2.1. General
In the standardization of a drug, organoleptical, morphological, anatomical, physicochemical, phytochemical, (qualitative and quantitative) and chromatographical methods are used.
Morphological and anatomical characters play a vital role in crude drug standardization. Morphological characters involve size, arrangement, venation, texture, surface characters, markings and hardness of the plant materials.
As stated by Metcalfe and Chalk (1957), microscopical methods are often necessary to establish the botanical identity of commercial samples of medicinal plants, timbers, fibers etc. and may play an important part in checking adulteration and substitution. It involves longitudinal and transverse sectional views of the parts of the drug.
Plant based crude drugs whose botanical identity is not known are identified based on their morphological and anatomical characters. Park et al. (1995) studied the market samples of ‘Man Byung Cho'. Based on morphological and anatomical characters of leaf midrib and leaf lamina, he concluded that they belong to the leaves of Rhododendron brachycarpum and R. brachycarpum var. roseum. Yamaji et al., (1993) after studying the anatomical characters of flower stalk and xylem vessels of rhizome established that the drug 'Spang-RtziDo-Do' is evolved from Pterocarpus hookeri. Mehrotra and Sharma (1984) analyze the various market samples of the Ayurvedic drug 'Sappan' and compare with its genuine drug Caesalpinia sappan. Using morphological and anatomical parameters, they establish the genuineness of the drug.
Organoleptical characters play an important role in the identification of crude drugs. In this method, the color, fractures, taste and smell of the drugs are characterized. Chakraborti et al. (1988) studied the stem barks of Strychnos nux-vomica and S. potatorum, and distinguished the authenticity of 'Nux-vomica' bark from other barks. Shah and Khanna (1961) distinguish the fruits of Embelia ribes with its grayish black color and warty surfaces with that of E. robusta, which has reddish, longitudinally wrinkled surface and more prominent calyx with five sepals.
Powder microscopy is another parameter used to identify and distinguish the drug from its substitutes and adulterants. For example, Patel and Satakopan (1979) distinguish Saraca asoka bark from its adulterants by the analysis of the powder and put fourth a key for the identification of the 'Asoka' bark powder. Srivastava and Srivastava (1988) identified the adulterants of Catharanthus roseus, by the analysis of powdered drug.
Physicochemical and phytochemical studies include ash value, solubility and extractive values and qualitative and quantitative analysis of phytochemicals. Satakopan and Thomas (1970) distinguish the leaves of Adhatoda vasica and its adulterant Ailanthus excelsa based on the palisade ratio and anatomical characters of leaf and petiole.
Mucuna cochinchinensis is the adulterant of the Unani drug 'Karanj' (Genuine: Pongamia Pinnata). Hasmi and Singh (1997) established it with the use of fluorescence analysis and other pharmacognostical parameters.
Quantitative estimation of major phytochemical constituents of a drug is another parameter. Liu et al. (1993) estimate the alkaloid content of three species of Phellodendron and distinguish them with one another based on quantity of alkaloid and texture and color of the herbs.
Geographical and climatic factors influence the percentage of active constituent of a medicinal plant. Dadun et al. (1992) concluded that the percentage of alkaloids content of Ephedra sinica is dependent upon the season and geographical locality in which the plant is grown. Dragur and Menary (1992) proved that the oil yield of Olearia phlogopappa is higher in summer months than in autumn. Mallavarupu et al., 1995, estimate the essential oil content of Cinnamomum zeylanicum grown in different localities. It was found that the plants grown in Hyderabad are having more oil percentage than that in the plants grown in Bangalore.
Khatoon et al. (1993) used TLC finger printing technique and identify that the market samples 'Ratanjot' is derived from Arnebia nohilis.
Asif and Shafiullah (1993) analyzed 175 herbal drugs with Infrared spectrum and evolved a method for checking the purity of herbal drugs. Mitra et al. (1976) after careful pharmacognostical study stated that no difference was found in the macro and micro characters in rhizome of red and white varieties of Nelumbo nucifera. Abdurahman et al. (1996) tried to distinguish the two varieties of lrvingia gabonensis based on their pharmacognostical characters.
Scanning electron microscopical (SEM) studies are also useful in the identification of crude drugs even at the variety level. Mehrotra and Shome (1993) were able to differentiate the red flowered variety with the white flowered variety of N. nucifera, by SEM studies of petals.
DNA fingerprinting is a latest tool to identify the crude drugs. Cao et al. (1996) prove that the drug 'Ku-Di-Dan' sold in Taiwan, Hongkong and Macan markets is derived from Elephantopus scaber L. With the use of artibitry primed polymerase chain reaction and random amplified polymorphic DNA he is able to pinpoint its botanical identity.
2.2. Cissus quadrangularis L.
Morphological features of C. quadrangularis, specimen collected during floristic surveys in various geographical areas have been described (Gamble, 1967, Hooker, 1978, Gileslal and Livingstone, 1978, Matthew, 1983 and Nair and Henry, 1983). Kirtikar and Basu (1980) also describe the species along with its medicinal uses. Though, in all the above works, most of the morphological characters described are in consonance, discrepancy occurs with regard to nature of tendril and season of flowering. Matthew (1983) observed that tendrils are forked or not and stout and flowers are seen throughout the year. Kirtikar and Basu (1980) described the tendrils long and slender. Flowering season of C. quadrangularis, was observed as July - December (Gileslal and Livingstone, 1978).
Metcalfe and Chalk (1957) record anatomical characters of species belonging to Vitaceae. Presence of pearl glands, mucilage cells and anomalous secondary thickenings are observed. Madan and Nair (1959) described the anatomical characters of stem of C. quadrangularis. Janardhanan et al. (1981) studied epidermal and stomatal cells of C. quadrangularis. They observed the occurrence of pearl glands on vigorously growing young stem, leaves and tendrils.
It is to be noted that in all the botanical studies on C. quadrangularis, occurrence of varieties or variants has not been recorded. However, Kumbhojkar et al. (1991) in their ethnobotanical work has noted the occurrence of round-stemmed and flat-stemmed cultivars of C. quadrangularis. A specimen of C. quadrangularis, with wingless stem (round-stemmed) has been collected by Srinivasan and deposited at Botanical Survey of India (BSI), Coimbatore (MH NO.63656 and 68684). .
Siddha literatures are abounding with various types of 'Pirandai' (Vernacular name of C. quadrangularis. In a compilation of vernacular names of herbs and trees, Mathayan and Chitraputtran (1987) recorded three types of 'Pirandai' viz. 'Naanmugappirandai' or 'Sadurappirandai' (four angled or square stemmed) as C. quadrangularis, 'Pulippirandai' (acrid taste) as C. setosa and 'Sempirandai' (red colored) as C. vitiginea. Murugesamudaliar (1988) has mentioned seven types of 'Pirandai'. They are 'Olaippirandai' (two sided), 'Uruttuppirandai' (round), 'Muppirandai' (three sided), 'Sadurappirandai' (four sided), 'Kalippirandai', 'Teempirandai' (sweetish), and 'Pulippirandai' (acrid taste). Prema (1989) has mentioned ten types of 'Pirandai'. They are 'Naanmugappirandai' (four sided), 'Muppirandai' (three sided), 'Coppirandai' 'Kalippirandai', 'Kaattuppirandai' (wild), 'Sempirandai' (reddish), 'Teempirandai' (sweetish), 'Pulippirandai' (acrid taste), 'Naatuppirandai' (available in the plain) and 'Pirandai'.
Warrier et al. (1994) noted that two-sided variety of C. quadrangularis, is found in gardens. Singh and Arora (1978) mentioned that 'Birandai' is edible and 'Marol' is non-edible. Stem, stem juice, young shoots and total ash from young shoots C. quadrangularis, are medicinally useful (Chopra et al. 1956, Kirtikar and Basu, 1980 and Anonymous, 1976).
Ethnobotanical uses of the herb for bone fracture, intestine worm control, whooping cough and as curries have been compiled by Anonymous (1903), Quisumbing (1951), Ahmed (1957), Mahato and Mahato (1996), Burkil (1966), EI. Hamidi (1970), Kurup et al. (1979), Pal (1980), Rao (1980), Anonymous (1986) Jain (1989), Reddy et al. (1989), Sarin (1989), Bhat et al. (1990), Nagarjun and Rao (1990) Khan et al. (1991), Goel et al. (1994) and Girace et al. (1994).
Fracture healing mechanism of the herb is studied by Prasad and Udupa (1963, 1970 and 1972), Udupa and Prasad (1964 and 1984) Udupa et al. (1965), Prasad et al. (1970), Chopra et al. (1976) and Pradhan (1994). Other pharmacological actions of the plant are studied by Das and Sanyal (1964), Subbu (1968, 1970 and 1971) and Balachandran et al. (1991).
LD 50 of extract of aerial parts of C. quadrangularis was observed as 1000 mg/kg
(Bhakuni et al., 1969) and that of stem extract was 681 mg /kg (Dhawan et al., 1980).
Sivasamy et al. (1991) studied the positive effect of mutagenic activity of the fruit of C. quadrangularis, against various strains of Salmonella typhimurium. Alcoholic extract of aerial parts of C. quadrangularis was tested for insecticidal property against Musca domestica and Tribolium castaneum, but found inactive (Atal et al., 1978). Antimolluscidal activity of leaf extract (alcoholic) was tested against Biomphalaria pfe~fferi and Bulinus truncatus but with negative results (Abdel-Aziz et al., 1990).
Vitamin C concentration of shoots and leaves of, C. quadrangularis, is quantified as 39 mg/100 gm and that of fresh juice as 471 mg/100 gm (Anonymous, 1950). Presence of steroids was identified by Sen (1964 and 1966). Triterpenoids were isolated by Bhutani et al. (1984), Pluemjai and Saifah (1986) and Gupta and Venna (1990 and 1991).
2.3. Madhuca longifolia
Morphological characters of Madhuca longifolia, val. latifolia and M. longifolia, var. longifolia are described in various Floras and treatises of medicinal plants. (Mukerji, 1953, Anonymous, 1962, Gamble, 1967, Gileslal and Livingstone, 1978, Kurup et al. 1979, Kirtikar and Basu, 1980,Matthew, 1983, Henry et al., 1987 and Jain, 1996). The two varieties show variation in the color of the bark, number of anthers and seeds. However, there is no conformity in the descriptions of these characteristic features in the various treatises. Bark of Bassia latifolia is described as gray colored and that of B. longifolia as dark yellowish gray in Gamble (1967), but as dark colored in M. latifolia and dark brown in M. longifolia in Mukerji (1953), Anonymous, (1962) and Kirtikar and Basu (1980). Number of anthers in M. latifolia is variously given, 16 anthers roughly in 3 rows (Matthew, 1983), and 20-30 in 3 series (Mukerji, 1953, and Kirtikar and Basu, 1980). In M. longifolia also the number of anthers are given as 18 in 2 series (Matthew, 1983), and 16-20 in 2 rows (Mukerji, 1953 and Kirtikar and Basu, 1980). Discrepancies also exist in the descriptions as to the number of seeds in a fruit, 2 in M. latifolia and one in M. longifolia Matthew, 1983) and 1-4 in M. latifolia and 1-2 in var. longifolia (Mukerji, 1953 and Kirtikar and Basu, 1980).
Anatomical characters of the members of the family Sapotaceae are described by Metcalfe and Chalk (1957). Bishayee and Bhattacharya (1992) studied the plants associated with Madhuca sp., in Birbhum district of West Bengal. Literature on Siddha medicine mentions 3 types of 'Illuppai' (Tamil vernacular name for Madhuca sp.) namely 'Illuppai', 'Seemaiillupai' (exotic) and 'kaattuilluppai' (wild) (Prema, 1989). Madhuca sp. is medicinally and commercially useful. The plant parts like stem bark, corolla lobes, seeds and seed oil are used in diabetes, burns, scalds, bronchitis, rheumatism, cough, piles, galactagogue skin diseases, tonsillitis, stomach-ache, aphrodisiac and respiratory diseases and have laxative, insecticidal and piscicidal properties (Mukerji, 1953, Chopra, et al., 1956, Anonymous, 1962, Kurup et al., 1979, Kirtikar and Basll, 1980, Anonymous, 1986, Murugesamudaliar, 1988, Warrier et al., 1994, Jain, 1996 and Hill, 1996). According to Tribal Co-operative Marketing Development Federation of India Limited, the production of oil from seeds of 'Mahua' is 171 MT/year in India (Josh, 1993). Hulagu, (1993) used the oil cake of Madhuca sp., as fertilizer and found that it is moderately immobilized.
Ethnobotanical studies on M. longifolia and M. latifolia are reported by Jain and Suri, 1980, Ainslie, 1984, Chandra et al., 1985, Hajra, 1987, Sarin, 1989, Lahankar, 1991, Sikarwar, 1992, Jain and Shall, 1993, BuIll, 1994, Goel, et al., 1994, Singh, 1994a, Nigam and Misra, 1994, Sinha, 1994, Maheswari, 1995, Bajpayee and Dixit, 1996, Banerji and Pal, 1996, Goud and Pullaiah, 1996, Khanna et al., 1996, Reddy et al., 1996, Saini, 1996, Samwatsar and and DiwaiYi, 1996 and Sharma, 1997. Singh (1994 b) suggested the methods for natural conservation of M. longifolia which have ethnic'va1ue in Santhal Parganas of Bihar.
Dolui (1988 a and b) found that the fat prepared from oil of Madhuca sp. is a promising suppository base in drug preparation. Alam et al. (1983) substituted the flowers of Madhuca instead of 'Dhataki' flowers in the preparation of 'Mustakarishta'. Saxena et al. (1992) identified that the oil of M indica is one of the ingredients of 'Kubja Prasavini Taila'. Uniyal (1993 a) found that 'Sthal Madhuk' derived from M indica. Uniyal (1993 b) identified that 'Madhok' is having two types, 'Mahua' is the M indica and 'Madhupuspi' is Aisandra hutyracae.
The oil of Madhuca sp. contains oleic acid, palmitic acid, linoleic acid and myristic acid, seeds contain morwin and flowers have invert sugars and cane sugar (Mukerji, 1953). Triterpenoids are identified from seed kemals (Mitra and Awasthi, 1962), nut shells and fruits (Awasthi and Mitra, 1967) and in trunk barks (Awasthi and Mitra, 1968) of M. latifolia. Flavonoids are isolated from fresh leaves (Subramanian and Nair, 1972) saponins from defatted seeds (Hariharan et al., 1972) and leaves (Banerji et al., 1985) and sterols from seed oil (Bhargava and Singh, 1958 and Singh, 1959) of M. latifolia. Bhatnager et al. (1972) identified triterpene esters and oleanolic acid and palmitate from leaves and Kitagawa et al. (1978) identified a 'Mi saponin C' from seed kernels of M. longifolia. Chemical and biological aspects of polysaccharides from flowers of M indica are reported by Rao (1992).
The fluoride content in fruits of M. longifolia is estimated to be 0.2 ppm (Nandha, 1972). Daniel et al. (1978) quantified the tannins (4.86%) in leaves. Gopalan et al. (1984) estimated the calcium, phosphorus, vitamin C, iron and carotene in the flower of
M. longifolia as 45 mg, 22 mg, 101 mg, 40 mg and 307 mg/lOO gm respectively. The seed cake of M. latifolia contains marwin (Murthy et al., 1991). Atal et al. (1978) reported that the stem bark of M. indica is devoid of tannins.
Atal et al. (1978) found that the extract of stem bark of M. longifolia have anti-insecticidal property against housefly. Oil of Madhuca sp. has anti-insecticidal activity against Callosohruchus cinensis (Chander and Ahmed, 1986 and Ali et al. 1983) and
C. macultatus (Jadhav and Jadhav, 1984). The oil is also active against Meloidogyne incognita (Lanjewar and Shukla, 1986) and for common pests of various crops and ornamentals (Sounderrajan and Nimbisan, 1993). Katole et al. (1996) reported that, 'Mahua' oil is ineffective against citrus black fly nymphs when compared to monocrotophos, phosalare and neem oil.
Oil cake of Madhuca sp. are found to be highly deleterious to the nematodes like Meloidogyne incognita, Rotylenchulus reniformis and Tylenchorhynchus hrassicae (Alam et al., 1982), control the population of R. reniformis (Patel and Patel, 1992), inhibit gall formation of M. incognita (Goswami and Vijayalakshmi, 1986) inhibit Meloidogyne sp. population (Alam, 1989) and when applied with porate 10G inhibit Meloidogyne sp. population (Sundarababu and Vadivelu, 1989). Padmanaban and Daniel (1993) used the oil cake of M.indica against grubs of arecanut cockchafer and Leucopholis lepidophora and found active at 2440.81 kg/ha, but inactive for larval emergence from the egg sacs of cyst nematodes when compared to neem and saw dust (Devi and Gupta, 1995). Mahajan et al. (1994) found significant raise in organic and inorganic constituents of fresh water crab, Paratelpusa jacquemontii (Ruthdum) when exposed to defatted oil cakes of M indica.
Seed oil of M. latifolia is inactive against Marophomia phaseolina in cowpea (Ratnoo and Bhatnagar, 1993) and seed kernal is inactive against yellow mosaic virus in black gram (Mariappan et al., 1987). Stem bark extract of M. indica is inactive against Ranikhet disease virus and vaccinia virus. (Bhakuni et al., 1969).
Stem bark extract of M indica is inactive against Bacillus suhtilis, Staphylococcus aureus, Salmonella typhi, Escherichia coli and Agrohacterium tumefaciens. It has no antifungal properties against Candida alhicans, Cryptococcus neoformans, Trichophyton mentagrophytes, Microsporun canis and Aspergillus niger (Bhakuni et al., 1969). The methanolic extracts of flowers, leaves, stem and stem bark of M. longifolia have been reported to possess antibacterial activity against Bacillus anthracis, B. pumilus, B. suhtilis, Salmonella paratyphi, Vihrio cholerae, Xanthomonas campestris and X malvacearum (Trivedi et al., 1980). Pasmer and Datta (1988) reported that the oil of M. latifolia has synergistic action with malathion when malathion and oil were tested at 1:1 and 1:5 levels. Alam et al. (1984) studied the microflora of corolla lobes and found that the microbe responsible for fermentation is from external sources.
50% alcoholic extract of stem bark of M indica reveal hypotensive activity and devoid of diuretic and anticancer property and LD 50- was 1000 mglkg i.p. in albino mice. (Bhakuni et al. 1969). Seed saponins of M. longifolia are reported to have spermicidal activity at 2.0% concentration (Shetty et al., 1976) and anti-inflammatory property (Yamahara et al., 1979). However, Banerji et al. (1979) reported that seed saponin of M. latifolia has spermicidal activity at 0.03% concentration. Seed saponion has no effect on cardiovascular activity and haemolytic activity (BaneIji et al., 1981) but have cholinergic activity (Mulky and Gandhi, 1977). Leaf saponins are found to have no spermicidal or spasmolytic activity (BaneIji, et al., 1985). Pyne et al. (1981) reported that the seed cake of M. indica had no ill effect in the health of milk murrah buffaloes when added up to 15% in their diet, but Cherian et al. (1996) found that it is unpalatable to rats. Varma and Singh (1978) suggested to use desaponified seed cake as cattle feed.
Rout and Das (1993) made tissue culture studies on M. longifolia var. latifolia. He successfully transferred the regenerated plantlets to soil.
2.4. Basella alba
Morphological characters of Basella alba L. (Syn. B. rubra) are described by Anonymous (1948), Sharma (1961) Gamble (1967), Kirtikar and Basu (1980), Hooker (1978), Mattllew (1983) and Warrier et al. (1994). All these treatises, B. alba and B. rubra are treated as synonyms and describe the color of the flowers as white or red. Henry et al. (1987) treated them as same species but of different varieties. According to them the green variety is B. alba var. alba and purple variety is B. alba var. rubra.
Anatomical characters of the family Basellaceae are described by Metcalfe and Chalk (1957) and that of B. rubra by Sharma (1961). Murugesamudaliar (1988) and Prema (1989) mention the presence of two types of 'Kodippasali' - white colored vine and red colored vine.
B. alba is economically useful as food and dye and in medicine. It is used as diuretic, aphrodisiac and antipyetic and in gonorrhoea, balanitis, urticaria, constipation of children and pregnent women, leprosy, dysentery, laxative, ulcer, and burns (Anonymous, 1948, Chopra et al., 1956, Gamble, 1967, Kirtikar and Basu, 1980, Anonymous, 1986, Murugesamudaliar, 1988, Warrier et al., 1994, Jha, 1996, Saini, 1996).
Etlmobotanical uses of B. alba for cold, rheumatism, boils and blisters, dysentry and constipation are recorded by Singh and Anand (1994), Viswanathan (1995), Gurib-Fakinl et al. (1996) and Kham1a, et al. (1996b).
Alam (1981) reported that B. alba is a host of the root-knot nematode Meloidogyne incognita. Suriachandraselvan and Narayanasamy, (1988) have successfully controlled the potato virus 'Y' infection in chilli with the leaf extract of B. rubra.
Atal et al., (1978) found that the alcoholic extract of B. rubra have anti-insecticidal activity against Musca domestica and Tribolium casteneum.
B. rubra contains proteins, calcium, iron, vitamin A, B and B2 (Anonymous, 1948) and devoid of tannins (Atal et al., 1978).
