Natural antioxidants present in several medicinal plants are responsible for inhibiting the harmful effects of oxidative stress. These plants contain polyphenols and flavonoids that act as free radical scavengers and reduce oxidative stress and may be an alternative remedy to cure various harmful human diseases. This study aims to quantify the total phenolic and flavonoid contents (TPC and TFC) and antioxidant properties of methanol extracts of fruits, seeds, and bark of an important medicinal and aromatic plant, Zanthoxylum armatum collected from wild and cultivated populations in Nepal. TPC was determined by Folin–Ciocalteu colorimetric method using gallic acid as standard, and various concentrations of the extract solutions were measured at 760 nm. TFC was calculated by aluminum chloride colorimetric assay. Quercetin was used as standard, and the absorbance was measured at 510 nm. The antioxidant potential of the different extracts was estimated by DPPH free radical scavenging assay, and the absorbance was measured at 517 nm. The highest TPC value was 226.3 ± 1.14 mg GAE/g in wild fruits, and the lowest was 137.72 ± 4.21 mg GAE/g in cultivated seeds. Similarly, the highest TFC value was 135.17 ± 2.02 mg QE/g in cultivated fruits, and the lowest was 76.58 ± 4.18 mg QE/g in cultivated seeds. The extracts showed variable antioxidant properties. The fruits exhibited excellent antioxidant properties with IC50 values of 40.62 μg/mL and 45.62 μg/mL for cultivated and wild fruits, respectively. Similarly, the IC50 values of the bark were 63.39 μg/mL and 67.82 μg/mL, respectively, for cultivated and wild samples. And the least antioxidant capacity was shown by the seeds extract with IC50 values of 86.75 μg/mL and 94.49 μg/mL for wild and cultivated seeds, respectively. The IC50 value of the standard ascorbic acid was 36.22 μg/mL. Different extracts of Z. armatum contain considerable amount of phenols and flavonoids, including antioxidant properties, suggesting the potential use of this species in pharmacy and phytotherapy as a source of natural antioxidants.

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Variation in the composition of leaf essential oils of Zanthoxylum armatum in relation to altitude and soil chemistry was analyzed in Nepal. The essential oil was extracted by Clevenger apparatus and the components were analyzed through GC-MS. The results showed that: Yield of the essential oil obtained from the hydro-distillation of dried leaves ranged from 0.16% to 0.50%. GC-MS analysis revealed total of 17 compounds in the essential oil from the dried leaves of Z. armatum from different altitudes and populations (wild and cultivated). The three major components, linalool, limonene and undecan-2-one, present in higher proportion in all the samples were analyzed. Other components tridecan-2-one, myrcene, cinnamate(E)-methyl and alpha-bergamotene were also identified in most of the samples but in lower proportions. The highest number of components (12) was identified from the wild samples collected from 1990 m and the lowest number (7) was from 1060 m (wild) and 2030 m (cultivated populations). Linalool, a major component was found in higher amount (64.88%) at the lowest altitude of 1060 m. Results of the present study showed that the altitude, soil chemistry, and growing conditions significantly affect the essential oil components in Zanthoxylum armatum.

Crude methanol extracts of fruits, seeds, and bark of Zanthoxylum armatum were investigated in vitro for antimicrobial activities against 9 different bacterial strains: Bacillus subtilis, Enterococcus faecalis, Proteus vulgaris, Pseudomonas aeruginosa, Salmonella typhi, Shigella dysenteriae, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus epidermidis using agar well diffusion method, and the MBC values were determined. Only 5 bacteria, i.e., Bacillus subtilis, Enterococcus faecalis, MRSA, Staphylococcus aureus, and Staphylococcus epidermidis exhibited antibacterial properties against the different extracts. The fruit and seed extracts showed activities against 5 bacteria, while the bark extract was active against 2 bacteria only (MRSA and Staphylococcus aureus). Staphylococcus aureus was found to be more susceptible for all the extracts compared to other strains. The maximum ZOI of 20.72 mm was produced by fruits (wild) and 18.10 mm (cultivated) against Staphylococcus aureus. Likewise, the least ZOI of 11.73 mm was produced by seeds (wild) and 11.29 mm (cultivated) against Escherichia faecalis. Similarly, the lowest MBC value of 0.78 mg/mL was obtained for fruit extracts against MRSA, 1.56 mg/mL for fruits, seeds, and bark extracts against Bacillus subtilis, MRSA, and Staphylococcus aureus, and highest value of 50 mg/mL for fruits and seeds extracts against S. epidermidis. The fruits, seeds, and bark extracts of Z. armatum exhibited remarkable antibacterial properties against different pathogenic bacteria causing several diseases, which suggests the potential use of this plant for treating different bacterial diseases such as skin infection, urinary tract infection, dental problems, diarrhea, and dysentery.

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The common method of propagation is through seeds but seed germination in Zanthoxylum armatum is very low due to the presence of hard seed coat, which might be a great hurdle for large scale production of plantlets. So an attempt was made in this study to see the effect of different growth hormones, their concentrations and different rooting media on the rooting and sprouting of Z. armatum. The stem cuttings of Z. armatum were treated with two types of auxins namely Indole-3-Butyric Acid (IBA) and Naphthalene Acetic Acid (NAA) at different concentrations (2000 ppm, 3000 ppm and 5000 ppm), while the untreated cuttings were used as control. The cuttings were planted in three different rooting media: sand, neopeat and mix (containing a mixture of sand, soil and vermin-compost). The completely randomized design was used for the experiment. The total number of stem cuttings of Z. armatum used in the experiment was 1080 for 18 treatments in three replicates (20 cuttings per treatment x 18 treatments x 3 replicates). The experiment was set up in controlled greenhouse conditions at Dabur Nepal Private Limited Nursery, Banepa, Kavre District. The parameters evaluated were root length, shoot length and number of roots per cutting. The collected data were analyzed statistically using R-program with Agricola. Least significant difference (LSD) and Duncan multiple Range Test (DMRT), as mean separation technique was applied to identify the most efficient treatment in the rooting and shooting behavior of Z. armatum (Gomez and Gomez, 1984). Hormone concentration and growth media significantly affected the rooting and shooting ability of Z. armatum stem cuttings. IBA was found to be more effective than NAA. Neopeat medium was better than sand and mix media. The highest number of roots (6.5) and root length (11.6 cm) were recorded under IBA 5000 ppm in neopeat medium.

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