PHARMACOGNOSTIC AND PRELIMINARY PHYTOCHEMICAL EVALUATION OF THE LEAVES OF CRINUM LATIFOLIUM L.HTML Full Text
PHARMACOGNOSTIC AND PRELIMINARY PHYTOCHEMICAL EVALUATION OF THE LEAVES OF CRINUM LATIFOLIUM L.
J. Solanki*1, A. Dhiman 1, A. Nanda 1 and A. Dhankhar 2
Department of Pharmaceutical Sciences, Maharshi Dayanand University 1, Rohtak-124001, Haryana, India
Suresh Gyan Vihar University 2, Jagatpura, Jaipur - 302025, Rajasthan, India
Crinum latifolium L. is a rosette-like herb showing promising results as rubefacient, tonic, for treatment of allergic disorders, benign prostatic hyperplasia, cancer and others. The present study deals with pharmacognostic and preliminary phytochemical evaluation of leaves of Crinum latifolium (Amaryllidaceae). The pharmacognostical studies were carried out as organoleptic, microscopic and physical parameters such as moisture content, ash value, fluorescence analysis and extractive value were determined. For phytochemical evaluation, Crinum latifolium leaves were subjected to successive solvent extraction using petroleum ether, chloroform, hydroalcoholic solution and water. These extracts were then screened for presence of different chemical constituents. Thin layer chromatography (TLC) of the tested extracts was also performed to determine the active principles. These studies are useful in identification and chemical characterization of Crinum latifolium and to explore its phytochemical and pharmacological potential.
|Keywords:Crinum latifolium L.,
INTRODUCTION: Crinum latifolium L. (Amaryllidaceae), popularly known as Sudarshan, has been used in Asian folk and traditional medicine 1 in the treatment of serious health conditions like prostatitis adenoma, benign prostate enlargement 2, uterine fibroids, hypoxia, inflammation, detoxification, tissue regeneration, hormone balancing, to enhance cell-mediated immunity and an effective T-lymphocyte activator 3.
Leaf juice is used for earache, rheumatic pain, and sprain. These activities are attributed to the presence of different chemical principles such as Amaryllidaceae alkaloids 4 lycorine, 2-epilycorine, pyrrolophenan- thridine alkaloids, 2-epilyocorine and 2-epipancrassidine 5, carbohydrates, glycosides, proteins & amino acids etc. and thus imparting significant role in medicines. But insufficient data is found in literature regarding pharmacognostic and physiochemical characteristics which can further help in betterment of its medicinal activity.
Hence, the present study was designed to evaluate the pharmacognostic aspects of leaves including morphology, histology and quantitative microscopy, determination of various physicochemical parameters, fluorescence analysis, phytochemical screening and TLC of different leaf extracts of Crinum latifolium L.
MATERIALS AND METHODS: Collection and authentication of plant material: The leaves of C. latifolium L. were collected from herbal garden, Maharshi Dayanand University, Rohtak, Haryana, India in december 2010 and authenticated by Dr. J.P. Yadav, Department of Biosciences, M.D. University, Rohtak and a voucher specimen was deposited in Herbarium of Pharmaceutical Sciences, M.D.University (voucher specimen number DPS 0016).
Morphological Features: The morphological features of leaves such as presence of foreign organic matter, color, odor, size, shape and taste were studied 6.
Microscopic features: Microscopic characteristics were studied under the following headings-
- Microscopy of intact drug as well as powdered drug characteristics
- Quantitative microscopy
Powder Characteristics: The powder microscopy was performed using standard methods and characteristic features were recorded 7.
Quantitative Microscopy: Various leaf constants such as stomatal number, stomatal index, palisade ratio, vein islet number and vein termination number were determined followed by official methods 8.
Physicochemical Parameters: Physicochemical parameters, i.e., percentage of ash values, extractive values, percentage of moisture content and fluorescence analysis were performed according to recognized methods 9.
Determination of Ash Values: The ash of the leaf powder was prepared by incinerating the powder in muffle furnace and ash value was determined 10.
Determination of Extractive Values: The leaves were shade dried, coarsely powdered and then were extracted successively with petroleum ether, chloroform, hydro alcoholic solution (70:30) and finally with chloroform water using hot extraction technique. The extractive values were determined in percentage w/w 9.
Fluorescence Analysis: Fluorescence analysis of crude drug powder was carried out in various solvents and reported.
Preliminary Phytochemical Screening: Different extracts were then screened for the presence of the various organic chemical constituents in the leaves of C. latifolium L 12.
TLC Fingerprint Profile: Thin layer chromatography of different extracts was performed and the Rf values were determined 13.
RESULTS AND DISCUSSION:
Morphological Characteristics: Crinum latifolium L. is a rosette-like herb that arises from an underground bulb. It is a stout perennial herb 2 m in height. Leaves are long, linear, acute apex, entire margin, ligulate type 14. Flowers are white in color and arranged in umbel. The morphological characteristics of Crinum latifolium L. are listed in table 1.
TABLE 1: MORPHOLOGICAL CHARACTERISTICS OF FRESH CRINUM LATIFOLIUM L. LEAVES
|Blade length||> 36 inches|
Histological features of Crinum latifolium L. leaves.
Microscopy of Crinum latifolium L. leaves.
Powder characteristics: The powder is green in color and contains diacytic stomata, fibers, epidermal cells with stomata, pitted vessels, acicular calcium oxalate crystals and starch grains (Figure 4-10).
Quantitative Microscopy: The leaf constants were determined following the standard methods 9 and results have been furnished in table 2.
TABLE 2: LEAF CONSTANT VALUES OF CRINUM LATIFOLIUM L. LEAVES
|Stomatal Number (upper surface)||3 - 6||4.3|
|Stomatal Number (lower surface)||4 - 7||5.3|
|Stomatal index (upper surface)||6.90 - 11.67||9.15|
|Stomatal index (lower surface)||6.45 - 11.51||8.97|
|Palisade ratio||3 - 6||4.2|
|Vein islet number||2 - 4||2.6|
|Vein termination number||3 - 5||4.3|
Physicochemical Parameters: The various physicochemical parameters were determined and are represented in table 3.
TABLE 3: PHYSICOCHEMICAL PARAMETERS OF CRINUM LATIFOLIUM L. LEAVES
|Acid insoluble ash||9.0|
|Water soluble ash||18.0|
|Petroleum ether soluble extractive||2.5|
|Chloroform soluble extractive||5.0|
|Water soluble extractive||31.5|
*average of three readings
Fluorescence Analysis: The results of fluorescence analysis are presented in Table 4.
TABLE 4: FLUORESCENCE ANALYSIS OF CRINUM LATIFOLIUM L. LEAF POWDER
|Treatment||Normal light||Under UV light|
|254 nm||366 nm|
|Dry powder||Brownish green||Dark brown||green|
|Powder + 5% NaOH||yellowish||Greenish dark brown||Dark brown|
|Powder + 5% KOH||Light yellowish||Light greenish brown||Light brown|
|Powder + 5% FeCl3||Yellowish green||Dark brown||Dark black|
|Powder + conc. H2SO4||Reddish brown||Dark black||Dark black|
|Powder + dil NH3||Light orange||Dark brown||Yellow|
|Powder + conc. HCl||Dark brown||Dark black||Dark black|
|Powder + conc. HNO3||Orange||yellow||Dark yellow|
|Powder + iodine solution||Greenish brown||Dark black||Dark brown|
|Powder + 5% HCl||Yellowish brown||Dark brown||Dark brown|
|Powder + 5% H2SO4||Yellowish brown||Dark brown||Yellowish green|
|Powder + dil HNO3||Brownish orange||green||Dark green|
|Powder + Na2CO3||Yellowish brown||Yellowish brown||yellow|
|Powder + alc. KOH||Light yellow||Dark yellow||Yellowish orange|
|Powder + NH4OH||Dark orange||Dark brown||yellow|
|Powder + 1% KMnO4||Dark brown||Dark brown||Dark black|
|Powder + AgNO3||Yellow||Light yellow||Light yellow|
Preliminary Phytochemical Screening: Preliminary phytochemical screening of different extracts shows the presence of alkaloids, proteins, carbohydrates, anthraquinone glycosides, cardiac glycosides, gums & mucilages and tannins. The results are depicted in Table 5.
TABLE 5: PRELIMINARY PHYTOCHEMICAL INVESTIGATION OF DIFFERENT EXTRACTS OF CRINUM LATIFOLIUM L. LEAVES
|Group of Phytoconstituents||Extract|
|Gums & mucilages||_||+||+||+|
|Fats & oils||_||_||_||_|
|Tannins & phenolics||_||_||+||+|
|Proteins & amino acids||+||+||+||+|
TLC Fingerprint Profile: The thin layer chromatography of different extracts was performed using the following mentioned solvent systems and results are shown in table 6. The detection of developed plates was done under UV at 366 nm.
TABLE 6: THIN LAYER CHROMATOGRAPHY OF DIFFERENT EXTRACTS OF CRINUM LATIFOLIUM L. LEAVES
|Extract||Mobile phase||No. of spots||Color of spot||Rf value|
|Petroleum ether||Toluene : ethyl acetate( 8 : 1 )||3||a) orangeb) orange
|Chloroform||Toluene : ether : cyclohexane(5 : 2 : 1)||3||a) yellowb) orange
|Hydro-alcoholic||n-butanol : acetic acid: water( 4 : 1 : 5)||5||a) light brownb) light yellow
c) light blue
d) light brown
e) very light brown
|Aqueous||Methanol : chloroform(2 : 3.5)||2||a) yellow F*b) yellow F*||0.270.67|
*F refers to fluorescence
CONCLUSION: The various morphological, microscopic and physicochemical standards developed in this study will help for botanical identification and standardization of drug in crude form. Further, the authentic plant material can be explored for its pharmacological and phytochemical potential on the basis of its phytochemistry.
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J. Solanki*, A. Dhiman , A. Nanda and A. Dhankhar
Department of Pharmaceutical Sciences, Maharshi Dayanand University, Rohtak-124001, Haryana, India
17 August, 2011
28 September, 2011
24 November, 2011
01 December, 2011