The document discusses precision measurements using UV/Vis spectroscopy. It describes calibrating pipettes and volumetric flasks, then taking absorbance readings of a methyl orange solution using the calibrated equipment. Statistical analysis found pipettes had lower standard deviations, and thus higher precision, than volumetric flasks. The document also details determining the concentration of salicylate in a face wash using a reaction with Fe(III) ions and developing a calibration curve. A Job plot method was used to find the reaction stoichiometry. Data from repeated measurements was statistically analyzed to check if results came from the same population.
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This study is done to access the chemical stability of the candidate compound in the pharmaceuticals.
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New RP HPLC method for the simultaneous estimation of sulbactum and ceftriaxo...SriramNagarajan19
A simple and selective LC method is described for the determination of Sulbactum and Ceftriaxone tablet dosage forms. Chromatographic separation was achieved on a C18 column using mobile phase consisting of a mixture of mixture of 60 volumes of 20mM Phosphate buffer pH 3.5: 40 volumes of Acetonitrile (60:40 v/v) with detection of 210 nm. Linearity was observed in the range 30-70 µg /ml for Sulbactum (r2 =0.9998) and 60-140µg /ml for Ceftriaxone (r2 =0.9983) for the amount of drugs estimated by the proposed methods was in good agreement with the label claim. The proposed methods were validated. The accuracy of the methods was assessed by recovery studies at three different levels. Recovery experiments indicated the absence of interference from commonly encountered pharmaceutical additives. The method was found to be precise as indicated by the repeatability analysis, showing %RSD less than 2. All statistical data proves validity of the methods and can be used for routine analysis of pharmaceutical dosage form.
STUDY OF MPS UNDER STRESSED CONDITIONSvivatechijri
This study is done to access the chemical stability of the candidate compound in the pharmaceuticals.
Usually, it is performed at the preliminary stage in the process of drug development. Forced degradation/ stress
testing is performed under accelerated environment. The experimental conditions cause the candidate compound
to degrade under extreme conditions like acid and base hydrolysis, peroxide oxidation, photo-oxidation and
thermal stability to identify the resultant degradation products. This helps to establish degradation pathways and
thus intrinsic stability of a drug substance. The stability of product describes shelf life and storage conditions and
helps in the selection of appropriate formulations and their suitable packaging. This is compulsory for regulatory
documentation. The commonly used analytical approach for FDS is HPLC with UV and/ or MS but these
techniques consume a lot of time and not provide high resolution to confirm the precise detection of degradation
products. Use of UPLC with photodiode array and MS analysis supports the identification of degradation
products and also reduces the time needed to evolve stability indicating methods.
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Introduction Part Precision Measurements using Spectroscopy Aim.pdf
1. Answer: Introduction Part 1: Precision Measurements using UV/Vis
Spectroscopy Aim
Answer:
Introduction
Part 1: Precision Measurements using UV/Vis Spectroscopy
Aim
This lab work was meant to foster understanding in:
Use of an air-displacement piston pipette
Use of an analytical balance
Calibration of glassware
4.To demonstrate the influence of volume measuring equipment on precision
Introduce the concept of propagation of uncertainty
Statistical Analysis Of Data
The science of measurement is known as metrology. Mass, length, area, volume, time,
pressure, and other weights and measurements are all part of physical metrology. The
CSIRO National Measurement Laboratory, NATA (National Association of Testing
Authorities), ISO9000, and other regulating agencies such as the FDA (Federal Drug
Administration) in the United States are the bodies that supervise weights and measures in
Australia (Jorio and Dresselhaus, 2016).
The link between a sequence of measurements, especially whether they show the same or
comparable results, is referred to as precision. Precision is sometimes used interchangeably
with accuracy, which characterizes the connection between a measurement and a
predetermined 'actual' value, such as a standard reference material.
Because no two measurements are ever precisely the same, analysts often refer to
2. measured values as having a degree of uncertainty, which refers to the range of possible
values on each side of the observed value that the measurement is most likely to fall into.
In a data collection, precision is expressed as the standard deviation, range, or variance. The
width of the curve in a Gaussian distribution plot represents the standard deviation, which
is a measure of how tightly a collection of results are grouped around the mean.
Part 2: Quantitative determination of Salicylate via Reaction with Fe(III)
Aim
This practical exercise reinforces knowledge in:
Preparation of calibration curves
Quantitative analysis using a complexometric reaction
Determining reaction stoichiometry using a Job plot
Spectroscopic analysis is an important method for identifying and quantifying various
compounds. This experiment teaches you how to determine salicylate using electronic
absorption spectroscopy in the visible portion of the spectrum.
Salicylate has a variety of applications, hence it's found in a variety of items. Salicylic acid is
the most prevalent metabolite of aspirin, and it's used to treat acne, warts, and other skin
conditions (https://www.facebook.com/Drugscom, 2020).
Salicylic acid reduces the loss of skin cells from hair follicles, which helps to cure acne.
These cells are usually the ones that obstruct pores and cause pimples. Salicylic acid also
has a keratolytic (peeling) action, which promotes the shedding of dead cells. This makes it
easier to remove a thin layer of skin and helps to unclog pores. Salicylic acid at higher
concentrations is used in wart therapy to soften the wart and induce an immunological
response against the human papillomavirus, which causes wart production.
Salicylic acid's diverse medicinal uses need the development of analytical procedures for its
measurement. Gas-liquid chromatography (GLC), ultraviolet spectroscopy, and fluorescence
spectroscopy were among the techniques used. Colorimetric or visible spectrophotometry,
on the other hand, is the most extensively used approach in clinical labs. To measure
salicylate in a commercial product, a variation of this approach will be used throughout the
trial (face wash). The second stage of the technique employs spectrophotometry to look into
the chemical composition of the reaction that results in the colored product under
investigation.
Measurement Principles
3. Beer's Law says that a compound's absorbance is proportionate to its concentration
(A=εbc). We may use this linear connection to create a calibration curve by collecting
absorbance values for samples of known concentration at a certain wavelength, ideally the
max, which is the wavelength where maximum absorption occurs. The linear regression
equation that results allows us to calculate the concentration of an unknown material by
measuring its absorbance at the same wavelength. The inability of salicylate and salicylic
acid to absorb visible light poses an experimental obstacle. However, when iron (III) ions
are added, a brightly colored species emerges:
With a basic spectrophotometer, you can readily identify the complex, allowing you to
measure salicylate in unknown materials. All salicylate will be protonated under the acidic
experimental circumstances, as illustrated in the chemical equation above. The coefficients
and subscripts x and y are indicated in the chemical equation above. The method of
continuous variation (also known as Job's method) will be used in the second part of this
experiment to determine these numbers for the dominating complex. Several solutions
containing various amounts of salicylate and Fe3+ will be created for this technique. The
total moles of both reagents will stay constant while the quantity of each reactant is altered.
The solution with the highest absorbance at max is the one with the most stoichiometry.
Materials And Methods
The method was adapted as described in the Analytical Chemistry 1 Practical Manual.
Briefly, for the first part of the experiment, the air-displacement piston pipette and 10.0 mL
volumetric flask to be used for the experiment were firstly calibrated. For the pipette, it was
done by pipetting 1.00mL(1000uL) of water into a 20.0mL vial and recording the weight
displayed on the balance. This was repeated by adjusting the scale on pipette until the
balanced displayed 1.00g (which was at 1.009uL on the pipette and then 4 more times after
that to ensure the correct volume was being dispensed. The volumetric flask was then
calibrated by placing it on the balance and filling it to the line with water to record the
weight displayed. This was repeated 4 times.
After the calibration of the pipette and the volumetric flask, the 1000uL of the stock solution
that was provided (methyl orange in water) was transferred to the 10.0 mL volumetric flask
using an air-displacement piston pipette and then diluted to volume with distilled water.
The absorbance of this solution was measured at 473nm on the UV/Vis spectrometer and
the value recorded. This was repeated four times using the same pipette and volumetric
flask so that there were 5 absorbance values recorded in total at the end.
For the second part of the experiment, at first, for the spectrophotometric determination of
salicylate in acne medication, five standard solutions of sodium salicylate in deionised water
were prepared as following- 1. A 100ml volumetric flask was used to prepare an initial
stock solution of 100mM (0.100 M) of sodium salicylate. The mass of sodium salicylate
needed for this was calculated to be 1.60g and accurately weighed out on the balance.
4. By dilution in 10mL volumetric flasks, standards of 20.0mM, 30.0mM, 40.0mM and 50.0mM
were made and another 100mL of a 10mM standard in water was also made.
A sample of the acne face wash solution was collected and the suggested quantity of the
salicylate on the label is noted that is 2.00%.
In 5 separate test tubes, 20.0uL of each standard and of the acne face wash was transferred
using an air-displacement pipette. Then 10.0mL of acidic 10.0mM ferric nitrate solution was
added to each test tube and mixed well.
After the spectrophotometer was blanked using the ferric nitrate solution, λmax was
determined from an absorbance spectrum (400-700nm) collected with the most
concentrated standard and then the absorbance values were recorded for each of the
standards at the determined λmax.
Within the second part of the experiment, to examine the reaction stoichiometry of the
reaction between Fe (III) and salicylate, 20.0mL of 10.0mM acidic ferric nitrate, 50.0mL of
dilute nitric acid (60.0mM) and the 10.0mM sodium salicylate solution made earlier were
used to prepare solutions for spectrophotometric analysis by pipetting the correct amount
of each solution into a vial as suggested in the table below. After that, the vials were labelled
and 4.00mL of dilute nitric acid was added to each vial to make a total of 5mL. Using the
dilute nitric acid to blank the spectrophotometer, the absorbance values were recorded at
λmax for each solution.
Results
Part 1
A table of Calibration of Pipette and Volumetric Flask
Run
Pipette
Volumetric Flask
9. A job plot for data from part 2
A table showing statistical analysis of the unknown sample.
Unknown
λmax (nm)
absorbance
Concentration (mM)
wt% (g/100g solution)
1
546
0.082
24.31
0.3893
20. 0.01269
t Critical two-tail
2.570582
Discussion
When the quantified value of salicylate is converted to g/L, a value of 3.9% is noted. This is
contrary to the literature value of 2%. This is due to errors both in measurements carried
out and the inherent uncertainty of the apparatus.
In the comparison of two data sets in part 3, it is found that the data indeed come from the
same population but vary widely and their standard deviations are quite different. This can
be attributed to random errors.
Questions:
Part 1
From your data, which dilution method offers the best precision? Discuss the possible
reasons with reference to the statistical results obtained and sources of error and
uncertainty.
Precision from standard deviations show pipettes to possess a lower standard deviation
(0.003725) compared to that of volumetric flaks (0.02838). Precision can also be deduced
from the uncertainty results. The pipette offers an experimental uncertainty (0.001665)
lower than that of the volumetric flask (0.01269). Pipetting methods are thus considered
more precise in producing more reproducible results. These uncertainties can either be
grouped as systemic or random errors.
Compare and contrast the experimental and the theoretical uncertainty associated with the
air-displacement pipette. Comment on potential differences.
The theoretical uncertainty is observed to be higher when compared to the experimental
uncertainty. This can be inferred to experimental uncertainty not only accommodating for
21. systemic errors but also random errors.
Why is it important to use the same pipette and glassware when completing the 5 repeat
measurements for each solution?
This helps to keep systemic errors due to apparatus changes to a minimum. Every
apparatus will give a different uncertainty, for success in the experiment the value of
uncertainty by apparatus should remain constant.
Part 3
In your Discussion, answer the following questions;
a) Were there any outliers in the data sets? Show the values that identify if there are or
aren’t outliers.
The dataset did not contain any outliers as shown by no value going below Gcritical in the
Grub’s test.
Was your standard deviation similar to your lab partners? Show the values that identify if
the standard deviations are or aren’t the same.
F-test gives a good discerning platform as it compares the variances in the data. It is
hypothesised that the standard deviations are similar. It can thus be observed that;
F>Fcritical. leading to rejection of the hypothesis.
Was your data from the same population as your lab partners? Show the values that identify
if they are from the same population.
The null hypothesis is that the samples are from the same population. This is evaluated by
using a t-test. It can be observed that tsample