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March 29, 2024, 07:30:47 pm

Author Topic: Need help interpreting graph  (Read 3056 times)

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jinx_58

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Need help interpreting graph
« on: June 23, 2022, 06:40:32 pm »
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Howdy all.

I hope you all are doing well.

I need help analysing and interpreting a graph -> liquid chromatogram. For context, I am doing my IA3 and this is my RQ: Does understanding the metabolites of lidocaine show the effectiveness of its anaesthetic properties?

The graph is attached separately to the whole report. I understand that the graph shows a blank plasma sample that was spiked by lidocaine and MEGX (a metabolite), but how does it help me to answer my research question?

Any help will be appreciated as the draft is due at 11:59pm tomorrow.

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Billuminati

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Re: Need help interpreting graph
« Reply #1 on: June 23, 2022, 08:54:51 pm »
+2
Howdy all.

I hope you all are doing well.

I need help analysing and interpreting a graph -> liquid chromatogram. For context, I am doing my IA3 and this is my RQ: Does understanding the metabolites of lidocaine show the effectiveness of its anaesthetic properties?

The graph is attached separately to the whole report. I understand that the graph shows a blank plasma sample that was spiked by lidocaine and MEGX (a metabolite), but how does it help me to answer my research question?

Any help will be appreciated as the draft is due at 11:59pm tomorrow.

 -jinx_58

They just took this particular chromatogram to confirm that there are no peaks with the same retention times as the species of interest. This might be caused by some other chemical present in the plasma and may cause an overestimation in concentration of your desired species. Since there aren’t any such interfering peaks, we conclude that the analytical technique is selective (formal definition: relatively free from interference, any changes in signal is purely due to changes in analyse concentration)

This journal is an analytical chem journal so the article is mainly demonstrating that this analytical technique is suitable for analysis of your drug, so it doesn’t have much to do with your research question. What you could say is that the effectiveness of the drug may vary based on any physical conditions the patient has. BTW, usually the longer it takes for the anaesthetic to breakdown or diffuse away, the more effective it is. Breakdown is signalled by high metabolite concentrations
« Last Edit: June 23, 2022, 09:08:19 pm by Billuminati »
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jinx_58

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Re: Need help interpreting graph
« Reply #2 on: June 23, 2022, 09:40:32 pm »
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Thank you!

I have some questions:
BTW, usually the longer it takes for the anaesthetic to breakdown or diffuse away, the more effective it is. Breakdown is signalled by high metabolite concentrations

Is there a source for this, so I can explain it in my rationale?

Also, what is considered a "high" metabolite concentration?

If I put my analysis in here, can you guide me on what to fix?

Thank you,
 - jinx_58

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Re: Need help interpreting graph
« Reply #3 on: June 23, 2022, 10:12:58 pm »
+3
Thank you!

I have some questions:
Is there a source for this, so I can explain it in my rationale?

Also, what is considered a "high" metabolite concentration?

If I put my analysis in here, can you guide me on what to fix?

Thank you,
 - jinx_58

“Is there a source”: Do some research on pharmacokinetics and pharmacodynamics, I learned it from my biochem/molecular bio units. The example our prof used is thiopentone which isn’t very effective as it diffuses too quickly and doesn’t concentrate at the site of injection

“What is “high””: It’s relative to other drugs eg if you inject equal quantity of drugs X and Y and after 5 minutes there are 5 concentration units of drug X metabolite and 10 concentration units of drug Y metabolite, then drug Y probably has shorter effect times than drug X since it diffused or was broken down

I can look at it if your school allows it, but the caveat is that I didn’t do QCE and am not familiar with your marking rubric
VCE 2016-2018

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jinx_58

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Re: Need help interpreting graph
« Reply #4 on: June 24, 2022, 07:10:42 pm »
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I know this isn't much, but this is my analysis for this graph so far:
•Graph B, a liquid chromatogram spiked with lidocaine and MEGX
•Blood plasma samples spiked with lidocaine and monoethylglycinexylidide (MEGX)
•MEGX has greater peak, hence greater amount of MEGX formed in blood, showing that lidocaine metabolises in blood

I don't what else to add lol. I didn't fully understand the report either so that doesn't help. Should I look for another source to analyse?

Attached is the QCAA analysis and interpretation marking rubric.

Also, when in reports, it says n = x, x being a positive integer, what does n stand for/mean?

Thank you,
 -jinx_58
« Last Edit: June 24, 2022, 07:35:27 pm by jinx_58 »
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Re: Need help interpreting graph
« Reply #5 on: June 24, 2022, 07:53:22 pm »
+3
I know this isn't much, but this is my analysis for this graph so far:
•Graph B, a liquid chromatogram spiked with lidocaine and MEGX
•Blood plasma samples spiked with lidocaine and monoethylglycinexylidide (MEGX)
•MEGX has greater peak, hence greater amount of MEGX formed in blood, showing that lidocaine metabolises in blood

I don't what else to add lol. I didn't fully understand the report either so that doesn't help. Should I look for another source to analyse?

Attached is the QCAA analysis and interpretation marking rubric.

Also, when in reports, it says n = x, x being a positive integer, what does n stand for/mean?

Thank you,
 -jinx_58

This article itself isn’t very suitable for your investigation since it’s on analytical chem. Analytical chem is more to do with identifying + quantifying chemicals and instrumentation, it doesn’t tell you much about the chemicals themselves. You need to be discussing the chemical themselves according to your criteria sheet. However, I’d think the study they continue to allude to throughout this article might be good since you can research on the steps of pharmacokinetics and how drugs are chemically modified by the body, with regards to the structure and the implications on its water solubility (important for excretion).

BTW “n =“ refers to sample size in scientific literature
« Last Edit: June 24, 2022, 07:56:23 pm by Billuminati »
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jinx_58

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Re: Need help interpreting graph
« Reply #6 on: June 24, 2022, 08:07:09 pm »
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Thank you for the advice.

Quick question: does a greater peak indicate greater concentration of a particular chemical?

- jinx_58
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Re: Need help interpreting graph
« Reply #7 on: June 24, 2022, 08:13:33 pm »
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Thank you for the advice.

Quick question: does a greater peak indicate greater concentration of a particular chemical?

- jinx_58

Yes, peaks with larger peak areas show larger concentrations. A lot of people think it’s peak height which isn’t correct
VCE 2016-2018

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jinx_58

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Re: Need help interpreting graph
« Reply #8 on: June 24, 2022, 08:21:22 pm »
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Okay, so with graph above, would you say that the MEGX has a greater peak area than the lidocaine peak?

Thank you,
- jinx_58
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Re: Need help interpreting graph
« Reply #9 on: June 24, 2022, 09:04:02 pm »
+2
Okay, so with graph above, would you say that the MEGX has a greater peak area than the lidocaine peak?

Thank you,
- jinx_58

It doesn’t matter as it’s not a chromatogram for a freshly prepared plasma sample. They added both chemicals (of known concentration) to stored plasma artificially so you can’t really say lidocaine has metabolised.
« Last Edit: June 24, 2022, 09:07:04 pm by Billuminati »
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jinx_58

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Re: Need help interpreting graph
« Reply #10 on: June 24, 2022, 09:27:05 pm »
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Oh lol. Just did a full on analysis and evaluation for that source. Thank you for telling me early before a submitted absolute rubbish.

If I chuck up my analysis of my other graphs on here, could you please give me feedback?

Thank you so much for your guidance,
 -jinx_58
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Re: Need help interpreting graph
« Reply #11 on: June 24, 2022, 10:21:51 pm »
+1
Oh lol. Just did a full on analysis and evaluation for that source. Thank you for telling me early before a submitted absolute rubbish.

If I chuck up my analysis of my other graphs on here, could you please give me feedback?

Thank you so much for your guidance,
 -jinx_58

If it’s allowed then yes
VCE 2016-2018

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jinx_58

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Re: Need help interpreting graph
« Reply #12 on: June 24, 2022, 10:48:43 pm »
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I'm pretty sure its allowed.

Thanks in advance,
 -jinx_58

Evidence 1:
The above graphs show the individual predicted concentrations of the metabolites of lidocaine against the observed concentration in nanomolars (nM). The observed concentrations were retrieved from 50 µL blood plasma samples of a rat, in solutions consisting of 0.3% lidocaine. A trend observed was, there are no observed concentrations of GX between 0nM and 1nM, as it is a derivative of MEGX. In the graph showing the concentrations of MEGX (monoethylglycinexylidide), the smallest concentration is around 0.07nM, whereas the largest concentration is around 6000nM. In the graph showing the concentrations of GX (glycylxylidide), the smallest concentration is approximately 0.085nM and the largest concentration is around 4000nM. This shows that MEGX has a greater concentration in the blood than GX. As GX was not used up fully, it can be inferred that GX was used as a blocker, in which it blocks signals from the pain receptor signals to the brain. MEGX fully metabolises lidocaine in the blood as it has a greater concentration. The effectiveness of an anaesthetic is determined by the length of time it takes for the drug to breakdown or diffuse away. This breakdown is signalled by high metabolite concentration. As the concentration of MEGX was relatively high, lidocaine can be considered an effective local anaesthetic. Therefore, understanding the metabolites of lidocaine show the effectiveness of its anaesthetic properties.

Evidence 2:
Evidence 3 depicts a table showing the concentrations of lidocaine and its metabolite, MEGX in the blood plasma of cows (bovine), horses, dogs, pigs (swine), sheep and birds (avian). The serum of 10 cows were analysed, and there was generally a larger concentration of MEGX – 108 ± 9 ng/mL - than lidocaine, which was 97 ± 5 ng/mL. Similarly, from serum of the 5 horses that were analysed, the concentration of MEGX was 105 ± 11 ng/mL, larger than lidocaine, 99 ± 2 ng/mL. Furthermore, from the dog serum analyzation of 5 dogs, the concentration of lidocaine was comparatively significantly lower, 100 ± 1 ng/mL, than MEGX, at 113 ± 3 ng/mL. The swine serum had similar concentrations of lidocaine and MEGX concentrations of 100 ± 1 ng/mL and 100 ± 7 ng/mL, respectively. The sheep serum consisted of 100 ± 2 ng/mL of lidocaine and 104 ± 9 ng/mL of MEGX. Moreover, from the blood plasma samples of 5 avian, there was MEGX concentration of 102 ± 6 ng/mL and a lidocaine concentration of 98 ± 2 ng/mL. From the serums and blood plasma samples of these 6 animals, it can be concluded that MEGX had a greater concentration than lidocaine. This showed that lidocaine can be considered an effective local anaesthetic. Hence, the effectiveness of lidocaine’s anaesthetic properties can be shown through understanding the metabolites of lidocaine.
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Re: Need help interpreting graph
« Reply #13 on: June 25, 2022, 12:12:45 am »
+2
I'm pretty sure its allowed.

Thanks in advance,
 -jinx_58

Evidence 1:
The above graphs show the individual predicted concentrations of the metabolites of lidocaine against the observed concentration in nanomolars (nM). The observed concentrations were retrieved from 50 µL blood plasma samples of a rat, in solutions consisting of 0.3% lidocaine. A trend observed was, there are no observed concentrations of GX between 0nM and 1nM, as it is a derivative of MEGX. In the graph showing the concentrations of MEGX (monoethylglycinexylidide), the smallest concentration is around 0.07nM, whereas the largest concentration is around 6000nM. In the graph showing the concentrations of GX (glycylxylidide), the smallest concentration is approximately 0.085nM and the largest concentration is around 4000nM. This shows that MEGX has a greater concentration in the blood than GX. As GX was not used up fully, it can be inferred that GX was used as a blocker, in which it blocks signals from the pain receptor signals to the brain. MEGX fully metabolises lidocaine in the blood as it has a greater concentration. The effectiveness of an anaesthetic is determined by the length of time it takes for the drug to breakdown or diffuse away. This breakdown is signalled by high metabolite concentration. As the concentration of MEGX was relatively high, lidocaine can be considered an effective local anaesthetic. Therefore, understanding the metabolites of lidocaine show the effectiveness of its anaesthetic properties.

Evidence 2:
Evidence 3 depicts a table showing the concentrations of lidocaine and its metabolite, MEGX in the blood plasma of cows (bovine), horses, dogs, pigs (swine), sheep and birds (avian). The serum of 10 cows were analysed, and there was generally a larger concentration of MEGX – 108 ± 9 ng/mL - than lidocaine, which was 97 ± 5 ng/mL. Similarly, from serum of the 5 horses that were analysed, the concentration of MEGX was 105 ± 11 ng/mL, larger than lidocaine, 99 ± 2 ng/mL. Furthermore, from the dog serum analyzation of 5 dogs, the concentration of lidocaine was comparatively significantly lower, 100 ± 1 ng/mL, than MEGX, at 113 ± 3 ng/mL. The swine serum had similar concentrations of lidocaine and MEGX concentrations of 100 ± 1 ng/mL and 100 ± 7 ng/mL, respectively. The sheep serum consisted of 100 ± 2 ng/mL of lidocaine and 104 ± 9 ng/mL of MEGX. Moreover, from the blood plasma samples of 5 avian, there was MEGX concentration of 102 ± 6 ng/mL and a lidocaine concentration of 98 ± 2 ng/mL. From the serums and blood plasma samples of these 6 animals, it can be concluded that MEGX had a greater concentration than lidocaine. This showed that lidocaine can be considered an effective local anaesthetic. Hence, the effectiveness of lidocaine’s anaesthetic properties can be shown through understanding the metabolites of lidocaine.

I only asked if it’s allowed since some schools see this as receiving unauthorised assistance

You contradicted yourself there, high [metabolite] usually means short effective time span which makes an active drug less effective. For prodrugs (drugs activated by metabolism) the opposite is true. From the way you described the lidocaine —> MEGX —> GX pathway it would seem that lidocaine is a prodrug, not an active drug.

I’m not sure if this is expected by the task, but you seem to be summarising the data a little too much without providing a lot of interpretation

Your criteria sheet asks for properties and structure of the organic molecules involved, you should talk about the structures more since right now there’s very little discussion about them eg you mentioned blockers, how does it block the target? What functional groups do the actual blocking?
« Last Edit: June 25, 2022, 12:17:15 am by Billuminati »
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2018: Methods [37], French [38], Chem [40], English [44]

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2019-2021: Bachelor of Biomedical Science at Monash (Scholars), minoring in Chemistry

GAMSAT September 2021: 65/67/86, 76 overall (98th percentile)

2022: Chilling

2023+: Transfer to teaching degree

jinx_58

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Re: Need help interpreting graph
« Reply #14 on: July 12, 2022, 07:00:33 pm »
+1
Thank you Billuminati for the feedback. It is very greatly appreciated.

I checked with my teacher, and he said I don't need to talk about the properties and structure for my investigation, as it is too complex to go into.

With regards to the high metabolite stuff, can I say that there is a high concentration of X metabolite? Or am I contradicting myself?

Thank you,
 -jinx_58
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