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Chemical Composition, Minerals and Vitamins Analysis of Lyophilized et International Journal on Emerging Technologies 10 (4): 137-144(2019) ISSN No. (Print): 0975-8364 ISSN No. (Online): 2249-3255 Chemical Composition, Minerals and Vitamins Analysis of Lyophilized Wheatgrass Juice Powder Nandita Thakur 1, Harcharan Singh Dhaliwal 1 and Vivek Sharma 2 1Department of Biotechnology, Akal College of Agriculture, Eternal University, Baru Sahib-173101 (Himachal Pradesh), India. 2Department of Botany, Akal College of Basic Sciences, Eternal University, Baru Sahib-173101 (Himachal Pradesh), India. (Corresponding author: Vivek Sharma) (Received 17 August 2019, Revised 26 October 2019, Accepted 02 November 2019) (Published by Research Trend, Website: www.researchtrend.net) ABSTRACT: Wheatgrass is known as significant source of vitamins, minerals, carbohydrates, enzymes, chlorophyll and polyphenols. Lyophilization technique helped to preserve the heat sensitive constituents and enhanced shelf-life of product. The lyophilized wheatgrass juice powder subjected to use for chemical, nutritional and antioxidant analysis. Chlorophyll, the main constituent of wheatgrass was analyzed as 7.46 mg/g and essential elements such as Fe, Mg, Zn, K and Mn having higher concentration in wheatgrass juice powder observed by AAS. The presence of vitamins B1, B2, B3, B4, B6, B10 and C detected at 254nm by following RP-HPLC-PDA method and their assistance to enhance antioxidant potential determined with DPPH assay. The presence of these amazing phytoconstituents in wheatgrass makes it one of the healthiest remedy to cure minor ailments to chronic disorders. Keywords: Lyophilization, wheatgrass, chlorophyll, vitamins, minerals, anti-oxidant. I. INTRODUCTION and powder. Such kind of antioxidant rich products have anti-ageing properties and protect us from chronic Cereal grasses have great importance in various diseases like cancer, Alzheimer [7]. The balanced diet cultures of world because of health saving benefits for enriched with minerals, vitamins and proteins needed to all wellbeing. Wheatgrass, one of them has played regulate the body functioning in adequate manner. Lack important role as dietary supplement in most of the of any nutrient lead toward malfunctioning of organs and countries. The 7-10 days germinated wheat seedlings caused chronic diseases. The wheatgrass known as are called as wheatgrass that recognized as detoxifying complete nutrient package that enclosed important agent which maintained the alkalinity of blood. minerals such as magnesium, zinc, selenium and Wheatgrass enriched with chlorophyll content that vitamins like vitamin A, E, C and polyphenols like ferulic actively suppressed the metabolic functioning of acid, cinnamic and gallic acid etc., [8]. The presence of carcinogenic compounds [1]. The presence of flavonoids and apigenin like inflammation reductants polyphenols, vitamins like β-carotene, ascorbic acid and were assisted to reduce the severe abdominal pain and chlorophyll pigments revealed the antioxidant rectal bleeding in case of ulcerative colitis disease [9]. potentiality i.e. high at sprouting stage [2]. Different Our study based on analysis of active constituents or changes observed during germination period due to the metabolites such as vitamins, minerals and antioxidant impact of physical parameters. The beneficial primary by following methods of HPLC (High Performance and secondary metabolites such as minerals, vitamins, Liquid Chromatography), AAS (Atomic Absorption polyphenolic and antioxidant compounds have been Spectroscopy) and spectrophotometer precisely. synthesized in wheatgrass. Varying proportions of K/Ca caused alteration in anti-oxidative mechanism of II. MATERIAL AND METHODS phytoconstituents. Elevating concentration of K A. Material enhanced the polyphenolic components and vitamins that improved the quality of product [3]. Schnabel Wheat grains ( Triticum aestivum L.) of best quality were recognized as pioneer of wheatgrass who familiarized it taken from local market for experimentation and surface as beneficial medicinal therapy for human beings. sterilized by using 0.1% NaOCl. After proper washing of According to him the nutritive quality of cereal grasses wheat grains with distilled water, cultivation was carried depended on their developmental and growth levels [4]. out in plastic trays (Dimensions: length-49.0 cm, width- Daily intake of wheatgrass reduced the risks of 28.0 cm and depth-2.5 cm) at laboratory scale under chemotherapy and need of medicines in breast cancer controlled temperature and light conditions. The salts of suffering patients [5]. The presence of poly-phenols in Hi Media and analytical grade solvents were utilized for wheatgrass assisted to revert the consequences of analysis. reactive oxygen species and limited the chances of B. Wheatgrass Juice Extraction and Lyophilization cancer like disease [6]. Due to increasing demand of On tenth day wheatgrass was cut with sterilized knife antioxidant rich food, the wheatgrass became familiar to and after washing immediately took for extracting juice. all and available in different forms like drinks, capsules Thakur et al., International Journal on Emerging Technologies 10(4): 137-144(2019) 137 Extracted juice was preserved into deep freezer and Freshly formulated 2mL DPPH solution was added to then lyophilized by using Martin Christ-Lyophilization the 1mL extract at different concentrations and unit. The process carried out in three phases i.e., incubated in dark for 30 minutes at room temperature. freezing, primary drying and secondary drying. The After incubation absorbance was Staken at 517nm by lyophilized Wheatgrass Juice Powder (WGJP) was using ascorbic acid as standard. The DPPH solution further used for experimental studies. was prepared in 95% ethanol. The antioxidant potential of extracts determined by using following formula: C. Pesticide Analysis The GC-MS based pesticide analysis of wheatgrass DPPH radical scavenging activity (%age) = juice powder was accomplished by using GC-MS {(A Control – ASample ) / (A Control )} × 100 (Thermo Finnegan) Trace GC ultra and Polaris Q The A Control indicated the absorbance of ethanol + DPPH equipment. The oven temperature of GC was initially whereas A Sample is the absorbance of plant extract + held at 80°C for 1 minute, then 200°C @ 25°C/min for 2 DPPH. minute to 230°C @ 2°C/min for 1 minute and finally its 280°C @ 2°C/min for 10 minutes. The PTV-LV G. Elemental Analysis (Programmable temperature vaporizing large volume The beneficiary elements were analyzed with the help of AAS (Atomic Absorption Spectrometer of Agilent injector) temperature was 50°C and 40-450 amu acquired mass range along with helium as a carrier gas. Technology). The 500 mg powdered sample was digested on digesting unit by using HNO 3 and HF (4:2) D. Proximate Composition Analysis as digesting chemicals. After that volume of digestive The pH, moisture and ash content of WGJP samples were prepared up to 50mL by using double (Wheatgrass juice powder) were analyzed by following distilled water for elemental analysis. Stock solutions of AOAC methods [10]. Acidity, crude fat, crude protein desired mineral standards were prepared in the range of and crude fiber were determined by methods of 0.5 ppm to 50 ppm for calibration and then analyzed the Ranganna [11] and total soluble solids (TSS) analyzed samples after filtration [18]. by using Abbe’s refractometer [12]. Anthrone assay used to analyze carbohydrates [13], phenol-sulphuric H. Vitamin Analysis by HPLC acid method for total sugar content [14] and DNSA (3, 5- Vitamin B-complex (B1, B2, B3, B4, B6 and B10) and Dinitrosalicylic acid) method for reducing sugars [15] by vitamin C were analyzed by RP-HPLC-PDA (Reversed observing color intensity at absorbance (Abs) 630 nm, phase-high pressure liquid chromatography with 490 nm and 575 nm spectrophotometrically. photodiode array detector) method in various extracts. Carbohydrate and sugar content analyzed as GluE Chemicals Used: The HPLC grade solvents such as (glucose equivalent) mg/g dry weight of extract by using acetonitrile, methanol and water were acquired from calibration curve of standard glucose. Merck, Life Science Pvt. Ltd. Mumbai, India and the standards of vitamins from Sigma-Aldrich, USA. E. Chlorophyll and Carotenoid Estimation Sample Preparation: Constituents are extracted from Chlorophyll and carotenoid content was estimated by sample by sonication using methanol: water (80:20) as following the method of Arnon with slight modifications extracting solvents. The filtered extracts were dried by The 100mg of powdered sample was dissolved in using rotary vacuum evaporator at 45 °C.ۜ Now dried .[16] 80% acetone and sonicated properly. After sonication extracts were immersed in methanol (HPLC grade) and supernatant was filtered through Whatmann filter paper filtered by 0.22 µm filter. Different concentrations of no. 1. Repeated the procedure 2-3 times and made final samples were prepared for estimation. Stock solutions volume 10 mL. Then the absorbance of supernatants of vitamin standards were prepared as 1.0mg/2mL in was determined at 663 nm, 645 nm and 470 nm by methanol: water (50:50 v/v) solvents. For making using UV/Visible spectrophotometer. The amount of standard curve concentration range of 1.0 µg/mL to pigments was determined by using following formulas: 5.0 µg/mL was taken. Chlorophyll a (mg/g) = (12.7 × A 663 ) – (2.69 × A 645 )/w × V HPLC Instrumentation and experimental conditions: The HPLC based vitamin analysis of extracts was Chlorophyll b (mg/g) = (22.9 ×
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