The present study was focussed to carry out the detailed pharmacognostic studies in terms of organoleptic characters, physicochemical parameters, macroscopical, microscopical characters and phytochemical screening for major groups of compounds and other WHO recommended parameters for standardization of the leaves of (Mill.) family Simaroubaceae. Phytochemical screening of methanolic and aqueous extracts of leaves of Ailanthus altissima demonstrated the presence of Carbohydrates, flavonoids, tannins, cardiac glycosides, phenols, saponins, diterpenes, fats and oils. However alkaloids and anthraquinone glycosides were found absent in all the extracts. Information obtained from phytochemical and pharmacognostical analysis can be used as markers in the identification and standardization of this plant for monograph development and as herbal remedy.
Keywords: Tree of heaven, Phytochemical, Pharmacognostic analysis, Bioactive compounds, Ailanthus altissima

Bashir, Andleeb

Bhat, Zulfiqar Ali

Farooq, Saeema

Mohi-ud-din, Roohi

Journal of Drug Delivery and Therapeutics (JDDT)

Bashir et al                                                                                                             Journal of Drug Delivery & Therapeutics. 2019; 9(2-s):179-183 ISSN: 2250-1177                                                                                  [179]                                                                                 CODEN (USA): JDDTAO Available online on 15.04.2019 at http://jddtonline.info Journal of Drug Delivery and Therapeutics Open Access to Pharmaceutical and Medical Research © 2011-18, publisher and licensee JDDT, This is an Open Access article which permits unrestricted non-commercial use, provided the original work is properly cited Open  Access                                                                                                                                                     Research Article Pharmacognostic Standardization and Phytochemical Evaluation of Ailanthus altissima (Mill.) Swingle leaves Andleeb Bashir, Roohi Mohi-ud-din, Saeema Farooq, *Zulfiqar Ali Bhat Department of Pharmaceutical Sciences, School of Applied Sciences and Technology, University of Kashmir, Hazratbal, Srinagar-190006, J&K, India  ABSTRACT The present study was focussed to carry out the detailed pharmacognostic studies in terms of organoleptic characters, physicochemical parameters, macroscopical, microscopical characters and phytochemical screening for major groups of compounds and other WHO recommended parameters for standardization of the leaves Ailanthus altissima (Mill.) of family Simaroubaceae. Phytochemical screening of methanolic and aqueous extracts of leaves of Ailanthus altissima demonstrated the presence of Carbohydrates, flavonoids, tannins, cardiac glycosides, phenols, saponins, diterpenes, fats and oils. However alkaloids and anthraquinone glycosides were found absent in all the extracts. Information obtained from phytochemical and pharmacognostical analysis can be used as markers in the identification and standardization of this plant for monograph development and as herbal remedy. Keywords: Tree of heaven, Phytochemical, Pharmacognostic analysis, Bioactive compounds, Ailanthus altissima.  Article Info: Received 19 Feb 2019;   Review Completed 25 March 2019;   Accepted 27 March 2019;   Available online 15 April 2019 Cite this article as: Bashir A, Mohi-ud-din R, Farooq S, Bhat ZA, Pharmacognostic Standardization and Phytochemical Evaluation of Ailanthus altissima (Mill.) Swingle leaves, Journal of Drug Delivery and Therapeutics. 2019; 9(2-s):179-183     http://dx.doi.org/10.22270/jddt.v9i2-s.2484        *Address for Correspondence:  Zulfiqar Ali Bhat, Department of Pharmaceutical Sciences, School of Applied Sciences and Technology, University of Kashmir, Hazratbal, Srinagar-190006, J&K, India   INTRODUCTION Description Ailanthus altissima commonly known as smoke tree or tree of heaven belongs to family simaroubaceae. The plant is locally known as Handoon or Alamthar in Kashmir. A. altissima is a medium-sized tree that reaches heights between 17 and 27 metres with a diameter at breast height of about 1 metre 1. The bark is smooth and light grey. The twigs are stout, smooth to lightly pubescent, and reddish or chestnut in colour. They have lenticels as well as heart-shaped leaf scars with many bundle scars around the edges. The buds are finely pubescent, dome shaped, and partially hidden behind the petiole 2. The branches are light to dark gray in colour, smooth, lustrous, and containing raised lenticels that become fissures with age. The ends of the branches become pendulous. All parts of the plant have a distinguishing strong odour that is often likened to peanuts, cashews or rotting cashews 2,3,4. The leaves are large, odd- or even-pinnately compound, and arranged alternately on the stem. They range in size from 30 to 90 cm in length and contain 10-41 leaflets organised in pairs, with the largest leaves found on vigorous young sprout. When they emerge in the spring, the leaves are bronze then quickly turn from medium to dark green as they grow 5. The rachis is light to reddish-green with a swollen base. The leaflets are ovate-lanceolate with entire margins, somewhat asymmetric and occasionally not directly opposite to each other. Each leaflet is 5 to 18 cm long and 2.5 to 5 cm wide. They have a long tapering end while the bases have two to four teeth, each containing one or more glands at the tip. The leaflets' upper sides are dark green in colour with light green veins, while the undersides are a more whitish green. The petioles are 5 to 12 mm long. The flowers are small and appear in large panicles up to 50 cm in length at the end of new shoots. The individual flowers are yellowish green to reddish in colour, each with five petals and sepals 1,2. They appear from mid-April in the south of its range to July in the north. A. altissima is dioecious, with male and female flowers being borne on different individuals. Male trees produce three to four times as many flowers as the females, making the male flowers more conspicuous. Furthermore, the male plants emit a foul-smelling odour while flowering to attract pollinating insects. Female flowers contain ten (or rarely five through abortion) sterile stamens (stamenoides) with heart-shaped anthers. The fruits grow in clusters; a fruit cluster may contain hundreds of seeds. The seeds borne on the female trees are 5 mm in diameter and each is encapsulated, Bashir et al                                                                                                             Journal of Drug Delivery & Therapeutics. 2019; 9(2-s):179-183 ISSN: 2250-1177                                                                                  [180]                                                                                 CODEN (USA): JDDTAO appearing July through August, but can persist on the tree until the next spring 6. Distribution A. altissima is native to subtropical/warm temperate climates but is able to invade climates ranging from cool temperate to tropical. In South Africa it invades forest margins, roadsides and riverbanks in cool, moist regions unlike other members of the genus Ailanthus, it is found in temperate climates rather than the tropics. The tree was first brought from China to Europe in the 1740 and to the United States in 1784. It has become an invasive species due to it’s due to its ability both to colonise disturbed areas quickly and to suppress competition with allelopathic chemicals. It is considered a noxious weed in Australia, United States, New Zealand and many countries of easten and southern Europe 7. Traditional uses Ailanthus altissma has been traditionally used in Chinese folk medicine as an astringent, antispasmodic, antihelminthic, parasiticide and narcotic 8,9. Fresh stem bark was used to treat diarrhoea and dysentery; root bark for heat ailments, epilepsy, and asthma; fruits as an emmenagogue and to treat ophthalmic diseases; and leaves as astringent used in lotions for seborrhoea and scabies 8. Recent pharmacological studies indicate a broad array of possible uses of quassinoids, for example to treat malaria. Epstein–Barr-virus infection and HIV infection. In addition, Ailanthus is used for homoeopathic remedies 10,11,12,13. MATERIALS AND METHODS  Plant Material Collection  In the present study, the fresh leaves of the plant Ailanthus altissima were collected from Botanical Garden, University of Kashmir, Srinagar, Kashmir, at an altitude of 1585m in the month of August. The plant was identified and authenticated by Prof. Akhtar H. Malik, KASH Centre for Biodiversity and Taxonomy University of Kashmir under specimen voucher number 2322-KASH. A sample specimen of collected material was deposited in herbarium for future reference. Macroscopic and microscopic analysis  Macroscopic and microscopic analysis of the leaves of Ailanthus altisimma was studied according to the method mentioned in the Trease and Evans pharmacognosy 14,15.  Preparation of extracts  After collection and authentication, leaves of the plant material were shade dried and powdered. The powdered sample was then passed through sieve no. 40 and subjected to extraction. A weighed quantity of the powdered drug was extracted in a soxhlet with methanol. Aqueous extract was prepared by decoction method. The extracts were evaporated to dryness under reduced pressure and controlled temperature 40-50ºC 12. The extracts were then kept in desiccators to remove remaining moisture, and finally stored in air tight containers at 4oC for further use.  Physicochemical analysis The leaves were shade dried and powdered for the determination of physicochemical parameters like ash value, extractive values, Flourescence analysis study of powdered drug material reagents was carried out to observe the color reactions. The drug was treated with various chemical reagents and exposed to visible and ultra violet light to study their fluorescence behaviour16,17.  The methanolic and aqueous extracts so obtained were subjected to preliminary phytochemical screening. Phytochemical studies were performed to identify the presence of various constituents according to the standard methods 18,19,20. RESULTS AND DISCUSSION Macroscopic characters The leaves are large, odd or even pinnately compound, and arranged alternately on the stem. They range in size from 30 to 90 cm in length. The leaflets are ovate-lanceolate with entire margins, somewhat asymmetric and occasionally not directly opposite to each other.  Microscopic characters The microscopic characters of the leaves reveals the presence of epidermal cells, collenchyma, spongy mesophyll, palisade cells, covering trichomes, glandular trichomes, xylem and phloem vessels (Figure 1). The powder microscopy of leaves reveals the presence of epidermal cells, paracytic stomata, palisade cells, covering trichomes and crystals of calcium oxalate (Figure 2).  Figure 1: Transverse section of leaf of Ailanthus altissima showing epidermis, collenchyma, spongy mesophyll, Xylem vessels, phloem vessels, covering trichrome, glandular trichrome and ground tissue   Bashir et al                                                                                                             Journal of Drug Delivery & Therapeutics. 2019; 9(2-s):179-183 ISSN: 2250-1177                                                                                  [181]                                                                                 CODEN (USA): JDDTAO     (a)                                                                         (b)                                                                             (C) Figure 2: Microscopic structures of leaf of Ailanthus altissima (a): Epidermis with palisade cells and calcium oxalate crystals; (b): Paracytic stomata; (c): Covering trichomes  Physicochemical analysis Various physicochemical parameters like ash value, extractive values, Flourescence analysis were determined and are shown in Table- 1, 2 and 3 respectively. Table 1: Extractive values of metanolic and aqueoua extracts of Ailanthus altissima Extractive value Solvent Leaves Cold extractive value Petroleum ether 0.3325% Chloroform 0.8625% Methanolic 3.1325% Ethylacetate 1.72% Aqueous 3.865% Hot extractive value Petroleum ether 18.764% Chloroform 20.325% Methanolic 21.376% Ethylacetate 19.487% Aqueous 23.235% Successive extractive value Petroleum ether 0.448% Chloroform 0.986% Methanolic 3.436% Ethylacetate 1.698% Aqueous 3.423%  Table 2: Ash values and other physicochemical constants of leaves of Ailanthus altissima Physico-chemical Parameters Values pH 1% solution pH 10% solution 4.7 4.1 Loss on drying at 105°C 6.651% Swelling index 2 Total Ash value (% w/w) 4.37% Foaming index    Table 3: Flourescence analysis of leaves of Ailanthus altissima Treatment Day light Ultraviolet (254nm) Short wavelength Ultraviolet (266nm) Long wavelength Powder as such Green  Green Light green Powder with distilled water Green  Green  Light green Powder with Iodine Blackish green  Light green  Fluorescent green Powder with FeCl3 Dark green  Blackish green Green Powder with Picric acid Green  Greenish black  Fluorescent green Powder with NaoH Green  Green  Light green Powder with CHCl3 Fast green  Fast green  Green Powder with Ethylacetate Green  Light green  Greyish green  Powder with Concentrated HNO3 Light green  Orange  Black Bashir et al                                                                                                             Journal of Drug Delivery & Therapeutics. 2019; 9(2-s):179-183 ISSN: 2250-1177                                                                                  [182]                                                                                 CODEN (USA): JDDTAO Powder with concentrated Hcl Green  Greyish green  Brown Powder with K2Cr2O7 Orange  Black  Fluorescent green Powder with Pet. ether Green  Green  Light green  Powder with Methanol Green Dark green  Green Powder with Concentrated H2SO4            Black  Black  Dark green Powder with ammonia Green  Light green  Light green Powder with concentrated Hcl and distilled water Green  Light green  Green Powder with concentrated H2SO4 and distilled water Black  Black  Brown Powder with concentrated HNO3 and distilled water Light green  Brown Black Powder with GAA Brown  Olive green  Light green Powder with alcoholic KOH Green  Light green  Light green  Phytochemical Screening In the preliminary phytochemical screening of methanolic and aqueous extracts of Ailanthus altissima secondary metabolites like carbohydrates, tannins, flavonoids, phenols, phytosterols, proteins, saponins, diterpenes, cardiac glycosides, anthraquinones and alkaloids were tested. Phytochemical screening of methanolic and aqueous extracts of leaves of Ailanthus altissima demonstrated the presence of Carbohydrates, flavonoids, tannins, cardiac glycosides, phenols, saponins, diterpenes, fats and oils. However proteins, alkaloids and anthraquinone glycosides were found absent in all the extracts and phytosterols were also found absent only in aqueous extract. (Table-4).  Table 4: Preliminary phytochemical analysis of metanolic and aqueoua extracts of Ailanthus altissima Tests Inference Extracts   Methanolic Aqueous Carbohydrates Molisch’s test Violet ring + + Fehling’s test Brick red ppt. + + Benedict’s test Orange red ppt. + + Tannins 5% FeCl3 test Yellow color + + Lead acetate test White ppt. + + Gelatin test White ppt. + + Flavonoids Shinoda test Pink color + + Alkali reagent test Intense yellow color Which becomes colorless on addition of dil. acid  +  + Lead acetate test Yellow color ppt. + + Phenols 1% FeCl3 Bluish color + + Phytosterols Salkowski test Golden yellow ring at junction + _ Libermann’s test Brown ring at junction + _ Proteins Xanthoproteic test Yellow color _ _ Biuretic test Blue color _ _ Saponins Foam test Foaming + + Ftoth test Frothing + + Diterpenes Copper acetate test Emerald green color + + Cardiac glycosides Keller killiani test Brown ring at junction + + Legal test Pink color + + Alkaloids Mayer’s test Cream ppt. _ _ Hager’s test Yellow ppt. _ _ Dragendroff’s test Orange ppt. _ _ Wagner’s test Reddish brown ppt. _ _    Bashir et al                                                                                                             Journal of Drug Delivery & Therapeutics. 2019; 9(2-s):179-183 ISSN: 2250-1177                                                                                  [183]                                                                                 CODEN (USA): JDDTAO CONCLUSION  From the present study, it was concluded that Ailanthus altissima showed the presence of a number of active secondary metabolites such as carbohydrates, tannins, flavonoids, phenols, phytosterols, saponins, diterpenes, cardiac glycosides. From the results, it was noted that the extracts of Ailanthus altissima were found to be a rich source of variety of active secondary metabolites. The leaf constants also play the major role for the standardization of plant and one of the backbone of microscopical evaluation. Information obtained from above study can be used as markers in the identification and standardization of this plant or herbal drugs which provide the healthy and maximum potent drug in the market.  CONFLICT OF INTEREST  We declare that we have no conflict of interest. REFERENCES 1. Miller JH.  Ailanthus altissima. In Russell M. Burns and Barbara H. Honkala (eds.), Silvics of North America, hardwoods. 1990; 2:654. 2. Miller JH. Tree-of-heaven. Nonnative invasive plants of southern forests: a field guide for identification and control. USDA Forest Service/UNL Faculty Publications.2003. 3. Hu SY.  Ailanthus altissima. Arnoldia. 1979; 39 (2):29-50. 4. Anonymous. Tree of Heaven Ailanthus altissima. Division of forestry, Ohio Division of forestry. 2015; 12-16. 5. Miller JH. Nonnative invasive plants of southern forests. USDA Forest Service/UNL Faculty Publications. 2006; 103. 6. Henderson L. Alien Weeds and Invasive Plants. Plant Protection Research Institute Handbook No. 12. 2001. 7. Adamik KJ, Brauns FE. Ailanthus glandulosa (Tree of heaven) as a pulpwood Part II. Tappi. 1957; 40(7):522-527. 8. Burrows, George E, Ronald JT. Toxic Plants of North America. 2011; 147-254. 9. Haisey RM. Potential of ailanthone, an allelochemical from A. Altissima, as a natural product herbicide. InBook of abstracts of the 24th ACS national meeting, Las Vegus. 1997; 7-11. 10. Kang TH et al. Ailanthus altissima Swingle has anti-anaphylatic effect and inhibits inflammatory cytokine expression via suppression of nuclear factor-kappa B activation. In vitro cellular and Developmental Biology-Animal. 2010; 46(1):1708-11. 11.  Bown D. The Royal Horticulture Society encyclopedia of herbs and their uses. Dorling Kindersley Limited.1995. 12.  Che YH. Handbook of Chinese Herbs and Formulas, Ed. Institute of Chinese Medicine, Los Angels. 1995. 13. Evans WC, Evans D, Trease GE. Trease and Evans Pharmacognosy. 16th ed. Saunders/Elsevier; 2009. 14. Trease GE, Evans WC. Pharmacognosy. International edition. W.B.Saunders. 2008; 2(3):538-544 15. Farooq S, Mohi-ud-din R, Bhat ZA, Preliminary phytochemical screening of Iris kashmiriana Baker collected from Budgam, Kashmir, India., Journal of Drug Delivery and Therapeutics. 2019; 9(1-s):121-124. 16. Raja WY, Bhat ZA, Chashoo IA. Pharmacognostic and Phytochemical characteristics of Ailanthus altissima (Mill.) Swingle stem and root bark:A comparative study. Pharmacognosy journal.2017; 9(5):668-73. 17. Anonymous, Quality control methods for medicinal plant materials, World Health Organization, Geneva, 1998; 8:25-28. 18. Harborne   JB, Phytochemical   methods:   A   guide   to   modern techniques of plants analysis, Chapman & Hall.  London, Ltd., 1998; 1-188. 19. Kokate CK, Purohit AP, Gokhale SB. Pharmacognosy. 22nd ed. Nirali Prakash, Pune,India. 2003. 594. 20. Jigna P , Sumitra VC, In-vitro antimicrobial activity and phytochemical analysis of some Indian medicinal plants, Turkish Journal of Biology, 2007; 31:53-58.          

Pharmacognostic Standardization and Phytochemical Evaluation of Ailanthus altissima (Mill.) Swingle leaves

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Pharmacognostic Standardization and Phytochemical Evaluation of Ailanthus altissima (Mill.) Swingle leaves

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