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Table of Contents
ORIGINAL ARTICLE
Year : 2019  |  Volume : 2  |  Issue : 3  |  Page : 135-140

Pharmacognostic standardization and physicochemical evaluation of Caesalpinia crista L. root for quality control assessment


1 Department of Pharmacognosy, School of Pharmaceutical Sciences, Sardar Bhagwan Singh Post Graduate Institute of Biomedical Sciences and Research, Dehradun, Uttarakhand, India
2 Department of Pharmaceutical Sciences, Ch. Devi Lal College of Pharmacy, Jagadhri, Haryana, India

Date of Web Publication1-Jul-2019

Correspondence Address:
Bhuwan Chandra Joshi
Assistant Professor, School of Pharmaceutical Sciences, Sardar Bhagwan Singh Post Graduate Institute of Biomedical Sciences and Research, Balawala, Dehradun - 248 001, Uttarakhand
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/JNSM.JNSM_61_18

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  Abstract 


Background: Caesalpinia crista Linn. (Caesalpiniaceae) have been used widely as ethnomedicinal practices and traditional medicine for its medicinal values. Conventionally, the plant was used for the treatment of bladder stone, fever, leucorrhea, as a tonic for rheumatism and backache; there is no published data on standardization of the plant for its authenticity, quality, and purity. Aim: The present study was to scientifically establish a pharmacognostic profile of C. crista root as per standard procedures. Materials and Methods: The investigation deals with comprehensive pharmacognostical parameters were carried out by organoleptic, microscopic characters, physicochemical, and phytochemical studies. Results: The present study provides the general organoleptic, microscopic characteristics of the root. Physicochemical studies revealed total ash (9.20 ± 0.17), acid insoluble ash (1.25 ± 0.11), water-soluble ash (3.1 ± 0.05), alcohol-soluble extractive (11.54 ± 0.46), and water-soluble extractive (12.66 ± 0.24), respectively. Preliminary phytochemical screening of root revealed the presence of carbohydrates, proteins, alkaloids, triterpenoids, glycosides, flavonoids, tannins, diterpenes, glycosides, and saponins. Conclusion: These studies will provide referential information for the correct taxonomical identification, standardization, and establishing monograph of C. crista.

Keywords: Caesalpinia crista (Linn.), pharmacognostic analysis, physicochemical standards, phytochemical analysis


How to cite this article:
Upadhyay P, Joshi BC, Sundriyal A, Mukhija M. Pharmacognostic standardization and physicochemical evaluation of Caesalpinia crista L. root for quality control assessment. J Nat Sci Med 2019;2:135-40

How to cite this URL:
Upadhyay P, Joshi BC, Sundriyal A, Mukhija M. Pharmacognostic standardization and physicochemical evaluation of Caesalpinia crista L. root for quality control assessment. J Nat Sci Med [serial online] 2019 [cited 2019 Sep 15];2:135-40. Available from: http://www.jnsmonline.org/text.asp?2019/2/3/135/250748




  Introduction Top


Plants are used as medicine to maintain human health.[1] Plants are a valuable source of a wide range of secondary metabolites, which are used as pharmaceuticals, agrochemicals, flavors, fragrances, colors, biopesticides, and food additives.[2] According to the World Health Organization (WHO), 80% of the rural population in the developing countries depend on traditional medicines to meet their primary health-care needs.[3] The Indian Materia Medica includes about 2000 drugs of natural origin, and most of them are derived from different traditional system and folklore practices.[4] However, there are large numbers of plants, which have not been mentioned in these reports, in spite of their usage in the folk and traditional medicinal systems. Authentication and standardization are prerequisite steps while considering source materials for herbal formulation in any system of medicine.

Caesalpinia crista (Linn.) belonging to family Caesalpinioideae is a prickly shrub widely distributed throughout the tropical and subtropical regions of Southeast Asia; also found throughout the hot, humid part of India.[5],[6] It is commonly known as Karaunj, Latakranja and chiefly found in eastern sea cost from Orissa; western sea cost from Konkan southwards; Sunderbans in West Bengal; in Bhopal, and Himachal Pradesh. Conventionally, this plant was used for the treatment of gynecological disorders, skin diseases, constipation, piles, and ulcers.[7] The whole plant including roots, stem, bark, leaves, fruits, seeds, and nuts have medicinal properties. The leaves of plant are reported to be used as uterine tonic, as liver tonic; as emmenagogue; root cures leukorrhea, intermittent fevers, as tonic for the treatment of rheumatism; bark is used as a febrifuge and externally in inflammation; seeds are considered as liver tonic, febrifuge, anthelmintic, and antiblennorrhagic.[8],[9],[10] kernels are used as a tonic in intermittent fevers, asthma, and colic;[6] seed is used to treat asthma.[5],[10],[11] The plant elaborates different classes of organic compounds of medicinal importance including proteins, alkaloids, saponins, triterpenoids, diterpenoids, and flavonoids.[12],[13] Plant is also reported to have anthelmintic activity, anti-inflammatory activities, nootropic activity, antioxidant activity, anticancer activity, hepatoprotective Potential,[14] antidiabetic activity,[15] anti-amyloidogenic activity,[16] antimalarial activity,[17] and cytotoxic activity.[18],[19] As per literature survey and scientific data revealed that no systematic pharmacognostical parameter had been carried out on the root of C. crista Linn. Hence, the objective of the present study is to evaluate various pharmacognostic parameters such as macroscopy, microscopy, physicochemical, and phytochemical evaluations of the plant.


  Materials and Methods Top


Chemicals and instruments

Petroleum ether (CDH), Chloroform (CDH), ethyl acetate (Loba chem.), ethanol (Loba chem.), phloroglucinol (CDH), glycerin (CDH), glacial acetic acid (CDH), ferric chloride (CDH), lead acetate (CDH), hydrochloric acid (Rankem), potassium hydroxide (Himedia), sulfuric acid (Loba chem.), Nitric acid (Loba chem.), Wagner's reagent (CDH), Hager's reagent (CDH), Mayer's reagent (CDH), Dragendorff's reagent (CDH), Millon's reagent (CDH), Molisch's reagent (CDH), Barfoed's reagent (CDH), Biuret reagent (CDH), Benedict's reagent (CDH), and ninhydrin reagent (CDH) all chemicals and solvents used in the study were of analytical grade. Microscope (OLYMPUS, India), Soxhlet extractor (Perfit) Water bath (Perfit).

Plant material

C. crista (Linn.) plant roots were procured along with authentication certificate from Dr. K. Madhava Chetty, Department of Botany, Sri Venkateswara University, Tirupati, Andhra Pradesh, India. The voucher specimen number was (Reference: 1229). Plant drug was shade dried (<40°C), coarsely powdered and stored in airtight container.

Macroscopic and microscopic examinations

The macroscopic study was done by using a simple microscope. The plant root was macroscopically examined for color, odor, taste, shape, size, texture characteristics. Microscopic study of root was carried out by preparing of freehand sections; cleared with 5% KOH solution then stained with concentrated hydrochloric acid-phloroglucinol (1:1) and mounted with 50% glycerine solution. Photographs were obtained of the section. The same procedure was followed for microscopic studies of the powdered material of C. crista root.[20],[21]

Physiochemical analysis

Physicochemical parameters were determined as per guidelines of the WHO 2002. A physiochemical parameter such as moisture content by loss on drying method, ash values (total ash, acid insoluble ash, and water-soluble ash), and extractive values (water soluble and alcohol soluble extractives) was determined using the powdered drug.[22]

Loss on drying

Accurately weighed 1 g powder of root was placed in a china dish then dried in an oven at a temperature of 100°C for an hour. The powder was weighed again and compared with the original weight of the powder. The loss on drying was calculated by the following expression.[23]

Percentage loss on drying = Weight loss (g)/Weight of sample (g) × 100.

Fluorescence analysis

Fluorescence analysis of root powder was treated with various chemical and observed exclusively to different wavelengths of ultraviolet (254 nm and 365 nm) and visible light for observing characteristic color presentation.[24]

Extraction of plant material

Successive solvent extraction scheme was used for the preparation of different extracts. The coarsely powdered root sample (30 g) was extracted with Petroleum ether (40°–60°C) using Soxhlet apparatus. The extract was concentrated with rotary-evaporator and transferred to a preweighed china dish and dried in a vacuum desiccator. The marc obtained was then air dried and used for further extraction with chloroform followed by ethyl acetate and ethanol. The dried extracts were placed in desiccators for further studies and yield was calculated.

Preliminary phytochemical screening

The preliminary phytochemical screening was qualitatively tested for the presence of phytochemical as per described standard methods.[25],[26]

Statistical analysis

All the results were calculated in triplicates. The represented data areas mean ± standard error of the mean Microsoft Excel 2013 (Microsoft Corporation, Redmond, Washington), is used for statistical analysis.


  Results and Discussion Top


From the past few years, medicinal plants have played an important role in preventing and curing diseases as well as a source for the development of lead compounds. It becomes extremely important to make an effort toward standardization of the plant material to be used as medicine. The pharmacognostic standardization is the major and reliable criterion for identification and authentication of crude drug. The parameters used in the pharmacognostic studies are very necessary for the determination of quality and purity of plant material.[1] The process of standardization can be achieved by stepwise physiochemical studies.

As per the WHO, the organoleptic and microscopic characteristics of a crude drug are the starting step toward establishing its identity and purity.[22]

Macroscopic examination

The C. crista (Linn.) root is hard and brittle; the surface is light brown. Roots are branched with several rootlets; thickness varying with age; short fracture exposing rough surface which is pale yellow in color, characteristics odor, and has a bitter taste [Table 1].
Table 1: Macroscopic and organoleptic characters of Caesalpinia crista L. root

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Microscopic examination

The microscopic characteristics of the root transverse section revealed the following outlines such as cork with several layers of the thin-walled cell, flat, polygonal parenchymatous cells with a brownish-red matter which are saturated with suberin. Followed by thin-walled epidermal cells, cortex comprises an elongated parenchyma cell containing starch grains. Phloem consisting of sieve tube, a bundle of fibers with calcium oxalate crystal; entire central portion occupies by xylem with prominent vessels; biseriate medullary rays; and pith lie of parenchyma cell was very small shown in [Figure 1].
Figure 1: Transverse section of Caesalpinia crista root. (a) stained root section (b) stained pith section; where Xy: Xylem, Ed: Epidermis, Ck: Cork, Co: Cortex, Ph: Phloem, Mr: Medullary ray, Mxv: Medullary xylem vessel, Pi: Pith, (MG, ×100)

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Powder characteristics

Microscopic observation of C. crista (Linn.) root indicated the presence of cork cells, cork cells with cell inclusion, parenchyma cell, fiber, lignified xylem vessels, and prismatic calcium oxalate crystal were shown in [Figure 2].
Figure 2: Powder microscopy of Caesalpinia crista Linn. root where (a) Calcium oxalate crystal, (b) Fiber, (c) Lignified xylem vessels, (d) Lignified cells, (e) Pitted xylem vessels, (f) Cork cells, (g) Cork cells with cell inclusion, (h) Parenchyma cell) (MG, ×100)

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It was found that morphological characteristics such as color, odor, taste, size, and shape are useful in gross identification. It is therefore considered as a primary screen in the qualitative assessment of crude drugs. Microscopic characterization is a reliable tool for identification of plant, a small fragment of crude drugs, and the detection of authentic plant or adulterants and substituents.[27] The use of various reagents or dyes which stains particular cells helps to distinguish cellular structures depending on their chemical nature.

The powdered drug was evaluated for its physicochemical parameters such as moisture content, total ash, acid insoluble ash, and different extractive values.

Physicochemical parameters

The physicochemical constants can use as a valuable source of information and used in evaluating the purity, quality of crude drugs.[28] Total Ash values of the drug give an idea about inorganic composition such as earthy matter and other impurities present along with the drug. The total ash, water-soluble ash, and acid insoluble ash were found to be 9.20 ± 0.17, 3.1 ± 0.05, and 1.25% ± 0.11% w/w, respectively. These values were found to be reasonably low indicating low contamination. This shows the normal quality of the drug. The extractive values give an idea about phytoconstituents of crude drugs. However, plant root extractive values were found to be 11.54% ± 0.46% w/w alcohol soluble and 12.66% ± 0.56% w/w water soluble extractives, respectively, given in [Table 2]. Comparing the water-soluble and alcohol-soluble extractive values of the C. crista, it was concluded that the water extractive values were higher than alcohol indicated the presence of more amount of water-soluble contents in the plants.
Table 2: Physicochemical parameters of Caesalpinia crista L. root

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The total ash values and extractive values are useful in the identification and authentication of the plant material. Ash values of drug give an idea about the earthy matter, inorganic composition, and other impurities such as carbonates, phosphates, silicates, and silica present along with the drug. Extractive values are used to evaluate the nature of chemical constituents present in crude drug and also to help in the estimation of specific constituents soluble in a particular solvent.[27],[29]

These ash and extractive values can be efficiently used for the standardization of herbal drugs. The results obtained from the study could be utilized as a reference for setting limits for the reference standards for the quality control and assurance of drug.

Loss on drying

Loss on drying is the loss in weight in %age w/w. It determines the amount of volatile matter of any kind (including water) that can be driven off under the conditions specified (desiccators or hot air oven). The loss on drying of root powder was 5.34% ± 0.42% w/w which seems to be lower than that necessary to support the growth of microorganisms such as mold, bacteria, yeast, and fungi to bring the changes in the chemical compositions of the crude drugs.[30]

Fluorescence analysis

Fluorescence analyses of root powder were conducted with different reagents, and fluorescence was observed under ultraviolet (UV)-254 nm and UV-366 nm. The results were compared with their respective observations in visible light and observations were represented in [Table 3].
Table 3: Observations of Caesalpinia crista L. root powder under visible and ultra violet (254 nm and 366 nm) light

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Fluorescence analysis of the root powder helps in the qualitative investigation which can be used as a reference data for the identification of adulterants.[31] Fluorescence is a significant phenomenon exhibited by various chemical constituents present in plant material. The fluorescence color is specific for each compound.

Extraction of plant material

Successive solvent extracts of plant root were subjected for calculating their % yield, color, and consistency respectively given in [Table 4].
Table 4: Percentage extractive value and characterization of Caesalpinia crista L. root

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Preliminary phytochemical screening

Preliminary phytochemical screening gives a general idea regarding the nature of the different chemical constituents of the plant. The qualitative preliminary phytochemical screening of C. crista (Linn.) root was mentioned in [Table 5]. C. crista reveals the presence of steroids/terpenoids, saponins, flavonoids, tannins, and phenolics in different extracts. Phytochemical evaluation and chemo-profiling are useful for the quality assessment of plant materials.[25] It is a valuable step in the detection of the bioactive principals present in the medicinal plants and may lead to drug discovery and development.
Table 5: Preliminary phytochemical screening of Caesalpinia crista L. root

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  Conclusion Top


In the present study can serve as great useful information on correct identification and standardization of the plant material while evaluating the organoleptic, microscopic studies, physicochemical parameter as well as investigating its phytoconstituents which would help in the investigation of its possible therapeutic actions. In addition, the results of the present study could be useful for the drafting of a monograph of the C. crista (Linn.).

Acknowledgment

We express our sincere thanks to Shri S.P. Singh Honorable Chairman, Sardar Bhagwan Singh Post Graduate Institute of Biomedical Sciences and Research, Balawala India for providing the facilities.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

  [Figure 1], [Figure 2]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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