Pharmacognostic Study and Development of Quality Control Parameters for Different Parts of Lavatera Cashmeriana


 Lavatera cashmeriana Camb. is an endemic and important medicinal plant species of Kashmir Himalaya. It is widely used locally and is sold as crude drug in Kashmir markets due to its various medicinal properties. However, despite its traditional usage there is limited information pertaining to pharmacognostic characterization of L. cashmeriana. So, the present study was carried out to investigate the pharmacognostical and physicochemical profile of different parts of L. cashmeriana. The measures taken for pharmacognostical characterization were organoleptic study, macroscopy, anatomy, powder microscopy, foreign matter analysis, ash values, loss on drying, swelling index, foaming index, ph values, fluorescence analysis and extractive yield. Organoleptic and macroscopic studies revealed that L. cashmeriana is a perennial herb with very thick branched tap root system; orbicular, palmate, slightly cordate at base, upper surface of leaves with dense simple, fascicled or stellate hairs, lower surface densely stellate hairy, lobes ovate or obtuse, stipule linear lanceolate; axillary, solitary or in terminal sub-racemes inflorescence; fruit discoid mericarps and seeds glabrous, dark brown. Anatomical studies and powder microscopy revealed some important diagnostic features in the plant species. The pharmacognostic profile studies generated helpful data to set up standards for the plant. The present study for the first time endows with an inclusive pharmacognostic profiling of L. cashmeriana, thereby, acting as a platform for accurate identification, authentication and development of quality control parameters of the species. Data obtained may be used as a standard for future studies.


INTRODUCTION
Plants have dole out human beings as a natural resource for curing diseases and therapies since earliest times. The medicinal herbs have gained interest because of its extensive use and less side effects. An enormous amount of evidences have been collected to show vast potential of medicinal plants used in various traditional systems (Prabhavathi et al., 2016). Therefore, there is a need to appraise phytoconstituents obtained from traditional medicines, based on a range of phytochemical screening, pharmacological and analytical methods (Patil et al., 2010). Furthermore, there is need to standardize the plants and its parts to be utilized as a medicine. The process of standardization can be acquired by step wise i.e. pharmacognostic and phytochemical studies. Proper identification and quality declaration of beginning material is a noteworthy march to assure reproducible eminence of herbal medicine which will facilitate us to establish its safety and effectiveness (Prasanth et al., 2016). The pharmacognostical study not only offers the authentication but also quality, purity and standard of the plant drug.
Various methods are used to establish pharmacognostical standards which include microscopy and macroscopy, physicochemical parameters, extractive values, fluorescence analysis and heavy metal analysis. These parameters in turn can help to ensure the quality of the drug.
Lavatera cashmeriana Camb. (Kashmir mallow or wild hollyhock) a member of family Malvaceae, is a perennial herb which inhabits forest clearings, shrubberies, wet meadows, and sunny rocky slopes. It is a medicinal plant broadly used in conventional folk remedy such as, abdominal disorders and renal colic (Kaul, 2010), flowers for common cold and mumps (Malik et al., 2011) and seeds as antiseptic etc. (Dar et al., 2002). Four protease inhibitors viz LC-pi I, II, III and IV were purified from seeds of Lavatera cashmeriana which inhibited trypsin, chymotrypsin and elastase in vitro (Rakashanda et al., 2013). L. cashmeriana extract also showed antibacterial activity (Rakashanda et al., 2012). The decoction of flowers of L. cashmeriana mixed with leaves of Salix alba is given to cure the skin irritation in pregnant women (Ganie et al., 2013).
Inspite of its tremendous medicinal importance a lesser amount of information is available on the standardization parameters of L. cashmeriana. Therefore, the present work is an attempt to provide comprehensive report on the quality control and standardization parameters of L. cashmeriana. In this connection, different parts (leaves, stem, root, flowers and seeds) of the plant were examined. The methods used to establish pharmacognostical standards include microscopy and macroscopy, physicochemical parameters, extractive values and fluorescence analysis.
These parameters in turn can help to ensure the quality of the drug and be useful for the compilation of a suitable monograph for its proper identification.

Preparation for pharmacognostic studies
Healthy and disease free plants of L. cashmeriana were collected from Aharbal area of Jammu and Kashmir. The collected specimens were identified and deposited in Kashmir University Herbarium (KASH) under voucher number 2038-KASH. The plant collections were made quite judiciously throughout the course of the present study. The plant materials were fragmented into different parts (root, stem, leaves, flowers and seeds) and dried under shade at room temperature for 15-20 days. After shade drying, the plant materials were pulverized to coarse powder using grinder and stored under proper conditions for future use. The pharmacognostic studies were carried out on different parts (root, stem, leaves, flowers and seeds) separately.

Organoleptic evaluation
It refers to the evaluation of plant material by color, odour, taste, shape, texture etc. Different dried parts of L.

Macroscopic evaluation
Fresh and healthy plants of L. cashmeriana were assessed for their external characteristics.

Anatomy
Transverse sections of fresh materials of different parts of L. cashmeriana were cut with the help of sharp blades. Peels were obtained from fresh leaves by forceps. Different sections/peels were stained with safranine and observed under microscope and photographed.

Powder microscopy
For the analysis of plant powder, pinch of fine powder is taken in a test tube and boiled in chloral hydrate solution for few minutes. A few drops of powder were smeared on a slide mounted with phloroglucinol followed by few drops of concentrated HCl (Anonymous, 1998). The prepared slides were then observed under a microscope and photographed.

Macroscopic and organoleptic description
The

Anatomy
The anatomical studies of different parts of L. cashmeriana revealed presence of various diagnostic features as depicted in Fig. 2(a-e).

Powder microscopy
The result of powder microscopy of different parts of L. cashmeriana revealed many important features which are illustrated in the Fig.3 (a-m).

Physicochemical parameters
The results attained from various physicochemical parameters including ash value, foreign matter, moisture content, swelling index, foaming index, pH in different parts of L. cashmeriana are presented in Table ( (Pandey and Tripathi, 2014).
Organoleptic assessment can be made by means of sense organs, which offer the simplest as well as quickest means to ascertain the distinctiveness and clarity to make sure quality of a particular drug (Chanda, 2014). According to WHO, the macroscopical and microscopical description of a medicinal plant is the first step towards ascertaining the identity and the degree of purity of such material (Pandey and Tripathi, 2014). The microscopic study encompasses powder microscopy as well as anatomical studies of different parts of the plants under study. The physicochemical parameters are also important for the standardization and quality control of herbal drugs. Estimation of extractive values determines the amount of the active constituents in a given amount of plant material when extracted with a particular solvent. The extractions of any crude drug with a particular solvent yield a solution containing different phytoconstituents. It also hints whether the crude drug is exhausted or not (Chanda, 2014;Tatiya et al., 2012).
Fluorescence analysis is a vital pharmacognostic factor. Some components illustrate fluorescence in the visible range in daylight. In many natural products the ultra violet light produces fluorescence whichdoes not visibly fluoresce in daylight. If substance themselves are not fluorescent, they may often be changed into fluorescent derivatives or decomposition products by applying different reagents. In this way crude drugs are often assessed qualitatively and it is an essential parameter for pharmacognostic estimation of crude drugs (Chanda, 2014;Zhao et al., 2011).

CONCLUSION
The study may present a base for further accomplishments towards generating understanding about medicinal plants of Kashmir Himalaya. The pharmacognostic studies are the first step towards ascertaining the identity and the degree of purity of herbal materials. The pharmacognostic analysis is not reported previously in this plant species which makes this first report providing complete pharmacognostic profile of L. cashmeriana and hence will be useful for correct identification and authentication of the species for future studies.