Comparative Study of Various Plants of Piperaceae Family Commonly Used in Ayurvedic Formulations

A.K. Meena¹*, M.M. Rao¹, V. Nageswara Rao³, Komalpreet¹, M.M. Padhi² and Ramesh Babu²

¹National Institute of Ayurvedic Pharmaceutical Research, Patiala-147001, Punjab.

²Central Council for Research in Ayurveda and Siddha, Janakpuri, Delhi-110058.

3 National Institute of Ayurveda, Jaipur.

ABSTRACT:

The present article attempts to compare TLC and physico-chemical parameters of P. nigrum Linn., Piper cubeba Linn., Piper longum Linn. and Piper chaba Hunter belonging to common family piperaceae. Each of them is considered to have huge medicinal value in Ayurveda, Sidhha and Unani traditional medicines. Since ancient times, these drugs are used according to their medical value. Investigation of such traditionally used medicinal plants is thus valuable on two levels, firstly, as a source of potential chemo therapeutic drugs and secondly, as a measure of safety for the continued use of medicinal plants.

The present paper attempts to evaluate the physicochemical parameters like pH, Loss on drying at 105°C, Water soluble extract, Alcohol soluble extract, Total Ash, Acid insoluble ash and thin layer chromatography. The study revealed specific identities for crude drug taken which will be useful in identification and control to adulterations of the raw drug.

KEYWORDS: Piperaceae, Physico-chemical, Herbal drugs, TLC, Ayurvedic medicine

INTRODUCTION:

The genus Piper Linn. is estimated to contain over 1000 species which are distributed mainly in tropical regions of the world. Several species have great economic and cultural importance and are used as foods, medicines, stimulants, antiseptics and antioxidants There are four important plants used from family piperaceae are Piper nigrum Linn., Piper cubeba Linn., Piper longum Linn. and Piper chaba Hunter¹. Piper nigrum is the most important cultivated species due to its economic value. Geographically, it is confined to Western-Ghats of South India². Piperine is an active component in Piper nigrum and contributes to its pungency. Piper nigrum is reputed in the local system of medicine of India, Latin America and West-Indies for its multidimensional medicinal properties³. Secondary metabolites from Piper nigrum play defensive role against infections by microbes, insects and animals. Another important component of pepper volatile oil is pipene, which is a famous odorants. Piper nigrum is anti-microbial, anti-mutagenic, a free-radical scavenger, immuno-modulator, anti-tumor, anti-depressant, anti-apoptotic, anti-metastatic, anti-thyroid, hepatoprotective, immunostimulator, anti-diarrheal and anti-spasmodic. Piper nigrum was reported to treat pulmonary diseases, fever, cold, colic disorder and gastric conditions. The following biologically important Phytochemical have been extracted from Piper nigrum plants: alkaloids, amides, propenyphenols, lignans, neolignans, terpenes, steroid, kawapyrones, piperolides, chalcones,

dihydrochalcones, brachyamide⁴, dihydropipericide, 4-dihydroxy-6(N-ethyamine), benzamide, (2E, 4E)-Neicosadienoyl pereridine, N-transferuloyltryamine, N-formylpiperidine, guineensine, (2E , 4E)-N-5[(4- Hydroxyphenyle)-pentadienoyl] piperidine, (2E,4E)-N isobutyldecadienamide), (2E,4E)-N-isobutyleicosadienamide, (2E,4E,8Z)-N isobutyl-eicosatrienamide, (2E,4E)-Nisobutyloctadienamide, piperamide, piperamine piperettine, pipericide, piperine, piperolein, trichostachine, sarmentine, sarmentosine, tricholein, retrofractamide. The major components of the essential oil obtained from the aerial parts of Piper nigrum were glulol, -pinene, -caryophyllene and -terpinene. Piperine was the first amide to be isolated from piper species.

Piper cubeba (in Indonesia known as kemukus), is a plant, native to Java and Borneo that produces spicy berries (cubeb berries). It is now also cultivated in several other tropical areas, including East Africa. In Indonesia Piper cubeba is valued as a medicinal plant⁵. Only three groups of secondary metabolites have been reported from the berries of Piper cubeba, i.e. alkaloids, lignans and terpenoids (essential oil). Piperine is an abundant alkaloid in the berries of this species. Twenty four lignans have so far been reported from Piper cubeba. Cubebin has been shown to possess anti-inflamatory, analgesic and trypanocidal activities. Yatein is also an interesting lignan due to its biological activity and its function as a biosynthetic precursor of deoxypodophyllotoxin and podophyllotoxin that are well known for their anticancer properties. Methanol and water extract of Piper cubeba berries have been shown to display an inhibitory effect against the hepatitis C virus⁶. Anti-inflammatory, antioxidant, anti-allergic and analgesic activities of Piper cubeba have been studied using chemically-induced edema and arthritis in vivo⁷.

Piper longum Linn. is the accepted source of the drugs Pippali and Pippalimulam throughout the country. Pippali is the dried ripe fruits; Pippalimulam is the roots of this plant⁸. The plant is a dioecious slender aromatic climber with perennial woody roots, or a perennial creeping under shrub. Branchlets erect, glabrous with swollen nodes; roots clasping at nodes, which help to get attached to the host trees; leaves alternate, ovate, cordate, apex acute to acuminate, margin entire, glabrous. The male and female plants are morphologically very similar till the formation of spikes. Mature female spikes, known as long pepper are shorter and thicker than the male spikes. Fruit spikes cylindrical, oblong, berries red or black when ripe, globose with aromatic odour and pungent taste. This plant shows different pharmacological activity like antibacterial activity, Antiallergic activity, Antitumour activity, antitubercular activity, Antifertility activity, Vaso-dilating activity⁹, anti-giardial and immuno-stimulatory activity. This plant is also used in treatment of intestinal disorders, hepatoprotective effect, chronic bronchitis, cough and cold and bronchial asthma in children¹⁰.

The plant Piper chaba Hunter, which is called chab in Hindi, chavya in Kannada, chavyam in Malayalam, Kankala in Marathi, Cavya in Sanskrit, Cavikai in Tamil, Chaikama in Telugu, belongs to the family piperaceae. It is renowned in the Indian Ayurvedic system of medicine, in the folklore of Africa, Latin America and the West Indies as well as in the Chinese herbal medicines. The plant Piper chaba is an annual/perennial shrub, cultivated in India, Malaysia and Bangladesh. In Bangladesh and India, the decoction of the roots of Piper chaba Hunter (Fam: Piperaceae) is used in cough and catarrh, inflamation of the nose, throat, larynx and bronchi, constipation, worms, colic, tympanities, dyspepsia, diarrhoea, gastritis and renal diseases, acute and chronic gonorrhoea, gleet and cystitis, visceral enlargements and externally boils, piles, paralysis, toothache, earache and painful eye affections^{11, 12}. Extracts of Piper chaba have also been found to exhibit anti-diarrhoeal and diuretic activities¹³.

MATERIALS AND METHODS:

Samples were washed in running water and air-dried. The samples were studied for physicochemical evaluation. Powder of the samples was used for chemical analysis. Physicochemical studies like Total ash, Acid insoluble ash, Loss on drying at 105°C, extractive values and TLC were carried out as per the API, WHO and AOAC guidelines^14-16.

RESULTS AND DISCUSSION:

Physico–chemical parameters of the samples are tabulated in Table 1. For Piper nigrum Linn. The pH value of 10% aqueous solution was found to be 7.04. Loss on drying at 105°C in sample was found to be 14.99% w/w. Analytical results showed ash value of 4.84% w/w. The amount of acid insoluble ash present in the sample was 0.74 % w/w. The water soluble extractive value & alcohol soluble extractive value were found to be 7.76 and 8.49%w/w respectively. For Piper cubeba. Linn. The pH value of 10% aqueous solution was found to be 4.48. Loss on drying at 105°C in sample was found to be 10.45 % w/w. Analytical results showed ash value of 3.63 % w/w. The amount of acid insoluble ash present in the sample was 0.73 % w/w. The water soluble extractive value & alcohol soluble extractive value were found to be 12.92 and 10.05% w/w respectively. For Piper chaba Hunter. The pH value of 10% aqueous solution was found to be 5.65. Loss on drying at 105°C in sample was found to be 16% w/w. Analytical results showed ash value of 10.21 w/w. The amount of acid insoluble ash present in the sample was 3.57%cw/w. The water soluble extractive value & alcohol soluble extractive value were found to be 6.6% and 1.22 %w/w respectively. For Piper longum Linn. The pH value of 10% aqueous solution was found to be 6.09. Loss on drying at 105°C in sample was found to be 10.09% w/w. Analytical results showed ash value of 4.20%w/w.

1. TLC profile of Piper longum Linn. (Toluene: Ethyl acetate: Formic acid:: 5:1.5: 0.5v/v), 2. TLC profile of Piper chaba Hunter (Toluene : Ethyl acetate :: 5: 2 v/v), 3. TLC profile of Piper nigrum Linn. (Toluene : Ethyl acetate :: 7.5: 2.5 v/v), 4. TLC profile of Piper cubeba Linn. (Toluene : Ethyl acetate :: 7.5: 2.5 v/v)

Table 1. Physico-chemical Parameters of various plants of Piperaceae family

S. No.	Parameters	Piper nigrum Linn.	Piper cubeba Linn.	Piper chaba Hunter	Piper longum Linn.
1.	pH (10% w/v aqueous solution)	7.04	4.48	5.65	6.09
2.	Ash value (% w/w)	4.84	3.63	10.21	4.20
3.	Acid-insoluble ash (% w/w)	0.74	0.73	3.57	0.29
4.	Water-soluble extractive (% w/w)	7.76	12.92	6.6	7.82
5.	Alcohol soluble extractive (% w/w)	8.49	10.05	1.22	6.86
6.	Loss on drying at 105°C (% w/w)	14.99	10.45	16	10.09

Table 2. TLC profile of Pippali (Piper longum Linn.)

S. No	254 nm		366 nm		After derivatization in visible light
	Colour	Rf.	Colour	Rf.	Colour	Rf.
1.	Pale green	0.24	Pink	0.26	Blue	0.24
2.	-	-	Pale pink	0.32	-	-
3.	-	-	Pale pink	0.37	-	-
4.	Pale green	0.40	Pale blue	0.47	Grey	0.40
5.	Green	0.53	Blue	0.53	-	-
6.	Green	0.58	Pale pink	0.58	Blue	0.61
7.	Pale green	0.63	Blue	0.66	Bluish grey	0.63
8.	Pale green	0.66	Pink	0.71	Bluish grey	0.71
9.	-	-	Pale pink	0.76	Dark blue	0.95

Table 3. TLC Profile of Chavya (Piper chaba Hunter)

S. No	After derivatization in visible light
	Colour	Rf.
1.	Grey	0.37
2.	Yellow	0.42
3.	Bluish grey	0.48
4.	Blue	0.53

Table 4. TLC Profile of Piper nigrum Linn.

S. No	ʎ 254 nm		After Derivatisation with Iodine in visible light		After Derivatisation with Vanillin sulphuric acid in visible light
	Colour	Rf.	Colour	Rf.	Colour	Rf.
1.	Violet	0.04	Yellow	0.04	Grey	0.16
2.	Violet	0.18	Yellow	0.12	Purple	0.22
3.	Brown	0.36	Yellow	0.16	Yellow	0.33
4.	Violet	0.47	Yellow	0.33	Purple	0.37
5.	Violet	0.54	Yellow	0.39	Purple	0.41
6.	Violet	0.59	Yellow	0.45	Pink	0.45
7.	Violet	0.72	Yellow	0.54	Green	0.64
8.	Violet	0.97	Yellow	0.72	Violet	0.70
9.	-	-	-	-	Pink	0.81
10.	-	-	-	-	Green	0.95

Table 5. TLC Profile of Piper cubeba Linn.

S. No	After Derivatisation with Iodine in visible light		After Derivatisation with Vanillin sulphuric acid in visible light
	Colour	Rf.	Colour	Rf.
1.	Yellow	0.04	Purple	0.14
2.	Yellow	0.10	Purple	0.20
3.	Yellow	0.56	Purple	0.24
4.	Yellow	0.60	Purple	0.34
5.	Yellow	0.71	Purple	0.42
6.	Yellow	0.82	Purple	0.56
7.	Yellow	0.93	Purple	0.60

The amount of acid insoluble ash present in the sample was 0.29%w/w. The water soluble extractive value & alcohol soluble extractive value were found to be 7.82% and 6.86 %w/w respectively.

TLC of an ethanol extract has been developed in various mobile phases and observed under UV 254 nm, 366 nm and after derivatisation in visible light. The results are given in Figure 1 and Table 2, 3, 4and 5.

CONCLUSION:

The samples were studied and described physicochemical parameters and TLC studies. These parameters will be useful in authentification and identifying the adulterants and quality control of raw drugs. Sample exhibits a set of diagnostic characters, which will help to identify the drug in dried condition.

It has been concluded from this study of various species that estimation of physicochemical parameters like pH, Loss on drying at 105°C, Water soluble extract, Alcohol soluble extract, Total Ash, Acid insoluble ash and thin layer chromatography profile (TLC) is highly essential for raw drugs or plant parts used for the preparation of compound formulation drugs. The periodic assessment is essential for quality assurance and safer use of herbal drugs.

ACKNOWLEDGEMENT:

The authors are very grateful to Director General CCRAS, New Delhi and for providing encouragement and facilities for carrying out this work. Authors acknowledge the secretarial assistance rendered by Ms. Rekha in the preparation of this paper.

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Received on 23.07.2010

Accepted on 04.08.2010

© A&V Publication all right reserved

Research Journal of Pharmacognosy and Phytochemistry. 2(5): Sept.-Oct. 2010, 407-410