Determination of Thiamine Hydrochloride in Commonly used food Materials-Rice by UV-Spectrophotometric Methods

 

Senthilraj. R¹*, Harikrishnan. N², Swetha.V³, Kavitha. S⁴, Gous Basha. T⁵, Mohamed Nabis A⁶

¹Associate Professor, Faculty of Pharmacy, Dr. M.G.R Educational and Research Institute

(Deemed to be University), Velappanchavadi, Chennai 600077 Tamilnadu.

²Associate Professor, Faculty of Pharmacy, DR. M.G.R Educational and Research Institute

(Deemed to be University), Velappanchavadi, Chennai 600077 Tamilnadu.

³Assistant Professor, Department of Pharmaceutics, Karpagam College of Pharmacy, Coimbatore –32

⁴Faculty of Pharmacy, Dr. M.G.R Educational and Research Institute

(Deemed to be University), Velappanchavadi, Chennai 600077 Tamilnadu.

⁵Assistant Professor, Faculty of Pharmacy, Dr. M.G.R Educational and Research Institute

(Deemed to be University), Velappanchavadi, Chennai 600077 Tamilnadu.

⁶Faculty of Pharmacy, Dr. M.G.R Educational and Research Institute (Deemed to be University), Velappanchavadi Chennai 600077 Tamilnadu.

 

ABSTRACT:

Vitamins and their coenzymes roles are indispensable for normal functioning of biochemical metabolic pathways. The water soluble vitamin thiamine plays an important role in energy metabolism and its continuous intake is very essential. The source of vitamins can be obtained from many foods like milk, legumes, peas and rice and which is depends on availability, cost. Although most part of the south Indians especially in Tamilnadu the people predominantly depends on rice as a major source of food. So in order to identify the recommended dietary allowance of thiamine the following rice samples were collected from provisional store in Chennai. The collected five rice samples were Deluxe, Bapatla, Karnataka, Rupali ponni rice and Basmati rice. The concentration of thiamine present in the rice was estimated by UV Spectrophotometrically at 246 nm using 0.1M hydrochloride as blank. In order to measure the actual content the high heat and pressure not used and washing is not used. Among the tested 10gm of rice samples, Rupali ponni rice has127 µg which is very less quantity and Bapatla, Deluxe, Karnataka ponni rice has 420 to 430 µg. The high content of was present in Basmati rice-983 µg. Anyhow normal consuming rice quantity is more sufficient to meet Recommended Daily Allowance- RDA level but more washing of rice, prolong boiling will reduce the amount of thiamine content in rice during usage.  

 

 

 

INTRODUCTION:

Vitamins plays a major role in human health especially water soluble vitamin Thiamine, which act as a coenzymes for a series of enzymes in different metabolic pathways and it is needed for the production of ATP.^1,2 The continuous supply of vitamin from food is inevitable because which are organic compounds cannot be synthesized by the human beings.^3,4

 

All water soluble vitamins are easily excreted from the body and other factors also influence the thiamine level such as age, consumption of alcohol, barbiturates, and parenteral administration of glucose solutions increase the need for thiamine.⁵ Further Thiamine needed for reduce the risk of several impacts such as endothelial vascular diseases, lipid profile, retinopathy, and neuropathy in the Diabetes Mellitus and severe thiamine deficiency causes heart disease, beriberi.^6-10 Due to the above fact, daily continuous intake of vitamins is very important.^11-14 Generally major sources of thiamine in the diet are available in whole grain-rice, cereals, green vegetables, and beef.^15,16 Even though many food materials act as good source of vitamin thiamine, the food usage in particular area is depends on number of factors such as availability, cost and seasonal variability etc.^17-19 Due to that people in particular area cannot be possible to use all types of food as a source for procuring vitamins in a single time. Further predominately for carbohydrate source most part of the south India depends on rice. Hence in this study most principally Rice used as food source area of Tamilnadu and Chennai region was selected.^{20, 21} Rice samples were taken from provisional store and its vitamin thiamine availability was measured by UV-Spectrophotometrically to know whether it meets the recommended daily allowance.  

 

MATERIALS AND METHODS:

The absorbance measurements are carried out using Shimadzu UV-160 UV Visible spectrophotometer. The analytical grade of all chemicals used and all dilutions and solutions were prepared from double distilled water. Thiamine Hydrochloride (THC) was obtained from Sigma-Aldrich, Hydrochloric Acid (HCl) from SD Fine, Whatman No.1 filter paper, Rice samples (100gm per sample) obtained from different rice seller in Chennai city and all each samples were mixed together.

 

Preparation of 0.1N HCl:

In a cleaned and dried 1000 ml volumetric flask 100 ml of double distilled water was taken. Carefully added about 8.3 ml of Concentrated Hydrochloric acid with continuous stirring. Add more about 700 ml of double distilled water, and allowed to cool to room temperature. Finally the remaining volume 1000 ml filled with double distilled water. The solution was kept one hour and mixed thoroughly before use.

 

Calibration Curve of THC:

The range of concentration of 0.5-10µg/ml Standard solution of THC was prepared and the absorbance was measured at 246 nm against 0.1N HCl used as blank. By using the standard calibration curve the Unknown concentration was determined.

 

Determination of Thiamine content:

The collected rice sample was weighed accurately and particle size of the Rice is reduced with mortar and pestle and then size reduced into flour. Rice flour 10gm was weighed and shifted into a beaker and 0.1 N HCL 90 ml was added. The mixture was heated for 30 minutes at 100 °C in a water bath with continuous stirring. After cool, the solution was filtered with Whatman filter paper. The filtrate was transferred to a 100 ml volumetric flask and diluted with 0.1 N HCL to the marker boundary. All the rice samples absorbance was performed by triplicates in a same time.^22-27

 

RESULTS AND DISCUSSION:

The images of rice samples collected from the Chennai city was given in the Fig:1. The collection is based on their common usage by the people for food material preparations.

 

Fig: 1 Rice samples which is commonly used in Chennai city

 

Determination of Maximum Wavelength:

The maximum wavelength was confirmed, before to the determination of THC. By using UV Spectrophotometer at wavelength 200-400 nm measurements were made by reading the absorbance of THC standard solution. The measurements of results obtained at 246nm maximum absorbance. This 246nm of wavelengths is used for the determination of THC on the rice sample.

 

Calibration Curve:

 

Table: 1 absorbance value for standard calibration curve

S. No	Concentration µg/ml	Absorbance at 246nm
1	1	0.122
2	2	0.251
3	4	0.506
4	6	0.788
5	8	1.009
6	10	1.269

 

Graph: 1 Standard calibration curve-Thiamine

 

Table: 2 Absorbance values of collected Rice samples at 246nm

S. No	Name of the Rice	Absorbance at 246nm
1	Bapatla ponni rice	0.554±.003
2	Basmati rice	1.252±.003
3	Deluxe ponni rice	0.539±.001
4	Karnataka ponni rice	0.537±.002
5	Rupali ponni rice	0.162±.002

 

Table: 3 Unknown concentration of Thiamine after interpretation from the calibration graph

S. No	Name of the Rice	Thiamine µg/10gm
1	Bapatla ponni rice	435
2	Basmati rice	983
3	Deluxe ponni rice	423
4	Karnataka ponni rice	421
5	Rupali ponni rice	127

 

According to Beer-lamberts law in the concentration range 1to10 μg/ml shows good linear relationship. The calibration curve as mentioned in the graph no.1 indicates the regression equation y = 0.1275x- 0.0012 and the correlation coefficient (r) = 0.9993 and the calibration curves obtained correlation coefficient value close to one. So it shows there is a strong correlation was found between the concentrations of standard-Thiamine solution and its absorbance at 246nm.^28,29

 

Determination of Thiamine content:

The Thiamine content on the selected Parboiled rice samples after extraction of rice using 0.1N HCl. The result of absorbance value is shown in Table 2 and based on the absorbance value the unknown concentration was determined by after interpretation with standard calibration curve was give on Table 3. Among the tested 10gm samples Rupali Ponni Rice has very less thiamine content that is 127 µg but the highest content 983 µg was found in Basmati Rice. The remaining rice samples Deluxe, Bapatla and Karnataka Ponni has 421 to 435µg. In this study rice samples were not washed and not heated at higher temperature and the results indicates the actual maximum content of thiamine present in the rice samples because mostly water soluble vitamins are easily eluted from the food materials during washing and furtherer amount may reduce while boiling at higher temperature.

 

Table: 4 Quantity of selected rice needed to be taken to meet RDA of Thiamine

S. No	Name of the rice	Recommended Daily Allowance (RDA)gm
1	Bapatla ponni rice	30
2	Basmati rice	15
3	Deluxe ponni rice	30
4	Karnataka ponni rice	30
5	Rupali ponni rice	100

 

The table 4 indicates that the RDA of Thiamine of the various selected rice samples. Among the selected sample, people to meet RDA, less quantity of Basmati rice is sufficient and similarly 30 gm of Bapatla, Deluxe and Karnataka ponni rice is required but Rupali Ponni rice is needed more quantity (100gm).

 

Even though the people are consuming daily more than the above mentioned quantity of rice, due to washing and high boiling conditions in cooking water soluble vitamins are easily solubilized.

 

CONCLUSION:

Commonly eaten type, parboiled rice has significantly more thiamine than white rice. As per WHO guidelines RDA of thiamine is 1.2mg. The above study concluded that all the selected rice has sufficient quantity of thiamine only under no washing and mild cooking conditions.

 

ACKNOWLEDGMENTS:

The authors are thankful to management of Dr. M.G.R Educational and Research Institute, Chennai, and management of Karpagam College of Pharmacy, Coimbatore for providing facilities required for the successful completion of the research work.

 

AUTHORS CONTRIBUTIONS:

All the authors have contributed equally.

 

CONFLICT OF INTEREST:

The authors declared that they have no conflict of interest.       

 

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Received on 14.05.2020           Modified on 14.08.2020

Accepted on 19.09.2020         © RJPT All right reserved