How do different concentrations (0.5%, 1%, 1.5%, 2%, and 2.5%) of acetone solution affect the membrane integrity of Beta vulgaris L. over 15 minutes as measured by the pigment absorbed by beetroot cells detected through a vernier colourimeter at wavelength 470 nm?

How do different concentrations (\(0.5\%, 1\%, 1.5\%, 2\%\), and \(2.5\%\)) of acetone solution affect the membrane integrity of Beta vulgaris \(L\). over 15 minutes as measured by the pigment absorbed by beetroot cells detected through a vernier colourimeter at wavelength 470 nm?

In the biomedical world, drugs are evolving to be more effective. Various methods have been explored to enhance the absorption and permeation of biologically active components found in pills. These methods make the drugs more soluble and improve the absorption and spread of drugs. Lipid-based drug delivery systems, known as 'pharmacosomes', have been proven to do this. They form when drugs with active hydrogen combine with phospholipids, creating amphiphilic structures. The collaboration between drugs and phospholipids serves to improve the properties of the medication, ultimately enhancing its bioavailability and optimizing drug delivery (Semalty). This efficacy arises from certain kinds of hydrogen atoms (e.g. \(-\mathrm{COOH},-\mathrm{OH},-\mathrm{NH} 2\)) found in solvents (Semalty). This is because they have the unique ability to facilitate movement across various biological barriers such as cell membranes, tissues, and cell walls in organisms (Supraja). One of the drugs that contains these hydrogen atoms is acetone.

Acetone is an organic solvent, they can alter the lipid structure of membranes to make them more permeable. Acetone molecules act as disruptors, causing a reorienting of the phospholipid bilayer by pushing apart the lipid heads (Posokhov). All this contributes to an increase in membrane fluidity. This heightened fluidity implies a reduction in overall stability since the individual lipid molecules can move more easily. As membrane fluidity increases, so does membrane permeability. This is because a more fluid membrane allows molecules to move more freely through the lipid bilayer. Furthermore, acetone can dissolve lipids due to its chemical properties and interactions with lipid molecules. As lipids are naturally hydrophobic, acetone intervenes by inserting itself between their hydrocarbon chains, disrupting lipid-lipid interactions (Posokhov). This interference reduces lipid cohesion, ultimately decreasing their membrane integrity.

In the pharmaceutical industry, acetone is used as a solvent in both pharmaceutical and commercial preparations, especially in the production of pills to ensure the effectiveness of the drug (Acetone: What Is It and Where Is It Used). They often use acetone in small concentrations in preparations of certain drugs as manufacturers prioritise minimising the use of solvents in drug production for safety reasons. Solvents offer no therapeutic value, and their potential toxicity can hasten product decomposition. Regulatory guidelines establish maximum permissible levels of residual solvents (RS) in pharmaceutical products (Grodowska). Therefore, Class 3 solvents, such as acetone, have daily exposure allowances of 50 \(\mathrm{mg}(0.5\%)\) or less in the final product (Grodowska). Examples of medications using acetone as solvents include Allegra-D 24, Azulfidine EN-tabs, Dexmethylphenidate Hydrochloride, and Methylphenidate Hydrochloride. (Acetone: What Is It and Where Is It Used?). This is because solvents like acetone are used in the reaction step of chemical synthesis and their primary purpose is to enhance the solubility of the reactants, thereby promoting reactivity and allowing for control over reaction rates and product quality. When solutes are introduced into a solvent, it disrupts the cohesive forces that bind crystalline and liquid solutes together. This disruption effectively increases the reactivity of the solutes. (Grodowska). By choosing and manipulating the properties of these solvents, the rate of the reaction can be controlled. Knowing how different concentrations of acetone affect our membrane permeability can help me understand the optimal acetone concentration used in drugs that ensures effectiveness but also complies with the limits on residual solvents to prevent potential adverse effects on our cells.

To measure how different concentrations of acetone affect the membrane, beetroot cells will be used. Beetroot cells are a type of eukaryotic cells and consist of a double layer made up of phospholipid and protein molecules. The phospholipid bilayer is a thin and semi-permeable barrier separating the cell's internal environment from its external surroundings. It allows small, nonpolar, hydrophobic molecules to pass while resisting large, hydrophilic, polar molecules. The semi-permeable property can be attributed to its amphipathic properties, with the hydrophobic tails on the inside and the hydrophilic heads on the outside. In beetroots, there's an abundant red pigment known as betacyanin, located within the large central vacuoles of beet cells. These vacuoles are enclosed by a membrane, also known as the tonoplast. (Vodopich). As long as the cell and its membranes remain intact, the betacyanin stays within the vacuoles. However, if these membranes undergo stress or damage, betacyanin can seep through both the tonoplast and phospholipid bilayer, causing its surroundings to turn red. (Vodopich). As a result, to test the effect of different concentrations of acetone, we will test the amount of pigment released by the beetroot cells when it is exposed to acetone. When it is exposed to acetone, the membranes should start to become more permeable and pigment is released, indicating the membrane integrity of beetroot has compromised.

• Vernier Colorimeter
• Distilled water
• Stopwatch or Timer
• 15 cm Ruler (\(\pm 0.05 \mathrm{~cm}\))
• \(2 x\) glass cuvettes
• Test tube rack
• 6x rubber stoppers
• Tweezers
• Gloves
• 1 x 700 mL glass bottle for disposal
• 1.1 cm cork borer \((\pm 0.05 \mathrm{~cm})\)
• 6x test tubes
• 7 Malaysian Beetroot (Beta vulgaris L.)
• 6x 100mL beaker (\(\pm 5 mL\))
• Goggles
• 25 mL graduated cylinder \((\pm 0.025 mL)\)
• \(110 \mathrm{~mL} 0.5\%\) acetone
• \(110 \mathrm{~mL} 1.0\%\) acetone
• Apron
• 1x glass pipette
• Pencil
• Plastic cutting board
• Marker
• Knife
• Fumehood
• \(110 \mathrm{~mL} 1.5\%\) acetone
• \(110 \mathrm{~mL} 2.0\%\) acetone
• \(110 \mathrm{~mL} 2.5\%\) acetone
• Paper towels

To determine which wavelength to use, I placed a cuvette containing the solution after the beetroot had been submerged in distilled water for 15 minutes and checked which of the wavelengths yielded the highest absorbance, which was 470 nm with 0.234. In addition to that, the results from the pre-trials shouldn't exceed the range of 0.05 to 1.0 because according to the user manual. Thus, the concentrations and duration had to be tested to make sure they fell within that range, the concentrations used for the pre-trials were \(0.5\%, 2\%, 5\%\), and \(10\%\) and the duration used was 15 minutes. During the first pre-trial, \(0.5\%\) acetone solution yielded the highest absorbance which was 0.325 and \(10\%\) yielded the second lower absorbance at \(0.154.0\%\) had the lowest absorbance which was expected. Due to this, another pre-trial was conducted, and the results were also abnormal, with \(0\%\) having 0.255 absorbance, \(0.5\%\) having 0.457 absorbance, \(2\%\) having 0.203 absorbance, and \(10\%\) having 0.217 absorbance. Due to the abnormal results, the experiment was transferred to a water-based solution, because there might be a chemical interaction with beetroot pigment and acetone. The beetroot was first submerged into the acetone solution for 15 minutes and then it was transferred into distilled water to let the pigment from the beetroot leak out. However, transferring the beetroot into 40 mL of distilled water didn't produce reliable results as it was outside the range of 0.05 to 1.0, it gave absorbance values of around 0.02 to 0.04. For the third pre-trial, I realised the solution had been drawn from different parts of the test tube, so an adjustment was made by switching to a longer pipette and shaking the test tube before drawing the samples. This modification was prompted by the observed variation in pigment distribution within the solution, with a significant difference between the upper and lower portions of the test tube. Consequently, the results showed an increase in absorbance as the concentration increased, but the \(2\%\) and \(10\%\) yielded similar results, therefore, I decided to only go up to \(2.5\%\).

INDEPENDENT: Concentration of acetone (\(0\%, 0.5\%, 1\%, 1.5\%, 2\%, 2.5\%\))

DEPENDENT: Membrane integrity of beetroot as measured by the pigment absorbed

Exposure to and inhalation of acetone can lead to eye irritation, headaches, nausea, drowsiness, and dizziness (LabChem). Therefore, goggles should be worn to protect the eyes and contact lenses should be removed. Furthermore, using a ventilator is important to further prevent exposure to acetone vapour. When not in use, acetone should be stored in a tightly closed container and in a fume hood. In addition, wearing protective clothing like gloves & aprons and washing your hands can help minimise exposure to acetone.

A knife and cork borer will be used to cut slices of beetroot. Because of the sharp edges, the blade should be pointed away from your body and a cutting board should be used. When using the cork borer, place the beetroot on a flat surface and insert the cork borer vertically, don't hold the beetroot in your hand.

No animals or humans are required for my experiment, hence there are no ethical issues.

Acetone is not toxic to aquatic organisms and it is not expected to bioaccumulate in food chains. Furthermore, it is biodegradable in the soil under both anaerobic and aerobic conditions and it is biodegradable in water (NHS Greater Glasgow and Clyde). However, it is important to prevent material from entering drains and watercourses. When hazardous waste is mixed with other types of hazardous materials, there may be a risk of pollution or cause problems for the management of waste (NHS Greater Glasgow and Clyde). Therefore, transferring acetone into suitable containers for disposal and soaking used beetroots in water before disposal can reduce the potential harm acetone can cause to the environment.

1. Turn on the Fume Hood, and make sure it is working by holding a tissue paper in there. If the tissue paper is moving, then it works. This is to make sure there is proper ventilation (Safety)
2. Get 7 Malaysian beetroots from the same store (CV: Type of Beetroot)
3. Rinse the beetroot under tap water for 30 seconds and remove the ends of the beetroots (CV: Rinse)
4. Using a 1.1 mm cork borer and cutting board, cut out 6 slices of beetroot. Make sure the beetroot is on a flat surface when inserting the cork borer, the cork borer should be inserted vertically to prevent injuries. (CV: Beetroot Size) (Safety)
5. Using the back end of a pencil to remove the beetroot from the cork borer. Remove the skin at the ends and cut them using a knife so they are 3 cm in length (CV: Beetroot Size)
6. Make sure you have 6 pieces of beetroot cylinders (one for each concentration and one for controlled)
7. Rinse the beetroot for 5 seconds under distilled water to get the excess pigments out (CV: Rinse)
8. Transfer the beetroot to a paper towel
9. Gently dap the beetroots until it's dry to remove excess pigments. Then using a tweezer transfer them into the test tubes. One beetroot in one test tube. Make sure you have one test tube just for distilled water to be used as a control.
10. Put on your gloves, goggles, and apron (Safety)
11. Move the test tubes with beetroot to the fume hood where your different acetone concentrations (\(0.5\%, 1\%, 1.5\%\), \(2\%\), and \(2.5\%\)), distilled water, vernier colourimeter, 2 glass cuvettes, 6100 mL beakers, and 25 mL graduated cylinder should be.
12. Do one trial for all concentrations at once instead of doing all the trials for a certain concentration, so the order will be \(0\%, 0.5\%, 1\%, 1.5\%, 2\%\), and \(2.5\%\).
13. A 25 mL graduated cylinder was used to measure 15 mL of the acetone solution, which was poured into each test tube. For the control, measure 15 mL of distilled water using the 25 mL graduated cylinder. (Do this one at a time) (CV: Volume)
14. Once you poured 15 mL of acetone or distilled water into the test tube, start your timer immediately.
15. Put a stopper on top of the test tube.
16. Before pouring 15 mL of acetone into the next test tube, use a stopwatch to measure a 3-minute time lag between each test. Repeat steps 13 to 15 for each concentration. (CV: Duration)
17. As the timer is running for the beetroot, pour distilled water into an empty glass cuvette. Once it reaches the top, put on the cap and gently dap the outside so it is dry.
18. Hold onto the ribbed sides and put the cuvette with distilled water inside the vernier colourimeter with the clear sides placed where the arrows are. (CV: Cuvette)
19. Press the button with 470 nm wavelength and click the 'cal' button. The red light should stop before you put anything else inside the calorimeter. The reading should be set to 0.000.
20. Once the timer runs out for the first one, shake the test tube twice. Make sure the beetroot inside touches both ends of the test tube when you shake it. (CV: Pipette)
21. Open the other glass cuvette that isn't inside the calorimeter.
22. Use a long pipette and place it inside the test tube. Make sure the pipette reaches the end of the test tube which should be right next to the beetroot. (CV: Pipette)
23. Squeeze the pipette to get the liquid
24. Release the pigment collected by the pipette into the glass cuvette
25. Repeat steps 22 to 24 until the glass cuvette is filled.
26. Seal the glass cuvette with a cap
27. Remove the cuvette with distilled water from the colourimeter and replace it with the glass cuvette with the solution. Make sure the cuvette is dry by dapping it with a paper towel before you put it in
28. Hold on to the ribbed sides and make sure the clear sides go in where the arrows are. Close the slider.
29. Wait 1 minute for the reading to stabilise and record the absorbance to its corresponding acetone concentration on Google Sheets or on a piece of paper.
30. Remove the cuvette in the calorimeter and put in the 'blank'
31. Press the 'cal' button to calibrate it again and make sure the absorbance is at 0.000
32. Dispose of the acetone from the test tube and glass cuvette into the 700 mL empty glass bottle (Environment)
33. Use a tweezer to take the beetroot out and place it in a clean and empty 100 mL beaker. Each concentration should have its own beaker for the beetroot, so the beetroot from the \(0\%\) acetone concentration would go into the beaker for the beetroot that was soaked in the \(0\%\) acetone concentration.
34. Rinse the glass cuvette that had the acetone concentration inside with distilled water before testing the absorbance for the other concentrations.
35. Repeat steps 20 to 34 for each concentration (\(0\%, 0.5\%, 1\%, 1.5\%, 2\%, 2.5\%\))
36. When you have repeated this at least 5 times for each concentration and taken pictures, soak the beetroot in tap water for at least 5 minutes before you dispose of it in the trash can. (Environment)
37. Repeat steps 12 to 36 for each trial, you should have 7 trials in total for each concentration.

Each time the beetroot was submerged in the solution, it became visibly paler and the pigment of beetroot in the solution was quite apparent. The change in colour was more pronounced in the test tubes with higher concentrations of the solution. In addition to the colour change, the physical appearance of the beetroot also transformed, the beetroot began to look shrivelled and wrinkly, particularly in the test tubes with higher solution concentrations. Leaving them out to dry made the beetroot paler and their rough and shrivelled texture was more pronounced. Furthermore, in test tubes with higher concentrations of the solution, the pigment was more intense. However, with certain trials like 4 and 5, the pigment released from the beetroot was a lot higher than in other trials. Consequently, the absorbance of pigment in these trials was notably higher than the absorbance at the same concentration observed in other trials.

When using an online calculator for outliers, a single outlier is detected at the value of 0.633, which exceeds the upper quartile value of 0.621 (Outlier Calculator). The value 0.633 was the absorbance at \(2.5\%\) for trial 5. Trial 5 had absorbance values that were significantly above the absorbance values at other concentrations. For example, at \(0.5\%\) concentration, it had an absorbance value of 0.471 compared to the values from other trials which ranged from 0.116 to 0.272. This might be due to the different levels of pigment in the different beetroots.

With reference to Figure 1, there isn't a distinct difference between the mean pigment absorbed across the different concentrations. For \(0\%, 68\%\) of the data ranges from 0.13 and 0.234, there is less variability in pigment absorbed for this concentration compared to others, and therefore, it is more reliable. The data is more concentrated around the mean. For \(0.5\%, 68\%\) of the data ranges from 0.142 to 0.37. For \(1\%, 68\%\) of the data ranges from 0.138 to 0.422. For \(1.5\%, 68\%\) of the data ranges from 0.17 to 0.48. For \(2\%, 68\%\) of the data ranges from 0.218 to 0.546. For \(2.5\%, 68\%\) of the data ranges from 0.262 to 0.564. Whereas for these concentrations, the standard deviation was all above 0.1, so there is more variability in the pigment absorbed and therefore less reliable. The error bars significantly overlap. Furthermore, the range between the raw data was above 0.1 with the highest at 0.441 and the lowest at 0.132. This suggests there isn't a statistically significant relationship between the pigment absorbed by the different concentrations of acetone, so the different concentrations of acetone doesn't have a significant effect on the membrane integrity of beetroots.

When plotting all the data points into a histogram, it showed a right-skewed distribution instead of a symmetrical, bell-shaped curve which tells us that most of the values cluster towards the lower end (Histogram Maker). Furthermore, the mean exceeds the median. This provides clear evidence that the dataset does not fit a normal distribution. As a result, using a parametric statistical test such as the \(t\)-test or ANOVA can be problematic as these tests rely on the assumption that the data is normally distributed. For this reason, a Spearman's rank correlation coefficient test will be used to measure the strength of the relationship between the concentration of acetone and absorbance.

My null and alternative hypotheses will be stated below.
\(H_{0}=\) There isn't a monotonic association between the concentration of acetone and the pigment absorbed by beetroot cells
\(H_{1}=\) There is a monotonic association between the concentration of acetone and the pigment absorbed by beetroot cells

With reference to Figure 1, the linear regression demonstrated a positive monotonic relationship between the concentration of acetone and the pigment absorbed by the beetroot cells. From Figure 1, the slope was 0.0902 and there is a correlation of 0.992. This indicates that for each unit increase in the concentration of acetone, the average pigment absorbed by beetroot cells is expected to increase by approximately 0.0902 units. The y-intercept at 0.194 indicates that at \(0\%\) acetone solution, the absorbance of pigment by beetroot cells will be 0.194. Furthermore, the correlation coefficient, which measures the strength and direction of the linear relationship between the two variables, was found to be 0.992. This high correlation value suggests a strong positive linear relationship between the concentration of acetone and the average pigment absorption by beetroot cells and \(99.2\%\) of the data can be explained by the linear trendline. To confirm this, a Spearman Rank correlation coefficient will also be calculated, the calculations can be seen in table 4. The result of this was 0.554 and the degree of freedom was 33. The degrees of freedom are calculated by subtracting two from the number of raw data we have which will be 33. Because the Spearman Rank value of 0.554 is greater than the critical value of 0.291 (Pearson Education Limited) at a p-value of 0.05 determined by the degrees of freedom, the null hypothesis is rejected and the alternative hypothesis that there is a monotonic association between the concentration of acetone and the pigment absorbed by beetroot cells is accepted. This can also be seen on the graph, where the highest concentration (\(2.5\%\)) had the highest average absorbance (0.413) and the lowest concentration (\(0\%\)) had the lowest average absorbance (0.182). In addition to that, the pictures of the beetroot after it has been submerged in the acetone solution further support this hypothesis, with the beetroot at higher concentrations being visibly paler than the beetroots at lower concentrations. Therefore, we can conclude there is a moderate positive relationship between acetone concentration and pigment absorption.

As a result of my statistical test, linear regression, and qualitative data, it can be concluded that there is a strong correlation between increases in acetone concentration and the membrane permeability of the beetroot (Beta vulgaris L.). However, the overlap between the error bars in the graph suggests that the absorbance at different acetone concentrations may not be statistically significant. When error bars overlap, it implies that the means of absorbance for different concentrations are within this overlapping range, making it challenging to confidently assert that there is a substantial effect of acetone concentration on beetroot membrane permeability.

From my observations and data analysis, it can be concluded that different concentrations of acetone doesn't have a significant effect on the membrane integrity of the beetroot. Even though the strength of the relationship between the increasing concentrations and pigment absorbance, the difference between the means weren't statistically significant. The positive monotonic relationship between these two variables can be attributed to the solvent properties of acetone. When acetone molecules are present, they cause the lipid molecules in a membrane to spread out because the acetone molecules act as spacers, pushing the lipid heads apart in the upper part of the membrane (Posokhov). This spreading effect increases the average space for each lipid molecule in that region. Moreover, in the lower part of the membrane, the tails of the lipids also become less crowded or dense (Posokhov). This increases the membrane permeability as there is more space between the lipid molecules, allowing the pigment in the beetroot to leak out. Therefore, the error bars from my experiment could be due to procedural or methodological errors.

My results is not consistent with Gabriela Dyrdal's experiment where they investigated how exposure to different organic solvents (Methanol, Ethanol, Acetone, DMF, DMSO, Nujol) affects the growth of certain microorganisms (E. coli DH5a, B. subtilis, and S. cerevisiae D273) (Dyrdal). They tested different concentrations of organic solvents, \(0\%, 4.8\%\), \(9.1\%\), and \(20.0\%\). All of the tested microorganisms were found to be susceptible to the organic solvents they were treated with, exposure to these solvents hindered the growth and development of the microorganisms. This suggests that changes in cell membrane fluidity induced by these solvents play a role in their inhibitory effects on growth. Based on their qualitative and quantitative data for acetone, as they increased the acetone concentration, the Saccharomyces cerevisiae and Escherichia coli decreased in size and at \(20\%\), the cell disintegrated completely (Dyrdal). As they increased concentrations of acetone, they concluded that "more dramatic changes resulting in breaking of the membrane structure were found when the microbes were left to grow in the presence of acetone" (Dyrdal). Dyrdal's research demonstrated that increasing concentrations of acetone resulted in more pronounced inhibitory effects on microbial growth. These changes in cell structure are indicative of alterations in membrane properties. Thus, Dyrdal's results indirectly support the idea that changes induced by different concentrations of acetone can affect membrane integrity through the decease in size and the disintegration of cells under certain conditions. However, her experiment uses higher concentrations of acetone compared to me which might have contributed to more extreme results.

The high degree of variability, as indicated by the large standard deviation of absorbance for each acetone concentration is a limitation in my conclusion about the relationship between increasing acetone concentration and pigment absorbed. This suggests that the results have a high degree of variability. This large standard deviation could be due to using different beetroots for each trial and using different parts of the beetroot. Even though it was the same type and bought from the same store at the same time, the pigments in the beetroot can vary. Beetroot samples with low initial pigment content may be less sensitive to changes in acetone concentration, potentially leading to smaller differences in pigment absorption between treatment groups. Beetroot samples with high initial pigment will naturally absorb more pigment when placed in the acetone solution. Furthermore, different parts of the beetroot had visible differences in colour intensity, meaning they also started off with different pigment concentrations. To improve on this, using beetroot pieces of similar mass can help decrease the variability. Measuring the mass of the beetroot before putting it in the solution can help indirectly account for the initial pigment content. Beetroot samples with higher initial pigment content are likely to have a greater mass. Furthermore, using the same part of the beetroot (eg. centre, edge) across each trial and concentration can further decrease the variability in pigment content, but this can lead to an increase in waste.

The current method does not include a water bath for the acetone solution, the temperature was controlled by experimenting in the same room. However, this isn't accurate as the room temperature can change due to various factors such as changes in heating or cooling systems, ventilation, and human activities. Since two different bottles of acetone solution were used for each concentration, it was difficult to ensure they were at the same temperature. We were only able to do it qualitatively by touching the bottle. Temperature affects the rate at which molecules move and interact. Higher temperatures generally result in increased molecular motion and kinetic energy. Higher temperatures can lead to faster penetration of acetone into the cell membranes. This can potentially increase the rate at which pigment leaks out of the beetroot cells. At lower temperatures, the phospholipids within the membrane have lower energy levels, causing them to move less and maintain a tighter, more rigid structure in the membrane. However, at higher temperatures, these phospholipids gain more energy, leading to increased movement and a looser packing arrangement within the membrane, which results in higher permeability. A water bath can be easily regulated using a thermometer to ensure that the temperature remains constant throughout the experiment. The water bath will be kept at 37 Celsius to replicate the conditions in the human body.

Acetone is volatile which can cause it to evaporate from solutions rapidly, making it challenging to maintain a solution of a definite concentration (LabChem). Even though a rubber stopper was used, the concentration of acetone might still vary and evaporation can still occur. Different concentrations may evaporate at different rates, leading to varying effects on the beetroot cells' membrane integrity over time. To minimise the impact of this, wrapping the test tube and rubber stopper with a cling wrap can help create a more effective seal, reducing the potential for acetone evaporation and providing a more stable environment for maintaining a consistent solution concentration throughout the experiment.

Another procedural error in this experiment was a change in colorimeter in the middle due to a technical issue. Given that the experiment aimed to assess the impact of varying acetone concentrations on membrane integrity by measuring the pigment absorbed using the colorimeter, any alteration in the instrumentation introduces another variable that can affect the absorbance value. When a different colorimeter is introduced, even if it's of the same model, there will be slight variations in calibration, sensitivity, or performance between instruments. These differences may lead to discrepancies in the way each colorimeter measures and records absorbance values. Therefore, maintaining the consistency and reliability of instrumentation throughout the experiment is crucial.

This experiment focuses solely on small concentrations of acetone, due to this the experiment fails to capture the full spectrum of potential membrane integrity changes in Beta vulgaris L. Small concentrations of acetone may not elicit significant changes in membrane integrity. The experiment, therefore, might underestimate the potential impact of acetone on beetroot. Furthermore, this experiment only used beetroot cells so they may have different responses to acetone compared to other cell types. Beetroot cells are plant cells and possess a rigid cell wall. This structural feature distinguishes them from animal cells and cells of other organisms lacking a cell wall. The cell wall acts as a barrier that provides structural support to the cell. It can impede or modify the penetration of external substances, such as acetone, into the cell. This could influence the rate at which acetone interacts with the cell membrane. Animal and human cells have different membrane compositions and structural characteristics that could result in different responses to acetone compared to beetroot cells. Therefore, we cannot generalise the findings of this experiment for animal and human cells. We also cannot guarantee it will follow the same trend for higher concentrations of acetone.

This investigation illustrates a positive trend between increasing acetone concentration and the integrity of beetroot cells. However, the use of acetone in the biomedical field primarily revolves around studying its effects on human cells and tissues. Therefore, to apply the findings from this experiment to the biomedical context, researchers might need to modify the experiment to test the impact of increasing acetone concentration on animal cells rather than plant cells. This modification would involve using synthetic or natural animal cells as the test subjects instead of beetroot cells. Animal cells are often more relevant for biomedical research because they closely resemble human cells in terms of structure and function. By conducting experiments on animal cells, researchers can gain insights into how acetone affects cellular integrity in a context that is more directly related to human biology. Furthermore, different organic solvents such as ethanol, DMSO, and N N-Dimethylacetamide could be tested as these are also commonly used in the biomedical industry (Koç). These solvents possess distinct properties, including differences in polarity and chemical reactivity. This can provide a deeper understanding of which types of solvents may be more damaging or less damaging to cell membranes. This knowledge can help inform the selection of solvents for specific applications and enhance drug delivery systems.

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Figure 1
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Table 1
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Table 2
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Table 3
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Table 4
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