Biochemical Characterization of γ-Glutamyl Transpeptidase from Bacillus altitudinis IHB B1644 and Its Application in the Synthesis of l-Theanine.

An extracellular γ-glutamyl transpeptidase (GGT) produced from Bacillus altitudinis IHB B1644 was purified to homogeneity employing ion-exchange chromatography. GGT comprised two subunits of 40 and 22 kDa determined by SDS-PAGE. The maximum enzyme activity was optimal at pH 9 and 37 °C. The purified enzyme was stable from pH 5-10 and <50 °C. Steady-state kinetic studies revealed a Km value of 0.538 mM against γ-GpNA. For substrate specificity, GGT showed highest affinity for l-methionine. The inhibitors' effect demonstrated that serine or threonine and tryptophan residues are essential for enzyme activity. l-Theanine production was optimized by employing a one-variable-at-a-time approach with 60-65% conversion rate. The final reaction consisted of 20 mM l-glutamine, 200 mM ethylamine hydrochloride, and 10 U mL-1 enzyme concentration at 37 °C in Tris-Cl (50 mM, pH 9) for 5 h. l-Theanine was purified using a Dowex 50W X 8 hydrogen form resin and confirmed by HPLC and 1H NMR spectroscopies.

Targeted UHPLC-QTOF-IMS based metabolite profiling for bioactive compounds in Rosa webbiana wallich ex royle: An unexploited native from western himalayas.

The current study focused on the tissues of wild Rosa webbiana from different altitudes of Indian Western Himalayas for vitamin C content, total phenolics, flavonoids, total sugars, and antioxidant potential. To date, there are very few studies on underutilized tissues viz. fruits, leaves and stem of Rosa webbiana growing in the higher altitudes. The targeted UHPLC-QTOF-IMS illustrated the phenolics fingerprinting of tissue extracts. Twelve bioactive compounds were detected with quercetin, kaempferol, and their derivatives dominantly in stem and leaves. The results have revealed that fruits possessed the highest vitamin C, and sugar contents (960, and 191.6 mg/100 g, respectively). Compared to other tissues, leaves showed the highest total phenolics as well as best results in vitro assays employed to assess antioxidant potential. The antioxidant activity showed a positive correlation with total phenolics. A significant variation in total phenolics, total flavonoids, and antioxidant potential was mainly attributed to oxidative stress on plants due to altitude difference, and secondary metabolite production. Taken together, the underutilized tissues of Rosa webbiana could be exploited as a promising, low-cost resource of phenolic compounds in food, cosmetic, and pharmaceutical industries. Our study will pave the way to developing food products from Rosa webbiana, a natural source for health-conscious people.

Mechanistic studies on polyphenol rich fractions of Kangra tea by HPTLC and NMR for their antioxidant activities.

Abstract: The antioxidant activity in tea is largely driven by its polyphenolic content, however, the antioxidant reaction mechanism and the compounds involved are not well characterized. Therefore, in this study, we performed in-depth profiling of the antioxidant reaction mechanism of Green Tea (GT), Black Tea (BT), and their polyphenolic fractions with free radical using state-of-the-art analytical techniques. The polyphenol enriched fractions from GT and BT were isolated using column chromatography. Catechins were isolated and characterized by diverse spectroscopic techniques. Samples were screened for their antioxidant activity by HPTLC and further evaluated using a spectrophotometer. The free radical reactions with GT, BT, enriched fractions viz, GT Polyphenols (GTP), BT Polyphenols (BTP), and isolated catechins were studied using the 13C NMR technique. The highest polyphenol content was found in GTP (795.4 ± 0.012 mg/g) whereas GT (321.0 ± 0.028 mg/g) showed maximum flavonoids content. Individual catechins isolated from GTP were EGCG, ECG, EGC, EC and C. Antioxidant activity followed the order EGCG > ECG > EGC > EC > GTP > C > BTP > GT > BT. In GT, the antioxidant reaction mechanism showed single electron and H-transfer in all catechins, which involved the transformation of the hydroxyl group to the carbonyl group. Whereas in BT theaflavins, conversion of the benzotropolone ring to the six-membered ring was observed. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05297-w.

Amino Appended Coumarin Composites for Pb2+ Capturing in Aqueous Medium.

The C1-C3 receptors were synthesized by using coumarin and amines viz., 1-butylpiperazine (1), cis-myrtanylamine (2), and 3-methyldiphenyl amine (3) at room temperature without using harsh conditions. All the probes show beautiful and strong binding with Pb2+ ions among all the tested essential elements of human body. The binding is clearly seen and confirmed in UV-visible, NMR and HPLC studies. Also, all the substituted amines (1-3) are well known bioactives viz., piperazine as anthelmintic, cis-myrtanyl use for cannabinoid receptor (CB2) antagonists, 3-methyldiphenyl is used in probes for selective detection of explosive nitroaromatic compounds further increases their sensitivity for use as Pb2+ sensor. As they are already well in use for research on human body metabolomics their future introduction as sensors in the human body for lead toxicity is highly favourable.

Pb2+ sensing by coumarin sulphonamide hybrids in aqueous medium.

Metals play an important role in various metabolic activities in the human body, but above desired concentrations, a role reversal occurs that causes deadly outcomes viz., cancer. Metals cannot be cracked down and are non-biodegradable. It is the bioaccumulation of toxic metals inside the biomatrices, that further intensifies the research on different means of metal detoxification from different matrices. Among heavy toxic metals lead is a brutal carcinogen that requires pitiless sensors for its capturing. The use of heterocycles for metal sensing in supramolecular chemistry is preferred due to the strong chelation they offer to toxic metals. The C1-C3 probes were synthesized and studied for their Pb2+ binding ability. All the probes were prepared by treating bromoacetyl coumarin with camphor sulphonamide, 5-dimethylamino-1-naphthalene sulphonamide, and methyl-2-amino-sulphonyl benzoate at room temperature. The probes show selective binding with Pb2+ ions in aqueous acetonitrile among different tested metal ions viz., Cu2+ , Zn2+ , Ni2+ , Mn2+, and Pb2+ ions as shown in ultraviolet (UV)-visible, nuclear magnetic resonance (NMR), and high-performance liquid chromatography (HPLC) studies. These sulphur-containing probes bind very well with Pb2+ ions by offering selectivity in binding positions that capture lead ions at their minimum possible concentration.

Statistical optimization of culture conditions of mesophillic gamma-glutamyl transpeptidase from Bacillus altitudinis IHB B1644.

Microbial gamma-glutamyl transpeptidase (GGT) is a key enzyme in production of several γ-glutamyl compounds with food and pharmaceutical applications. Bacterial GGTs are not commercially available in the market owing to their low production from various sources. Thus, the study was focused on achieving the higher GGT production from B. altitudinis IHB B1644 by optimizing the culture conditions using one-variable-at-a-time (OVAT) strategy. A mesophillic temperature of 28 °C, agitation 200 rpm and neutral pH 7 were found to be optimal for higher GGT titre. Among the medium components, the monosaccharide glucose served as the best carbon source over disaccharides, and yeast extract was the preferred organic nitrogen source over inorganic nitrogen sources. The statistical approaches (Plakett-Burman and response surface methodology) were further employed for the optimization of medium components. Medium composition: 0.1% w/v glucose, 0.3% w/v yeast extract, 0.03% w/v magnesium sulphate, 0.20% w/v potassium dihydrogen phosphate and 2.5% w/v sodium chloride with inoculum size (1% v/v) was suitable for higher GGT titres (449 U ml-1). Time kinetics showed the stability of enzyme up to 96 h of incubation suggesting its application in the industrial use. The proposed strategy resulted in 2.6-fold increase in the GGT production compared to that obtained in the unoptimized medium. The results demonstrated that RSM was fitting to identify the optimum production conditions and this finding should be of great importance for commercial GGT production.

Selective binding of Ni2+ and Cu2+ metal ions with naphthazarin esters isolated from Arnebia euchroma.

Naphthazarin esters (C1-C4) isolated from the roots of Arnebia euchroma are found as skilled dual chemosensors for Ni2+ and Cu2+ among Pb2+ , Na2+ , K2+ , Hg2+ , Mg2+ , and Ca2+ metal ions. C1-C4 esters exhibited a red shift of 54 nm with Ni2+ and 30 nm with Cu2+ metal ions in absorption. There is a formation of red-shifted bands between 517 and 613 nm in the absorption spectrum of C1-C4 sensors on binding with Ni2+ and Cu2+ ions. The addition of Ni2+ and Cu2+ ions to sensors C1-C4 stimulates a remarkable color change from reddish pink to purple and light blue, respectively. These color changes can be identified with the naked eye. The significant downfield shifts of CO and OH peaks in nuclear magnetic resonance (NMR) spectrum confirm the chelation as binding mechanism. With ultraviolet-visble and NMR studies, it is found that C1-C4 esters possessed notable selectivity and sensitivity toward Ni2+ and Cu2+ over other metal ions.

Correction to: Diet supplemented with phytochemical epigallocatechin gallate and probiotic Lactobacillus fermentum confers second generation synbiotic effects by modulating cellular immune responses and antioxidant capacity in aging mice.

Authors of the original article have observed an inadvertent error in their manuscript post-publication.

Diet supplemented with phytochemical epigallocatechin gallate and probiotic Lactobacillus fermentum confers second generation synbiotic effects by modulating cellular immune responses and antioxidant capacity in aging mice.

PURPOSE: In the present study, we systematically identified and evaluated a synbiotic combination of phytochemical epigallocatechin gallate (EGCG) and probiotic bacteria in amelioration of immunosenescence and oxidative stress in aged mice. METHODS: Inhibitory effects of EGCG against different bacterial species were evaluated in vitro, followed by analysis to identify potential combination of EGCG and probiotic bacteria against alleviation of oxidative and inflammatory stress ex vivo. The best synbiotic combination, vis-à-vis prebiotic and probiotic supplementation alone, was then evaluated in aged Swiss albino mice for modulation of various immunological and antioxidative parameters. RESULTS: EGCG strongly inhibited the growth of pathogenic microbes as compared to probiotic bacteria. A combination of EGCG with probiotic Lactobacillus fermentum (LF) provided evidence of additive effects in the amelioration of oxidative and inflammatory stress-induced cell death. In vivo study revealed that combined supplementation of LF and EGCG significantly enhanced neutrophil oxidative index, CD3+ cell numbers and activation status, Th1/Th2 cytokines in splenic supernatants as well as liver Nrf-2 expression in comparison with treatments with LF or EGCG alone. The combined application of EGCG and LF did not simply result in additive or synergistic effects in relation with individual treatments. CONCLUSION: These observations suggest that EGCG could be considered as a potential prebiotic that can offer second generation synbiotic health beneficial effects for the alleviation of some of the deleterious aspects of immunosenescence and aging.

Chrysin isolated from Pyrus pashia fruit ameliorates convulsions in experimental animals.

Objective: The traditional use of the ethanolic extract of the fruit of Pyrus pashia (EPP) as a potential anticonvulsant was validated using experimental animal models. Furthermore, the anticonvulsant activity of isolated chrysin was investigated against experimental animal models to draw a possible therapeutic mechanism of EPP. Additionally, the safety profile of chrysin was evaluated to explore the possible therapeutic alternative in the management of epilepsy. Method: The anticonvulsant activity in terms of duration of onset of hind limb tonic extension and convulsion of standardized EPP was evaluated against maximal electroshock (MES) and pentylenetetrazole (PTZ) model of experimental epilepsy respectively. Furthermore, the anticonvulsant activity and electrophysiological properties of chrysin was investigated in addition to antioxidant activity against PTZ-induced convulsion in experimental animals. Moreover, the neurotoxic profile of the chrysin was assessed in terms of duration of movement and running in photoactometer and rotarod apparatus, respectively. Results: EPP (100, 200, and 400 mg/kg) exhibited significant anticonvulsant activity against an acute model of MES and PTZ-induced convulsions in experimental animals. Furthermore, chrysin (2.5, 5, and 10 mg/kg) also exhibited significant anticonvulsant activity against PTZ-induced convulsions in rats. In addition, chrysin did not exhibit sedative-like behavior in experimental rodents. Discussion: EPP could be considered as a potential and alternative therapeutic option in the management of epilepsy.

Epigallocatechin gallate suppresses premature senescence of preadipocytes by inhibition of PI3K/Akt/mTOR pathway and induces senescent cell death by regulation of Bax/Bcl-2 pathway.

The phytochemical epigallocatechin gallate (EGCG) has been reported to alleviate age-associated immune disorders and organ dysfunction. However, information regarding the mechanistic role of EGCG in the suppression of cellular senescence is limited. The present study thus assessed the effects and underlying mechanisms of EGCG in the inhibition of senescence as well as its potential to selectively eliminate senescent cells (senolytics) using 3T3-L1 preadipocytes. Premature senescence was established in cells by repeated exposure of H2O2 at a sub-lethal concentration (150 µM). H2O2 treated cells showed characteristic senescence-associated features including increased cell size, senescence-associated ß-galactosidase activity (SA-ß-gal), development of senescence-associated secretory phenotype (SASP), activation of reactive oxygen species (ROS) and pathways, DNA damage as well as induction of cell cycle inhibitors (p53/p21WAF1/p16INK4a). In addition, a robust activation of PI3K/Akt/mTOR and AMPK pathways was also observed in H2O2 treated cells. Presence of EGCG (50 and 100 µM) showed significant downregulation of PI3K/Akt/mTOR and AMPK signaling along with the suppression of ROS, iNOS, Cox-2, NF-κB, SASP and p53 mediated cell cycle inhibition in preadipocytes. In addition, EGCG treatment also suppressed the accumulation of anti-apoptotic protein Bcl-2 in senescent cells thereby promoting apoptosis mediated cell death. Our results collectively show that EGCG acts as an mTOR inhibitor, SASP modulator as well as a potential senolytic agent thereby indicating its multi-faceted attributes that could be useful for developing anti-aging or age-delaying therapies.

l-Theanine: An astounding sui generis integrant in tea.

l-theanine (l-Th), a non-protein amino acid present in tea, is a valuable nutraceutical product with unique health benefits and used as an additive in food industry. l-Th enhances the umami taste but its use is limited due to its inadequate production. Different extraction approaches from tea shoots, chemical synthesis to microbial transformation have been tried to meet its demand. In vitro, in vivo as well as clinical studies have shown its positive effect in regulating CNS disorders. l-Th has become choice ingredient in CNS active products due to its anti-stress and neuroprotective role in dementias particularly in retrogression of Alzheimer's. l-Th biochemically modulates various anti-neoplastic agents by increasing their bioavailability in tumour cells. The review, is an effort to condense the recent research on l-Th highlighting its biological resource, plausible role in tea plant, production approaches, its physiological role on human health and future prospects.

Anthocyanins enriched purple tea exhibits antioxidant, immunostimulatory and anticancer activities.

Purple coloured tea shoot clones have gained interest due to high content of anthocyanins in addition to catechins. Transcript expression of genes encoding anthocyanidin reductase (ANR), dihydroflavonol-4-reductase (DFR), anthocyanidin synthase (ANS), flavonol synthase (FLS) and leucoantho cyanidin reductase (LAR) enzymes in three new purple shoot tea clones compared with normal tea clone showed higher expression of CsDFR, CsANR, CsANS and lower expression of CsFLS and CsLAR in purple shoot clones compared to normal clone. Expression pattern supported high content of anthocyanins in purple tea. Four anthocyanins (AN1-4) were isolated and characterized by UPLC-ESI-QToF-MS/MS from IHBT 269 clone which recorded highest total anthocyanins content. Cyanidin-3-O-ß-d-(6-(E)-coumaroyl) glucopyranoside (AN2) showed highest in vitro antioxidant activity (IC50 DPPH = 25.27 ± 0.02 µg/mL and IC50 ABTS = 10.71 ± 0.01 µg/mL). Anticancer and immunostimulatory activities of cyanidin-3-glucoside (AN1), cyanidin-3-O-ß-d-(6-(E)-coumaroyl) glucopyranoside (AN2), delphinidin-3-O-ß-d-(6-(E)-coumaroyl) glucopyranoside (AN3), cyanidin-3-O-(2-O-ß-xylopyranosyl-6-O-acetyl)-ß-glucopyranoside (AN4) and crude anthocyanin extract (AN5) showed high therapeutic perspective. Anthocyanins AN1-4 and crude extract AN5 showed cytotoxicity on C-6 cancer cells and high relative fluorescence units (RFU) at 200 µg/mL suggesting promising apoptosis induction activity as well as influential immunostimulatory potential. Observations demonstrate potential of purple anthocyanins enriched tea clone for exploitation as a nutraceutical product.

Stress-Tolerant Viridibacillus arenosi Strain IHB B 7171 from Tea Rhizosphere as a Potential Broad-Spectrum Microbial Inoculant.

Viridibacillus arenosi strain IHB B 7171 identified based on 16S rRNA gene sequence produced colony forming units (cfu/ml) ranging from 3.3 × 104 to 1.2 × 1010 under pH 5-11, 2.2 × 102 to 1.4 × 1010 for temperature 5-40 °C, 2.4 × 102 to 1.1 × 1010 for PEG 6000 10-30%, 2.2 × 102 to 1.4 × 1010 for 2.5-10% NaCl, 3.1 × 103 to 1.7 × 109 for 2.5-7.5 mM CaCl2, 2.2 × 102 to 1.4 × 107 for 2.5-7.5 mM AlCl3, and 3.2 × 102 to 1.2 × 107 for 2.5-7.5 mM FeCl3. The activities of plant growth-promoting attributes with the increasing acidity, desiccation and salinity ranged from 408 to 101, 20 to 8, 14 to 5 µg/ml P-liberated from tri-calcium phosphate, aluminium phosphate and iron phosphate, 20-9% siderophore units, 14-4 µg/ml IAA and 190-16 α-ketobutyrate h/mg protein ACC-deaminase activity. Plant height, leaf number, and leaf weight on treatment with bacterial inoculum showed an increment of 9.5, 17.6, 54.5 and 31.0% in tea seedlings, respectively. The bacterium also enhanced plant height and yield by 10 and 13% in pea and 2.8 and 13.9% in wheat. The results exhibited stress-tolerance and plant growth-promoting activities by the strain under stressed growth-conditions with potential as a broad-spectrum plant growth-promoting rhizobacterium.

Consumption of green tea epigallocatechin-3-gallate enhances systemic immune response, antioxidative capacity and HPA axis functions in aged male swiss albino mice.

The present investigation assessed the potential of green tea phytochemical epigallocatechin-3-gallate (EGCG) in alleviating age-associated aberrations in immunity, hypothalamus-pituitary-adrenal (HPA) axis and redox homeostasis using 16 months old male Swiss albino mice. Four groups of animals (n = 6 per group) were supplemented with either aqueous EGCG at 25, 50 and 100 mg/kg/animal or vehicle control for 6 weeks. A concurrent analysis of CD4+ and CD8+ lymphocytes in splenocytes, differential leucocyte population, T cell differentiation markers in peripheral blood mononuclear cells (PBMCs), neutrophil functions, immunoglobulins profile in intestine, circulatory HPA axis hormonal levels as well as inflammatory and oxidative stress in the liver was performed. We observed a remarkable increase in plasma dehydroepiandrosterone (DHEA) levels of 100 mg EGCG fed animals while eosinophils and monocytes counts in blood increased. EGCG consumption increased the fraction of CD3+CD8+ cells in splenocytes and CD28 expression on PBMCs. The immunoglobulins profile revealed decreased production of secretory IgA, IgE and IgG1/IgG2a ratio. Liver extracts showed increase in superoxide dismutase activity and total antioxidant capacity while lipid peroxidation along with inflammatory markers (IL-6 and TNF-α) decreased. Our results collectively show that EGCG consumption during aging strengthens systemic immunity by enhancing cellular immune response and simultaneously attenuating antibody response aided by an increase in adrenal DHEA production. Thus, consumption of green tea may be beneficial in alleviating some of the deleterious aspects of aging and immunosenescence in elderly.

Screening and purification of catechins from underutilized tea plant parts and their bioactivity studies.

Comparative investigation of major phytoconstituents was performed from various parts of tea plant viz. apical bud, subtending 1st-5th leaf, stem, coarse leaves, flowers, fruits and roots. From the results of comparative RP-HPLC-DAD analysis it was found that underutilized tea parts especially coarse leaves, flowers and fruits contains abundant amount of phenolics (17.5%) and catechins (4-5%). From these underutilized tea plant parts the catechins were extracted and purified and then screened for their anticancer, immunomodulatory effect and antimicrobial activity against food borne pathogens. The results showed that tea fruit extract exhibited higher toxicity against oral cancer cells and also promotes proliferation of mice splenocytes. The results of antimicrobial studies revealed the inhibitory effect of these extracts against both gram positive and gram negative bacteria. These investigations clearly demonstrated that the underutilized tea plant parts could act as economical and sustainable bioresource of functionally active constituents which further lead to the development of new cost-effective nutraceuticals and other formulations.

Towards biological plausibility of electronic noses: A spiking neural network based approach for tea odour classification.

The paper presents a novel encoding scheme for neuronal code generation for odour recognition using an electronic nose (EN). This scheme is based on channel encoding using multiple Gaussian receptive fields superimposed over the temporal EN responses. The encoded data is further applied to a spiking neural network (SNN) for pattern classification. Two forms of SNN, a back-propagation based SpikeProp and a dynamic evolving SNN are used to learn the encoded responses. The effects of information encoding on the performance of SNNs have been investigated. Statistical tests have been performed to determine the contribution of the SNN and the encoding scheme to overall odour discrimination. The approach has been implemented in odour classification of orthodox black tea (Kangra-Himachal Pradesh Region) thereby demonstrating a biomimetic approach for EN data analysis.

Studies on quality of orthodox teas made from anthocyanin-rich tea clones growing in Kangra valley, India.

Recently anthocyanin-rich purple tea varieties have been developed. The quality of these new purple tea varieties developed in Kangra valley was assessed, and compared with the quality of tea from standard Kangra clone. Purple tea shoots (PL) recorded higher amount of polyphenols compared to standard green tea shoot (GL) while epigallocatechin gallate (EGCG) recorded higher levels in GL. Higher levels of theaflavins were recorded in orthodox black tea from purple shoots (BTP) compared to black tea (BT) made from green shoots. Both theanine and caffeine recorded higher levels in GL. Volatile flavour profiles of these teas showed qualitative and quantitative differences. Aroma extract dilution assay showed higher dilution factors in BTP than BT. Orthodox teas from purple shoots exhibited higher antioxidant activity compared to standard black tea. Strong correlation of total quality scores with aroma and infusion colour was observed. Tea from anthocyanin-rich cultivars can become specialty teas with high antioxidant activity.

Fractionation and identification of minor and aroma-active constituents in Kangra orthodox black tea.

The aroma constituents of Kangra orthodox black tea were isolated by simultaneous distillation extraction (SDE), supercritical fluid extraction and beverage method. The aroma-active compounds were identified using gas chromatography-olfactometry-mass spectrometry. Geraniol, linalool, (Z/E)-linalool oxides, (E)-2-hexenal, phytol, ß-ionone, hotrienol, methylpyrazine and methyl salicylate were major volatile constituents in all the extracts. Minor volatile compounds in all the extracts were 2-ethyl-5-methylpyrazine, ethylpyrazine, 2-6,10,14-trimethyl-2-pentadecanone, acetylfuran, hexanoic acid, dihydroactinidiolide and (E/Z)-2,6-nonadienal. The concentrated SDE extract was fractionated into acidic, basic, water-soluble and neutral fractions. The neutral fraction was further chromatographed on a packed silica gel column eluted with pentane and diethyl ether to separate minor compounds. The aroma-active compounds identified using gas chromatography-olfactometry-mass spectrometry were 2-amylfuran, (E/Z)-2,6-nonadienal, 1-pentanol, epoxylinalool, (Z)-jasmone, 2-acetylpyrrole, farnesyl acetone, geranyl acetone, cadinol, cubenol and dihydroactinidiolide. AEDA studies showed 2-hexenal, 3-hexenol, ethylpyrazine, (Z/E)-linalool oxides, linalool, (E/Z)-2,6-nonadienal, geraniol, phenylethanol, ß-ionone, hotrienol and dihydroactinidiolide to be odour active components.

Comparative studies for screening of bioactive constituents from various parts of Incarvillea emodi.

A comparative study was performed on various parts (shoots, roots and flowers) of Incarvillea emodi. The alcoholic extracts of different parts were fractionated with solvents of different polarity and studied for the determination of total polyphenol content and total antioxidant potential. Furthermore, we have isolated major iridoid glucosides from the dried flowers of I. emodi followed by the comparative cytotoxicity studies of these iridoids against five different human cancer cell lines. The results have demonstrated that ethyl acetate fraction of all parts have higher phenolic content (167.87-294.31 mg/g as gallic acid equivalent) and higher total antioxidant potential (252.95-384.64 mg/g as trolox equivalent). The results of in vitro cytotoxicity studies have indicated that boschnaloside (2) possesses promising anticancer potential against three human cancer cell lines, THP-1, A-549 and PC-3, which belong to leukaemia, lung and prostate cancers, respectively, while plantarenaloside (1) expressed relevant cytotoxic activity against THP-1 cell lines of leukaemia.