Elemental Analysis of Paeonia emodi and Punica granatum by Atomic Absorption Spectroscopy

During present study Paeonia emodi and Punica granatum were selected for elemental analysis due to its wide application in the indigenous medicinal system and the importance of its chemical constituents. The plant samples were digested with concentrated Nitric acid (HNO3), fo llowed by concentrated Perchloric acid (HClO4). The transparent solutions of these samples were analysed by Atomic Absorption Spectrophotometer. The analysis for various elements in the sampled medicinal plants indicated that Sodium, Potassium, Calcium, Magnesium, Copper, Zinc, Iron, Cobalt, Manganese, and Lead were present in all samples of both plant species in different concentration. In Paeonia emodi Calcium is found in high concentration (66.26 mg/L) while Cobalt is in low concentration (0.039 mg/L). Similarly in Punica granatum Potassium concentration is maximum (55.19 mg/L) while Cobalt concentration is min imum (0.031 mg/L).


Introduction
Ethnobotanical information on med icinal plants and their uses by indigenous cultures is useful in the conservation of traditional cultures, biodiversity, for co mmunity health care and drug development [1]. The authentic knowledge of the uses of medicinal plants passed on fro m one generation to another, after refining and additions [2]. The folk recipes are prepared either fro m the whole p lant or fro m their different organs, like leaves, stem, bark, root, flo wer, seed, prop roots etc and also fro m their secondary product such as gum, resins and latex [3]. Plant based recipes have been in use in the amelioration of various ailments ranging fro m common cold to cancer [4]. Plants supply the body with minerals, vitamins and certain hormone precursors in addition to protein and energy [5,6]. Relat ively h igh levels of essential elements, such as Fe, Mn, Zn, and Ca, have been demonstrated to influence the retention of to xic elements in animals and human beings [7,8].
In recognition of the important ro le that major and trace elements play in health and d isease of human body, in the building up and restoration phenomenon, progress has occurred in this area of health sciences during the last few years [3].
Elements research has definitely been part of this exp losion of scientific knowledge [9]. Trace elements are required in very small quantities fo r p lant life [10]. The quantitative estimation of various element concentrations is important for determining the effectiveness of the medicinal p lants in treating various diseases and also to understand their pharmaco logical action [4]. The imbalance in hu man health has been linked with the excess or deficiency of elements in soils, water, plants and animals [3]. The continuous intake o f d iets that are excessively h igh in a particular element can influence changes in the functioning, forms, activit ies of some organs or concentrations of such element in the body tissue and fluids can rise above the permissible limit [11].
District Battagram is located in the Western Himalayan Province between 34°25′ and 34°58'N and 72°54′ and 73°15'E with a total area of 1,301 km 2 [12,13]. District Battagram is bounded by the Kohistan valley in the north, by Siran valley in the east, by the Konsh and Agror valleys in the south and by the Black mountain and river Indus in the west [2]. The d istrict consists of two sub-divisions i.e. Battagram and Allai. District Battagram can be categories into agricultural land, wasteland, forest and alpine meadows [14,15].
District Battagram is mostly mountainous ranging in altitude from 525 m at Thakot to 4690 m above mean sea level at Sukaisar. The area is generally rough and mountainous having variable slopes from gentle to precipitous. The climates varying fro m sub tropical at the base of the hills to "alpine" conditions prevailing in the higher reaches. Majority of the people are dependent on agriculture as a first source of livelihood, fo llo wed by pasture animal husbandry. They grow rice, maize, wheat, red beans and vegetables. All small streams coming fro m the sub valleys join two main streams granatum by Atomic Absorption Spectroscopy Nandiar Khuwar and Allai Khuwar in District Battagram to join River Indus [2,13,15,16].
The diversity and distribution of Paeonia emodi and Punica granatum were d iscussed in District Battagram [13], their trad itional medicinal uses [2] and conservation status were also discussed [17]. The comparative determination of trace elements fro m Alliu m sativum, Rheum australe and Terminalia chebula were also recently analysed [5]. The Physical and Chemical Properties of Punica granatum [18] and the chemical constituents of Paeonia emod i [19] were also studied. Keeping in mind the wide application Paeonia emodi and Punica granatum in traditional medicine were therefore analysed for further elemental contents.

Materials and Methods
The field survey was conducted during the year 2010 to collect the info rmation and specimens of Paeonia emodi and Punica granatum fro m different parts of District Battagram for their element analysis. The fresh root of Paeonia emodi and fruit of Punica granatum were cleaned and oven dried to a constant weight at 60°C for 72 h. A fter dry ing, the plant materials were ground into fine powder using an electric grinder. The powder o f both plant species were weighted 0.5g separately and put in a separate 100ml conical flasks and 30ml nitric acid (HNO 3 ) were added. Each flask were placed on magnetic stirrer heater in fu me hood for four hours at 250Cº and then colour solution were changed to milky solutions which were cooled for 10 minutes and then 15ml concentrated Perchloric acid (HClO 4 ) were added and heat the solutions until colourless solutions were obtained. The colourless solutions were filtered to remove the impurities for trace elements analysis by Atomic Absorption Spectroscopy.

3.1.3.Tradit ional uses
The infusion of d ried flower is used in diarrhea. Rhizo me is used to increase milk production in livestock, also used as tonic.

Punica granatum Linn
Punica granatum commonly known as Pomegranate and is locally called as Narsaway belongs to family Punicaceae.

3.2.2.Description
Deciduous shrub to small t ree up to 6 m h igh, with thin smooth grey bark. Leaves are mostly opposite, short-petiolate, blades oblong-elliptical, up to 8 cm long. Flowers showy and up to 6 cm broad, b isexual, 5-8 petals, reddish and up to 2.5 cm long, numerous stamens surrounding a conspicuous hypanthial tube, he flowers usually occuring terminally or in axils. Fruit a red spherical berry up to 13 cm broad, with a leathery rind enclosing nu merous seeds surrounded by edible juicy, tart, b right red "kernels". Fl. Apr.-May.

Trad itional Uses
A decoction of seed is used to treat syphilis. Its ju ice of seeds is used to treat jaundice and diarrhea. Juice o f the fru it is used to treat jaundice and diarrhoea. The rind of the fruit is ground in water and drunk every morning by diabetics. The fruit together with the juice of Cynodon dactylon leaves is used for runny noses and colds. The juice of the flo wers is used to treat nose bleeds. The fruit pulp and the seed are a stomachic. The root and stem bark have astringent and anthelmintic properties.

Elemental Anal ysis
The analysis for various elements in the sampled me- In present study it was observed that the concentration of Potassium is ma ximu m in Punica granatum (55.19 mg/ L) and the concentration of Cobalt is minimu m (0.031 mg/ L).

Discussion
The elements play both curative and preventive role in combating diseases [5]. There is a vast scope to explo it the preventive medicinal aspects of various elements [20].
Sodiu m (Na) involves in the production of energy, transport of amino acids and glucose into the body cells, the average concentration Na was 1.235mg/L in Paeonia emodi and 1.279mg/ L in Punica granatu m.
Potassium (K) is helpful in reducing hypertension and maintaining card iac rhythm. In the hu man body, Potassium play vital role in many physiological reactions and their deficiency or excess can affect human health [21]. The average concentration of Potassium was 33.38mg/L in Paeonia emodi and 55.19mg/ L in Punica granatum.
Calciu m (Ca) overco me the problems of high blood pressure, heart attack, premenstrual syndrome, co lon cancer and keeping the bones strong and reduces the risks of osteoporosis in old age [3]. The average concentration Ca was 66.26 mg/ L in Paeonia emodi and 1.650mg/ L in Punica granatum.
Magnesium (Mg) imp roves insulin sensitivity, protect against diabetes and its complicat ions and reduce blood pressure [5]. The average concentration Mg was 9.525mg/L in Paeonia emod i and 3.721mg/L in Punica granatu m.
Copper (Cu) p lay impo rtant role in treat ment of chest wounds and prevent inflammation in arthritis and similar diseases. The average concentration Cu was 0.329mg/ L in Paeonia emodi and 0.231mg/ L in Punica g ranatum.
Zinc (Zn) deficiency may contribute to arrested sexual maturation, growth retardation and hair loss, delayed wound healing and emot ional disturbance [3]. The average concentration of Zn was 0.956 mg/ L in Paeonia emod i and 0.430mg/ L in Punica granatum.
Iron (Fe) is an essential mineral to prevent anaemia and cough associated with angiotensin-converting enzyme (ACE) inhibitors. The average concentration Fe was 2.667mg/ L in Paeonia emodi and 0.466mg/ L in Punica g ranatum.
Cobalt (Co) is necessary in very small amounts in all mammals and is used to treat several different types of cancer in hu mans and treat anaemia but the intake of high amount can cause heart diseases. The average concentration Co was 0.039mg/L in Paeonia emodi and 0.031mg/ L in Punica granatum.
Manganese (Mn) can help to assist the body in metabolizing protein, help the diabetic also metabolize carbohydrates and in treating diabetes. The average concentration Mn was 0.082mg/ L in Paeonia emodi and 0.033mg/ L in Punica granatum.
Lead (Pb) is toxic metal and non-essential element for human body as it causes a rise in blood pressure, kidney damage, miscarriages and subtle abortion, brain damage, declined fertility of men through sperm damage, d iminished learning abilities of children and disruption of nervous systems [3,11]. The average concentration Pb was 0.735mg/ L in Paeonia emodi and 1.119mg/ L in Punica g ranatum.
The environ mental factors including at mosphere and pollution, season of collection sample, age of plant and soil conditions in which plant grows effect the concentration of elements as it varies from p lant to plant and region to region [5].

Conclusions
In this study 10 elements were determined in Paeonia emodi and Punica granatum. A mong the various elements detected in different medicinal p lants in different concentration is there fo re used in the treat ment of various diseases. The data obtained in p resent study will be helpful in the synthesis of new modern drugs with various combinations of plants which can be used in the cure of many diseases ethno med icinally. Ho wever, mo re detailed analysis of chemical composition of these medicinal p lants is required.