Liquid chromatography has been used to track the concentration of remdesivir (a broad-spectrum antiviral drug, structure shown at right) in COVID patients undergoing experimental treatments. Intensity The authors provide the following details regarding standard solutions preparation: HN CN HO OH NH2 Remdesivir (RDV) stock solution (5000 µg/mL) was prepared by dissolving RDV drug powder using the mixture of DMSO: MeOH (30:70 v/v). The RDV working standard solutions for calibration and quality controls were prepared using methanol in concentrations of 100, 10, 1, 0.1, 0.01 µg/mL. 1, 2.5, 5, 7.5, 10, 25, 50, 75, 100, 250, 500, 1000, and 5000 ng/mL sample solutions were prepared freshly by spiking calibration standard solutions into the blank human plasma samples for method calibration. a) What type of calibration method is being described? Why do you think the authors chose this method as opposed to another? b) Based on the details provided in part a, describe an appropriate method blank for this analysis. 25000 20000- (b) Spiked (a) Unspiked The authors provide the following details regarding the chromatographic analysis of remdesivir: A C18 column (150 x 4.6 mm, 3 um stationary phase particles) was utilized for the separation of RDV. The mobile phase A used was 0.05% (v/v) formic acid in ultrapure water, and mobile phase B was 100% ACN with isocratic elution (A: B) 52:48 % with the total analysis time of 10 min. The column flow rate was set at 0.5 mL/min with 35 °C as the ideal column temperature, and the sample injection volume was 5 μL. 15000- 10000 5000- 0 RDV/RT 5.65 min (b) www (a) T T T 2 3 4 5 6 7 8 9 10 Retention time (minutes) c) What is one parameter you could change to decrease the retention time of RDV? Explain the theory that supports why this would work. d) What is one parameter you could change that would decrease the peak width of RDV? Explain the theory that supports why this would work.
Liquid chromatography has been used to track the concentration of remdesivir (a broad-spectrum antiviral drug, structure shown at right) in COVID patients undergoing experimental treatments. Intensity The authors provide the following details regarding standard solutions preparation: HN CN HO OH NH2 Remdesivir (RDV) stock solution (5000 µg/mL) was prepared by dissolving RDV drug powder using the mixture of DMSO: MeOH (30:70 v/v). The RDV working standard solutions for calibration and quality controls were prepared using methanol in concentrations of 100, 10, 1, 0.1, 0.01 µg/mL. 1, 2.5, 5, 7.5, 10, 25, 50, 75, 100, 250, 500, 1000, and 5000 ng/mL sample solutions were prepared freshly by spiking calibration standard solutions into the blank human plasma samples for method calibration. a) What type of calibration method is being described? Why do you think the authors chose this method as opposed to another? b) Based on the details provided in part a, describe an appropriate method blank for this analysis. 25000 20000- (b) Spiked (a) Unspiked The authors provide the following details regarding the chromatographic analysis of remdesivir: A C18 column (150 x 4.6 mm, 3 um stationary phase particles) was utilized for the separation of RDV. The mobile phase A used was 0.05% (v/v) formic acid in ultrapure water, and mobile phase B was 100% ACN with isocratic elution (A: B) 52:48 % with the total analysis time of 10 min. The column flow rate was set at 0.5 mL/min with 35 °C as the ideal column temperature, and the sample injection volume was 5 μL. 15000- 10000 5000- 0 RDV/RT 5.65 min (b) www (a) T T T 2 3 4 5 6 7 8 9 10 Retention time (minutes) c) What is one parameter you could change to decrease the retention time of RDV? Explain the theory that supports why this would work. d) What is one parameter you could change that would decrease the peak width of RDV? Explain the theory that supports why this would work.
Chemistry
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ISBN:9781305957404
Author:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
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Chapter11: Properties Of Solutions
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Transcribed Image Text:Liquid chromatography has been used to track the concentration of remdesivir
(a broad-spectrum antiviral drug, structure shown at right) in COVID patients
undergoing experimental treatments.
Intensity
The authors provide the following details regarding standard solutions
preparation:
HN
CN
HO
OH
NH2
Remdesivir (RDV) stock solution (5000 µg/mL) was prepared by
dissolving RDV drug powder using the mixture of DMSO: MeOH (30:70
v/v). The RDV working standard solutions for calibration and quality
controls were prepared using methanol in concentrations of 100, 10, 1,
0.1, 0.01 µg/mL. 1, 2.5, 5, 7.5, 10, 25, 50, 75, 100, 250, 500, 1000, and 5000 ng/mL sample solutions
were prepared freshly by spiking calibration standard solutions into the blank human plasma samples
for method calibration.
a) What type of calibration method is being described? Why do you think the authors chose this
method as opposed to another?
b) Based on the details provided in part a, describe an appropriate method blank for this analysis.
25000
20000-
(b) Spiked
(a) Unspiked
The authors provide the following details regarding the chromatographic analysis of remdesivir:
A C18 column (150 x 4.6 mm, 3 um stationary
phase particles) was utilized for the separation
of RDV. The mobile phase A used was 0.05%
(v/v) formic acid in ultrapure water, and mobile
phase B was 100% ACN with isocratic elution
(A: B) 52:48 % with the total analysis time of
10 min. The column flow rate was set at 0.5
mL/min with 35 °C as the ideal column
temperature, and the sample injection volume
was 5 μL.
15000-
10000
5000-
0
RDV/RT 5.65 min
(b)
www
(a)
T
T
T
2
3
4
5
6
7
8
9
10
Retention time (minutes)
c) What is one parameter you could change to decrease the retention time of RDV? Explain the
theory that supports why this would work.
d) What is one parameter you could change that would decrease the peak width of RDV?
Explain the theory that supports why this would work.
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