Uji In Vivo (Bioavaibilitas Obat)

Summary

The transcript describes an experimental procedure conducted on rabbits, involving the administration of sodium citrate and paracetamol, blood sampling, and the use of various laboratory equipment like centrifuge tubes, syringes, and a UV-Vis spectrophotometer. The experiment measures the effects of these substances over time, with blood samples taken at regular intervals, mixed with trichloroacetic acid, and analyzed for absorbance. The goal seems to be assessing the substance's impact, likely on rabbit blood, through detailed chemical and spectral analysis.

Takeaways

• 😀 The experiment involves the use of various tools and materials such as centrifuge tubes, syringes, and a vortex mixer.
• 😀 Sodium citrate (0.125 mL) is used in the experiment for blood collection from the rabbit.
• 😀 Blood is collected through the marginal vein of the rabbit's ear, with the process repeated at multiple time intervals: 5, 10, 20, 30, 45, 60, and 75 minutes.
• 😀 Trichloroacetic acid (0.625 mL) is added to the blood samples during the experiment.
• 😀 Paracetamol is injected (0.14 mL) into the rabbit's ear at the 5-minute mark.
• 😀 After each blood collection, samples are vortexed for 5 minutes to ensure homogeneity.
• 😀 Paracetamol tablets are administered orally to the rabbit using tweezers.
• 😀 A centrifuge is used to process the blood samples at a speed of 2000 rpm for 10 minutes.
• 😀 Phosphate buffer medium is used to rinse the measuring flask and adjust the volume of the sample.
• 😀 The absorbance of the sample is measured using a UV-Vis spectrophotometer to analyze the results of the experiment.

Q & A

What are the main materials used in the experiment?

The main materials used in the experiment include a centrifuge tube, a dropper pipette, a vortex machine, a syringe, paracetamol tablets, sodium citrate, trichloroacetic acid, and a UV-Vis spectrophotometer.

What is the purpose of using sodium citrate in the experiment?

Sodium citrate is used to prevent blood clotting. It is injected into the rabbit's ear vein before blood collection to ensure the blood remains in a liquid state during analysis.

How is the blood collected from the rabbit in this experiment?

Blood is collected from the marginal vein in the rabbit's ear. A syringe pre-filled with sodium citrate is used to withdraw 0.5 milliliters of blood at various time points (5, 10, 20, 30, 45, 60, and 75 minutes).

What is the role of trichloroacetic acid in the procedure?

Trichloroacetic acid is used to precipitate proteins from the blood samples. It is added to the blood samples and then vortexed to ensure a homogeneous mixture.

Why is the blood sample vortexed during the experiment?

Vortexing is done to mix the blood and trichloroacetic acid thoroughly, ensuring the reaction is complete and the sample becomes homogeneous for further processing.

What is the significance of measuring absorbance using a UV-Vis spectrophotometer?

The absorbance is measured using a UV-Vis spectrophotometer to analyze the concentration of specific compounds in the supernatant after centrifugation. This helps in quantifying substances in the blood samples.

How is the sample prepared before measuring the absorbance?

After vortexing and centrifuging, the supernatant is collected and transferred to a measuring flask, which is then filled with phosphate buffer medium up to the mark line. This solution is then homogenized before absorbance measurement.

At what speed is the centrifuge set during the experiment?

The centrifuge is set to 2000 rpm for 10 minutes to separate the components of the blood samples after they have been treated with trichloroacetic acid.

What role does paracetamol play in this experiment?

Paracetamol is injected into the rabbit in this experiment, possibly to observe its effects on the blood or to test the pharmacokinetics and metabolism of the compound in the rabbit's system.

What happens after the centrifugation process is completed?

After centrifugation, the supernatant is carefully removed and placed into a measuring flask, where it is diluted with phosphate buffer medium. This preparation allows for the subsequent absorbance measurement.