Spectrophotometry - determination of arginine

In summary, the student is trying to determine the amount of L-arginine in a capsule by using spectrophotometric determination. He followed the instructions provided, but the results were disastrous. He is worried that something went wrong, and he needs help.
  • #1
herminatorr7
1
0
Hi everyone!

I'm making a research and I want to figure out how much L-arginine is in a capsule, which is said to contain 0,5 grams of L-arginine. Basically I want to check if the nutrition values on the lebel of the product are right.

I decided to try with spectrophotometric determination (there is a short abstract of this method - http://www.sciencedirect.com/science/article/pii/0308814684901109).

In the description it writes that I need thymol (0,02 % in 1N alkali), sodium hypobromite (2%: 0,68 ml of bromine in 100 ml of 5 % sodium hydroxide) and arginine solution (0,1%). After I have everything I have to put into a 5 ml standard volumetric flask an aliquot of arginine solution (0,2-2,6 ml), thymol (2 ml) and sodium hypobromite (0,1 ml) and then I mix them and diluted to the mark with distilled water. The absorbance should be measured at 440 nm. And than the amount of arginine is deduced from a standard calibration curve.

That is what is in the description.
- I put 0,02 g of tymol in 100 ml 1 M NaOH
- I put 0,1 g of chemical arginine in 100 ml distilled water
- I put 5 grams of NaOH in 100 ml distilled water and added 0,68 ml of bromine

Than I tried to make samples for calibration curve. I made five samples, I followed the instructions. In one sample I gave 0,2 ml of arginine solution, in one 0,8 ml, in one 1,2 ml, in one 2 ml and in the last I gave 2,6 ml of arginine solution.
I'm worried because colours in the volumetic flasks at the end didn't go from light to dark. The darkest one was the one which contained 0,8 ml od arginine. Well that makes no sense to me... And results on the spectrophotometer were disastrous. There was no line.

Have you have any idea what could have gone wrong?
I am a high school student so I really need some help there.
Sorry for my English..

Thank you!
 
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  • #2
</code>It sounds like you may not have measured the concentrations correctly. When you are measuring the concentration of your solution, make sure to use accurate measurements. For example, when combining 0.02 g of thymol with 100 ml of 1M NaOH, make sure that you are using the right amount of thymol and the right amount of NaOH. Also, make sure you are using a calibrated measuring device when measuring out the solutions.When preparing your calibration curve, make sure that you are using the same volume of each sample. For example, if you are using 0.2 mL of arginine solution in one sample, use the same amount of arginine solution in all other samples.Finally, make sure that you are measuring the absorbance at the correct wavelength. The wavelength for arginine is usually around 440 nm, so make sure that your spectrophotometer is set to the correct wavelength.I hope this helps! Good luck with your research!
 

What is spectrophotometry and why is it used in the determination of arginine?

Spectrophotometry is a technique used to measure the amount of light absorbed by a substance. It is commonly used in the determination of arginine, which is an amino acid found in proteins. This method is preferred because it is non-invasive and can provide accurate and precise measurements of arginine levels.

How does spectrophotometry work in the determination of arginine?

In spectrophotometry, light is passed through a sample containing arginine. The amount of light absorbed by the sample is then measured using a spectrophotometer. The more arginine present in the sample, the more light is absorbed. This allows for the quantification of arginine levels in a sample.

What are the benefits of using spectrophotometry in the determination of arginine?

Spectrophotometry is a highly sensitive and accurate method for determining arginine levels. It is also non-invasive, meaning it does not require any physical contact with the sample. Additionally, spectrophotometry is a relatively quick and cost-effective method compared to other techniques for measuring arginine levels.

What factors can affect the accuracy of spectrophotometric measurements for arginine?

Some factors that can affect the accuracy of spectrophotometric measurements for arginine include the purity of the sample, the concentration of the arginine in the sample, and the wavelength of light used for measurement. It is important to carefully prepare the sample and calibrate the spectrophotometer to ensure accurate results.

Can spectrophotometry be used for the determination of arginine in different types of samples?

Yes, spectrophotometry can be used to determine arginine levels in various types of samples, including biological samples such as blood, urine, and tissue samples. It can also be used for environmental samples, food samples, and pharmaceuticals. However, the sample preparation and measurement conditions may vary depending on the type of sample.

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