Spatio-Temporal Variations in Nutritional and Medicinal Properties of Ocimum basilicum collected from Soon valley, Punjab, Pakistan. Abdul Ghani, Muhammad Nadeem, Ramsha Aflak, Maria Yameen, Muhammad Abu Bakar, Mujahid Hussain Abstract: The present study was conducted to analyze and investigate the nutritional and medicinal properties of Ocimum basilicum collected from four different regions of Soon valley. The selected sites were Kenhatti Gardens, Uchali Jheel, Khabeki Jheel and Khura. Elemental contents were studied for Ocimum basilicum and Atomic Absorption Spectrometry was used for determination of some elements (Na, K, Ca, Mg, Zn, Mn, Fe, Cd, Pb, Ni and Cu). Proximate attributes including moisture contents, ash contents, fiber contents, fat contents, protein contents and carbohydrate contents were analyzed. Some biochemical parameters like Chl a, Chl b, Total Chl, Alkaloids, Flavonoids and Oxalates were also determined. After conduction of experiment result was collected and then analyzed. The result was then compared with results of other scientists which either shows variations as well as similarities among parameters of elemental analysis, biochemical attributes and proximate composition. . These differences in results are mostly due to spatiotemporal variations, soil texture, temperature, water composition, nutritional status, soil composition and other environmental factors. The elemental contents present in Ocimum basilicum collected from Kenhatti Gardens shows maximum values of macro and micro elements as compared to species collected from other sites. Ocimum basilicum taken from Khabeki Jheel showed maximum percentage of phytochemicals like alkaloids, oxalates and Flavonoid as compared to collected from other three sites (Kenhatti Gardens, Uchali Jheel and Khura). Proximate analysis revealed that Ocimum basilicum collected from Uchali Jheel was rich in nutrients as compared to collected from other three sites. Results revealed that elemental; biochemical and proximate values of this medicinal plant should be analyzed and progressed further which will be helpful for production of nutritional and phytochemical products and new medicines used to cure various diseases. Introduction: Therapeutic plants have turn out to be the center of powerful study in terms of justification of their habitual uses through the purpose of their authentic pharmacological properties. Almost two million years ago, the father of medicine Hippocrates declared about 400 therapeutic plants and advised, “Let food be your medicine and let medicine be your food. “This illustrates the requirement for current medicine and science structure to turn its concentration to plants so that we once again find a new medication that might treat AIDS, cancer, diabetes and various infection, diseases and circumstances.” Ocimum basilicum is widely present in Soon valley. It is a scented plant commonly known as “Tulsi” and belongs to Lamiaceae family. It has notched and ovate leaves. Its plant is straight, 13-34 inch long, stems are hairy, and leaves are simple with purple and green colour and are fragrant. Roots, stems, fruit and leaves of this plant are used as medicine by people of Soon Valley and related areas. It grows in dry areas and shows good antimicrobial activities (Bayati and Mola, 2008). It is used against treatment of various diseases like cough, headache, urination problems etc (Neychev et al., 2005). It contains large amount of essential oils, eugenol, β-elemene, carbohydrates and anti-oxidants. Various chemical compounds used for medicinal purposes are taken from plants due to the occurrence of alkaloids, glycocides, flavonoids etc. Anti-inflammatory agent like, saponins, flavonoids, triterpenes, fatty acid and carotenoids are main reasons of using plant components as medicine (Mors et al. 2000). Materials and Method: Area of Study Survey was conducted to Soon valley, Punjab, Pakistan for collection of Ocimum basilicum plants for determination of Phytochemistry, proximate analysis and elemental composition from four selected sites (Khabeki Jheel, Uchali Jheel, Kenhatti Gardens and Khura). Plant Sample collection: Plants were collected from four different areas. Leaves and stems were obtained from various regions of Soon valley for analysis process. Replicates of Ocimum basilicum plants were collected randomly. Samples were first washed with distilled water and then dried properly in sunlight and shade. After that process, plants were crushed into fine powder and then stored at 30°C after wrapping and labeling properly and then used for further analysis. Estimation of mineral contents: Samples were dried in shade and ground into powder before applying of wet digestion method. 10 ml of HNO3 and 0.5 g of sample was taken in digestion flask and held it overnight. Digestion was done on hot plate by adding 5 ml per chloric acid into solution of tested material. When transparent solution was formed then condensed water was added to make 100 ml of standard solution for elemental analysis. Soil digestion: 50 g of soil sample was dissolved in 200 ml of condensed water. After proper stirring, formed solutions were kept for 1 day. Whatman filter paper was used to filter solutions for further analysis and to gain extract. Atomic Absorption Spectrophotometer: Solution of plant sample was run in Spectrophotometer so that average arc for every metal will obtain. Calculations of elemental contents were done by standard curve with the help of AOAC method 1998. Micro and macro nutrients (Sodium, Potassium, Calcium, Magnesium, Zinc, Manganese, Iron, Cadmium, Lead, Nickel and Copper) were calculated through this method. Proximate Analysis Ground plant samples were further used for proximate investigation. AOAC (2006) was used for determination of nutrients through proximate study. This analysis was conducted in Food Science Department, Sargodha University. Specific procedures were adopted for all parameters during analysis. Moisture contents (%), Ash contents (%), Fiber contents (%), Fat contents (%), Protein contents (%) and Carbohydrate contents (%) are the parameters used for analysis. Biochemical parameters: Estimation of Chlorophyll contents: Arnon method was used to determine amount of chlorophyll from plant sample. Fresh foliage weighing 0.1 g was scratch, extracted and then mixed with 5 ml of 80% acetone. Put it in dark for 12 hours. Visible Ultra Violet Spectrophotometer was used to calculate assimilation of extract at 663 nm and 645 nm. Chlorophyll contents were calculated by given formula. Chlorophyll “a” (mg/g fresh Weight) = {12.7 (ΔA 663 – 2.69 (ΔA 645)} x Volume/1000 x Weight. Chlorophyll “b” (mg/g fresh Weight) = {22.9 (ΔA 645 – 4.68 (ΔA 663)} x Volume/1000 x Weight. Total Chlorophyll (mg/g fresh Weight) = {20.2 (ΔA 645 – 8.02 (ΔA 663)} x Volume/1000 x Weight. Whereas, ΔA = Absorption at definite wavelength Statistical analysis: ANOVA (Statistical analysis of variance) of records was done through the help of SSPS and MS excel. At 0.05 possible points, importance between means was confirmed by Steel et al. (1997). Results: Medicinal plants are mostly utilized for manufacturing of various medicines to cure different disorders. Various metabolites are present in these plants which play vital role in development, growth and defense in plants (Akhtar et al., 2009). Because of difference in water and soil texture from site to site, these metabolites also varied among different plants. Metabolites may be therapeutic, progressive or dangerous to flora. Metabolites may be disease specific or used against various diseases (Nwachukwu et al., 2010). Table 1. Proximate analysis data of Ocimum basilicum collected from four sites of Soon valley. Site Moisture % Ash % Fiber % Fat % Protein % Carbohydrate Kenhatti Garden 9.93 19.64 41.84 15.75 26.47 47.82 Uchali Jheel 7.64 20.11 42.86 18.33 22.92 51.51 Khabeki Jheel 8.63 22.46 41.93 17.27 30.15 48.22 Khura 9.1 18.54 43.72 12.41 33.44 44.94 Table 2. Elemental concentration of Ocimum basilicum collected from four sites of Soon valley. Site Na(mg/g) K (mg/g) Ca (mg/g) Mg(mg/g) Zn (mg/g) Fe (mg/g) Ni (mg/g) Kenhatti Garden 521.62 520.56 316.6 292.26 52.61 2.53 5.39 Uchali Jheel 518.4 515.34 320.66 294.65 55.6 3.25 6.38 Khabeki Jheel 513.3 521.13 323.82 295.34 58.37 3.11 11.92 Khura 512.37 516.63 315.29 291.54 54.76 1.49 7.37 Table 3. Physiological attributes concentration of Ocimum basilicum collected from four sites of Soon valley. Site Chl a Chl b Total Chl. Alkaloids Oxalates Flavonoid Kenhatti Garden 2.61 3.85 6.46 8.24 2.75 2.81 Uchali Jheel 3.41 2.97 6.38 9.98 0.71 1.26 Khabeki Jheel 3.31 2.44 5.75 6.90 1.48 0.96 Khura 4.88 1.78 6.66 7.56 2.91 2.63 Discussion: Some herbs are utilized in manufacturing of flavours, herbal teas, healthy food, fragrances and colour ingredients etc (Singh, 2015). Medicinal plants contain many antioxidants like lignin, flavonoid, alkaloids and tannins etc. which minimize injuries that results in better life and food values (Altemimi et al., 2017). Medicinal flora possesses a variety of phytochemicals which are protective against herbivores (Hassan, 2011). These phytochemicals protect humans against many disorders (Shrivastava et al., 2010). Ocimum basilicum results regarding amount of moisture and carbohydrate content was similar with results discussed by Sarfraz et al. (2011). Various samples of O. basilicum overcome different concentration of moisture and fats having values from 7.64% to 9.93% and from 12.41% to 18.33% respectively. However, O. americanum showed varied range of moisture contents (12.72 ± 0.11%). The variation among our result may be for the reason that of difference in seasonal variations, soil structure as well as growing attributes. Results collected about ash content were in association with those declared by Hussain et al. (2010). Proximate analysis showed that value of ash content varies from 18.54% to 22.46%. Results distinguished for fiber contents and fat contents were almost analogous with results of Sarfraz et al. (2011) and Ojo et al. (2014). The range of fiber concentration was from 41.84% to 43.72%. But, Hussain et al. (2010) concluded 36.6% concentration of fiber in O. minimum leaves. The divergence in results is due to different species. Our results about protein content were in association to the results given by Jabeen et al. (2010). O. basilicum showed diverse amount of protein content (22.92% to 33.44%) in different samples of plant. The results collected for chlorophyll contents, alkaloid and oxalate content were a little different from results by Ojo et al. (2014) and Deshpande et al. (2013). The plant samples for O. basilicum enclosed diverse values of oxalate content, chlorophyll content and alkaloid content which ranges from 0.71 mg/g to 2.91 mg/g, 2.61% to 4.88% and 6.90% to 9.98% respectively. The variation in results is due to difference in carbohydrate contents, environmental variations, composition, soil structure and temperature. The elemental status exposed significant variation between different elements. Results collected for concentration of Ca and Na was associated to results of Ojo et al. (2014) and Ghani et al. (2016). The O. basilicum has dissimilar values of Ca in order from 315.29 mg / g to 323.82 mg / g and Na varies from 512.37 mg / g to 521.62 mg / g. However, concentration of sodium composed in Peganum hermla was 71ppm. The potassium contents in O. Basilicum recorded was from 515.34 mg / kg to 521.13mg / kg while amount of Mg was from 291.54 mg / g to 295.34 mg / g. Results about concentration of K and Mg of O. basilicum were in harmony by means of Ghani et al. (2012). The dissimilarity in our results is due to variation in spatiotemporal difference, environmental conditions and soil texture. Results regarding Fe and Zn concentration were associated to those collected by Deshpande et al. (2013) in O. basilicum. The amount of Fe and Zn ranges from 1.49 mg / g to 3.25 mg / g and from 52.61 mg / g to 58.37 mg / g respectively. Our conclusion about concentration of Cd was in association with the results given by Hussain et al. (2010). In O. basilicum the value of Cd recorded was from 4.71 mg/g to 9.13 mg / g. Results collected for quantity of Zn were close to the result of other researchers. Results recorded for Mg were high than the results of Erum et al. (2012). The concentration of Mn in of O. basilicum was about 9.47 mg / g to 18.49 mg / g. The small unlikeness in provided data is possibly due to variation in growth circumstances, seasonal changes, water properties, soil structure and temperature etc. References: Aletmimi, A., Lakhssassi, N., Baharlouei, A., Watson, D. and Lightfoot, D. A. (2017). Phytochemicals: Extraction, Isolation and Identification of Bioactive Compounds from plant Extarcts. Plants, 6(4): 42. Deshpande, J. D., Mohini, M. J. and Purushottam, A. G.(2013). Zinc: The trace element of major importance in human nutrition and health. International Journal of Medical Science and Public Health, 2(1): 1-3. Jabeen, S., Shah, M. T., Khan, S. and Hayat, M. Q. (2010). "Determination of Major and Trace Elements in Ten Important Folk Therapeutic Plants of Haripur Basin, Pakistan". Journal of Medicinal Plants Research, 4(7): 559-66. Neychev, V. K., and Mitev, V. I. (2005). The aphrodisiac herb Ocimum basilicum does not influence the androgen production in young men. Journal of ethnopharmacology, 101(1): 319-323. Srivastava, R., Rurum, A. M., Mishra, P., Shehu, I. A. and Rajak, C. (2018). Preliminary phytochemical investigation and TLC fingerprint profile of Amaranthus herbs with nutraceutical potential. The Pharma Innovation Journal, 7(5): 224-229. Al-Bayati, F. A. and Al-Mola, H. F. (2008). Antibacterial and antifungal activities of different parts of Tribulus terrestris L. growing in Iraq. Journal of Zhejiang University Science B, 9(2):154-159. Erum, S., Ambreen, F., Naeemullah, M., Masood, S., Qayyum, A. and Rabbani, M. A. (2012). Genetic divergence in Amaranthus Collected from Pakistan. The Journal of Animal & Plant Sciences, 22(3), 653-658. Ghani, A., Nadeem, M., Ahmed, M. M., Hussain, M., Ikram, M. and Imran, M. (2016). Spatial Variation In Nutritional And Elemental Profile Of Mako (Solanum nigrum) Collected From Different Thehsils Of District Mianwali, Punjab, Pakistan. Science International, 28(6): 5251-5255. Hassan, L. G., Hassan, S. W., Hashim, T., Umar, K. J. and Sani, N. A. (2011). Detrimental Of Nutritive Values Of Garden Cress (Lepidium sativum L.) Leaves. Bayero Journal of Pure and Applied Sciences, 4(2): 18-23. Hussain, A. I., Anwar, F., Shahid, M., Ashraf, M. and Przybylsk, R. (2010). Chemical composition, antioxidant and antimicrobial activities of essential oil of spearmint (Mentha spicata L.) from Pakistan. Journal of Essential Oil Research, (22): 78-84. Ojo, A., Abiodun, O. A., Odedeji, J. O. and Akintoyese, O. A. (2014). Effects of Drying Methods on Proximate and Physico-chemical Properties of Fufu Flour Fortified with Soybean. British Journal of Applied Science and Technology, 4(14): 2079-2089. Singh, B., Borthakur, S. K. and Phukan, S. J. (2014). A survey of ethnomedicinal plants utilized by the indigenous people of Garo Hills with special reference to the Nokrek Biosphere Reserve (Meghalaya), India. Journal of Herbs, Spices and Medicinal Plants, 20: 1-30. Sarfraz, Z., Anjum, F. M., Khan, M. I., Arshad, M. S. and Nadeem, M. (2011). Characterization of Basil (Ocimum basilicum L.) parts for antioxidant potential. African Journal of Food Science and Technology, 2(9), 204-213.