Pharmacognostical Evaluation of Indigofera glandulifera Stem

R. Vijaya Bharathi¹, C. Vamsadhara², G. Sumathi³ and K. Rajendran⁴

¹Department of Pharmacognosy, Madras Medical College, Chennai, India.

²Tamil Nadu Medical Services, Tamil Nadu, India

³Department of Microbiology, Madras Medical Collage, Chennai, India.

⁴Department of Pharmacognosy, College of Pharmacy, 7^th of April University, Al- Zawia, Libya.

ABSTRACT:

This article presents an identity based pharmacognostical study of the stem of the crude drug Indigofera glandulifera Linn. (Papilionaceae). Morphoanatomy of the stem was studied using light and confocal microscopy, World Health Organization guidelines on quality control methods for medicinal plant materials. The physico-chemical, morphological, histological parameters presented in this paper may be proposed as parameters to establish the authenticity of I. glandulifera and can possibly help to differentiate the drug from its other species

KEYWORDS: Pharmacognostical, Indigofera glandulifera, stem, Papilionaceae

INTRODUCTION

Indigofera glandulifera Linn. (Papilionaceae) is a sub-shrub and grows up to 60 cm in height. The plant is distributed in India, Himalayas, Sri Lanka, South Eastern Asia, China, Japan, Malaysia and Australia. Seeds are used as astringent, aphrodisiac, tonic and restorative. The plant is used in rheumatism, lumbago, general debility after delivery, seminal weakness and leucorrhoea. A decoction of the seeds is useful for the relief of pain in the back and waist¹.

In spite of the numerous medicinal uses attributed to this plant, there are no pharmacognostical reports on the stem of this plant. Hence, the present investigation deals with the pharmacognostical evaluation of the stem of I. glandulifera. The study includes morphological and anatomical evaluation and determination of physico-chemical constants of Indigofera glandulifera.

MATERIALS AND METHODS:

Plant material:

The plant I. glandulifera had collected from Chennai, Tamil Nadu, India, during the months of September 2008. The botanical identity of the plant was confirmed by Dr. P. Jayaraman, Botanist, Plant Anatomical Research Centre, Chennai, Tamil Nadu.

Macroscopic and microscopic analysis:

The macroscopy and microscopy of the stem were studied according to the method of Brain and Turner². For the microscopical studies, cross sections were prepared and stained as per the procedure of Johansen³. The micropowder analysis was done according to the method of Brain and Turner⁴ and Kokate⁵.

Physico-chemical analysis:

Physico-chemical analysis i.e. percentage of ash values and extractive values were carried out according to the official methods prescribed by Indian Pharmacopoeia⁶ and the WHO guidelines on quality control methods for medicinal plant materials⁷. Fluorescence analysis was carried out according to the method of Chase and Pratt⁸ and Kokoski et al⁹.

RESULTS AND DISCUSSION:

Macroscopic features:

The plant is a sub-shrub and grows up to 60 cm in height. Young branches are grey-glabrescent; leaves- trifoliolate, leaflets oblanceolate, 1.5 cm long 30 mm broad, lower surface pubescent, gland dotted, lease cuneate, margins entire, apex obtuse; inflorescence- dense raceme, 6-10 flowered, calyx-5 sepals, gamosepalous, sepal lobes setaceous, corolla-flame colored, pod-linear, 4 angled, narrowly winged.

Microscopic features:

Young stem measuring 1.5 m in diameter was studied. It consists of a broken epidermis of narrow rectangular cells (Fig. 1 a). In matured stem the periderm is deeper in position and consists of three or four layers of tabular cells (Fig. 1 b). On the outer and inner part of the periderm are narrow cortical layers. Secondary xylem cylinder is thick comprising of xylem fibres and vessels. The xylem cylinder is corroded by wide masses phloem, so that xylem is seen in broken irregular patches. Primary xylem is in continuous radial rows around the pith. The vessels are thin walled, narrow, circular and diffuse in distribution. The vessels are 20-45 µm wide. Pith is wide and parenchymatous cells are large, thin walled, angular and compact (Fig. 1 c and 1 d).

Ads- Adaxial side; Co- cortex; Ep, Epidermis; Gt- Ground tissue; Pe- Periderm; Pi- Pith; Ph- phloem; Px-Primary xylem; Sph- Secondary phloem; Sx- secondary xylem.

Powder characteristics:

The stem powder is light brown in colour with a characteristic odour and astringent taste. The vessels are thin walled, narrow, circular and diffuse in distribution. The vessels are 20-45 µm wide (Fig 2 a). Vascular elements are seen with bordered pits. Parenchymatous cells are polyhedral in shape, thick walled and compact, some of the cells are filled with dense tannin content (Fig. 2b). Lignified fibres are often seen associated with vessels (Fig.2 d) and non- lignified fibres are rarely seen (Fig. 2c). It occurs either single or in groups of 4-10 fibres and measure 460- 570-650 μm in length.

Fig. 2. Powder characters of I. glandulifera stem

(a) Vessels; (b) Parenchyma; (c) Non lignified fibres; (d) Lignified fibres

Physico-chemical constants:

Ash value of a drug gives an idea of the earthy matter or the inorganic composition and other impurities present along with the drug. The ash values (Table 1) of the powdered aerial part of I. glandulifera revealed a high concentration of sulphated ash. Extractive values are primarily useful for the determination of exhausted or adulterated drugs. The ethanol soluble extractive (Table 2) was high in I. glandulifera. The results of fluorescence analysis of the drug powder are presented in Table 3.

Table 1. Ash values of the stem powder of I. glandulifera

Parameters

Values % (w/w)

Total ash

Acid insoluble ash

Water soluble ash

Sulphated ash

09.40

0.70

1.36

11.69

Table 2. Extractive values of the stem powder of I. glandulifera

Parameters

Values % (w/w)

a) Water soluble extractive

b) Ethanol soluble extractive

c) Ether soluble extractive

3.27

6.23

1.08

CONCLUSION:

As there is no pharmacognostic/anatomical work on record for this much valued traditional drug, the present work was taken up with a view to lay down standards which could be useful to detect the authenticity of this medicinally useful plant. While discussing the purpose of systematic anatomy, Metcalfe and Chalk¹⁰ pointed out that any exercise that involves the identification of plant material when it is in a fragmentary or partly decomposed condition can be achieved by the method of comparative histology. Macro and micro morphological standards and physic-chemical profile discussed can be considered as identifying parameters to substantiate and authenticate the plant Indigofera glandulifera

REFERENCES:

1) Nadkarni (1976): Indian Materia Medica, In. Ayurvedic Unani and home remedies, vol. I, New Delhi: Popular Prakashan

2) Brain KR, Turner TD (1975a): The Practical Evaluation of Phytopharmaceuticals. Wright-Scientechnica, Bristol, pp 4-9.

3) Johansen DA (1940): Plant Microtechnique, McGraw Hill, New York, pp 182.

4) Brain KR, Turner TD (1975b): The Practical Evaluation of Phytopharmaceuticals, Wright-Scientechnica, Bristol, pp 36-45.

5) Kokate CK (1986): Practical Pharmacognosy, 1^st ed., Vallabh Prakashan, New Delhi, pp 15-30.

6) Indian Pharmacopoeia (1996): 4^th edn.,Vol. II, Government of India, Ministry of Health and Welfare, Controller of Publications, New Delhi, pp. A53-A54.

7) WHO/ PHARM/ 92.559/ rev.1 (1992): Quality Control Methods for Medicinal Plant Materials, Organization Mondiale De La Sante, Geneva, pp 9, 22-34.

8) Chase CR, Pratt RJ (1949): Fluorescence of powdered vegetable drugs with particular reference to development of a system of identification. J Am Pharmacol Assoc 38, 32-36.

9) Kokoski J, Kokoski R, Slama FJ (1958): Fluorescence of powdered vegetable drugs under ultraviolet radiation. J Am Pharmacol Assoc 47, 715.

10) Metcalfe CR, Chalk L (1981): Anatomy of the Dicotyledons. vol. I, Oxford: Clarendon Press.

Table 3. Fluorescence analysis of the stem powder of I. glandulifera

Treatment	Day light	UV light (254 nm)
Powder as such	Light green	Slightly brownish green
Powder + 1 N NaOH (Aqueous)	Bright greenish yellow	Slightly brownish green
Powder + 1 N NaOH (Alcoholic)	Bright greenish yellow	Light green
Powder + 1 N HCl	Green	Fluorescent green
Powder + Ammonia	Yellowish green	Fluorescent greenish brown
Powder + Iodine	Blackish green	Brownish green
Powder + FeCl₃	Green	Dark greenish brown
Powder + 1 N H₂SO₄	Slightly brownish green	Light brown
Powder + Acetic acid	Light green	Dark brown
Powder + 1N HNO₃	Greenish brown	Greenish brown