2. The experienced analyst who normally conducts these analyses fell ill and will be unable to analyze the urine samples for the drug in time for the sporting event. In order for the laboratory manager to assign a new analyst to the task, a "blind sample" experiment was done. a. The results for the blind sample experiment for the determination of Methylhexaneamine in a urine sample are shown in Table 1 below. Table 1: Results of blind sample analysis. Response factor (F) Analyst results Internal Standard Concentration 0.25 ug/ml 0.35 mg/ml Signals 522 463 Sample Analysis ? 1.05 ug/ml 15 ml 10 ml Original concentration Volume added to sample Total Volume Signals 25 ml 400 418 i. Provide justification why an internal standard was used in this analysis instead of a spike or external standard? ii. Determine the response factor (F) of the analysis. iii. Calculate the concentration of the internal standard in the analyzed sample. iv. Calculate the concentration of Methylhexaneamine in the analyzed sample. v. Determine the concentration of Methylhexaneamine in the original sample. b. Explain how the results from the blind sample analysis can be used to determine if the new analyst should be allowed to conduct the drug analysis of the athletes' urine samples. c. Urine is considered to be a biological sample. Outline a procedure for safe handling and disposal of the sample once the analysis is completed.

a.i) Justification of why an internal standard was used in this analysis instead of a spike or external standard:

An internal standard was used in this analysis instead of a spike or external standard because an internal standard is a compound that is similar to the analyte but is not present in the original sample. The use of an internal standard in analysis corrects the variation in response between sample runs that can occur with the use of an external standard. This means that the variation in the amount of analyte in the sample will be corrected for, resulting in a more accurate result.

ii) Response factor (F) of the analysis can be calculated using the following formula:

F = (concentration of internal standard in sample) / (peak area of internal standard)

iii) Concentration of the internal standard in the analyzed sample can be calculated using the following formula:

Concentration of internal standard in sample = (peak area of internal standard) × (concentration of internal standard in original sample) / (peak area of internal standard in original sample)

iv) Concentration of Methylhexaneamine in the analyzed sample can be calculated using the following formula:

Concentration of Methylhexaneamine in sample = (peak area of Methylhexaneamine) × (concentration of internal standard in original sample) / (peak area of internal standard)

v) Concentration of Methylhexaneamine in the original sample can be calculated using the following formula:

Concentration of Methylhexaneamine in the original sample = (concentration of Methylhexaneamine in the sample) × (total volume) / (volume of sample) = (concentration of Methylhexaneamine in the sample) × (25 ml) / (15 ml) = 1.67 × (concentration of Methylhexaneamine in the sample)

b. The results from the blind sample analysis can be used to determine if the new analyst should be allowed to conduct the drug analysis of the athletes' urine samples. The new analyst should be allowed to conduct the analysis if their results are similar to the results of the blind sample analysis. If their results are significantly different, this could indicate that there is a problem with their technique or the equipment they are using, and they should not be allowed to conduct the analysis of the athletes' urine samples.

c. Procedure for safe handling and disposal of the sample once the analysis is completed:

i) Label the sample container with the sample name, date, and analyst's name.

ii) Store the sample container in a refrigerator at 4°C until it is ready to be analyzed.

iii) Once the analysis is complete, dispose of the sample container according to the laboratory's waste management protocols. The laboratory should have protocols in place for the safe disposal of biological samples. These protocols may include autoclaving, chemical treatment, or incineration.

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