Pharmacognostic Profile of Ageratum conyzoides L Plant and Simplicia

The development of study on medicinal plants is growing rapidly. Especially the development of pharmacology and phytochemicals study based on indications of medicinal plants that have been used by some people with empirically tested properties. Many plants are considered as raw material for medicine so it is recommended for therapeutic use.1 The use of plants as traditional medicines and alternative medicines is increasingly in demand because it is relatively safer compared to synthetic drugs. It needs many studies to develop plant properties with various methods such as the study of ethnomedicine or ethnobotany, bio essay, and purification of bioactive compounds. Increasing the need for medicinal plants by the pharmaceutical industry and its own needs by 5-18% per year.2


INTRODUCTION
The development of study on medicinal plants is growing rapidly. Especially the development of pharmacology and phytochemicals study based on indications of medicinal plants that have been used by some people with empirically tested properties. Many plants are considered as raw material for medicine so it is recommended for therapeutic use. 1 The use of plants as traditional medicines and alternative medicines is increasingly in demand because it is relatively safer compared to synthetic drugs. It needs many studies to develop plant properties with various methods such as the study of ethnomedicine or ethnobotany, bio essay, and purification of bioactive compounds. Increasing the need for medicinal plants by the pharmaceutical industry and its own needs by 5-18% per year .2 The Asteraceae family consists of 1500 genera and 2500 different species. 3 Ageratum is one of the genera included in the Asteraceae family and consists of 30 species. 4 Ageratum conyzoides is called goatweed 5 and has been empirically used as a drug to treat boils and fever medications. 6 A. conyzoides is a tropical plant commonly found in the western and eastern regions of the African continent, in several regions of Asia and South America. The plant is a polymorphic aromatic weed. The leaves of Ageratum conyzoides are used as wounds therapy, 7 hemorrhoids, anti-dysentery, antibacterial, 8 anti-inflammatory, analgesic, antipyretic, antispasmodic, gastroprotective, antiulcer, insecticide, 9 antihelmintic and antimosquito. The results showed that many plants were recommended for medicinal purposes. 1 The use of plants for medicinal purposes and research can be used as a basis for reference in the discovery of active compounds as drugs. The recommendation is based on secondary metabolites produced by plants as natural organic chemical compounds. 10 Based on the content of chemical compounds in plants such as terpenoids, alkaloids, flavonoids, tannins, saponins, and steroids whose quality and quantity are sufficient so that they can promise therapeutic efficacy of medicinal plants. 4 Natural organic compounds can be used as marker compounds either in the synthesis or semi-synthesis processes so that they play an important role in the discovery and development of drugs based on therapeutic targets. Incorrect identification of natural products or herbal medicines causes its use is also wrong, 10 for example, the existence of vernacular names given to two or more completely different species. All of these problems can be solved by pharmacognostic studies in plants.

Macroscopic analysis
Fresh plants and simplicia were observed and performed organoleptically including shapes, colors, odors, tastes, surfaces, characters, textures, and bedding.

Microscopic analysis
Microscopic analysis was determined by preparing a thin section crosssection of leaves and stems. The incision was placed on a glass object and drops of water and covered with a deck glass then observed under a microscope with magnification (40X, 100X). Cells such as epidermal (cells upper and lower), xylem, phloem, collenchyma, and parenchyma were observed. All images were seen photographed and recorded for further analysis.

a. Drying shrinkage examination
One gram of powder that placed into a porcelain crucible has been preheated at 105ºC for 30 minutes and has been tamed. The powder was flatten in crushed porcelain. Then the powder was put into the oven, heated at a temperature of 105 0 C for 1 hour, removed and cooled in a desiccator. The results were weighed until a fixed weight and the dry weight was obtained.

b. Ash content analysis
One gram of powder was put into porcelain crucifix. The powder was put into a furnace temperature of 600ºC for 6 hours, put in the desiccator and weighed. Then the ash of the dried material content was determined. Information: A= Crucible weight empty B= Crucible weight + sample before incubation C= Crucible weight + sample after incubation

c. Determination of ash content which is not soluble in acid
The powder was boiled with 25 ml of dilute sulfuric acid for 5 minutes. The acid-insoluble portion was collected, filtered through an ash-free filter paper which was not soluble in acid to the dried material.

d. Determination of water soluble essence
Simplicia powder was weight carefully about 5 g. The powder was put in a clogged pumpkin, added of 100 mL of saturated chloroform water, shaked repeatedly for the first 6 hours, then settled down for 18 hours and then filtered. The filtrate of 20 mL was evaporated and then dried in a shallow cup that has been heated 105 0 and tapped. The remainder was heat at 105 0 until fixed weight. The value of % water-soluble extracts was determined.

e. Determination of ethanol soluble extract
Five gram of powder was put in the clogged pumpkin, added 100 mL of 95% ethanol, shaked repeatedly for the first 6 hours and settled down for 18 hours. The mixture was quick filtered to avoid ethanol evaporation. The filtrate of 20 mL was dried in a shallow flat-bottomed dish that has been heated 105 0 , tamed, and heat the remainder at 105 0 until the fixed weight. The value of % ethanol-soluble extract was determined.

f. Specific gravity
Sample of extract as much as 1% was diluted with ethanol in a pycnometer. The pycnometer was calibrated by setting the weight of the pycnometer and the weight of the water which heated at 25°C.
Temperature that used was set at around 20⁰C. Sample of extract were put in a pycnometer with a temperature of 25 0 C. The excess of liquid extract was removed and weighed. Specific gravity of extract was calculated employing the weight of the dilute extract divided by the weight of the extract and the weight of water. 11

g. Identification of chemical compounds in A conyzoides
Identification of essential oils using the steam distillation method, while for other compounds using chemical reagents such as FeCl3 to determine tannins, HCl 2N to determine saponins, Yodine 0.1 N to find carbohydrates and glycosides with FeCl3 and HCl addition. This chemical reagent will give a distinctive color or precipitate according to compounds contained in plants.

Macroscopic analysis
Ageratum conyzoides is a species that belongs to the family Asteraceae. This plant is a weed that grows wild, can be found around houses, gardens, and bushes. The performed organoleptically of plant including shapes, colors, odors, tastes, surfaces, characters, textures, and bedding were shown in Table 1 and Figure 1. The characteristics of plant was purple, bluish or white flowers. As shown in Figure 1, characteristic of A.conyzoides leaf was pale green, aromatic odor, spicy taste, hairy on both sides with cuneate base, 30-70 cm height, petiole 1.1-1.5 cm long and egg-shaped with the wide end at the base (ovoid) (Figure 1). We found the shape of A.conyzoides root was soft hairy (Table 1).

Microscopic analysis
Characteristics of fresh plants on the cross-section of the trunk were showed the presence of epidermis, collenchyma reinforcement tissue, sclerenchyma reinforcement tissue, parenchyma, transport beam, trichome gland (Figures 2 and 3), whereas trichome and stomata glands were found in leaves (Figures 3c and 3d). There were tissue fragments such as parenchymal tissue, parenchyma phloem, cork, trichome, mesophyll, parenchyma cortex, fibers, amylum, secretion cells and stomata in simplicia (Figures 3a and 3b).

Determination of quality of simplicia
Results of the Simplicia powder characterization including determination of water content, determination of total ash content and acid insoluble ash content, determination of drying losses and determination of water-soluble and ethanol extracts are presented in Table 2. Simplicia characterization of A.conyzoides was evaluated to determine simplicia specifications and clarify of the material under study because the origin of the growing environment affected the content of active compounds. As shown in Table 2, the highest value of water content was in the radix. The highest value of ash content was in the stem. The highest value of acid soluble ash content was in the radix. The highest value of water-soluble extract was in the folium. The highest value of ethanol-soluble extract was in the flower.

Determination of the specific gravity of extract
Powder of simplicia was extracted by maceration method using ethanol 96% for 3 x 24 hours. The filtrate was collected and evaporated with an evaporator and the value of yield was calculated ( Table 3). The specific gravity of extract on the roots, leaves, stems and flowers of A. conyzoides, each with the same specific gravity, while the highest yield is in the leaves.

Identification of chemical compounds of Ageratum conyzoides
Chemical constituent studies are supporting data on pharmacognitive profiles. In this study identification was carried out using chemical reagents that can change color or precipitate formation in accordance with the character of chemical compounds contained in the sample (Table 4). This research helps determine the compounds that are efficacious in plants. The compounds identified in A conyzoides are essential oils, tannins, saponins, carbohydrates and glycosides (Table 4).

DISCUSSION
Identification is an inseparable part of the standardization of medicinal ingredients. 12 Before the ingredients are put in Herbal Pharmacopeia, pharmacognostic standards and quality of Simplicia must be specified. Macroscopy standards, microscopy, and quality of Simplicia can be used as an identification parameter to authenticate medicines. 13 The Macroscopic character from the factory is very needed for the source of diagnostic parameters. 14 The microscopic method is one of the simple and inexpensive methods. This method can be used to identify plant specimens. Previous studies of leaf cross-sections have been reported. Standardization becomes an important measure to determine the quality, quantity, purity, and authenticity of plants. 14