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Lasbela. Uni. J. Sci. Tech., vol.3, pp. 10-13, 2014 ISSN 2306-8256 RESEARCH ARTICLE Analysis of Total Sugar and Free Amino Acids in Wheat Grains Samples from Five Different Regions of Balochistan Farida Behlil1*,Samiullah2, Muzaffar Khan1,Shafia Muzaffar1, Musarat Raiz1 and Shazia A Jan1 1. Department of Chemistry, Sardar Bahadur Khan Women University, Brewery Road Quetta, Pakistan. 2. Department of Chemistry, University of Balochistan, Sariab Road Quetta, Pakistan. Abstract:-Wheat is an important cereal enriched with starch, proteins and fibers. In this analysis five different samples of wheat grains grown in different ecological regions (Jaffarabad, Kohlu, Loralai, Quetta and Sibi) of Balochistan were studied for total sugar and free amino acids. These parameters result in differences between all the samples. Maximum concentration of both total sugar and amino acids were shown by wheat grain sample of Loralai while minimum concentration of total sugars were shown by the sample from Sibi and minimum concentration of amino acids were shown by sample from Jaffarabad. The differences in genotype and environmental conditions were considered to be responsible for the variation of total sugar and free amino acids concentration in wheat grain samples taken from different regions of Balochistan. Key Words: Wheat, total sugar, amino acids, Balochistan. (Tobeh and Jamaati-e-Somarin, 2011). Wheat grains consist of the sugars, amino acids, fats, vitamins, minerals and crude fibers. Sugars are the most important and chief component of wheat (Hussain and Qamar, 2007). The anthrone method for determination of sugars has been known for many years. Glucose is normally used to establish a calibration curve for the anthrone method and total sugars are expressed in terms of glucose (Cerning-Beroard, 1975; Kumar, 2011). Another component of wheat is amino acids which are the monomers of proteins. Amino acids in food components are present in free form or bound through peptide linkage or non-peptide bonded polymers. Analysis of the total amino acid in food components involve protein hydrolysis by a variety of methods that take into account variations in permanence of individual amino acids (Peace and Gilani, 2005). Ninhydrin is frequently used for the identification of free amino and carboxylic acid groups of proteins and peptides (Friedman, 2004). MATERIALS AND METHODS INTRODUCTION Wheat is a cereal grain that belongs to the genus Triticum. In 2010 it was considered as the third most abundant cereal plant after maize and rice. Pakistan occupies a very important position in wheat production. It is an important primary food, consumed 65% by human, 21% by animals, 8% as seeds and 6% used in industries. It is cultivated in all provinces of Pakistan (Noorka et al., 2009; Khan et al., 2010). Balochistan province is situated in the southwest of Pakistan. The climate of Balochistan varies from extreme cold winter to hot summers in highlands, plain regions have mild winter and summers are hot and dry (Khan and Aslam, 2004; Weiss et al., 2012). Amongst the environmental factors, temperature has a strong effect on the growth of wheat. Wheat growth consists of germination, requires cold temperature (3 ᵒC) while maturation process needs normal temperature (25-33 ᵒC). It takes about several weeks for the formation of grain The samples of wheat grains used in this analysis were collected from five different *Corresponding author: [email protected] 10 regions of Balochistan such as Jaffarabad, Kohlu, Loralai, Quetta and Sibi. These samples were washed and then dried in oven. For analysis these grain samples were grinded to form flour by using electrical grinder. To obtain standard graph, 0.5 ml of each standard leucine solution and 0.5 ml of each of the samples solution were taken in separate test tubes and their volume were made 1ml by using distill water. Then 1ml of 2% ninhydrine and 1ml of 10% pyridine solution were added to all test tubes. All the test tubes were heated in boiling water bath for 30 minutes and result was obtained in violet color. The solutions of all the test tubes were made the volume up to 50 ml by distilled water. The absorbance was recorded by UV-VIS spectrophotometer at 570 nm. For the preparation of sample solution 2 grams of flour of each sample was taken and dissolved in 75% solution of ethanol in separate test tubes and were filtered after 24 hours. Then these solutions were used for the analysis of total sugar and free amino acids by using following methods. RESULTS AND DISCUSSION ANALYSIS OF THE TOTAL SUGAR Wheat is a cereal plant cultivated largely than any other cereal in the world. It is planted in majority of sub-tropical regions and almost all the temperate countries of the world. In Pakistan it is cultivated in all the provinces. It is a primary dietary source in the world, containing all the basic nutrients like proteins, free amino acids, fats, carbohydrates and micronutrients.Wheat also play very important role in the economy of every country. The wheat nutritive value, grain yield, grain texture and size greatly depends upon the genotype and enviromental conditions. It depends upon soil composition, fertilizers, stress factors like humidity and drought.These factors are different even in the same country as well as in the same province. Pakistan is a country having areas ranging from very cold to very hot enviroment (Zeb et al., 2006). The analysis of total sugar is estimated by anthrone method (Cerning-Beroard, 1975). A stock solution of glucose (0.01%) was prepared. Glucose standards of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, and 1 ml and 84% Sulphuric acid, 0.2% Anthrone were also prepared. METHOD The glucose standards and 0.1 ml of each of the sample solution were taken in separate test tubes and their volume was made to 1ml by adding distilled water. 4 ml of 0.2% anthrone solution was then added to each test tube and stirred. The test tubes were heated in boiling water bath for (20) minutes and cooled. Blue green colour was observed in all the test tubes. The absorbencies of all the solutions (glucose standards and samples) were observed by UV-visible spectrophotometer at 620nm. The aim of the study was to analyze the chemical composition of wheat cultivated in five different regions of Balochistan i.e Jaffaraabad, Kohlu, Loralai, Quetta and Sibi. The difference was observed in the chemical composition of wheat samples from five different region of Balochistan. The maximum value (0.068 mg/2g) of free amino acid was shown by the sample from Loralai while minimum value (0.068 mg/2g) was shown by the sample from Jaffarabad. The samples from other regions also showed a marked difference in their free amino acid concentration as shown in table I. ANALYSIS OF TOTAL FREE AMINO ACIDS The analysis of total amino acids in wheat samples is taken by Ninhydrine method (Pandey, 1984). Standard solutions of Leucine with different concentration 1, 2, 4, 6, 8, and 10 mg were made. 10% Pyridine, and 2% Ninhydrine solutions were also prepared. METHOD 11 Similarly the maximum concentration of total sugar (0.09 mg/2g) was shown by the sample from Loralai and the minimum concentration of total sugar (0.048 mg/2g) was shown by the sample from Sibi. The samples from other regions also showed a marked difference in their total sugar contents as shown in table II. optimum temperature around 25-33°C which allow the grain to stores greater amount of sugar and other nutrients while the Sibi which is the hottest region provide high temperature during grain formation as a result decrease the time of grain maturation which affect the accumulation of sugar and other nutrients. Similar results were also conducted by Buriro et al., 2011 by using five different varieties of wheat grains. Temperature is also responsible for high amino acid concentration in wheat sample from Loralai and low concentration in wheat sample form Jaffarabad. The same result was also found by Balla et al, 2009 that the high temperature treatments at grain formation stage destroy the quality of wheat grains. But along the temperature change this difference may be due change in the salinity of the different areas of Balochistan which is the cause of formation of less sugar and other nutrients which was also determined by Rahman et al., 2009. The water availability and nutrients in the water also affect the chemical composition and other characteristics of wheat as the water stress also showed significant decline in wheat quality due decrease in nutrient availability in water as determined by Noorka et al., 2009. The type and amount of fertilizers is also responsible for this change in the concentration of amino acids and total sugar because fertilizers provide the basic elements required for the chemical parameters as the same result was obtained by the Sameen et al., 2002. Table I: Total free amino acids in wheat samples Regions Jaffarabad Absorbance (nm) 0.14 Total free amino acids (mg/2g of sample) 0.014 Sibi 0.21 0.044 Quetta 0.17 0.026 Kohlu 0.23 0.052 Loralai 0.27 0.068 Table II: Total sugars in wheat samples. Samples Absorbance (nm) Total sugar (mg/2g of sample) Jaffarabad 1.8 0.068 Sibi 1.50 0.048 Quetta 2.2 0.09 Kohlu 2.0 0.084 Loralai 2.3 0.099 Among the environmental factors, temperature is the most important contributing factor controlling wheat germination and wheat quality. Wheat consists of two phases; one is wheat germination which requires the temperature above 3 °C and another is wheat grain formation which require temperature 25-33 °C. The 25-33 °C temperature is required for grain maturation for several weeks. In the grain maturation the sucrose is converted into starch and the sugar of leaves, stems and other parts transfer to the grain and store in it (Tobeh and Jamaati-e-Somarin, 2011). The results of this analysis showed maximum sugar concentration in Loralai region because during grain formation in the month of March and April it provide REFERENCES Balla, K., Karsai, I., Veisz O. 2009, Analysis of the quality of varieties at extremely high temperature, DOI:10.1556/CRC.37.Suppl.2. Buriro, M., Oad, F. D., Keerio, M. I., Tunio, S., Gandahi, A. W., Hassan, S. W., Oad, S. M. 2011, Wheat seed germination under the influence of temperature regimes: Sharhad Journal of Agriculture, v. 27, p. 539- 543. Cerning-Beroard, J., 1975, A note on sugar determination by the anthrone method: Journal of Cereal Chemistry, v. 52, p. 857-860. Friedman, M., 2004, Applications of the ninhydrin reaction for the analysis of amino acids, peptides and protein to agriculturaland biomedical sciences: Journal of agricultural and food chemistry. v. 52, p. 385-406. 12 Hussain, S. S., Qamar, R. 2007, Wheat genomics: Challenges and alternative strategies, Proceedings Khan, A. M. and Aslam, M. 2004. Medicinal plants of Balochistan. Project on Introduction of Medicinal Herbs and Species as Crop. Ministry of Food, Agriculture and Livestock, Qarshi Industries (Pvt) Ltd. pp 3-44. Khan, M. R., Anjum, F. M., Zahoor, T., and Nawaz, H., 2009, Biochemical and technological characterization of Pakistani spring wheat: Pakistan Journal of Agricultural Science, v. 46, p. 271-279. Kumar, A., 2011, Determination of Total Carbohydrate by anthrone method. http://technologyinscience.blogspot.com/ Noorka, I. R., Rehman, S. U., Haidry, J. R., Khaliq, I., Tabassum, S., and Mueen-ud-din G., 2009, Effect of water stress on physico-chemical properties of wheat(Triticum aestivum L.): Pakistan Journal of Botany, v. 41, p. 2917-2924. Pandey, P. B., 1984, Economic Botany: J.V.College Baraut, 18. Peace, R.W., Gilani, G. S., 2005, Chromatographic determination of amino acids in foods: Journal of AOAC international, v. 88, p. 877-887. Rahman, M., Soomro, A. U., Haq, Z. M., and Gul, S., 2008, Effects of NaCl Salinity on Wheat (Triticum of the Pakistan academy of Science, v. 44, p. 305 327. aestivum L.) Cultivars: World Journal of Agricultural Sciences v. 4, p. 398-403. Sameen, A., Niaz. A., and Anjum, F. A., 2002, Chemical composition of three wheat varieties as affected by NPK doses: International Journal Agriculture and Biology, v. 4, p. 537-539. Tobeh, A., and Jamaati-e-Somarin, S., 2011, Low temperature stress effect on wheat cultivars germination: African Journal of Microbiology Research, v. 6, p. 1265-1269. Weiss, A., Mughal, A, M., and Zeb, M. A., 2012, Louis Kotzé and Stephen Morse (eds): Berkshire Encyclopedia of Sustainability, v. 9, p. 236-240. Zeb, A., Ali, Z., Ahmad, T., and Ab,dumanon, A., 2006, Physiochemical characteristics of wheat varieties growing in the same and different ecological regions of Pakistan: Pakistan journal of biological sciences v. 9, p. 1823-1828. (Received 24th July 2014, Accepted 3rdOctoberr 2014) Article can be viewed online at www.luawms.edu.pk 13