Nutritional analysis of Rhazya stricta collected from different sites of Kalabagh, Mianwali, Pakistan

Medicinal plants have constantly played significant parts in fields of culture, medicine and nutrition. Rhazya stricta is a small, upright perennial, poisonous shrub. Different plant parts have been used in medicine traditionally in contradiction of many diseases like diabetes, skin diseases, foot burning, stomach pain. The aim of research work was to study different nutrient composition in Rhazya stricta collected from different sites of Kalabagh, Mianwali. By using AOAC standard methods moisture content, fat content, fiber and protein content were checked. Proximate analysis showed plant besides having medicinal importance plant have moderate nutritional composition. There is little variation in nutrients of Rhazya stricta collected from different sites because of little variation in quality of soil and climatic conditions. It is concluded that medicinal plants should also be studied for their nutritional composition beside medicinal importance.


Introduction
Medicinal plants have constantly played significant parts in fields of culture, medicine and nutrition. Medicinal plants are considered as basic bio resources of traditional and modern medicines because they provide valuable bioactive compounds used in preparation of drugs [1]. Approximately, sixty percent of the world population and eighty percent of the developing countries population rely upon medicinal plants for their basic healthcare demands [2]. Over the centuries, humans have depended on plants for basic requirements such as food, shelter, clothing, and poisons for hunting or murder, hallucinogens, stimulants and medicines [3]. These plants also have been regarded important for research, transport, commercial purposes and income. According to an estimate, 12% of Pakistani flora is used to cure many diseases but attention has never been given to preserve or improve cultivation of these plants [4].For the physiological functions of body each plant has nutritional value and also has medicinally important phytochemicals. Bio chemicals sugars, proteins, fats and nutrients have vital role in supporting man needs for life [5]. Kalabagh is prominent for red Salt hills, Indus River and Kalabagh dam. Topography is combination of hills and semi-arid planes from underlying rocks limestone and sandstone detritus have given characteristics to the soil of area. Soil is fertile in quality, nearest to Indus River. Due to alluvial deposits from Indus and surrounding hills soil is loamy and area is semi-arid. Vegetation is unique due to diverse habitat, soil composition and topography. The forest range includes Kalabagh, Kundian and Kachha forest near Indus River. Mianwali and Kalabagh hills are facing the pressures of overgrazing because all kinds of livestock grazed in these hills. Since the area is hilly and plain type of land, overgrazing is more damaging in plains than hilly areas [6, 18,31]. Rhazya stricta is a small, upright perennial, poisonous shrub. The name Rhazya was given to species after the name Abu Bakr Mohammed bin Zakariya Ar-Razi a Muslim scientist and its Latinized name is Rhazes famous in Europe in Arabic, it is also known as "Harmal" in Urdu. Rhazya stricta is present in many areas of World and in sandy plains of Saudi Arabia. Rhazya is abundantly found in western Asia and North Western parts of sub-continent Different plant parts have been used in medicine traditionally in contradiction of many diseases like diabetes, skin diseases, foot burning, and stomach pain. Plant is used mostly in the form of powder in traditional medicine. Body pain and chronic rheumatism. Acne and pimples are treated by Powder of dried fresh leaves [7]. Proteins play a significant role in metabolic activities and act as the building block of all cells, bones and muscles. Amino acids serve as donors in the synthesis of non-protein and nitrogen-containing compounds, including nucleotides, heme, creatine, choline and other substances [8]. Carbohydrates Table 1).

Sample collection
Soil sampling was done at five sites. Ten samples were taken randomly from each site. Samples Z scheme was used for soil sampling. 3 replicates were taken from each site. Sample preparation Soil Sample was made by mixing 3 replicates from homogenized soil. To study % saturation paste of dry sample was made. Analysis of soil Soil texture was determined by Hygrometer method [12]. Electrical conductivity, pH and ions were checked according to [13,14].

Proximate analysis
Proximate analysis such as moisture content, ash content, crude protein, crude fiber, crude fat and carbohydrates was conducted by the methods described in [15]. Determination of moisture content 2g of each leaf sample was taken in a petri dish and fully dried in an oven at 100°C for about six hours. After drying, samples were cooled in a desiccator and weighed once more. Moisture content (%) was measured from the loss in weight of sample accordant with the formula: Moisture loss (g) = Original weight of sample (g) -Weight of dried sample (g)

Determination of Ash content
Ash content was estimated by keeping 2g of each sample in a pre-weighed crucible and burnt on low flame. Samples were incinerated in a muffle furnace for 6 hours at 550°C till a constant weight was attained. Crucibles were cooled in a desiccator and weighed once more. Ash content (%) was determined by subsequent formula: Ash (%) = Weight of ash (g) ×100 Weight of sample taken (g) Determination of crude protein Reagents used H2SO4, K2SO4, CuSO4, FeSO4, 40% NaOH, 4% boric acid and methyl red indicator. By applying Kjeldahl's method, organic nitrogen and crude protein content present in leaf samples were estimated. Initially 1g of each dried sample was digested in digestion flask with 20mL concentrated sulphuric acid in the presence of 5g digestion mixture [K2SO4 (90g) + CuSO4 (7g) + FeSO4 (3g)] for 3-4 hours on heating device until clear/green solution acquired. Digested material was diluted to 250ml and its 10ml was distilled with 10ml NaOH (40% solution) in micro Kjeldahl apparatus. Ammonia (NH3) thus released was assembled in a receiver having 10ml of 4% boric acid solution by using methyl red indicator (1 drop/10mL). For determination of nitrogen, the content in the receiver was titrated against 0.1N H2SO4 until a golden-brown endpoint obtained. Nitrogen %age was calculated by using following formula; Crude protein content (%) was measured by multiplying nitrogen percentage with factor of 6.25. Crude Protein (%) = % Nitrogen × 6.25 Determination of crude fat content Reagent used Petroleum ether. 2 g dried leaves sample, taken in a preweighed filter paper, was kept in extraction thimble of Soxhlet apparatus covered with a cotton plug. 250 ml petroleum ether was added in it and heated. Fat was extracted in Soxhlet extractor at a rate of 3-4 drops per second of petroleum ether for about four hours. Then, the samples were removed from the apparatus and dried in an oven or on a hot plate oven for 5-10 minutes at a temperature of 70-90°C. Samples were cooled in a desiccator and weighed. Formula used to estimate fat content present in a sample was; Crude Fat (g) = W1 -W2 Where, W1 = Weight of sample before fat removal W2 = Weight of sample after fat removal Crude fat content (%) was estimated by following formula; Fat (%) = Weight of fat in sample (g) × 100 Weight of sample (g)

Crude Protein % in Rhazya stricta
Analysis of variance of protein in Rhazya stricta was showed (Table 6). Maximum contents were observed at Site 4 (9.113 %) whereas minimum contents of protein were observed at Site 2 (8.15%). The decreasing order of crude protein content was S4>S5>S1>S2>S3 (Fig. 11