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Spectroscopy of Salicylates | Lab Report

The scenario is that a woman has been stopped by the police due to the condition of her driving. She tells the police that her driving was erratic due to trying to take the lid of a bottle of aspirin which she required for a severe headache. The police could not find the aspirin bottle within the car and the woman then told the police that it got thrown out of the vehicle. Serums samples were taken from the woman to do alcohol and drug tests.
Aspirin is an analgesic drug that provides pain relief without causing unconsciousness and anesthesia. Aspirin is used to help with pain, fever, osteoarthritis, inflammatory conditions, migraine headaches and many other things.
Salicylates are the main substance in aspirin and this is what is being tested for. To test for this substance visible spectroscopy will be used. Spectroscopy has a variety of methods that are the interaction between light and matter. For this practical the method used is visible spectroscopy via the use of a spectrophotometer. This equipment measures the amount of light absorbed by passing a beam of light through the sample and the amount of light is measured by a detector. This produces an absorbance value which can be used to calculate concentrations.
The aim of this test is to calculate the concentration of the serum samples to find out whether the levels of salicylic acid in these samples are toxic or not.
Methods and Materials The materials that were used are 0.2mg/ml Salicylic acid, Ferric Nitrate solution, 0.039M HNO3, water and a sample of the woman’s serum. The equipment used was a spectrophotometer, a graduated pipette and cuvettes.
To be able to find out the salicylic acid concentrations of the serum samples, six standard solutions had to be created first. Each standard solution was composed of a combination of H2O, salicylic acid, Ferric nitrate and HNO3 all totalling up to 2ml in each solution.

The components of the solutions seen in table 1 were placed into test tubes using a graduated pipette. The solutions were then mixed thoroughly and left for 5 minutes. After five minutes a sample of each solution was placed into cuvettes using the pipette. The spectrophotometer was then set a 540nm.Using solution 1 as the blank; the spectrophotometer was then set to zero. Every other standard solution was then inserted into the spectrophotometer to obtain absorption measurements using the blank as a zero reference.
Three 2ml samples of the woman’s serum were then created using a combination of the materials seen above.

The components of the serum solutions were then placed into test tubes at the volumes shown in table 2. These samples were then mixed thoroughly and left for five minutes. After five minutes the samples were then placed into cuvettes, and then the spectrophotometer. The spectrophotometer was again set at 540nm and the serum blank used as the blank. The other two serum samples were then placed into the machine and the absorption results recorded.
Results The measurements from the spectrophotometer where recorded and concentrations of Salicylic acid calculated for the standard solution. These concentrations then created a calibration curve to allow the serum sample concentrations to be discovered.

The concentrations in table 3 where calculated using C1V1=C2V2. Where C1 is 0.02mg/ml, the original concentration of salicylic acid, V1 is the volume of salicylic acid, C2 is the unknown concentration and V2 is 1ml, the total volume of H2O and Salicylic acid. For example standard 3 has the concentration 0.06mg/ml. From looking at table 1 it is seen that C1 is 0.02mg/ml andV1 is 0.3ml. it is then known that C2 is unknown and V2 is 1ml. The equation is then rearranged to C2 = C1V1/ V2 to produce C2=0.02mg/ml x 0.3ml / 1ml = 0.06mg/ml.
The concentration and absorption results are then put into a graph to create a calibration curve.
Calibration curve
The calibration curve shown in figure 1 was plotted from the results in table 3. This curve was then used to calculate the concentration of the serum solutions.
Concentration and absorbance results for the three serum samples

The results in table 4 where done as a duplicate to make the results more accurate and reliable but due to an error within the practical. An average was later calculated to correct the inaccuracy created by the error, but ideally the test should have been done with 3 samples.
Discussion Using the results from table 4 and the calibration curve from figure one it can be determined if the results agree with the drivers story. The average serum sample had a salicylic acid concentration of 0.116mg/ml. In relation to the standard solutions this result is quite high therefore the results confirm she did in fact take aspirin. From this it cannot be fully determined if the level is toxic or not.
Salicylic acid toxicity causes many bio-chemical that target no specific organ. An acute overdose would cause symptoms like nausea, vomiting and tinnitus. If the overdose was chronic there would be symptoms like confusion, fever, hypoxia, dehydration and metabolic acidosis. Other symptoms of toxicity are respiratory alkalosis, alkaline urine and headaches. Comparing the symptoms with the scenario some of the symptoms could explain the reasons for her erratic driving but there still could be other reasons.
Serum levels of salicylic acid provide if the ingested amount was therapeutic, toxic or chronic toxicity. A therapeutic level is 15-30mg/ml and a toxic level is 40-50mg/ml. Any serum level above 60mg/ml is a chronic level. To compare the concentration of the serum to toxicity levels the serum concentration must be converted to mg/dl. For this it is need to be known that there are 100 millilitres in 1 decilitre, therefore 0.116mg/ml needs to be multiplied by one hundred. The serum level is then 11.6mg/dl; this level is not a toxic level but does confirm that aspirin was taken at a possible therapeutic level. This confirms that the woman was telling the truth about taking aspirin but does not confirm why her driving was erratic.
Other test could also be done to test for salicylic acid, these as urine pH, a bedside ferric chloride test and arterial blood gases. Also high-performance liquid chromatography could be used but this requires time that would not be available in a serious toxicity. Salicylic acid has many related compounds, for example, benzoic acid, phenol, magnesium salicylate and aminosalicylic acid. Test for these could also be done to help determine the source.
Conclusion In conclusion the results from the visible spectroscopy show that the woman did not have salicylic acid toxicity even though she did take aspirin. As her levels where not toxic there has to be another reason for her driving. To confirm this more test could be done like the urine pH and the arterial blood gasses. Also test on the related compounds could be carried out to confirm is she did take aspirin or if the levels in of salicylate in her blood are from elsewhere. The next thing to do would be to test for alcohol and other drugs both legal and illegal. Other drugs to test for could be things like cocaine, opiates, amphetamines, marijuana, methadone, tramadol and oxycodone. These could be testing for either through the blood, urine or hair.

Reflective Essay NU-AGE

NU-AGE is a module that has had a profound influence of my understanding about the concept and self-perception about ageing. Having to learn how much more biology, lifestyle, clinical aspects, technology, living conditions and solutions can directly or indirectly alter how an individual age.
Ageism in one major concern that was given much attention to in the module. Ageism was thought be the direct action of our society towards the ageing population(reference). For the past century, the institutionalisation and the idea of making the older population redundant was a major concern to both the economy. Many had to opt for poor relief, begging, paid work when possible, charity, growing food, use of common rights and help from family. Unequal access to improved occupational and private pension have been a concern to how the ageing population had access to an income to support them through later life has contributed much to the of the negative concept of ageing. This led the population to have a stereotypical view of the prevalence of loneliness at older people that their prevalence of illness and disease were more likely higher and older workers are less efficient. Despite all these negative stereotypical views positive ideas and views have been acknowledged, but many recognise that the ageing population have a perceived superior wisdom from experience.
Health Nutrition and health has an undoubtedly profound aspect on how progression of ageing occurs. Research has shown that people in poorer parts of England lived on average 7 years less than those of in richer parts. This has been affiliated with the qualities of life the two regions face and the conditions they are assumed to face in such regions. The poorer regions are proposed to be subjected to adverse environment conditions, more stress and poorer diet. Richer regions are assumed to have the direct opposite qualities of those of the poorer regions. Research has proven that the genotype of an individual has a profound effect on the progression of ageing of an individual, such as having the Age-1 gene. All these factors are known to contribute oxidative stress, metabolic stress and inflammation which known to affect the trajectory of how ageing progresses and the prognostic implications in later life. It has been proven that diets low in calorific value and higher in unsaturated fats and omaga-3, such as the Mediterranean diet, showed marked decrease in the causes of mortality such as cardiovascular disease, neurodegenerative diseases and cancers. Physical activity has been a key player in the progression of how we age. Mental health has been shown to be related to how much an individual is involved in the arts and society. For example, risks of showing onsets of dementias have been shown to decrease with engaging in thought and social processes. (reference) This clearly shows us the ageing is plastic. This relates very strongly to the Clinical Ageing and Health module I took this year in Biomedical Sciences, iterating much of what was covered but in a simplified coherent manner.
Ageing affect individuals related to the ones ageing. Residents in the care homes haves generally shown that there are losses of formal association with members they have had earlier in life thus leading to increased prevalence to neurodegenerative disease and loneliness. This has been addressed by providing and involving the ageing population in the arts whereby bonding with those of a similar age, ethnicity and class which provides a strengthened emotional support, and social identities.
Wealth Engagement of the ageing population has been a paramount interest of discussion over the years. Establishments of capital development projects were one of the topics of interest that has been of great interest to me, especially one EX-BEX member shared his thoughts of how he thinks that life savings in the bank might not gain as much capital gains as when other forms of investment out-weighed the risks since equity has been rising for the past century.
Through this module, I have truly come to grasp of the global scale of what society is expecting from ageing, how it is perceived and what we as the younger generation can implement to address issues in relation to the current ageing population and to the younger generation to age in a graceful manner. I have learnt that ageing is not a concept to be worried about and that it should be regarded in a positive aspect to consider in life. Being an international student, I have come to experience thoughts and aspects of ageing in the UK and through other international students which I propose to implement in the Seychelles. Seychelles has had a graceful ageing population two decades ago, unfortunately the can no longer be said with confidence. Prevalence’s of age related diseases are rising and life-spans are decreasing. No strong investigation has been made to determine why such decline is occurring. I feel that it is my duty as a Seychellois to lead the way to investigate and support the ageing population into the aspect of graceful and healthy. My hope is to promote the Seychelles as an example to the world that ageing is in our control and we should not fear it. My dream is to see the ageing population of Seychelles to be healthy, active and vibrant thus defeating the ideology and perception of ageism.