome extracts for the â- asarone, indicated that it ranged from 2.2% to 7.2%. Our results also have revealed that there is no correlation between the ploidy status and the content of â-asarone. Indian accessions were found to have low to moderate levels of â- asarone content

CURRENT STATUS ON BIOLOGICAL ACTIVITIES OF ACORUS CALAMUS - A REVIEW

The review emphasis on the current status of the researches in Acorus calamus highlighting their number of useful biological activities. The Acorus calamus most extensively investigated phytochemically and pharmacologically. Number of bioactive constituents was identified and characterized from the leaves and rhizomes and their essential oils. Major chemical constituents identified are alpha and beta asarones which is responsible for therapeutic and medicinal properties of Acorus species. Several, recently published reports have revealed many newer useful bioactivities of leaves and rhizome extracts, essential oils and isolated chemical constituents such as anti-inflammatory, immunosuppressive, anti-adipogenic, antimicrobial, fungicidal, insulin sensitizing/antidiabetic, neuroprotective, wound healing, mitogenic, insecticidal, anthelmintic, allelopathic, antiepileptic, antispasmodic activities and inhibitor of acetylcholinesterase. This article highlights the various biological activities studied in A. calamus

Investigation of the Anti-Cancer Effects of B-asaron and Etoposide in MCF-7 Breast Cancer Cells

Currently, the options available for the treatment of various cancers including breast cancer, are associated with several limitations such as severe toxicity, drug resistance, poor prognosis, and high risk of recurrence. Therefore, there appears to be an increasing interest and necessity in investigating various phytochemicals from natural sources for a superior and safer alternative treatment strategy. The bioactive phytochemical alpha (alpha%253B) and beta (beta%253B)-asarone from Acorus calamus is a traditional medicine system that has been shown to have anti-tumor and chemo-inhibitory activities in numerous preclinical studies both in vitro and in vivo. Various experimental studies with human malignant cell lines and animal models have also confirmed the anti-tumor and anti-cancer activities of beta%253B-asarone. In this study, we aimed to investigate the anti-cancer effects of beta%253B-asarone alone or together with etoposide buy measuring cellular responses such as cell viablity, cell cycle arrest and apoptosis using breast cancer cell line MCF-7 cells. In order to get insight in to the mechanism, we also tested the expression of of NF-kappa%253BB %252F p65 activity and the expression of Bcl-2 family member pro-apoptotic Bax protein together with p53 and p21 activities in response to beta%253B-asarone alone or together with etoposide treatment. As a result, it was concluded that the use of beta%253B-asarone alone in breast cancer cells is effective in reducing cell viability, but when used together with Etoposide, it does not cause a synergistic effect. Here we suggest that that in particular activation of NF-kB%252Fp65 may be lead resistance to etoposide treatment

Metformin and asarone inhibit HepG2 cell proliferation in a high glucose environment by regulating AMPK and Akt signaling pathway.

Background: Metabolic dysregulation is one of the hallmarks of tumor cell proliferation. Evidence indicates the potential role of the 5′adenosine monophosphate-activated protein kinase (AMPK) and protein kinase B/Akt signaling pathway in regulating cell proliferation, survival, and apoptosis. The present study explores the effect of metformin HCl and the combination of α- and β-asarone on the proliferation of HepG2 cells in the presence of high glucose levels simulating the diabetic-hepatocellular carcinoma (HCC) condition. Results: The metformin and asarone reduced HepG2 cell viability in a dose-dependent manner and induced morphological changes as indicated by methyl thiazolyl tetrazolium (MTT) assay. The metformin and asarone arrested the cells at the G0/G1 phase, upregulated the expression of AMPK, and downregulated Akt expression in high glucose conditions as identified by the flow cytometry technique. Further, the upregulated AMPK led to a decrease in the expression of phosphoenolpyruvate carboxykinase-2 (PCK-2) and sterol regulatory element-binding protein-1 (SREBP-1). Conclusion: The anti-proliferative effect of metformin and asarone in the diabetic-HCC condition is mediated via AMPK and Akt pathway

Bioactivity of Sweet Flag (Acorus Calamus Linnaeus) Essential Oils Against Spodoptera Litura Fabricius (Lepidoptera: Noctuidae)

The study aims to determine the chemical compounds, toxicity, and antifeedant activity of sweet flag (Acorus calamus) essential oils against third instar larvae of Spodoptera litura. The study was conducted using a com-pletely randomized design (CRD) using various concentration of the essential oils (103, 2 × 103, 3 × 103, 4 × 103, 5 × 103 ppm). Mortality and antifeedant activity was observed 24 hours after treatment. Toxicity and anti-feedant activity values were 92.5% and 79.3%, respectively, with an LC50 value 586.96 ppm. Gas chromatog-raphy-mass spectrometry analysis showed that essential oil of A. calamus consists of five chemical compounds: methyl isoeugenol, 3.9-decadien-ol-1,3-methyl-6-(1-methylethenyl), 4-pentyl-1-(4propylcyclohexyl)1cyclohexene, γ-asarone and β asarone. Keywords: Acorus calamus, essential oils, mortality, antifeedant, Spodoptera lituraThe study aims to determine the chemical compounds, toxicity, and antifeedant activity of sweet flag (Acorus calamus) essential oils against third instar larvae of Spodoptera litura. The study was conducted using a completely randomized design (CRD) using various concentration of the essential oils (103, 2 × 103, 3 × 103, 4 × 103, 5 × 103 ppm). Mortality and antifeedant activity was observed 24 hours after treatment. Toxicity and antifeedant activity values were 92.5% and 79.3%, respectively, with an LC50 value 586.96 ppm. Gas chromatography-mass spectrometry analysis showed that essential oil of A. calamus consists of five chemical compounds: methyl isoeugenol, 3.9-decadien-ol-1,3-methyl-6-(1-methylethenyl), 4-pentyl-1-(4propylcyclohexyl)1cyclohexene, γ-asarone and β asarone

STANDARDIZATION AND COMPARATIVE EVALUATION OF AYURVEDIC POLYHERBAL GHRITA FORMULATION WITH MODERN EXTRACTION TECHNIQUE FOR EXTRACTION EFFICIENCY USING REVERSED PHASE-HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

Objective: Sarasvata ghrita (SG) is a polyherbal formulation in Ayurvedic Indian medicinal system, in which ghee is the main ingredient used for extraction. Ghee is 100% lipid, thus its regular use is limited, and there is a lack of quality control profile of SG. Thus, the objective of the study is to develop quality control method for standardization of SG and to analyze manufacturing process of SG and an effective method of extraction to extract phytoconstituents from herbs used in SG to overcome the limitation of SG.Methods: SG was processed as per the traditional method, whereas ethanolic extract (EE) and hydroalcoholic extract (HAE) were obtained by the conventional method and lipid-based extract (LE) was prepared by modern extraction method. SG and all extracts were standardized using newly developed high-performance liquid chromatography (LC) with respect to bebeerine, piperine, 6-shogaol, β-asarone, and chebulinic acid. All extracts were analyzed for pesticides, and heavy metal content by LC/mass spectrometry (MS/MS) and inductively coupled plasma/MS, respectively, screened for total polyphenols and flavonoids content, in vitro antioxidant potential, and for assessing its stability over time.Results: The better extraction was observed with maceration extraction using ethanol compared to ayurvedic method and LE method. All extracts were found to have a negligible amount of pesticide and heavy metals and found to be stable for 6 months under accelerated storage condition. Better polyphenols and flavonoid content and in vitro antioxidant potential were resulted in EE.Conclusion: EE showed a better potential in comparison with SG and LE