HPLC analysis, genotoxic and antioxidant potential of Achillea millefolium L. and Chaerophyllum villosum Wall ex. Dc.

BACKGROUND: Methanolic and chloroformic extract of Achillea millefolium and Chaerophyllum villosum were evaluated for HPLC analysis, genotoxic and antioxidant potential. MATERIALS AND METHODS: Genotoxic activity was carried out on human blood lymphocytes via comet assay and antioxidant activity was studied through DPPH method. RESULTS: The genotoxic potential of A. millefolium and C. villosum's methanolic and chloroformic extract was analysed using comet assay technique. Comet shaped human lymphocytes cells were observed when treated with different concentrations (50 mg/mL, 75 mg/mL, 100 mg/mL) of methanolic and chloroformic extract of both plants. Reading was taken on the basis of damaged DNA head and tail length. Greater the length of tail as compared to head, greater will be the damage and vice versa. Total comet score was obtained from A. millefolium subjected to different concentrations. After a time interval of 24 h both the extract showed dose dependant genoprotection with maximum genoprotectivity at 98.7 ± 12.7 and 116 ± 5.3 at 50 mg/100 mL for methanolic and chloroformic extract respectively. Similarly Total Comet score was obtained from C. villosum subjected to different concentrations of methanolic and chloroformic extract. After 24 h exhibited dose dependent genoprotection with maximum protectivity at 85.7 ± 22.0 and 101.7 ± 8.6 at 50 mg/100 mL for methanolic and chloroformic extract were determined. The antioxidant activity revealed that methanolic extract of A. millefolium showed highest antioxidant activity (84.21%) at 300 mg/ml after 90 min while the chloroformic extract of C. villosum exhibited highest (68.46%) antioxidant activity (59.69%) at 300 µg/ml after 90 min but less than the standard drug ascorbic acid (88.72%). Quantitative phytochemical screening revealed high percentage of alkaloids (27.4%), Phenols (34.5%), Flavonoids (32.4%) as compared to Tannins (12%) in methanolic extract of A.millefolium. While high percentage of alkaloids (31.4), Phenols (19.3%), Flavonoids (35.5%) as compared to Tannins (16.6%) in chloroformic extract of C. villosum. CONCLUSION: The present results showed that A. millefolium and C. villosum possess a number of important compounds and revealed genoprotective property which may be used to treat several genetic disorders such as alzeimer's disease in future (Grodzicki W, Dziendzikowska K, Antioxidants 9(3):229, 2020).

Micromorphological and phytochemical profiling of Delphinium suave Huth. from Hindukush range, Lower Dir Khyber Pakhtunkhwa Pakistan.

This is a very first attempt to study various parameters of a medicinal plant, Delphinium suave Huth. The plant is erect, geophytic, herbaceous, with tuberous root, trifid in a palmatipartite, strigose cuneate leaf and white spurred zygomorphic flower. The root was isodiametric phellem with single non-glandular trichomes. The stem revealed single-layered cuticle, multiseriate epidermis, cortex, pith ray and uniserate bowed non-glandular trichomes. The leaf was amphistomatic, showed tapering trichomes, prismatic crystals and ranunculaceous stomata with circumference 144.66-182.67 µm. Pollen grains in Light Microscopy (LM), were prolate, spheroidal trizonocolpate, isopolar, radiosymmetric, scabrate, elliptic and monads. Scanning Electron Microscope (SEM) pollen surface was scabrate, monad, size varied from 18.06 to 16.67 µm, colpus to inaperturate, tricolpate, ornamented, echinus, isopolar, isodiametric and circular. SEM roots showed sclerenchymatic tissues, stellate, glandular, non-glandular trichomes and crystals. The stem showed scalariform, pitted vessels, warty protuberances, unicellular, silicified, non-glandular trichomes. Leaves powder revealed, simple, unicellular, tapered headed, uniseriate, sessile, capitate, unbranched glandular, non-glandular, trichomes with crystals. Capitate, stellate, circular, unicellular, branchy trichomes were observed for the first time through SEM. Powder drug study of root, stem leaves through LM revealed different tissues. Preliminary phytochemical revealed alkaloids, anthocyanins, anthraquinones, coumarins, flavones, mucilages, saponins, steroids, terpenoids, volatile oils and proteins. GC/MS showed 36 compounds in roots, 33 in stem while 40 in leaves. Fluorescence analysis of roots, stem and leaves showed variations in color when treated with chemicals. This study will assist pharmacognostic exploration, authentication from adulterants/allied species for consistent quality, resulting in safe use, preservation and efficacy. RESEARCH HIGHLIGHTS: This was first attempt on pharmacognostic study on D. suave Huth. which could be used as a foundation for identifying and authenticating the specie from other allied species by these morphological, anatomical, GC/MS profiling, phytochemical analysis and fluorescence analysis.

Anatomical and histochemical characterization of some highly medicinal plants as a tool for quality control.

Medicinal plants are richest source of chemical compounds that can be used to evolve novel drugs. According to World Health Organization (WHO), in developing countries more than 3.5 billion people relay on herbal drugs for their primary health care. In the present study, an attempt was carried out to authenticate some selected medicinal plants (Fagonia cretica L., Peganum harmala L., Tribulus terrestris L., Chrozophora tinctoria L. Raf. and Ricinus communis L.) from family Zygophyllaceae and Euphorbiaceae using light and scanning electron macroscopic techniques. Macroscopic evaluation and comparative anatomy (Light Microscopy) of the root and fruits revealed great diversity in macro and microscopic features. Scanning electron microscopy (SEM) of root powder showed non-glandular trichomes, stellate trichomes, parenchyma cells and vessels. Fruits SEM exhibited non-glandular trichomes, glandular trichomes, stellate trichomes, peltate trichomes and mesocarp cells. Both macroscopic and microscopic evaluation plays a crucial role in the correct substantiation and validation of novel sources. These findings can serve as an essential source of information to accomplish the authenticity and to appraise the quality and purity of the herbal drugs in accordance to WHO guidelines. These parameters can be used to distinguish the selected plants from their commonly used adulterants. RESEARCH HIGHLIGHTS: In the current work, five plants (Fagonia cretica L., Peganum harmala L., Tribulus terrestris L., Chrozophora tinctoria L. Raf. and Ricinus communis L.) from Zygophyllaceae and Euphorbiaceae were investigated for macroscopy and microscopy (LM & SEM) for the first time. Macroscopic and microscopic evaluation disclosed great diversity in morphology and histology. Microscopy is the backbone of standardization process. Current study helped in correct identification and quality assurance of the plant materials. Their statistical investigation may possess great potency for plant taxonomists to further appraise the vegetative growth and tissue development which is requisite especially for fruit to enhance the yield of herbal drugs and their formulation. Further molecular studies, compounds isolation and characterization are required to deepen the knowledge of these herbal drugs.

Carduus edelbergii Rech. f. Mediated Fabrication of Gold Nanoparticles; Characterization and Evaluation of Antimicrobial, Antioxidant and Antidiabetic Potency of the Synthesized AuNPs.

Background: Due to the high expense, less effectiveness and more side effects of available synthetic medicine, the researchers and communities are focusing on phyto-based natural bioactive compounds, which are considered safer for the treatment of syndromes and chronic diseases. Aim: The current project was aimed to determine the phytochemicals constituents available in the aerial parts of methanol extract of Carduus edelbergii via GC-MS, fabrication of AuNPs mediated with the mentioned extract; characterization and evaluation of antimicrobial, antioxidant and antidiabetic potency of the synthesized AuNPs. Methods: Confirmation of green synthesis of AuNPs, functional groups responsible for the reduction in Au+, size and crystallinity, morphology and quantity of gold (Au) were carried out by Ultraviolet-Visible (UV-Vis) spectroscopy, Transform Infrared (FTIR) spectroscopy, Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD) and dispersive X-ray (EDX), respectively, whereas in vitro antioxidant characteristics were assessed by DPPH and ABTS assays. Wistar albino rats were used to test the anti-diabetic properties of the methanol extract and AuNPs. Results: GC-MS revealed that the diluted methanol extract of Carduus edelbergii consists of about 19 chemical constituents. Among the identified compounds, the 13-Docosenoic acid, methyl ester, (Z)­has the highest concentration (38.16%), followed by 9-Octadecenoic acid, methyl ester, (E)­(15.72%) and n-Hexadecanoic acid (15.07%). Methanol extract and its fabricated nanoparticles showed significant antioxidant and antimicrobial activities. In vivo antidiabetic study revealed a noteworthy (p < 0.05) decline in body weight and HDL and elevated concentration of blood glucose, bilirubin, creatinine, urea, triglyceride, VLDL, LDL, ALP, ALT and AST in diabetic control. The said changes were recovered significantly (p < 0.05) by treatment of diabetic rats with methanol extract (150 and 300 mg/Kg BW) and AuNPs of Carduus edelbergii (5 and 10 mg/Kg BW). Conclusion: The green synthesized AuNPs exhibit significant antioxidant, antimicrobial and antidiabetic characteristics.

Microscopic and pharmacognostic standardization of Astragalus scorpiurus Bunge.

The aim of the current study was to assess and compare the microscopic and pharmacognostic features of the selected parts of Astragalus scorpiurus Bunge. It is an annual and prostrate herb. Anatomy of leaves, stem, and root depicted a typical dicot structure. It will provide information regarding standardization and authentication. The powder drug study (Light Microscopy and Scanning Electron Microscopy) revealed the presence of xylem and phloem vessels, epidermal fragments, cortical cells, fibers, parenchyma tissue, calcium oxalate crystals, nonglandular trichomes, anomocytic stomata, and so on. It will be helpful in identification and quality control. Pollen and seeds micro-morphological features were observed through LM and SEM. These will play a vital role in the taxonomical identification of taxa and provide satisfactory information for taxonomists. Fluorescence analysis revealed different colors under visible and UV light. Extractive values were determined. Powder of the plant parts revealed high extraction in methanol followed by ethanol. Elemental and nutritional profiling were evaluated during two phenological periods that is, vegetative and postflowering period. There were significant phenological variations in mineral and nutritional status of the herb. These parameters are crucial for drug prescription and drug designing. These also provided information regarding nutraceutical and therapeutic value of this herb.

Micromorphology, pharmacognosy, and bio-elemental analysis of an important medicinal herb: Verbascum thapsus L.

The present study was conducted on characterization of morpho-anatomical, phytochemical, and bio-elemental analysis of root, stem, and leaf of Verbascum thapsus. Morphologically Verbascum is a biennial plant that flowers for a month and a half in mid- to late summer. Various organoleptic features of root, leaf, and stem were recorded. Anatomically the T. S of the root, stem, and leaf showed a typical dicot histological differentiation. Leaf possessed anomocytic stomata, crescent shape vascular bundles, and covered with long and stellate type trichomes while, stem contained collateral type of vascular bundles and a well-developed pith to store phytochemicals responsible for various pharmacological activities. The powder drug study through scanning electron microscopy revealed the presence of various types of tissues. Branched, tree like and stellate trichomes in root and leaf help in absorption and reduce loss of water. These anatomical features are responsible for the survival of the plant as biennial. Four macro elements (Na, K, Ca, and Mg) and seven microelements (Cr, Cu, Fe, Mn, Ni, Zn, and Cd) and their concentrations in ppm were also studied using Atomic Absorption Spectroscopy. Phytochemical screening of methanolic extract showed existence of various secondary metabolites, while mucilage and anthraquinones was not detected. The present study helps to understand the taxonomic identification of the plant based on morpho-anatomical features and throws the attention of the researchers to carry out the work for developing its various formulations, which can ultimately be beneficial for the human beings as well as animals.

Quantitative ethnobotanical appraisal of medicinal plants used by inhabitants of lower Kurram, Kurram agency, Pakistan.

OBJECTIVE: Medicinal plants are used for treatment of ailments throughout rural and urban areas of the world. Such use of plants varies from one region to another and is measured using quantitative techniques. The current research which was conducted from March to October 2015, is the first explorative study of medicinal plants used by inhabitants of lower Kurram, Kurram agency, Pakistan. MATERIALS AND METHODS: Field trips were done to 20 location of lower Kurram and information regarding medicinal use of plants was collected from the locals through semi-structured interviews. RESULTS: A total of 52 plant species that were reported by the people from the region, to have medicinal value, fall within 48 genera and 35 families. The family Asteraceae comprised most of these herbs (6 species) followed by the family Lamiaceae (4 species) and Solanaceae (3 species). Leaves (24.3%) and fruits (21.6%) were the frequently used parts in preparation of remedies. The reported plants were used for treatment of 50 ailments with most of these plants (35 species; 30.97%) being used for treatment of digestive problems. Seriphidium kurramensis had the highest relative frequency of citation (66.18) and use value (1.10). Fidelity level of Caralluma tuberculata and Artemisia scoparia for diabetes was (61.22) and (55.56), respectively. The highest fidelity level for malaria was reported for Artemisia absinthium (43.66) and S. kurramensis (40.00). CONCLUSION: The inhabitants of lower Kurram still practice medicinal plants and few of the plants were used for treatment of fatal diseases like malaria, hepatitis and blood cancer. Haphazard cutting of plants and overgrazing are major threats that can affect plants biodiversity.

Review - Glycyrrhiza glabra L. (Liquorice).

Medicinal plants are being used for treating various diseases. According to World Health Organization 80% of the world population depends on indigenous medicinal plant remedies. Herbal medicine employs fruits, vegetables, as dry materials or their extracts for the treatment of different diseases and health maintenance. Glycyrrhiza glabra (Liquorice) has been used in Europe since prehistoric times. It is well documented in written form starting with the ancient Greeks. Glycyrrhizin is the major active constituent obtained from liquorice roots, one of the most widely used in herbal preparations for the treatment of liver complaints. The plant is used as anti-inflammatory, spasmolytic, laxative, anti-depressive, anti-ulcer and anti-diabetic. The present review focuses Glycyrrhiza glabra distribution, ethno botany, ethno pharmacology, chemical constituents, medicinal uses, cultivation and trade. Plant requires a lot of attention as it has been reduced in population due to over-use in Baluchistan. The plant conservationists should consider this herb as priority species and should start its cultivation on the commercial scale to fulfill the requirements of the local markets and pharmaceutical industries as well as reduce the pressure on the wild plants.

Elemental, nutritional, phytochemical and biological evaluation of Hypericum perforatum Linn.

This study was carried out to study elemental, nutritional, phytochemical and biological evaluation of Hypericum perforatum collected from Swat in 2010. The elemental analysis showed that Ca was highest (5600 µg/g) in leaves and lowest (2500 µg/g) in flowers. The potassium was highest (840 µg/g) in fruit and lowest (80 µg/g) in leaves. Magnesium was highest (260 µg/g) in stem and lowest (200 µg/g) in flowers. Sodium was highest (4900 µg/g) in stem and lowest (4700 µg/g) in leaves and flowers. Copper was highest (26 µg/g) in stem and lowest (10 µg/g) in leaves. Iron was highest (5000 µg/g) in flowers lowest (1200 µg/g) in stem. Zinc was highest (80 µg/g) in flowers and lowest (46 µg/g) in stem. Nickle, cadmium and Cobalt were <5 µg/g for all plant parts. The nutritional analysis showed that the dry matter was in the range of (97.61%) in stem and (96.38%) in leaf, ash (5.43%) in flowers and (1.90%) in stem, crude protein (12.63%) in leaf and (6.15%) in stem, crude fibre (64.74%) in flowers and (13.0%) in leaf, ether extract (10.98%) in fruit and (1.88%) in stem and nitrogen free extract was (65.80%) in leaf and (10.98%) in flower, respectively. Hypericum perforatum did not show cytotoxic, insecticidal and antibacterial activity in vitro at different doses. The % activity was zero% in cytotoxic and insecticidal activities. However, H. perforatum plant parts revealed phytotoxic activity. The phytotoxic activity of leaf and fruit remained same (44.0%) at highest dose (500 µg/ml). The phytochemical screening showed the presence of mucilage, tannins, anthraquinones, saponins, fats and oils and proteins in all parts of the plant. Calcium oxalate was found in all parts except the fruit. Lignin and catechin was found in all parts except the leaf. Cutin was found only in stem and flower while chlorophyll was found only in stem and leaf. In various localities (Shartangaar, Panj Pali and Sharanko) of Swat fresh leaves were used while in Barani and Jaba fresh as well as dried leaves were used as stimulant, in fever, cough, diphtheria and as an anthelmintic only in Shartangaar.

Cytotoxic activity of plants of family zygophyllaceae and euphorbiaceae.

The methanolic and n-hexane extracts of studied plants showed significant toxicity to brine shrimps. The methanolic extract of Fagonia cretica had highest LD50 (117.72) value, while Peganum harmala showed low LD50 value (41.70) compared to n-hexane extract. The methanolic and n-hexane extracts of Tribulus terrestris showed similar LD50 values. The methanolic extract of Chrozophora tinctoria showed low LD50 value than the n-hexane extract. The methanolic extract of Ricinus communis showed highest LD50 value while the n-hexane extract showed lowest LD50 value. The LD50 value less than 100 was obtained for n-hexane extracts of Fagonia cretica, Peganum harmala and Ricinus communis. The n-hexane extracts of these plants also showed the highest toxicity as compare to methanolic extracts. The chemical constituents detected in the present investigation might be responsible for cytotoxic activity.

Ethnobotanical and antimicrobial study of some selected medicinal plants used in Khyber Pakhtunkhwa (KPK) as a potential source to cure infectious diseases.

BACKGROUND: Present investigation deals with antimicrobial screening of ten medicinally important plants used by the inhabitants of district Haripur, Khyber Pakhtunkhwa (KPK) for different infectious diseases. METHODS: Aqueous, n-hexane and ethanolic extracts of each plant were tested for their antimicrobial activity against both Gram positive and Gram negative strains of bacteria, as well as strain of yeast. Agar well diffusion and broth dilution methods were used to determine the antimicrobial activity of different plant extracts. RESULTS: The results indicated that all plants exhibited antimicrobial activity against one or more test pathogens. Interestingly, extracts of three plants showed strong and broad spectrum activity as compared to rest of the extracts which demonstrated the moderate activity. On the whole ethanolic extracts exhibited maximum antimicrobial effect than their corresponding aqueous and n-hexane extracts, when compared with standard antibiotics i.e., Streptomycin and Tetracycline. Among various extracts, only ethanolic extract of Azadirachta indica and aqueous and ethanolic extracts of Eucalyptus globulus and Bergenia ciliata and ethanolic extract of Punica granatum were found to have potentially promising activity against test microorganisms. CONCLUSION: Different plant extracts show promising antimicrobial activity justifying their usage in traditional medicines. This study will be continued to identify more plants with potential antimicrobial components.

Effect of cold water extracts of Acacia modesta Wall. and Glycyrrhiza glabra Linn. on Tribolium castaneum and Lemna minor.

The aim of the present study was to introduce an alternative way for insects control through biodegradable plants materials. The different cold water extracts dilutions of Acacia modesta and Glycyrrhiza glabra were tested against Tribolium castaneum. The extracts dilutions of both plants caused mortality of the Tribolium castaneum. ANOVA revealed that dilutions and plants were highly significant. The interaction between plants and dilutions was also significant at P < 0.05. Phytotoxic activity showed that dilutions of Acacia modesta and Glycyrrhiza glabra extracts significantly inhibited the growth of Lemna minor. ANOVA showed that dilutions of both plants extracts were significant at P < 0.05.