Herbal, botanical, or phytomedicine refers to the use of plants or plant extracts for therapeutic purposes. This kind of medicine has long been a vital part of human culture and healthcare systems, dating back thousands of years before modern medications [1]. Herbal medicine has long been used for therapeutic purposes in many historical societies, such as Native American healing techniques, traditional Chinese medicine, Indian Ayurveda, and the medical systems of the ancient Greeks and Egyptians. After discovering that some plants had medicinal properties, these societies developed intricate systems of herbal medicine to treat a variety of ailments [2]. Herbal medicine employs a vast range of plants, each with a unique chemical composition and potential health advantages [3]. In addition, a lot of people like how gentle it is and how much less likely it is to have negative effects than other traditional treatments. Despite the lengthy history of use of herbal medicine, it is important to use caution and seek the advice of experienced practitioners before using it. Herbal therapies may not be appropriate for everyone and may interfere with medicines, just like any other type of medicine [4]. Additionally, there is a wide range of scientific evidence about the safety and efficacy of particular herbal medicines, which emphasizes the need for more study and regulation in this area [5]. Known by most as the Philippine violet, Barleria cristata is a species of flowering plant endemic to Southeast Asia, specifically found in the Philippines, Malaysia, Indonesia, India, and Sri Lanka. It belongs to the family of flowering plants known as Acanthaceae, which comprises a wide variety of tropical and subtropical species. Herbal therapy has traditionally used Barleria cristata for a very long time. Traditional medicine uses parts of the plant, such as the leaves, stems, and roots, to treat a variety of illnesses. For example, Barleria cristata is used to treat skin conditions, respiratory infections, and digestive problems in traditional Ayurvedic medicine because it is said to have anti-inflammatory, antibacterial, and analgesic qualities. [6]

MATERIALS AND METHODS:

Collection and Authentication:

  The plant of Barleria cristata was collected from Neyveli, cuddalore district, Tamil Nadu during winter of November, 2023. The plant material was identified and authenticated as Barleria cristata (certificate NO. 722.18012401) by Dr. K.N. Sunil kumar R.O and HOD pharmacognosy, central Sidhha Research Institute, Government of India, Arumbakkam, Chennai-106. The fresh stems were used for the macroscopic and microscopic characters and histochemical studies.

Processing of raw material:

The gathered plant material underwent a thorough cleaning. Plants were dried for about a week in the shade. After it had completely dried, it was ground into coarse powder and sieved number 10. It was then kept in an airtight container and utilized to analyze Physico-chemical characteristics and Powder microscopy.

PHARMACOGNOSTICAL STUDIES

Macroscopy:

External feature of test sample was documented using Nikon D-5600 Digital camera.

 Microscopy:

The sample was kept for longer than 48 hours in fixative FAA. Using a sharp knife, thin transverse sections of the preserved specimens were cut, and the sections were dyed with 0.8% Safranin and 0.5% Astra Blue. Under bright field light, transverse slices were captured on camera using an Axiolab5 trinocular microscope coupled to a Zeiss Axiocam208 colour digital camera. A scale bar was used to show magnifications.

Powder microscopy:

 Once the powdered sample had been cleared with a saturated solution of chloral hydrate, a pinch was placed on a tiny slide using a drop of 50% glycerol. To verify that there were starch grains present, the sample was treated with an iodine solution. Under bright field light, characters were seen with a Nikon ECLIPSE E200 trinocular microscope attached to a Zeiss ERc5s digital camera. Diagnostic character photomicrographs were taken and recorded.

Histochemical tests:

Plant sections were treated following the standard procedures: 

1. Crystals:

Acetic acid was used to irrigate one end of the cover slip while the portion was submerged in water. Using a piece of filter paper at the other end of the cover slip, the water within was replenished while the microscope was being used. If no air bubbles developed, the experiment was repeated with concentrated HCl, and the presence of calcium oxalate crystal was indicated by the development of needle shaped calcium sulphate and the dissolution of crystals.

2. Fats, Fatty oils volatile oils and resins:

After adding one or two drops of Sudan-IV, the area was left to stand for a little while. Orange-red, pink, or red coloured globules indicated the presence of fatty oil components; red-hued irregular contents suggested resin.

3. Starch:

A drop of 2% iodine water solution was added - blue colour indicated starch.

4. Phenolic compounds:

A drop of alcoholic ferric chloride was added - bluish black coloured contents indicated phenolic compounds like flavonoids/ tannins etc

5. Mucilage:

A drop of ruthenium red was added - pink to red coloured contents indicated mucilage.

6. Lignified cell walls:

A drop of phloroglucinol was added to the section and allowed to stand for about 2 min or until almost dry. A drop of 50% HCl was added and observed over a cover-glass - cell walls-stained pink to cherry red indicating presence of lignin.

7. Suberized or cuticular cell walls:

A drop of Sudan red III added, and the cell walls turned orange-red or crimson, indicating the deposition of Suberin or cutin over the cell wall. The mixture was then let to stand for a few minutes, gently warmed if needed.

8. Alkaloids:

A drop of Wagner’s reagent was added - the presence of yellow to reddish brown coloured contents confirmed alkaloids [7-10].

9. Physico-chemical Analysis

Ash value, Extractive value, Loss on drying, Swelling index, Foaming index and Crude fiber content were among the Physico-chemical parameters that were measured in accordance with the official protocol and the WHO standards on quality control procedures for medicinal plant materials [11-15].

RESULTS AND DISCUSSION

Macroscopical characteristics

Colour:

Green to Brown.

Taste   :

Bitter, astringent or slightly pungent flavours

Odour :

Mild

Size     :

about 1-3 feet tall, diameter 0.5-2 cm

Shape  :

Slender, slightly ribbed or grooved, erect and cylindrical.

Description :

Stem is typically erect and cylindrical in shape. It is often slender and may have some branching as it grows taller. It is slightly ribbed or grooved especially when young. As the plant matures, the stems become woody at the base while remaining relatively flexible towards the top.

       
           
       
    Figure 1: Macroscopy of Barleria cristata stem

Microscopical characteristics

TS of stem is nearly circular shaped; outer layer is single layered epidermis covered by cuticle and bears several simple, multicellular covering trichomes and few glandular trichomes; cork layer formation can be seen in some places of epidermal layer; 5 to 6 layers of collenchymatous hypodermis is present followed by 2 to 3 layers of parenchymatous cortex; a ring of vascular bundle is present in the inner cortex; xylem is well developed with vessels, fiber, parenchyma, and xylem rays; phloem surrounds both sides of xylem; outer phloem found in more layers compared to inner phloem; few phloem fibers can be clearly seen in outer phloem; central portion of the section is occupied by parenchymatous pith with abundant of acicular crystals.

Diagrammatic TS of stem

       
           
       
    Detailed TS of stem

       
           
       
    Enlarged view of stem

       
           
       
    TS of stem under Polarized field

ACr - acicular crystals; Ck - cork; Col - collenchyma; E - epidermis; GrT - ground tissue; GT - glandular trichome; Hy - hypodermis; IPh - inner phloem; OPh - outer phloem; Pa - parenchyma; Ph - phloem; T - Trichome: V - vessel; VB - vascular bundle; Xy - xylem; XyR - xylem ray

Powder microscopy

The powder sample is dark green in colour with characteristic odour and tasteless; it shows the characters like simple covering trichomes, glandular trichomes, surface view of upper epidermis, surface view of lower epidermis with diacytic stomata, palisade cells, stem epidermis with multicellular trichomes, cortical parenchyma cells, vessels with simple and bordered pitted, reticulate, and spiral thickenings, acicular crystals from stem, starch grains and cell contents.

       
           
       
    Stem epidermis with multicellular trichome

       
           
       
    Epidermis with stomata from stem

       
           
       
    Cortical parenchyma cells

       
           
       
    Pitted vessel

       
           
       
    Reticulate Vessel

Spiral vessels

       
           
       
    Bordered pitted vessel

       
           
       
   Acicular crystals from stem

       
           
       
    Starch grains

       
           
       
    Cells with contents

Histochemistry

Stem

Cutin present on epidermis; alkaloids observed in phloem region; phenolic compounds found in cortex and vascular region; lignin detected in xylem; resin contents present in outer and inner cortical region; acicular crystals observed in pith parenchyma; mucilage absents in stem.