Mycotoxin detection in selected medicinal plants using chromatographic techniques.

Mycotoxins are toxic mycological products that when consumed, absorbed or inhaled cause sickness or even the death of humans. Therefore, the present study aimed to evaluate the contamination levels of mycotoxins (aflatoxins, AFB1 , AFB2 , AFG1 , AFG2 , and ochratoxin A, OTA) in selected medicinal herbs and shrubs using thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). A total of 15 samples of medicinal herbs and shrubs were selected. Among them, four samples were aflatoxin contaminated while two samples were ochratoxin A contaminated. The highest level of aflatoxin was detected in Justicia adhathoda (4,704.94 ppb) through HPLC (153.4 ppb) and through TLC, while the lowest level of aflatoxin was detected in Pegnum harmala (205.1 ppb) through HPLC. Similarly, the highest level of OTA was detected in Dodonia viscosa (0.53 ppb) through HPLC (0.5 ppb) and through TLC, while the lowest level was detected in J. adhathoda (O.11 ppb) through HPLC (0.4 ppb) and through TLC. The OTA concentration was very low, being negligible and below permissible limits. The present study concludes that there is a potential risk for the consumption of herbal decoctions. Therefore, regular monitoring and proper management of mycotoxins, including aflatoxins and OTA, in herbal medicines are needed to ensure the safety of herbal drugs to protect consumers.

Chromatographic analysis and antioxidant potency of the crude extract of Xanthium spinosum in various fractions.

Pharmacology experts place a high priority on therapeutic plants because the majority of pharmaceutical firms rely on medicinal plants as raw ingredients. Therefore, the potential bioactive components using gas chromatography-mass spectrometry analysis and antioxidant effects using DPPH free radical scavenging activity of various crude fractions of Xanthium spinosum were assessed. Gas chromatography-mass spectrometry analysis showed the presence of various bioactive compounds including benzenedicarboxylic acid (18.60%), 8-octadecenoic acid (4.86%), 11-octadecenoic acid and 10-octadecenoic acid in the crude methanolic extract, 1,2-benzenedicarboxylic acid, diisooctyl ester (14.42%), 1,2-benzenedicarboxylic acid, mono (2-ethylhexyl) ester (14.42%), 6-octadecenoic acid, methyl ester (7.56%), 8-octadecenoic acid, methyl ester (7.56%), 10-octadecenoic acid, methyl ester (7.56%) and hexadecanoic acid, methyl ester (6.55%) in the n-hexane extract, ethanal, 2-methyl-2-[4-(1-methylethyl)phenyl]-(3.02%), (+)-3-carene, 4-isopropenyl-(3.02%), 7H-indeno[5,6-b] furan-7-one, 4,4a,5,6,7a,8-hexahydro- (3.02%) and 2-[5-(2,2-dimethyl-6-methylene-cyclohexyl)-3-methyl-pent-2-enyl]-[1,4] benzoquinone (2.79%) in the chloroform extract and 1,2-benzenedicarboxylic acid, mono (2-ethylhexyl) ester (33.005%), 1,2-benzenedicarboxylic acid, diisooctyl ester (33.005%) and bis(2-ethylhexyl) phthalate (33.005%) in the ethyl acetate extract. Significant DPPH radical scavenging activity was exhibited by the chloroform fraction (43.37-88.65%) at all doses followed by the crude methanolic extract (36.02-83.75%) at all doses. In conclusion, different crude fractions of X. spinosum can be considered a rich source of pharmacologically active components that can be scoped for isolation and may be subjected to in-depth pharmacological study.

Pharmacological Activities of Rhododendron afghanicum; an Endemic Species of Khyber Pakhtunkhwa, Pakistan.

Majority of different kinds of metabolites having therapeutic characteristics are thought to be stored in medicinal plants. So, the present study was aimed to explore the crude extract of leaves and stem of R. afghanicum for phytochemical screening and various pharmacological activities. Toxicological studies at 100 mg/kg showed 60 % mortality where its safe dose level was 90 mg/kg. Phytochemical screening revealed the presence of alkaloids, glycosides, flavonoids and tannins in both extracts. Bacterial strains were susceptible to (RLEt) and (RLM) crude extracts except Staphylococcus aureus. RSM showed maximum anti-inflammatory activity (20.16 %) followed by RSEt (20.14 %) where lowest activity was displayed by RLEt (18.46 %). Phytotoxic activity showed a substantial dose-dependent phyto-inhibition of Lemna minor. An outstanding cytotoxic potential was displayed with LD50 values of 9.46 and 13.03 µg/ml in both stem extracts. RLEt demonstrated a dose-dependent pain relief at 30, 60 and 90 mg/kg which was 31 %, 40 % and 52 % respectively. A considerable spasmolytic action was observed by the shrinkage of jejunum muscle in albino mice. RLEt at 1000 ppm showed (17 mm) and RLM at 1000 ppm showed (16 mm) zone of inhibition against Aspergillus niger. These findings support and corroborate the traditional applications of R. afghanicum for treating digestive, analgesic and inflammatory ailments.

Combined application of hot water treatment and eucalyptus leaf extract postpones senescence in harvested green chilies by conserving their antioxidants: a sustainable approach.

BACKGROUND: Green chili is the predominant vegetable in tropical and subtropical regions with high economic value. However, after harvest, it exhibits vigorous metabolic activities due to the high moisture level, leading to a reduction in bioactive compounds and hence reduced shelf life and nutritional quality. Low temperature storage results in the onset of chilling injury symptoms. Therefore, developing techniques to increase the shelf life of green chilies and safeguard their nutritional value has become a serious concern for researchers. In this regard, an experiment was conducted to evaluate the impact of the alone or combined application of hot water treatment (HWT) (45 °C for 15 min) and eucalyptus leaf extract (ELE) (30%) on 'Golden Hot' chilies in comparison to the control. After treatment, chilies were stored at 20 ± 1.5 °C for 20 days. RESULTS: HWT + ELE-treated chilies had a significant reduction in fruit weight loss (14.6%), fungal decay index (35%), red chili percentage (41.2%), soluble solid content (42.9%), ripening index (48.9%), and reactive oxygen species production like H2O2 (55.1%) and O-2 (46.5%) during shelf in comparison to control, followed by the alone application of HWT and ELE. Furthermore, the combined use of HWT and ELE effectively improved the antioxidative properties of stored chilies including DPPH radical scavenging activities (54.6%), ascorbic acid content (28.4%), phenolic content (31.8%), as well as the enzyme activities of POD (103%), CAT (128%), SOD (26.5%), and APX (43.8%) in comparison to the control. Additionally, the green chilies underwent HWT + ELE treatment also exhibited higher chlorophyll levels (100%) and general appearance (79.6%) with reduced anthocyanin content (40.8%) and wrinkling (43%), leading to a higher marketable fruit (41.3%) than the control. CONCLUSION: The pre-storage application of HWT and ELE could be used as an antimicrobial, non-chemical, non-toxic, and eco-friendly treatment for preserving the postharvest quality of green chilies at ambient temperature (20 ± 1.5 °C).

Postharvest starch and sugars adjustment in potato tubers of wide-ranging dormancy genotypes subjected to various sprout forcing techniques.

The development of an efficient, safe, and environment-friendly technique to terminate tuber dormancy in potatoes (Solanum tuberosum L.) is of great concern due to the immense scope of multiple cropping all over the globe. The breakage of tuber dormancy has been associated with numerous physiological changes, including a decline in the level of starch and an increase in the levels of sugars during storage of freshly harvested seed potatoes, although their consistency across genotypes and various dormancy-breaking techniques have not yet been fully elucidated. The purpose of the present research is to assess the efficacy of four different dormancy-breaking techniques, such as soaking in 90, 60, or 30 mg L-1 solutions of benzyl amino purine (BAP) and 30, 20, or 10 mg L-1 gibberellic acid (GA3) alone and in the combination of optimized concentrations; cold pre-treatment at 6, 4, or 2 °C; electric shock at 80, 60, 40, or 20 Vs; and irradiation at 3.5, 3, 2.5, 2, 1.5, or 1 kGy on the tuber dormancy period and sprout length of six genotypes. Furthermore, the changes that occurred in tuber weight and endogenous starch, sucrose, fructose, and glucose contents in experimental genotypes following the application of these techniques were also examined. Overall, the most effective technique to terminate tuber dormancy and hasten spout growth was the combined application of BAP and GA3, which reduced the length of dormancy by 9.6 days compared to the untreated control, following 6.7 days of electric current, 4.4 days of cold pre-treatment, and finally irradiation (3.3 days). The 60 mg L-1 solution of BAP greatly reduced the dormancy period in all genotypes but did not affect the sprout length at all. The genotypes showed a weak negative correlation (r = - 0.4) (P < 0.05) of endogenous starch contents with dormancy breakage and weight loss or a moderate (r = - 0.5) correlation with sprout length, but a strong positive correlation (r = 0.8) of tuber glucose, fructose, and sucrose contents with dormancy breakage and weight loss. During 3 weeks of storage, sprouting commencement and significant weight loss occurred as tuber dormancy advanced towards breakage due to a reduction in starch and an increase in the sucrose, fructose, and glucose contents of the tubers. These findings could be advantageous for postponing or accelerating seed potato storage as well as investigating related physiological research in the future.

Isolation and characterization of peroxidase P7-like gene and Rab-GDI like gene from potential medicinal plants: A step toward understanding cell defense signaling.

Defensin genes form part of a plant's defense system and are activated when exposed to biotic or abiotic stress. They play a vital role in controlling many signaling pathways involved in various plant defense mechanisms. This research aimed to isolate and characterize novel defensin genes from selected medicinally important plants to explore their signaling mechanisms and defense associated roles for breeding. The DNA of Albizia lebbeck and Moringa oleifera was subjected to PCR amplification using gene-specific primers of defensin genes. Two novel defensin genes were isolated in each species, with sequence lengths of 300 bp in A. lebbeck and 150 bp in M. oleifera. In-silico analysis undertaken to retrieve and align their orthologous sequences revealed 100% similarity of the A. lebbeck gene with the Musa acuminate peroxidase P7-like gene and 85% similarity of the M. oleifera gene with the Manihot esculenta GDP dissociation inhibitor gene. The reliability, stability and physiochemical properties of homology models of these sequences was confirmed through online computational studies. This preliminary study confirmed the presence of novel genes with peroxidase P7 and Rab GDP dissociation inhibitor gene-like activity in A. lebbeck and M. oleifera, respectively, and their potential defense role in plants. Thus, the defensin genes of both species could be used in the synthesis of transgenic self-defensive plants with increased disease resistance and as potential candidates for improved crop production and thraputic formulation in the future.

Pharmacognostic standardization of Xanthium spinosum L. through scanning electron microscopy and analytical techniques.

The present study was intended to evaluate the pharmacognostic attributes including macroscopy, microscopy, physico-chemical and phytochemical screening of the aerial part of Xanthium spinosum, belonging to family Asteraceae. Morphologically, the leaf was covered with sparsely scattered non-glandular, multicellular hair on adaxial surface, while abaxial surface was reported with dense silvery furs. Leaf showed lamphystomatic nature bearing anomocytic stomata. Various other leaf constant values like vein islets number, vein termination number and palisade cell ratio were also observed and recorded as 4.36 ± 0.5, 3.36 ± 0.5, and 11.82 ± 2.04, respectively. Cross section of the stem showed typical dicot anatomy differentiated into a single epidermal layer followed by few rows of collenchyma cells, thin-walled parenchyma cells followed by closed and collateral vascular bundles having prephloemic sheaths and a central pith. Light and scanning electron microscopy of the powdered drug revealed a variety of tissues and cells fragments having different measurements. Fluorescence analysis of the fine powder treated with different reagents revealed a variety of colors implying as a sign for the presence of various biochemical constituents. Highest and lowest percent extractive value was found in methanol and distilled water which were 29.10% and 9.80%, respectively. Qualitative phytochemical screening revealed the presence of various secondary metabolites like flavonoids, phenols, tannins, glycosides, phytosterols, glycosides, saponins, and coumarins, while quantitative phytochemical screening detected maximum quantity of flavonoids in chloroform followed by methanol and ethyl acetate. The aforementioned studies have been reported first time for the said species, which may be helpful for the standardization of the current taxon.

Modulating response of Zea mays to induced salinity stress through application of nitrate mediated silver nanoparticles and indole acetic acid.

Nanotechnology has been amplified in different areas of science as well as agriculture in the present era. So, the present work was designed to evaluate the result of nitrate mediated silver nanoparticles (Nit-AgNPs) and indole acetic acid (IAA) on physio-biochemical features of the selected maize variety (Pahari white) under 40 and 80 mM salinity induction. Seeds were propagated in triplicates in earthen pots (18 cm inferior and superior inside diameter, 20 cm stature, and 2 cm breadth) filled with silt and soil (1:2) having 3.09-5.12 Electrical conductivity (EC), 6.8-7.3 pH, and 4-16% moisture contents. Scanning electron microscopy results showed the average particle size around 90 nm indicating a high surface area suitable for adsorption properties, agglomerated, roughly spherical, and were uniformly dispersed. Elemental quantification of biosynthesized AgNPs analyzed via energy dispersive X-ray spectroscopy showed a strong peak at 3.0 KeV along with the presence of elements K, N, O, and C. Results of Thermo-gravimetric Analysis (TGA)/Differential Thermal Analysis (DTA) showed endothermic major decline at 150-300°C, while exothermic peak at 300-400°C. The growth responses at 40 mM salinity concentration have been reduced representing from the least boundary of chlorophyll "a," "b," and peroxidase content, whereas; this adverse effect has been reduced by operation of Nit-AgNPs as separate treatment and in combination with IAA. From the current study, it has been concluded that salinity concentration at 80 mM adversely affected the values of osmolytes, protein, and superoxide dismutase whereas the maximum amplitude of proline reduced by the application of Nit-AgNPs as distinct treatment indicating that the plant behaves normal with the combined application of nanoparticles and IAA.

Phytochemical and pharmacognostic studies of Buddleja asiatica leaves.

Buddleja asiatica commonly known as "Booi" is a large deciduous shrub belongs to family Scrophulariaceae, traditionally used as antipyretic, analgesic, hypotensive, antimicrobial, anti-inflammatory, and antifungal agent recorded from essential oil obtained from leaves. The literature revealed that the plant has been widely used for many other purposes. The said plant has been analyzed through pharmacognostic techniques such as morphological, microscopic, and physio-chemical evaluations to keep the originality of the plant and to avoid adulteration. Morphologically, the plant is evergreen shrub, while organoleptic studies showed that the leaf has glabrous upper surface and tomentose lower surface, with slightly bitter taste and aromatic odor. Anatomically, the leaf showed typical dicot histological differentiation with hypostomatic nature having highest frequency (90%) of actinocytic stomata. The average stomatal number and stomatal index were 336 ± 39.5 and 30.01 ± 2.34, respectively. The palisade cell ratio, vein termination, and vein islet number were 9.2 ± 0.72, 10.2 ± 3.1, and 10.3 ± 3.3, respectively. Various tissue fragments have been observed during powdered drug analysis of the leaf. Preliminary phytochemical screening confirmed the presence of proteins, phenols, alkaloids, saponins, flavonoids, tannins, and glycosides. Fluorescence analysis in ordinary day light and under UV light along with extractive values was also analyzed. The above-mentioned studies that have been reported, for the first time, for the said plant may be significant to establish the pharmacognostic and phytochemical standards of the said species.

Marginal quality water arbitrated essential oil contents in metal hoarded flower petals of scented roses.

Climate change and the consequent alteration in agricultural circumstances enhance the susceptibility of fresh water use particularly in water-scarce regions. Marginal quality water reuse is a common alternative practice but possible perils of metal accretion in plant parts are mostly ignored. The present research aimed to probe the impact of treated wastewater (TWW) and untreated wastewater (UTWW) on metal accumulation in flower petals and their influence on essential oil contents of fragrant Rosa species (R. Gruss-an-teplitz, R. bourboniana, R. centifolia, R. damascena) in a peri-urban area of Faisalabad, Pakistan during January, 2017 to December, 2018. The mineral and chemical contents in canal water (CW) and TWW were less than recommended levels of national environmental quality standards (NEQS) for wastewater of Pakistan. The experimentally UTWW possessed higher electrical conductivity (EC), biological and chemical oxygen demand (BOD and COD), and some metals (Pb, Co, Cr) that were above the permissible levels. The experimental data revealed that except Cr other metals contents in the flower petals were less than the WHO recommended limits (for medicinal plants) under experimental irrigation regimes. Rosa centifolia and R. damascena possessed higher metal i.e. Zn, Cu, Pb, Cr, Co contents while Fe and Ni contents were higher in R. Gruss-an-Teplitz and R. bourboniana respectively. There were twelve constituents which were detected in essential oil by gas chromatography. Major constituents were phenyl ethyl alcohol, citronellol, geranyl acetate, γ- undelactone, methyl eugenol, and limonene whose share was 48.17%, 41.11%, 8.46%, 4.82%, 4.44%, and 4.15% respectively whereas concentrations of other 06 constituents were less than 3.7%. Phenyl ethyl alcohol, lion shared constituent of essential oil was found highest (48.17%) in R. Gruss-an-Teplitz whereas minimum level was recorded in R. damascena (28.84%) under CW. In contrast, citronellol (chief component of fragrance) was highest in R. damascena (41.11%) in UTWW while the lowest level was found in R. Gruss-an-Teplitz (17.41%) in CW. This study confirmed the variations in metal concentrations of Rosa species due to different absorbability of each metal in flower petals. It also indicates that wastewater did not affect the composition but there were quantitative differences in aroma constituents and chemical composition of essential oil.