Pharmacognostic and Physicochemical Studies of Garcinia lanceifolia Roxb. Var. Oxyphylla

 

Prakash Chandra Gupta¹*, Ashish Kar², Nisha Sharma¹, Nikunj Sethi¹, Prashant Pandey¹, Dipankar Saharia², Naba Kumar Goswami²

¹University Institute of Pharmacy, Chhatrapati Shahu Ji Maharaj University, Kanpur-208024, Uttar Pradesh India

²The Energy and Resources Institute (TERI), Northeastern Regional Centre, Guwahati-781036 Assam, India

 

ABSTRACT:

Garcinia lanceifolia Roxb. var. Oxyphylla (Planch. & Triana) Laness (Clusiaceae), is a small and tropical evergreen plant commonly known as “Rupahi-thekera” (Assamese). It is an important and endemic medicinal plant of Assam which has been used by various ethnic communities of North-East India to treat various disorders like dysentery, dyspepsia and jaundice. The plant is considered to be containing much medicinal value and is also eaten raw or made into pickles by the local people. However, detailed scientific information is not available to identify the plant material, in order to ascertain its quality and purity. Therefore the study of morpho-anatomical characters and physicochemical analysis of G. lanceifolia was undertaken to establish the pharmacognostic and phytochemical details about the plant. Macro and microscopical studies of leaf showed the presence of simple leaf, lanceolate in shape, acuminate apex and paracytic type of stomata. Bowl shaped arrangement of xylem in leaf, crescent form vascular tissue in petiole and well developed phloem in the form of ring in stem are some of the diagnostic features noted from anatomical study of the plant. Powder microscopy revealed the presence of fibers, spiral and pitted vessels. HPTLC finger printing of plant tried with solvent system chloroform and methanol (8:2) confirmed the presence of 09 spots with different R_f value under UV light 366λ.The fluorescent analysis under day light and UV light by treatment with different chemical reagents showed different colour. Phytochemical evaluation revealed the presence of glycoside, triterpenoids, saponins, tannins and flavonoids.

 

 

 

INTRODUCTION:

Garcinia lanceifolia Roxb.var. xyphylla (Planch. & Triana) Laness commonly known as “Rupahi-thekera” (Assamese), belonging to the family Clusiaceae is an important and endemic medicinal plant found in Assam. It is a small and tropical evergreen plant. The plant is glabrous and grows upto a height of 12 feet under the dense shade of other trees. Ripe fruits are eaten raw or dried and are considered to be effective in cases of diarrhea.

 

The fruits and gum resin, called "gamboge" as well as the oil and juice of the plant are used as medicine for fever, jaundice, diabetes and urinary problems^1,2. Sweet and mature fruits, young leaves and shoots which are slightly acidic in taste are cooked and eaten by the Karbi and Mishing tribes of Assam. The leaves of G. lanceifolia are used as stomachic, diuretic and the fruit is used as a cure for dysentery and diarrhoea. The leaves are also cooked as vegetables and made into pickles³. The fruits are acidic and dried fruits are used against dysentery in Assam⁴. Plants belonging to the genus Garcinia are known to contain bioactive compounds such as xanthones, biflavonoids, benzophenones, benzoquinones, and triterpenes. G. lanceifolia is reported to possess analgesic and anti-inflammatory activity, antimicrobial, antioxidant activity⁵. Correct identification and quality assurance of the starting materials is an essential prerequisite to ensure reproducible quality, which will provide safety and efficacy of herbal medicine. This study was undertaken to generate standardized data on various pharmacognostical, phyto and physico-chemical characteristics of the plant materials. The outcome of the present study will be helpful in identification, authentication and quality control of the plant materials.

 

MATERIAL AND METHODS:

Plant material:

Garcinia lanceifolia plants were collected from homestead garden of Ashrang village, Dimahasao district, Assam, India in the month of July 2017. The specimen was identified by Taxonomist, TERI-Northeastern Regional Centre, Guwahati and later specimen was confirmed in BSI, Shillong and voucher specimen was deposited in herbarium section of TERI-Guwahati for future reference.

 

Pharmacognostic study:

Fresh leaves, petiole and stem were taken for morphological and histological studies. Coarse powder (60#) was used to study microscopical characters, physicochemical parameters and phytochemical investigation. The detailed pharmacognostic studies of the plant leaf, petiole and stem were carried out according to well established method and procedure^6,7,8. The powder microscopy was performed according to the method of Khandelwal⁸.

 

Physicochemical constants:

Various physicochemical parameters like moisture content, ash values and extractive values were evaluated according to well established official method and procedure^8,9,10.

 

Fluorescence study:

Powdered leaf and fruit material were treated with various chemical reagents and exposed to visible, ultraviolet light (Short UV) to study their fluorescence behavior¹¹.

 

HPTLC studies:

For proper and meaningful utilization, it is important to have quality standards of material and for this quality standardization, HPTLC finger print profile of alcoholic extract of Garcinia lanceifolia (10 µL of 1 mg/mL) was developed. The HPTLC analysis was carried out on precoated silica gel 60 F₂₅₄ plate (Merck, India) with the help of Camag Linomat IV applicator. The plate was then eluted with solvent system in a CAMAG twin trough chamber up to a distance of 9 cm. After development, plate was dried and densitometrically scanned on a TLC scanner III at 366 nm using Wincats software (CAMAG, Switzerland) and peak area was recorded.

 

RESULT:

Pharmacognostic study:

Macroscopic Characters:

Macroscopically, the fresh leaf of Garcinia lanceifolia is 6 to 13 cm long, 2 to 3 cm in  width and petiole 1 to 1.3 cm long, lanceolate in shape, acuminate apex and green in colour (Fig. 1).  Fruits are small, ovoid, orange in colour and 6-8 seeded (Fig. 2).

 

 

Fig. 1. Macroscopic characterstics of G. lanceifoliaRoxb. leaf

 

 

Fig. 2. Garcinia lanceifolia Roxb.var. Oxyphylla

 

Microscopical characterstics:

Leaf microscopy:

Transverse section (T.S.) passing through midrib region shows dorsiventral shape. Upper and lower surface of the leaf consist of rectangular thin walled epidermis covered with thick cuticle followed by collenchymatous ground tissue. Palisade cell is 2-3 layered followed by several layer of spongy mesophyll. Midrib shows presence of 4-5 layered collenchyma below the upper epidermis and above the lower epidermis. Midrib region shows large vascular bundle covered with pericyclic fibres. Xylem is arranged in bowl shaped and surrounded by phloem. Xylem is consists of vessels, tracheids, fibers and xylem parenchyma (Fig. 3). Lower leaf surface of G. lanceifolia shows paracytic stomata (Fig. 4a), while stomata is absent in upper leaf surface. Leaf surface also shows the presence of veins, vein islet, vein terminations (Fig. 4b) and palisade cells (Fig. 4c). Leaf constant such as stomatal number, stomatal index, vein islets number, vein termination number were measured. The results are shown in Table 1.

 

 

Fig. 3. T.S. of G. lanceifolia leaf

 

Uep: Upper epidermis; Lep: Lower epidermis; Cu: Cuticle; Col: collenchymas; PF: Pericyclic Fibre; Xy: Xylem; Ph: Phloem

 

 

a: Stomata

 

 

b: Vein-islet and Veinlet termination

 

c: Palisade cells

Fig. 4. Leaf surface of G. lanceifolia

Table 1. Leaf constants of G. lanceifolia (at 100X)

S. No.	Parameters	Value (in 1 mm2 area)
1	Stomatal number, Upper surface	Nil
2	Stomatal number, Lower surface	24
3	Stomatal index, Upper surface	Nil
4	Stomatal index, Lower surface	16.21
5	Vein-islet number	2.0-3.0
6	Veinlet termination number	4.0-6.0
7	Palisade ratio	3.0-4.0 (per cell)

 

Petiole microscopy:

T.S. of petiole is somewhat circular in shape with two ridges at upper side; single layered epidermal cells covered with thick cuticle; ground tissue collenchymatous. Vascular tissue is arranged in crescent form in the center of petiole. Group of pericyclic fibers seen in vascular bundle region. Vascular bundle is composed of metaxylem, protoxylem and phloem (Fig.5).

 

Fig. 5. T.S. of G. lanceifoliapetiole

Ep: Epidermis; Col: collenchyma; PF: Pericyclic Fibre; Xy: Xylem; Ph: Phloem

 

Stem microscopy:

T.S. of stem almost circular with wavy outline. Single layered and thick walled epidermis covered with thick cuticle; hypodermis is collenchymatous followed by 2-3 layer of collenchymas; cortex is parenchymatous and pericyclic fibres are present in scattered form. Phloem is well developed in the form of ring and consists of sieve tubes, companion cells and phloem parenchyma. Xylem is present in the form of continuous ring and consists of vessels, tracheids, fibers and xylem parenchyma; vessels are arranged in radial rows. Medullary rays are distinct; centre portion is occupied by collenchymatous pith (Fig. 6).

 

Fig.6. T.S. of G. lanceifoliastem

Ep: Epidermis; Col: collenchymas; PF: Pericyclic Fibre; Xy: Xylem;

Ph: Phloem

 

 

Powder microscopic characters:

The powdered plant material is greenish in color; showing fragments of parenchyma, fibers, pallisade cells, epidermal celland vessels having scalariform thickening (Fig. 7).

 

Preliminary phytochemical screening:

Preliminary phytochemical screening mainly revealed the presence of glycoside, triterpenoids, saponins, tannins, flavonoids and steroids.

 

Physicochemical parameter:

Phsicochemical analysis of fruit viz. moisture content, ash value and extractive values are presented in Fig.8. The fluorescence analysis of leaf and fruit powder is recorded in Table 2.

 

HPTLC studies:

A densitometric HPTLC analysis was performed for the development of characteristic finger print profile which may be used as marker for quality evaluation and standardization of the drug. The preliminary HPTLC studies revealed that the solvent system chloroform: methanol (8:2) was ideal for the alcoholic extract and gave well resolved peaks of crude extract of G. lanceifolia (Fig.9).

 

 

 

 

Table 2: Fluorescence analysis of leaf and fruit powder of G. lanceifolia

Treatment	Observation under day light		Observation under U.V. light (254 nm)
	Leaf	Fruit	Leaf	Fruit
Powder as such	Pale green	Light brown	Green	Olive
Powder + 1N NaOH in Methanol	Mustard	Mustard	Bright green	Fluorescent green
Powder + 1N HCl	Tan	Yellow	Light green	Green
Powder + 1N NaOH in water	Mustard	Mustard	Green	Bright green
Powder + HNO3  (1:1)	Olive	Yellow	Dark brown	Fluorescent green
Powder + H2SO4  (1:1)	Tan	Yellow	Light green	Green

 

 

Fig.8. Results of physicochemical parameters of G. lanceifolia

 

 

Fig. 9. HPTLC profile and densitometric scanning of methanolic extract of G. lanceifolia

Solvent system: Chloroform: Methanol (9:1), Detection: Under UV light λ 366 nm

 

DISCUSSION:

Ethnomedically, the leaves and fruit of the plant were used by local people for  the treatment of various disease conditions without standardization. The standardization of a crude drug is an integral part of establishing its correct identity. Before any crude drug can be included in an herbal pharmacopoeia, pharmacognostic parameters and standards must be established. Microscopic method is one of the simplest and cheapest methods to starts with for establishing the correct identity of the source materials^12,13. The pharmacognostic standards of Garcinia lanceifolia are carried out for the first time in this study. Morphological and histological studies of the leaf, petiole and stem will enable to identify crude drug. Ash values and extractive values can be used as reliable aid for detecting adulteration. These studies help in identification of the plant materials. Correct identification and quality assurance of the starting materials is an essential prerequisite to ensure reproducible quality of herbal medicine which will contribute to its safety and efficacy^14,15. The presence of paracytic stomata, arrangement of xylem in bowl shape in leaf, crescent form vascular bundle in petiole and presence of xylem in the form of ring in stem are the some of the diagnostic features noted from anatomical study of plant. Preliminary phytochemical analysis indicated presence of triterpenoids, saponins, tannins, flavonoids and steroids. Extractive values are useful to evaluate the chemical constituents present in the crude drug and also help in estimation of specific constituents soluble in particular solvents. The fluorescent analysis under day light and UV light by treatment with different chemical reagents showed different colour. This analysis suggests that, leaves and fruit extract of G. lanceifolia probably contain active agent(s) and this provides the basis for their folkloric use as a cure for some human ailments. HPTLC fingerprint profile along with their R_f values were recorded, which would serve as a reference standard for the scientist engaged in research on the medicinal properties of plant.

 

CONCLUSION:

The present work was undertaken with an aim of pharmacognostic investigation of G. lanceifolia providing useful information, which could be useful to detect the authenticity of this medicinally useful plant. Pharmacognostic evaluation can be useful to substantiate and authenticate the drug.    

 

ACKNOWLEDGEMENT:

The authors wish to acknowledge the financial support provided by the Department of Biotechnology, Government of India (Grant sanction number BT/PR16665/NER/95/236/2015).

 

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

 

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Accepted on 28.11.2018        © RJPT All right reserved

Research J. Pharm. and Tech 2019; 12(2):706-710.