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Estimation of Whole Amino Acid in Wheat Varieties Both Its and Ils Condition in Bangladesh

M. H. H. Khan
Postharvest Technology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

S. Pervin
Postharvest Technology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

M. Miaruddin
Postharvest Technology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

M. M. Rahman
Postharvest Technology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

Abstract/ Executive Summary:

Grains of four released wheat varieties cultivated under both irrigated timely sown (ITS) and irrigated late sown (ILS) condition were collected from Wheat Research Centre, BARl, Gazipur to evaluate amino acid profile. The four varieties were BARl gom 25, BARl gom 26, BARl gom 27 and BARI gom 28. Amino acid profile were estimated by HPLC using CI8 column with UV detector at 254 nm wave length. Sixteen essential amino acids were found in four different wheat grains both ITS and Il. Scondition comparing with Standard amino acid (PEARCE). In this analysis it was found that Aspertic acid was absent in BARl Gom 27 and Lysine was also absent in BARl Gom 25 in both ITS and ILS condition. When all those four varieties were cultivated under ILS condition Cysteine was absent in wheat grain. Only Glutamic acid, Threoning and Proline reached higher chemical score than egg albumin.

Keywords: Wheat, Cultivation, BARI gom

Location: Bangladesh Agricultural Research Institute, Joydebpur, Gazipur, Bangladesh

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Wheat

Objectives:

To identify the estimation of whole amino acid in wheat varieties in Bangladesh

Methodology:

Amino acid standard,HPLC grade, 6N HCl and triethylamine (TEA), phenyl isothiocyanate (PITC) were from Pierce Chemicals (Rockford,IL, USA). HPLC grade Ammonium acetate was from Sigma (St. Louis, MO,USA). Acetonitrile (HPLC grade) was from Fisher Scientific(Fair Lawn, NJ, USA). Ultra-pure water (Millipore,Bedford, A, USA) of at least 18.2µ cm resistivity was used for the preparation of all reagents, eluents, and buffers. All were filtered through 0.2 µm membranes. Grains of four released varieties were collected from Wheat Research Centre, Joydebpur. Wheat seeds were initially grounded in a Micro grinder. For convenience, each sample of 1000 mg was defatted and stored in glass vial. Protein hydrolysis, precolumn derivatization, and HPLCanalysis were performed according to Heinrikson and Meredith (l984) with modifications. Hundred miligram samples were placed in each hydrolysis tube with 1000 µL of 6 N HC. Evacuation in 25-30 in-Hg pressure alternated with nitrogen flushing was performed at least three times and sealed. Samples were hydrolyzed at 108°C in lock heater for 24 h. After hydrolysis, samples were dried under vacuum not exceeding 45°C and stored in freezer under -20°C until derivatization and HPLC analysis. A 250-µL sample of amino acid standard mixture was also used. Both standard and hydrolyzed 100µL samples were neutralized by adding 20µL of a 2:2: 1 mixture of ethanol:water: TEA(v/v), and mixing well with a vortex stirrer. They were dried under vacuum. Derivatization was performed by adding 30µL of a mixture of 7:1:1:1 ethanol:water:TEA:PITC (v/v), and mixing well with a vortex stirrer. The reaction between PITC and the hydrolysate to produce phenylthiocarbamyI (PTC) amino acids was allowed to complete for 20 min at room temperature. Samples were then completely dried under vacuum and stored in a freezer under -20°C. Phenylthiocarbamyl amino acids in each sample and standard were dissolved from the dried matrix by vortex mixing with 1000/µL of 0.1 M ammonium acetate buffer. The fluid was then filtered through a 0.22-µm membrane. Samples were reconstitute done at a time due to the PTC amino acid sensitivity to light and ambient temperature. Ten microliters of sample were injected and analyzed with an HPLC system equipped with a column heater, auto-sampler, variable wavelength detector, and a data acquisition software controller. The reverse phase column used was C 18 (4.6x250 mm), dimethyloctadecylsilyl bonded 0.5 µm amorphous silica, with aninline column filter. The column temperature was maintained at 43°C. The PTC amino acids were separated andeluted by a gradient resulting from mixing eluents A and B. Eluent A consisted of 0.1 M Ammonium acetate, pH 6.4. Eluent B consisted of 46: 10:44 acetonitrile:Methanol:water (v/v). The flow rate was 1 mL/min throughout, and the gradient consisted of the following profiles: 100% A at start, 90% A at 15 min, 60% A at 30 min, 0 A at 50 min, 100 A at 56 min.

Source of Information: Annual Research Report, Postharvest Technology Division, BARI, Gazipur, 2013-2014

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Sixteen essential amino acids were found in four different wheat grains both ITS and ILS condition when compared with standard amino acid (PEARCE). In this analysis it was found that Aspertic acid was absent in BARl Gom 27 and Lysine was also absent in BARl Gom 25 in both ITS and ILS condition.

Knowledge Management: Journal