JOURNAL OF NATURAL REMEDIES DOI: 10.18311/jnr/2024/32921 Article Received on: 06.02.2023 Accepted on: 02.01.2024Revised on: 10.10.2023 1. Introduction Polyalthia longifolia (Sonn.) Thwaite is a flowering plant that belongs to the genus Polyalthia, and the Annonaceae family which falls within the Magnoliales order and Magnolids clade accordingly1. This plant originates from India, but it is grown in various parts of the tropics, including Ghana, Nigeria, and across West Africa for both ornamental and medicinal purposes. The plant is lofty, evergreen, and is mostly planted because of its effectiveness in alleviating noise2. The Annonaceae family belongs to the sub-class of Dialypetalae, which are primitive plants characterized by unclear boundaries between sepals, petals, and parts of fruit. Morphological features characteristic of the Annonaceae family include distichous leaf arrangement, cross-section of stem being striate, stems or twigs when exfoliated will produce a unique aroma, leaves having no stipules, aestivation of petal being valvate, ruminate endosperm, berries or drupe fruits, arillus seeds, and vessels with simple perforated fields3. Several plants species from the genus Polyalthia have been reported to possess various medicinal properties4,5, P. longifolia cv. pendula, grown in India, is known to be the most commonly used in traditional indigenous medicine6. Two distinct varieties of P. longifolia are known. One of them has spreading perpendicular RESEARCH ARTICLE Phytochemical Analysis and Elemental Contents of Varieties of Polyalthia longifolia (Sonn.) Thwaites Emelia Oppong Bekoe1*, Emmanuel Orman2, Michael Lartey3, Andrew Gordon4 and Tonny Asafo-Agyei5 1Department of Pharmacognosy and Herbal Medicine, School of Pharmacy, University of Ghana, Ghana; eoppongbekoe@ug.edu.gh 2Department of Pharmaceutical Chemistry, School of Pharmacy, University of Health and Allied Sciences, Ho, Ghana 3Department of Pharmaceutical Chemistry, University of Ghana, Ghana 4Department of Science Laboratory Technology, Accra Technical University, Accra, Ghana 5Department of Plant Development, Centre for Plant Medicine Research, Mampong Akuapem, Ghana Abstract Polyalthia longifolia (Sonn.) is a medicinal plant that belongs to the family Annonaceae, and it is distributed in the tropics. This plant is widely grown in West Africa for its ornamental and medicinal purposes. There are two varieties of P. longifolia which are commonly distinguishable by the direction of their branches. One has spreading perpendicular branches, and the other has drooping pendulous branches. Traditional herbal practitioners believe that one variety (P. longifolia cv. pendula) is more medicinal than the other. This study, therefore, sought to investigate the phytochemical components of the two varieties of P. longifolia by HPTLC, UPLC, and elemental analysis by ICP-EOS. No observable differences were found in the phytochemical and elemental profiles of these varieties that could help distinguish one from the other or could account for its supposed differences in medicinal properties. A total of 22 elements were detected in the samples of the two varieties of the plant. Qualitatively, the elemental content of both varieties was similar. Only Iridium was not detected in all samples. Heavy metals including As, Pb, Cd, and Hg had their levels above the recommended limits. Keywords: Elemental Content, Fingerprint, Phytochemical Analysis, Polyalthia longifolia *Author for correspondence 272 Phytochemical Analysis and Elemental Contents of Varieties of Polyalthia longifolia (Sonn.) Thwaites Journal of Natural Remedies | eISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 24 (2) | February 2024 branches and is generally known as the typical variety. The other variety which has drooping pendulous branches, is also commonly used as an avenue tree and is sometimes also known as P. longifolia cv. pendula6. This plant is used in traditional medicine across several Asian and African countries, with Ghana inclusive7,8 for the management of diseases such as malaria, fever, skin diseases, diabetes, hypertension and helminthiasis2,4,6,9,10. P. longifolia aqueous extract has been shown to lower the blood pressure, rate of respiration and glucose level in animal models9. The plant is reported to possess antibacterial, antifungal, antitumor, anti-ulcer and antioxidant properties, cytotoxic function and hypotensive effects2,10, hepatoprotective and anti-inflammatory, antimicrobial, hypoglycemic and anti-ulcer activities4,9,11. Polyalthia species have been shown to have various kinds of secondary metabolites, such as alkaloids, terpenes, and chalcone5, aporphine and azafluorene alkaloids, proanthocyanidins, β-sitosterol, and leukocyanidin, clerodane, and ent-helimane, diterpenoids. These compounds were isolated from the leaves, stem, and stem bark6. The clerodane diterpenoids and the alkaloids are known to contribute to its medicinal importance6. The aqueous leaf extract is known to contain terpenes, non-reducing sugars, flavonoids, resin, gums and mucilages, carbohydrates and fibre, low fat and phenols, as well as rich quantities of minerals10,12. The young leaves have been shown to contain 4% ash, 0.21% lipid, 25% fibre, 54% carbohydrate 9% protein, and 8% moisture while the mature leaves contained 10% protein, 5% ash, 0.26% lipid, 19% fibre, 9% moisture and 57% carbohydrates12. Quantitative analysis revealed that the young leaves contained slightly higher quantities of tannins and phenols but lower content of flavonoids as compared to the matured leaves. Both young and mature leaves showed appreciable quantities of minerals with the mature samples having higher concentrations of Na, K, Ca and Mg10. The essential oils of the leaf and stem bark of P. longifolia is exclusively composed of sesquiterpene derivatives, with allo-aromadendrene being in the highest content with 19.7%, followed by caryophyllene oxide, β-caryophyllene, β-selinene, α-humulene and ar-curcumene6. In Ghana, the two varieties of the plant are available. However, anecdotal reports have it that P. longifolia cv. pendula is more medicinally potent than P. longifolia. This study therefore sought to investigate the phytoconstituents and elemental composition of these two plant species in order to profer some scientific justification or otherwise to account for the postulated difference in medicinal properties. 1.1 Description Guatteria longifolia (Sonn.) Wall. and Uvaria longifolia Sonn. are synonyms often used for P. longifolia13. P. longifolia is a columnar, pyramid-like, tree: main stem straight, undivided, growing up to 12 m or more. Branches are slender, short, about 1 or 2 m long, glabrous, and pendulous. Leaves alternate, distichous, exstipulate, mildly aromatic, 7.5 - 23 by 1.5 - 3.8 cm. Leaves are shining, glabrous, tapering to a fine acuminate apex, narrowly lanceolate, margin markedly undulate, pinnately veined, leathery or ubcoriaceous, shortly petiolate; with petiole about 6 mm long. Flowers arise from branches below the leaves, 2.5 to 3.5 cm across, yellowish to green, in fascicles or shortly pendunculate umbels; petals 6, 2 seriate, flat, from a broad base, lanceolate, long accumulate, spreading; and sepals 3, broad, short, triangular, the tips reflexed. Stamens are many, cuneate; connective truncately dilated beyond the cells. Ovaries indefinite; ovules 1 or 2; style oblong. Ripe fruits ovoid, 1.8 to 2 cm long, numerous, stalked, glabrous, 1 seeded; stalk 1.3 cm long, short, glabrous. Seeds smooth, shining. Flowering and fruiting: February-June6. 2. Materials and Methods 2.1 Plant Sample Collection and Identification P. longifolia cv pendula (Voucher number 032020- 24), and P. longifolia (Voucher number 032020-24) samples leaves were harvested in March 2020 from the Kwabenya District (5.7021° N, -0.2446° W) and the University of Ghana campus (5.6539° N, -0.1859° W), both in the Greater Accra Region of Ghana. Plant materials were identified by Mrs. Gladys Schwinger, a Botanist at the Department of Plant and Environmental Sciences, University of Ghana, Legon. Two samples were harvested for each variety. The herbal material was thoroughly washed with distilled water and was air dried in a dust-free environment for 3 weeks (Figure 1). 273Bekoe et al., Journal of Natural Remedies | eISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 24 (2) | February 2024 Figure 1. Picture of P. longifolia cv. pendula and P. longifolia. 2.2 Extraction Pulverized plant material of 2 g each were extracted with 20 mL of water, ethanol 50 % v/v and petroleum ether at room temperature. Extraction of the materials were performed by ultra-sonication for 30 minutes in each solvent, and subsequently centrifuged at 8000 × g for 5 min. The supernatant were pooled together and concentrated low in vacuo at 40 oC and lyophilized. 2.3 High-performance Thin Layer Chromatographic (HPTLC) Analysis 2.3.1 Chemicals and Reagents All the chemicals were purchased in the highest quality available and used as received unless otherwise stated. Highly purified deionized water was freshly obtained from Millipore® Simplicity (Billerica, U. S. A.). 2.3.2 Sample Preparation and Application Ten microliters (10 μL) test solutions of 5 mg/L extracts dissolved in methanol were then applied to HPTLC plates of 20 × 10 cm dimensions of silica gel 60 F254 (Merck). The test solutions were applied as an 8 mm band with minimum of 11.4 mm distance between bands and 8 mm from lower edge of plate, and the bands dried. 2.3.3 Development A 20 × 10 cm Twin Trough Chamber (CAMAG, Muttenz Switzerland), saturated for 15 minutes with about 10 mL mobile phase in each trough was used. The developing distance was about 70 mm from the lower edge of the plate. The plates were then allowed to air dry and documented before and after derivatization. For the aqueous extracts, acetic acid-water-butanol (10:40:50) was used as the mobile phase. For the 50% ethanol extract, ethyl acetate-water-formic acid 90:5:5 (v/v) was used as the mobile phase, and for the petroleum ether extract, toluene-ethyl acetate 90:10 (v/v) was the mobile phase. 2.3.4 Reagent Preparation Reagents were prepared according to protocols described by Wagner and Bladt14. Anisaldehyde sulphuric acid, Natural Product Reagent (Naturstoff), and ferric chloride (FeCl3) reagents were used to detect the presence of tannins, flavonoids, fatty acids and sterols respectively. Anisaldehyde reagent was prepared by adding 20 mL of acetic acid, 10 mL of sulfuric acid, and 1 mL of anisaldehyde to 170 mL of ice-cooled methanol and well mixed. Naturstoff reagent was prepared by dissolving diphenylboryloxyethylamine in methanol to produce 1 % w/v solution. Iron-III- Chloride reagent was a 10 % w/v aqueous solution of FeCl3 in methanol. 2.3.5 Detection and Documentation The HPTLC analysis was performed on CAMAG TLC Visualizer 2 equipped with visionCATS software (version: 3.0) (Muttenz, Switzerland). The plates were examined both when underivatized and derivatized under white light, short UV λ 254 nm and long UV 366 nm. For the petroleum ether extracts, the plates were derivatized with anisaldehyde reagent and then heated at 100 °C for five min. For the ethanol extracts, the plates were derivatized with Naturstoff reagent and examination was performed under 366 nm for the presence of flavonoids. For the aqueous extracts, the plates were derivatized with ferric chloride reagent and examined under white light for the presence of tannins. 2.4 Ultra High Performance Liquid Chromatographic (UPLC) Analysis The UPLC analysis of ethanolic extracts of P. longifolia was performed to develop the fingerprint chromatogram for the presence of rutin, quercetin, 274 Phytochemical Analysis and Elemental Contents of Varieties of Polyalthia longifolia (Sonn.) Thwaites Journal of Natural Remedies | eISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 24 (2) | February 2024 myricetin, luteolin, and kaempferol. Instrumentation: Acquity™ UPLC, PDA eλ Detector, Quaternary Solvent Manager, Acquity UPLC H-Class manager, Sample Manager FTN Acquity UPLC, Milford, U.S.A. Stationary Phase: Acquity UPLC HSS T3, 1.8 µm, C18 2.1 × 100 mm (Waters, Milford, U.S.A.). Mobile phase: A: HCOOH (0.1%) in water, B: HCOOH (0.1%) in Acetonitrile; Gradient: t0 min A 95%, t1 min A 95%, t3 min A 90%, t7 min A 0%, t10 min A 0%; flow 0.5 mL/ min; injection volume 2µL; column temperature 400C; detection λ 210–400 nm. Reference standards (97% HPLC) for external calibration: rutin (Sigma-Aldrich, Taufkirchen, Germany), and kaempferol (Roth, Karlsruhe, Germany) were dissolved in methanol at concentrations of 1, 0.5, 0.25, 0.125, and 0.0625 mg/ mL. 2.5 Elemental Content Analysis by Inductively-coupled Plasma-optical Emission Spectrometric (ICP-OES) 2.5.1 Chemicals and Reagents All the chemicals used were purchased in the highest quality available. Highly purified water was freshly obtained from an Aquatron A4000D system (Barloworld Scientific, Nemours Cedex, France). 2.5.2 Standard Solutions The volumetric flasks for ICP-OES measurements were pre-treated with 2% v/v supra pure HNO3 and purified water to minimize adsorption effects. The standard solutions of the metals were diluted between 1 and 5,000 µg/L with 2% v/v HNO3 added from 1 g/L stock solutions of single-element standards. 2.5.3 Sample Preparation A microwave digestion system (Multiwave Go, Anton Parr GmbH, Austria) was used to digest the plant samples. The samples (approximately 0.1 g each) were placed in Teflon vessels and 4 mL of HNO3:HCl (3:1) was added to each. The vessels were heated at temperature programme as follows: 10 minutes heating to 100 oC and holding for 5 minutes, then heating from 100 oC to 150 oC for the next 10 minutes and holding for additional 5 minutes. Complete digestion was confirmed by decolourization of the sample solutions. The digests were cooled down to room temperature, transferred to 50 mL volumetric flasks, and made to volume with deionized water. A blank of HNO3: HCl (3:1) was used for the analysis. 2.5.4 Elemental Analysis by ICP-OES The elemental analysis was carried out using ArCos MV II (Spectro Ametek, Kleve, Germany) ICP-OES with axial plasma viewing. Gas flows were controlled by internal mass-flow controllers. A standard D-torch was employed. For sample introduction, the peristaltic pump of the system combined with a crossflow spray chamber was used under the following plasma conditions: 1200 W (RF-power), 13 L min-1 cooling gas, 0.8 L min-1, and 0.8 L min-1 nebulizer gas. 3. Results 3.1 Phytochemistry Profiling by HPTLC Upon HPTLC analysis of the extracts of both varieties of P. longifolia leaves, flavonoids were detected in the chromatograms (Figure 2). When underivatized, the spots quench fluorescence at λ 254 nm but a typical fluorescence at λ 366 nm was observed after derivatization with Naturstoff reagent. Phenol carboxylic acids (e.g. chlorogenic and caffeic acids) typically turn to blue bands while flavonols and flavones turn orange or yellow14. Two different profiles were detected for P. longifolia samples from different places. One sample of P. longifolia cv. pendula variety has a blue fluorescing spot at Rf ~ 0.30 and this is missing in both the other sample of similar variety and samples of the second variety. These samples originated from different places and could signal some sort of different chemotypes for the same species from different places. The corresponding video-densitometric profiles for the two types of profiles observed are shown in Figure 3. Comparing the densitometric profiles of the different channels for P. longifolia samples, only red channel profiles were similar for the samples. The green, blue and grayscale channel profiles all show the differences at the same Rf ~ 0.30. These observations are outlined in Figure 3. For samples belonging to the same variety of P. longifolia, similar profiles were seen irrespective of the origin, and this is evident in the densitometric profiles in Figure 3. With the exception of the profile with the distinct fluorescing band at Rf  0.3, the profiles developed with the use of Naturstoff reagent for 275Bekoe et al., Journal of Natural Remedies | eISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 24 (2) | February 2024 Figure 2. HPTLC profiles of the P. longifolia samples obtained after derivatization with Naturstoff reagent and detection at λ 366 nm. (A). P. longifolia cv. pendula (Tracks 2-5) and P. longifolia samples (Tracks 6-9). (B). Summary of video-densitometric profiles of the corresponding two different HPTLC profiles of the P. longifolia samples analyzed. all the samples (P. longifolia cv. pendula and P. longifolia varieties) were similar. Analysis of the petroleum ether extracts of both varieties of P. longifolia upon detection with anisaldehyde- sulphuric acid reagent, showed the presence of phenols, terpenes, sugars, and steroids by forming violet, blue, red, grey or green bands at day light (Figure 4D)14. Visual inspection of the HPTLC profiles showed they were similar, and this could be used to confirm the high level of similarity in phytochemical composition among the two different varieties of the plants. Iron-III-Chloride reagent revealed dark green and brown spots in the aqueous extract, as seen in Figure 5, indicating the presence of tannins14. Similarly, the pattern of bands observed were the same for the samples from the different varieties. This could imply that the tannins profiles in the two varieties were similar. 3.2 HPLC Fingerprint Chromatogram HPLC analysis also confirmed similar fingerprint chromatograms (Figure 6) of all samples of P. longifolia cv pendula and P. longifolia samples irrespective of the origin. The typical fingerprint chromatogram (Figure 7) of the two P. longifolia varieties showed 4 main peaks at retention times: 4.364, 4.478, 4.481 and 7.194 min. 3.3 Flavonoid Content Spiking samples of P. longifolia ethanol 50% extracts with reference flavonoids showed the possible presence of rutin and the absence of quercetin, myricetin, luteolin and kaempferol (Figure 8). 3.4 Elemental Content As seen in Table 1, the presence or absence of a total of 22 elements were detected in the samples of the two varieties of the plant. Qualitatively, the elements present in both varieties were similar. Out of the 22 elements investigated, only Iridium (Ir) was not detected among all samples. Regarding toxic heavy metals whose presence and contents are usually regulated, most of them (including As, Pb, Cd, and Hg) had their levels above the recommended limits. The exceptions were for chromium (Cr) and copper (Cu) where their median levels recorded for each variety were less than the recommended limits of 2 mg/kg and 276 Phytochemical Analysis and Elemental Contents of Varieties of Polyalthia longifolia (Sonn.) Thwaites Journal of Natural Remedies | eISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 24 (2) | February 2024 Figure 3. Densitograms of P. longifolia samples at the different channels. 150 mg/kg respectively. Comparatively, the levels of each of the elements in both varieties were comparable (p>0.05). The presence of the other elements detected demonstrate the additional benefits the plants possess with respective to their nutritional and medicinal values. Similar to previous results, the contents of all elements were similar (p>0.05). IQR – Interquartile range showing the lower quartile (Q1) and upper quartile (Q3); LOD – Limit of detection; Levels of elements in the two varieties were compared using the non-parametric Mann-Whitney test. Level of significance were determined at 95 % confidence level. NS – Not Significant. 4. Discussion The qualitative comparative study of the two varieties of P. longifolia by HPTLC revealed no distinguishable differences. In comparing the ethanol extracts of P. longifolia from various places, there was a high level of similarity. One sample which was different due to the presence of one spot could be said to be different from the other Polyalthia samples or a chemotype of P. longifolia. As reported in other literature, this study confirmed the presence of flavonoids and phenols (tannins)10,12 in both varieties of P. longifolia. P. longifolia is rich in flavonoids and based on the HPLC retention time, rutin which is suspected to be a constituent, has been reported to be an active constituent of this plant15. The high level of similarity from the chromatographic profiles is due to the similarity in the phytochemical compositions of the two plant varieties. Contrary to the accession that one of the varieties has more medicinal value than the other which is considered more for ornamental purposes. The observations from this study, shows that both varieties could be considered for medicinal uses. This assertion is further supported by the similarities of the elemental compositions. The physiological importance of these elements as outlined in several literature could also be thought to augment the medicinal importance of the plant16,17. Additionally, the phytochemical constituents also account for the medicinal properties. Phytochemical constituents such as flavonoid derivatives have shown to have antimalarial activity18. The flavonoids antioxidants also act to provide protection against free radicals that damage cells and tissues. In the treatment of high blood pressure, flavonoids have also been shown to have a potential to inhibit angiotensin converting enzyme in vitro19. Tannins promote healing of wounds, and are effective in diarrhea, colitis and peptic ulcers20. Phenolic such as catechins have an anti-hyperglycemic action, lowering both blood-glucose and normalizing insulin release21. Irrespective of the above-mentioned benefits, the presence of some heavy toxic heavy metals were also documented in high quantities. As, Cd, Hg and Pb exceeded their recommended limits. Many studies have demonstrated that reactive oxygen species production and oxidative stress play a key role in the toxicity and carcinogenicity of these four metals. These metals have a high degree of toxicity, and are ranked among the priority metals that are of great public health significance. They are all systemic toxicants with known activities to induce multiple organ damage, even at lower doses. These metals are classified as "known" or "probable" 277Bekoe et al., Journal of Natural Remedies | eISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 24 (2) | February 2024 Figure 4. HPTLC profile of petroleum ether extract of P. longifolia var pendula (tracks 13, 14) and P. longifolia (tracks 15, 16). Samples were derivatized with anisaldehyde sulphuric acid and documented: a. underivatized under λ 244 nm, and derivatized; b. white light c. λ 244 nm d. λ 366 nm. Figure 5. HPTLC profile of aqueous extract of P. longifolia cv. pendula (tracks 10, 11, 12) and P. longifolia (tracks 13, 14, and 15). Samples were derivatized with FeCl3 chloride and documented: a. white light, and b. λ 244 nm. 278 Phytochemical Analysis and Elemental Contents of Varieties of Polyalthia longifolia (Sonn.) Thwaites Journal of Natural Remedies | eISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 24 (2) | February 2024 Figure 6. HPLC overlaid chromatograms of P. longifolia cv pendula and P. longifolia samples. Figure 7. Typical UPLC fingerprint chromatogram of P. longifolia at λ 250 nm. 279Bekoe et al., Journal of Natural Remedies | eISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 24 (2) | February 2024 Figure 8. Overlaid UPLC chromatogram of 50% ethanol extract of P. longifolia with reference flavonoids at λ 250 nm; 1. Rutin, 2. Quercetin, 3. Myricetin, 4. Luteolin, 5. Kaempferol. Table 1. Elemental content of P. longifolia leaf samples Element Median (IQR) amount in P. longifolia cv pendula (mg/kg) Median (IQR) amount in P. longifolia (mg/kg) Limit (mg/kg) Al 217.2 (180.2–286.2) 216.6 (183.4–274.9) ns As 11.40 (7.881–16.59) 9.251 (7.220–23.54) ns 5 Ba 20.58 (11.81–31.00) 30.94 (1.875–31.72) ns Br 2320 (2244–2396) 2261 (2069–2452) ns Ca 10234 (7877–13072) 9541 (8193–11429) ns Cd 4.199 (0.000–36.46) 7.697 (0.000–57.06) ns 0.3 Cr 0.1836 (0.000–1.753) 0.000 (0.000 – 1.340) ns 2 Cu 6.070 (0.000–14.11) 9.739 (0.000–11.82) ns 150 Fe 434.4 (275.5–600.9) 302.7 (281.4–340.6) ns Hg 5.208 (4.666–5.751) 4.432 (2.257–6.608) ns 0.5 280 Phytochemical Analysis and Elemental Contents of Varieties of Polyalthia longifolia (Sonn.) Thwaites Journal of Natural Remedies | eISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 24 (2) | February 2024 human carcinogens22. Thus, and with such high levels reported, the safe use of this plant becomes questionable. However, this phenomenon is more reflective of the kind of anthropological and or industrial activities that occur around the collection sites of the plants. The main factor that affects the content of elements in medicinal plants are from the soil, effects of biological, geographical, and agrochemical activities, climatic conditions and he ability of some plants to specifically accumulate elements16. It is therefore necessary to strengthen the control measures applicable to medicinal plants, especially for those which are prepared to be orally administrable. Measures like Good Agricultural and Collection Practices should be encouraged to help control the presence and levels of some of these impurities. 5. Conclusion Irrespective of the minor morphological difference observed in the two varieties of P. longifolia, the HPTLC and UPLC analysis demonstrated similar phytochemical compositions in terms of flavonoids. As such, both varieties could be considered for medicinal uses. Similarly, they also possess nutritional values because of the high levels of essential elements. However, there are also high level of heavy metals in the analysed plant samples. For this reason, regulatory measures regarding the use of medicinal plants should be enhanced. There may also be the need to further investigate these plant varieties in terms of differences in other phytochemical components. 6. Funding The project was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – Project 423277515 (HE1642/12-1). 7. References 1. Chase MW, Christenhusz MJ, Fay MF, Byng JW, Judd WS, et al. Angiosperm Phylogeny Group. An update of the angiosperm phylogeny group classification for the orders and families of flowering plants: APG IV. Bot J Linn. 2016; 181(1):1-20. https://doi.org/10.1111/boj.12385 2. Sivashanmugam AT, Chatterjee TK. 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Element Median (IQR) amount in P. longifolia cv pendula (mg/kg) Median (IQR) amount in P. longifolia (mg/kg) Limit (mg/kg) Ir < LOD < LOD K 13878 (8479–18978) 15351 (10239–21786) ns Mg 1573 (1252–1886) 1452 (1118–1782) ns Mn 34.92 (26.40–41.96) 18.91 (6.783–31.78) ns Ni 18.69 (14.12–23.26) 11.76 (1.701–21.82) ns P 1474 (1305–1748) 1529 (1358–1725) ns Pb 26.77 (21.21 – 32.33) 35.49 (27.76 – 43.21) ns 10 S 2147 (1586 – 2832) 2188 (1923 – 2570) ns Sn 3.368 (0.000 – 6.736) 2.765 (0.209 – 5.321) ns Sr 56.21 (36.18 – 76.23) 44.52 (14.54 – 74.49) ns Ti 5.643 (4.748 – 5.918) 5.178 (1.513 – 7.288) ns Zn 15.12 (13.92 – 16.32) 14.50 (13.87 – 15.12) ns Table 1. Continued... 281Bekoe et al., Journal of Natural Remedies | eISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 24 (2) | February 2024 6. Katkar KV, Suthar AC, Chauhan VS. The chemistry, pharmacologic, and therapeutic applications of Polyalthia longifolia. 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