Renoprotective and therapeutic effects of newly water, ethanol, and butanol ginseng fractions in hypertensive and chronic kidney disease with L-NAME.

The present study investigated the protective and treatment effects of different ginseng fractions against L-NAME-induced renal toxicity in rats. The data obtained demonstrated that L-NAME significantly increased creatinine, urea, KIM-1, and lipocalin-2 levels in serum; and also increased renal MDA and eNOS levels compared with the control group. Three bioactive fractions were newly extracted from ginseng, analyzed by GC-MS analysis, and were examined for antimicrobial, prebiotic, and histological activities. All ginseng fractions improved such histological changes, as reflected by significant reductions in creatinine, urea, KIM-1, and LCN-2 levels in serum, and renal MDA and eNOS contents in tissue homogenate. The water ginseng fraction (WGF) has the highest prebiotic index of 4.7 toward Lactobacillus reuteri, and can improve the renal functions more than butanol ginseng fraction (BGF) and ethanol ginseng fraction (EGF). These three ginseng fractions significantly reversed L-NAME-induced depletion in the TNF-α gene expression level. Interestingly, WGF was able to improve the renal functions more than BGF and EGF. L-NAME led to alterations in the histological structure and functions of renal tissue of rats and ginseng supplementation could offer greater protection against these changes. Moreover, the WGF exhibited superior renoprotection properties when compared with the other two fractions: BGF and EGF, and the reference drug losartan.

The n-butanol fraction of the Xiao-Chai-Hu decoction alleviates the endocrine disturbance in the liver of mice exposed to lead.

ETHNOPHARMACOLOGICAL RELEVANCE: Lead is a toxic heavy metal that causes health risks globally. However, the mechanism of endocrine poisoning and detoxification of lead poisoning, especially in the liver, still needs to be studied. Xiao-Chai-Hu decoction (XCHD) is regarded as an antidote and an anti-hepatotoxic traditional prescription that has been recorded in the pharmacopeia of the People's Republic of China. AIM OF THE STUDY: The study aimed to probe the hepatoprotective activity of XCHD in the regulation of endocrine dysfunction in the liver and its molecular mechanism. MATERIALS AND METHODS: The mice from the Institute of Cancer Research (ICR) were exposed to different concentrations of XCHD and lead. Then, serum biochemical indices and liver pathology were analyzed. The key differential genes were detected by qRT-PCR and Western blot. RESULTS: According to the biochemical and histopathological analysis, XCHD-NBA was the most effective in attenuating lead-induced hepatotoxicity. From the transcriptome, we analyzed the key genes of XCHD-NBA in the regulation of lead toxicity, including Tubb2a, Stip1, Cyp4a12a, Cyp2c50, Ugt1a1, Cyp3a11, Cyp4a12b, Ahsa1, Cyp2c54, Tubb4b, Esr1, Hsp90aa1, Tuba1a, Tuba1c, and Hsph1. We also analyzed the main components of XCHD-NBA by LC-MS. Because of their extensive role in regulating the endocrine function, baicalin and glycyrrhizin were identified as the main active components of XCHD in regulating endocrine disorders caused by lead. CONCLUSIONS: Lead can disturb the endocrine regulatory process of the liver, while XCHD-NBA alleviates lead-induced liver injury by regulating the endocrine regulatory process.

Metabolic engineering for the production of butanol, a potential advanced biofuel, from renewable resources.

Butanol is an important chemical and potential fuel. For more than 100 years, acetone-butanol-ethanol (ABE) fermentation of Clostridium strains has been the most successful process for biological butanol production. In recent years, other microbes have been engineered to produce butanol as well, among which Escherichia coli was the best one. Considering the crude oil price fluctuation, minimizing the cost of butanol production is of highest priority for its industrial application. Therefore, using cheaper feedstocks instead of pure sugars is an important project. In this review, we summarized butanol production from different renewable resources, such as industrial and food waste, lignocellulosic biomass, syngas and other renewable resources. This review will present the current progress in this field and provide insights for further engineering efforts on renewable butanol production.

Phenolic Composition, Antioxidant and Anticancer Potentials of Extracts from Rosa banksiae Ait. Flowers.

Rosa banksiae Ait. (R. banksiae) is a traditional Chinese folk medicine and an ornamental plant. Most previous studies have focused on cultivation and utilization while there are few research papers on the pharmacological activity of R. banksiae. This study aimed to get a better understanding of R. banksiae by extracting polyphenols with fractionated extraction technology. The results showed that ethyl acetate phase (EAP) contained the most polyphenols, while water phase (WP) had the least. HPLC analysis indicated that rutin and luteolin-4'-O-glucoside existed in the EAP and butanol phase (BP), but quercetin was only detected in the EAP. Six phenolic compositions were not detected in WB. The antioxidant and anti-tumor abilities of the EAP and BP were excellent. The results revealed that R. banksiae possessed a great antioxidant capacity and was rich in polyphenols, thus indicating R. banksiae was suitable for being a natural antioxidant and an abundant source of polyphenols.

Matrix effects of the hydroethanolic extract and the butanol fraction of calyces from Physalis peruviana L. on the biopharmaceutics classification of rutin.

OBJECTIVES: The Biopharmaceutics Classification System (BCS) categorizes active pharmaceutical ingredients according to their solubility and permeability properties, which are susceptible to matrix or formulation effects. The aim of this research was to evaluate the matrix effects of a hydroethanolic extract of calyces from Physalis peruviana L. (HEE) and its butanol fraction (BF), on the biopharmaceutics classification of their major compound, quercetin-3-O-rutinoside (rutin, RU). METHODS: Rutin was quantified by HPLC-UV, and Caco-2 cell monolayer transport studies were performed to obtain the apparent permeability values (Papp ). Aqueous solubility was determined at pH 6.8 and 7.4. KEY FINDINGS: The Papp values followed this order: BF > HEE > RU (1.77 ± 0.02 > 1.53 ± 0.07 > 0.90 ± 0.03 × 10-5  cm/s). The lowest solubility values followed this order: HEE > RU > BF (2.988 ± 0.07 > 0.205 ± 0.002 > 0.189 ± 0.005 mg/ml). CONCLUSIONS: According to these results, rutin could be classified as BCS classes III (high solubility/low permeability) and IV (low solubility/low permeability), depending on the plant matrix. Further work needs to be done in order to establish how apply the BCS for research and development of new botanical drugs or for bioequivalence purposes.

A butanolic fraction from the standardized stem extract of Cassia occidentalis L delivered by a self-emulsifying drug delivery system protects rats from glucocorticoid-induced osteopenia and muscle atrophy.

We recently reported that a butanol soluble fraction from the stem of Cassia occidentalis (CSE-Bu) consisting of osteogenic compounds mitigated methylprednisone (MP)-induced osteopenia in rats, albeit failed to afford complete protection thus leaving a substantial scope for further improvement. To this aim, we prepared an oral formulation that was a lipid-based self-nano emulsifying drug delivery system (CSE-BuF). The globule size of CSE-BuF was in the range of 100-180 nm of diluted emulsion and the zeta potential was -28 mV. CSE-BuF enhanced the circulating levels of five osteogenic compounds compared to CSE-Bu. CSE-BuF (50 mg/kg) promoted bone regeneration at the osteotomy site and completely prevented MP-induced loss of bone mass and strength by concomitant osteogenic and anti-resorptive mechanisms. The MP-induced downregulations of miR29a (the positive regulator of the osteoblast transcription factor, Runx2) and miR17 and miR20a (the negative regulators of the osteoclastogenic cytokine RANKL) in bone was prevented by CSE-BuF. In addition, CSE-BuF protected rats from the MP-induced sarcopenia and/or muscle atrophy by downregulating the skeletal muscle atrogenes, adverse changes in body weight and composition. CSE-BuF did not impact the anti-inflammatory effect of MP. Our preclinical study established CSE-BuF as a prophylactic agent against MP-induced osteopenia and muscle atrophy.

Use of unconventional mixed Acetone-Butanol-Ethanol solvents for anthocyanin extraction from Purple-Fleshed sweetpotatoes.

Anthocyanins from purple-fleshed sweetpotatoes constitute highly valued natural colorants and functional ingredients. In the past, anthocyanin extraction conditions and efficiencies using a single acidified solvent have been assessed. However, the potential of solvent mixes that can be generated by fermentation of biomass-derived sugars have not been explored. In this study, the effects of single and mixed solvent, time, temperature, sweetpotato genotype and preparation, on anthocyanin and phenolic extraction were evaluated. Results indicated that unconventional diluted solvent mixes containing acetone, butanol, and ethanol were superior or equally efficient for extracting anthocyanins when compared to commonly used concentrated extractants. In addition, analysis of anthocyanidins concentrations including cyanidin (cy), peonidin (pe), and pelargonidin (pl), indicated that different ratios of pn/cy were obtained depending on the solvent used. These results could be useful when selecting processing conditions that better suit particular end-use applications and more environmentally friendly process development for purple sweetpotatoes.

Effect of Geographical Location and Type of Extract on Total Phenol/Flavon Contents and Antioxidant Activity of Different Fruits Extracts of Withania somnifera.

BACKGROUND: Withania somnifera (family solanaceae) is a well-investigated medicinal plant which is also called Indian ginseng due to its wide spectrum of medicinal properties. The contents and activity of the plant may vary depending on the habitat and part of the plant and the solvent used for extraction. The present study deals with the comparative chemical analysis and in vitro antioxidant activity of methanolic fruits extracts and its subfractions (in ethyl acetate, butanol and water) of W. somnifera collected from two different geographical locations. METHODS: In the present study, Withania somnifera fruits were collected from two different geographical locations (Uttarakhand and Rajasthan). The different fruit extracts were prepared and studied for total phenolic contents and total flavone contents. The in vitro antioxidant activity was assessed by DPPH free radical scavenging assay and peroxide scavenging assay. RESULTS: Methanol extract of W. somnifera Uttarakhand and ethyl acetate subfraction of W. somnifera Rajasthan showed the highest amount of Total Phenolic Contents (TPC). In W. somnifera Uttarakhand, ethyl acetate extract showed the highest amount of Total flavonoids while in W. somnifera Rajasthan, methanol extract was found to be the richest in flavonoids. Methanolic extract of W. somnifera Uttarakhand showed the highest free radical scavenging activity while in W. somnifera Rajasthan, the highest antioxidant activity was shown by the methanolic extract followed by butanolic extract, water extract and then ethyl acetate. In the peroxide scavenging assay of antioxidant activity, water extract of W. somnifera Uttarakhand showed the highest activity, while in W. somnifera Rajasthan, ethyl acetate extract showed highest scavenging activity. CONCLUSION: It was concluded that the geographical location exerts a vital effect on the presence of active constituents and also on the antioxidant potential of W. somnifera.

Evaluation of the chemical composition and antioxidant activity of extracts and fractions of Ocotea notata (Ness) Mez (Lauraceae).

The specie Ocotea notata (Nees & Mart). Mez is a tree with 5 meters high, that can be found in restinga regions in the Brazilian coast. This study describes a phytochemical investigation, total phenolic content and the antioxidant activity of extracts and fractions by DPPH and ORAC. Phenolic content revealed equivalent concentration between evaluated samples, similar to found in the leave extract (66.4 mEq GA/g). By DPPH, extracts and fractions showed effective concentration (EC50) lower than the standards Ginkgo biloba 761® (23.60 ± 0.64 µg/ml) and quercetin (6.06 ± 0, 92 µg/mL); for the ORAC method, ethyl acetate partition showed a value of 2.06 mmol Trolox equivalent g-1 better than obtained in Ginkgo biloba (1.03 ± 0.25 mmol.Trolox equivalent g-1. The butanol partition (0.52 mmol.Trolox equivalent g-1) and the aqueous residue (0.74 mmol Trolox equivalent g-1) have a lesser ORAC potential than ethyl acetate partition. The butanolic partition, investigated by LC-DAD-MS/MS and QTOF-MS, revealed six major compounds: miquelianin (1), isoquercitrin (2), quercitrin (3), kaempferol-3-O-pentose (4), afzelin (5) and isorhamnetin-glucuronide (6).

Production of bio-oil with reduced polycyclic aromatic hydrocarbons via continuous pyrolysis of biobutanol process derived waste lignin.

The fast pyrolysis of waste lignin derived from biobutanol production process was performed to determine the optimal pyrolysis conditions and pyrolysis product properties. Four types of pyrolysis reactors, e.g.: micro-scale pyrolyzer-gas chromatography/mass spectrometry, lab and bench scale fixed bed (FB) reactors, and bench scale rotary kiln (RK) reactor, were employed to compare the pyrolysis reaction conditions and product properties obtained from different reactors. The yields of char, oil, and gas obtained from lab scale and bench scale reactor were almost similar compared to FB reactor. RK reactor produced desirable bio-oil with much reduced yield of poly aromatic hydrocarbons (cancer precursor) due to its higher cracking reaction efficiency. In addition, char agglomeration and foaming of lignin pyrolysis were greatly restricted by using RK reactor compared to the FB reactor.

RIFM fragrance ingredient safety assessment, isobutyl alcohol, CAS Registry Number 78-83-1.

The existing information supports the use of this material as described in this safety assessment. Isobutyl alcohol was evaluated for genotoxicity, repeated dose toxicity, reproductive toxicity, local respiratory toxicity, phototoxicity/photoallergenicity, skin sensitization, and environmental safety. Data show that isobutyl alcohol is not genotoxic. Data on isobutyl alcohol provide a calculated MOE >100 for the repeated dose toxicity and reproductive toxicity endpoints. Data from read-across material isoamyl alcohol (CAS # 123-51-3) show that there are no safety concerns for isobutyl alcohol for skin sensitization under the current declared levels of use. The phototoxicity/photoallergenicity endpoints were evaluated based on UV spectra; isobutyl alcohol is not expected to be phototoxic/photoallergenic. The local respiratory toxicity endpoint was evaluated using the TTC for a Cramer Class I material and the exposure to isobutyl alcohol is below the TTC (1.4 mg/day). The environmental endpoints were evaluated; isobutyl alcohol was found not to be PBT as per the IFRA Environmental Standards, and its risk quotients, based on its current volume of use in Europe and North America (i.e., PEC/PNEC), are <1.

An Oviposition Stimulant for a Magnoliaceae-Feeding Swallowtail Butterfly, Graphium doson, from its Primary Host Plant, Michelia compressa.

Chemical examination of plant constituents responsible for oviposition by a Magnoliaceae-feeding butterfly, Graphium doson, was conducted using its major host plant, Michelia compressa. A methanol extract prepared from young leaves of the plant elicited a strong oviposition response from females. The methanolic extract was then separated by solvent partition into three fractions: CHCl3, i-BuOH, and aqueous fractions. Active substance(s) resided in both i-BuOH- and water-soluble fractions. Bioassay-guided further fractionation of the water-soluble substances by means of various chromatographic techniques led to the isolation of an oviposition stimulant. The stimulant was identified as D-(+)-pinitol on the basis of 13C NMR spectra and physicochemical properties. D-(+)-Pinitol singly exhibited a moderate oviposition-stimulatory activity at a dose of 150 µg/cm2. This compound was present also in another host plant, Magnolia grandiflora, in a sufficient amount to induce oviposition behavior of G. doson females. Certain cyclitols including D-(+)-pinitol have been reported to be involved in stimulation of oviposition by some Aristolochiaceae- and Rutaceae-feeding papilionid butterflies. A possible pathway of phytochemical-mediated host shifts in the Papilionidae, in which certain cyclitols could enact important mediators, is discussed in relation to the evolution of cyclitol biosynthesis in plants.

Chemoprotective effects of butanol fraction of Buchholzia coriacea (Capparidaceae) against type 2 diabetes and oxidative stress in male Wistar rats.

Recent studies have shown that Type 2 diabetes (T2D) in rats can result through a synergy that links obesity to insulin resistance and ß-cell dysfunction. The present study achieved T2D via high fructose (20%w/v, p.o.), streptozotocin single dose (40 mg/kg, i.p.) (HFSTZ) in rats. Also, chemoprotective potential of butanol fraction of Buchholzia coriacea (BFBC) was demonstrated. Control normal and diabetic untreated (HFSTZ-induced T2D) rats received CM-cellulose (1 mg/kg, p.o.). Diabetic rats received intragastric BFBC (20, 200, 400 mg/kg), glibenclamide (0.07 mg/kg), and BFBC (200 mg/kg) plus glibenclamide treatments, respectively. 2,2-Diphenyl-1-picrylhydrazyl, nitric oxide radical, hydroxyl radical scavenging activities, and α-amylase inhibition were assessed. After 2 weeks of treatments, blood glucose levels, lipid profiles, renal and liver function, serum insulin as well as in vivo oxidative stress biomarkers were assessed. BFBC shows highest antioxidants and α-amylase inhibitory activities in vitro HFSTZ-induced T2D produced hyperglycemia (P<0.05-0.001; F = 5.26-26.47), serum hyperinsulinemia (six-folds) plus elevated lipid peroxidation levels. Similarly, there were altered lipid profiles, liver and renal biomarker enzymes plus weight loss. BFBC administration alone or in combination with glibenclamide reversed T2D symptomatologies in treated animals, and improved body weights against control diabetic rats. In vivo antioxidant activities also improved while histological sections in treated rats show reduced tissue damage in pancreas, kidneys, liver, and heart, respectively. Oleic, stearic, 2-methyl-pyrrolidine-2-carboxylic, and n-hexadecanoic acids were present in BFBC in large quantities given GC-MS analysis. Overall, data from the present study suggest chemoprotective potentials of BFBC against HFSTZ-induced T2D rats.

[Antidermatophyte Activity of the Gentiopicroside-Rich n-Butanol Fraction from Gentiana siphonantha Maxim. Root on a Guinea Pig Model of Dermatophytosis].

BACKGROUND: Gentiana siphonantha Maxim. is a traditional medicine for the treatment of rheumarthritis, icterepatitis, pain and hypertension; it is rich in gentiopicroside with anti-inflammatory, antibacterial, and free radical-scavenging activities. This study was to evaluate the antidermatophyte activity of G. siphonantha on a guinea pig model in vitro and in vivo. Material andMethods: The antidermatophyte activities of 10 plants were tested by the broth microdilution method. Fractions and an extract of G. siphonantha were tested against Trichophyton mentagrophytes by the disc diffusion method. The morphological changes of T. mentagrophytes were observed. Component analysis of the n-butanol (n-BuOH) fraction was made by HPLC. Finally, the antifungal activity in an in vivo guinea pig model of dermatophytosis was examined. RESULTS: G. siphonantha had strong antidermatophyte activity with MIC50 values of 32-64 µg/mL. The n-BuOH fraction of G. siphonantha showed the most potent activity compared to the other fractions. After being exposed to the n-BuOH fraction at 80 and 160 µg/mL, the hyphae were distorted and collapsed. Gentiopicroside is the main active ingredient in the n-BuOH fraction of G. siphonantha. The lesion scores of the guinea pig model of dermatophytosis significantly declined in the 10% and 30% extract and positive control groups in comparison with the untreated control group. Periodic acid-Schiff and hematoxylin/eosin staining displayed similar results. CONCLUSION: The n-BuOH fraction of G. siphonantha demonstrated antidermatophyte efficacy in experimental dermatophytosis.

Nutrient modulation based process engineering strategy for improved butanol production from Clostridium acetobutylicum.

The present study demonstrates a process engineering strategy to achieve high butanol titer and productivity from wild type Clostridium acetobutylicum MTCC 11274. In the first step, two different media were optimized with the objectives of maximizing the biomass and butanol productivity, respectively. In the next step, attributes of these two media compositions were integrated to design a two-stage fed-batch process which resulted in maximal butanol productivity of 0.55 g L-1 h-1 with titer of 13.1 g L-1 . Further, two-stage fed-batch process along with combinatorial use of magnesium limitation and calcium supplementation resulted in the highest butanol titer and productivity of 16.5 g L-1 and 0.59 g L-1 h-1 , respectively. Finally, integration of the process with gas stripping and modulation of feeding duration resulted in a cumulative butanol titer of 54.3 g L-1 and productivity of 0.58 g L-1 h-1 . The strategy opens up possibility of developing a viable butanol bioprocess. © 2019 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2771, 2019.

In vitro antibacterial activity and phyto-chemistry of samples from the roots of Viola pilosa.

Anti-microbial activity and phytochemical analysis of samples from the roots of Viola pilosa was studied against six strains of bacteria. Data indicated that the tested bacterial strain differed in its sensitivity to the root extracts of Viola pilosa. The results showed that Xanthomonas campestris, Bacillus subtilis and S. aureus were more susceptible to butanol extracted fraction. S. aureus was totally resistant to aqueous extracted fraction at all concentrations and .Xanthomonas campestris and Bacillus subtilis were least susceptible to the same fraction at 0.5 mg disc-1. Similarly, ethyl acetate at 2 mg disc-1 concentrations was effective against Pseudomonas aeruginosa and Escherichia coli. Klebsiella pneumoniae was completely resistant to all the tested concentrations. Phytochemical analysis of the different solvent extracted samples suggested the presence or absence of different various metabolites including alkaloids, saponins, tannins, sterols, flavonoids, protein, carbohydrates and fats.

Industrial potato peel as a feedstock for biobutanol production.

Potato peel from a snack factory was assessed as possible feedstock for biobutanol production. This lignocellulosic biomass was subjected to various physicochemical pretreatments (autohydrolysis and hydrolysis with dilute acids, alkalis, organic solvents or surfactants) under different conditions of time, temperature and reagent concentrations, in order to favour the release of sugars and reduce the generation of fermentation inhibitors. Thereafter, the pretreated potato peel was treated enzymatically to complete the hydrolysis. Autohydrolysis at 140 °C and 56 min was the most effective pretreatment, releasing 37.9 ± 2.99 g/L sugars from an aqueous mixture containing 10% (w/w) potato peel (sugar recovery efficiency 55 ± 13%). The fermentability of the hydrolysates was checked with six strains of Clostridium beijerinckii, C. acetobutylicum, C. saccharobutylicum and C. saccaroperbutylacetonicum. C. saccharobutylicum DSM 13864 produced 2.1 g/L acetone, 7.6 g/L butanol and 0.6 g/L ethanol in 96 h (0.186 gB/gS), whereas C. saccharoperbutylacetonicum DSM 2152 generated 1.8 g/L acetone, 8.1 g/L butanol and 1.0 g/L ethanol in 120 h (0.203 gB/gS). Detoxification steps of the hydrolysate before fermentation were not necessary. Potato peel may be an interesting feedstock for biorefineries focused on butanol production.

Evaluation of Antiarthritic Activity of Butanol Fraction of Punica granatum Linn. Rind Extract Against Freund's Complete Adjuvant-Induced Arthritis in Rats.

Available conventional drugs produce several side effects; thus, a potent herbal drug is urgently required for the management of rheumatoid arthritis. The present investigation was performed to evaluate the antiarthritic activity of butanol fraction of Punica granatum Linn. rind methanolic extract (PGBF) against Freund's complete adjuvant (FCA)-induced arthritis in rats. An acute toxicity study of butanol fraction was conducted, accompanied by a study of its antiarthritic activity. Chromatography (thin layer chromatography and high-performance thin layer chromatography) analyses were also conducted. Phytochemical screening of the fraction was performed to confirm the presence of phytoconstituents. For antiarthritic activity, the active butanol fraction was administered at doses of 50 and 75 mg/kg body weight. The antiarthritic activity was evaluated by using biophysical paramaters (arthritic score, body weight, paw volume and joint diameter) and hematological parameters [red blood cell (RBC) count, white blood cell (WBC) count, hemoglobin (Hb) concentration and erythrocyte sedimentation rate (ESR)]. Dexamethasone (5 mg/kg) was selected as the standard. Phytochemical screening of butanol fraction showed the presence of steroids, tannins, flavonoids, irioid glycosides, and phenolic compounds. Acute toxicity studies suggested that butanolic fraction was safe up to a dose of 500 mg/kg. The data regarding biophysical and hematological parameters clearly indicated that the butanol fraction at a 75 mg/kg dose showed a more significant effect (**P<0.01 and ***P<0.001) than the 50 mg/kg dose. The antiarthritic potential of a butanol fraction of Punica granatum Linn. rind extract may be due to the presence of active phytoconstituents such as flavonoids, irioid glycosides, and phenolic compounds. Future studies will provide a new approach in relation to the antiarthritic activity of Punica granatum Linn., and the isolation of its active compound may eventually lead to the development of a new category of the antiarthritic agent.

Studies on the constituents of Helleborus purpurascens: analysis and biological activity of the aqueous and organic extracts.

In Southeast Europe, the ethnomedicinal use of Helleborus species has a very long tradition. Cardiac steroids (Hellebrin), cysteine-rich proteins (Hellethionins) and several steroidal saponins have been identified in these plants. Aim of the present work was to investigate the amino acid composition of native extracts from the root and rootstock of Helleborus purpurascens. The amino acids have been identified by the GC-MS technique on the previously derivatised (Phenomenex Faast Kit) extract samples by comparison with the mass spectra and retention-time of the standards. A remarkable finding was a relatively intensive peak attributed to the non-proteinogenic Pipecolic acid (Pic). A cyclisation of the derivatised glutamine was observed during the GC measurement and a mechanistic pathway is described. Samples of the extract and of some isolated fractions have also been tested on; altogether 12 cancer cell lines aimed to identify further potentially cytostatic components which should be less toxic than Hellebrin. The finding of one Hellebrin-free fraction (IC50 = 0.007 mg/L) with higher cytotoxicity than Hellebrin (IC50 = 0.008 mg/L) is remarkable.