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Salbutamol is almost exclusively metabolised by conjugation to a 4’-o-sulphate ester in the intestinal wall and liver. A second minor metabolite has been reported3-5. Side effects of salbutamol were reported to be minimal and well tolerated. One of the earliest instrumental techniques for analysis is UV-VIS spectroscopy. Many different types of materials can be characterized using UV-V is spectroscopy. FTIR is used to analyse the small molecules or complex molecules. Wide range of sample types such as solid, liquid and gas can be checked from about 4000-400 cm-1. Fourier transform infrared (FTIR) spectroscopy represents a modern and popular technique that addressed IR spectroscopy as a powerful and reliable analytical technique. The using the method of UV, FTIR and HPLC to find out the absorbance wavelength and percentage purity of salbutamol sulphate.
A sympathomimetic amine called salbutamol sulphate (SBS) is used as a bronchodilator to treat reversible bronchospasm. 2–4 mg, three–four times a day is the typical dosage1. Typically, it is given as a pill, capsule, syrup, or spray. In Turkey, there are just three brands of salbutamol sulphate tablets available for purchase. After oral treatment, salbutamol is well absorbed, reaching peak plasma levels in 1–4 hours (t max). Salbutamol is efficiently absorbed, but because of significant pre-systemic metabolism in the gut wall, its systemic bioavailability is only 50%.
Fig 1: Structure of salbutamol sulphate
MEDICINAL USE
Salbutamol is typically used to treat bronchospasm (due to any cause—allergic asthma or exercise-induced), as well as chronic obstructive pulmonary disease.
It is also one of the most common medicines used in rescue inhalers (short-term bronchodilators to alleviate asthma attacks).
As a β2 agonist, salbutamol also has use in obstetrics. Intravenous salbutamol can be used as a tocolytic to relax the uterine smooth muscle to delay premature labor.
UV SPECTROSCOPY
UV-VIS spectroscopy is among the oldest instrumental methods of analysis. UV-V spectroscopy can be used to characterize a wide variety of materials. The UV-Vis provides information based on the different responses of samples and the degree of transmission or absorption of a light beam with a range of wavelengths. Beer's law is a general law that provides a quantitative description of the radiant energy absorption by materials. Using and handling the UV-VIS spectrophotometer is easy.
Fig 2: UV spectroscopy
Application
Applications include impurity detection.
Clarification of organic molecules' structures.
Analysing quantitative qualitative evaluation.
Analysis of chemicals.
FTIR SPECTROMETER
Small or complex compounds can be analyzed using FTIR. Fourier transform infrared (FTIR) spectroscopy is a contemporary and widely used technology that addresses IR spectroscopy as a potent and dependable analytical method. It can examine a wide variety of sample kinds, including solid, liquid, and gas, from roughly 4000-400 cm-1. This spectroscopic method is most frequently employed in industry or by both organic and inorganic chemists.
Fig 3: Block diagram of FTIR
Application
High modes of vibration.
Lower symmetry of complexes.
Formation of chelates.
Miscellaneous examples.
HPLC
The analytical method known as High-Performance Liquid Chromatography (HPLC) is used to separate solutes according to the differences in their rates of elution within a chromatographic column. The distribution of solutes between the stationary phase and the mobile phase is crucial to this separation technique. A mobile phase reservoir, solvent delivery system, sample introduction device, column, detector, waste reservoir, connective tubing, and a computer, integrator, or recorder are the eight essential parts of HPLC equipment.
Fig 4: HPLC instrument
Application
The technique of isolating and purifying chemicals is known as preparative HPLC.
The level of solute purity and throughput—the quantity of compound generated in a given amount of time—are significant.
This is not the same as analytical HPLC, which focuses on learning more about the sample substance.
Identification, measurement, and resolution of a substance are among the information that can be obtained.
Objective
To evaluate the given drug.
To find the wavelength and absorbance of the drug (Salbutamol sulphate).
To find the functional group of the drug.
To check the percentage purity of given drug.
RESEARCH METHODOLOGY
Analysis of Salbutamol sulphate using UV- Spectrophotometer:
Name of Instrument:
UV- Spectrophotometer
Software: UV-WIN
Make: Lab India
SOP No. SA/LAB/07
Id. No. -SA/LB/UV/01
Mode of instrument
Spectrum Mode: obtains sample spectra using wavelength scanning.
Photometric Mode: measures the absorbance or transmittance at a single wavelength or at multiple wavelengths.
3.3 Procedure
Switch on the FTIR spectrophotometer & start Spectra Manager Software.
Weigh Kbr and triturate it in the mortar & pestle.
Prepare the pellet of Kbr with the help of hydraulic press.
Transfer it in pellet holder & place it in sample compartment.
Select "Background" from the "measure" menu of the spectra manager program. The "Background" window opens.
Click on the background scan and Background spectrum will be displayed. Now click on 'OK' so that it will scan background spectra.
Weigh KBr and Salbutmol sulphate, accurately transfer them into a mortar and pestle & triturate it.
Transfer this mixture into the disc plate to prepare pellet of sample using hydraulic press, ie, pressed pellet technique
Remove the previous KBr pellet from the sample compartment and place sample pellet disc plate into the sample compartment of FT-IR spectrophotometer. Click on Sample measurement. Enter sample details and click OK. Scan will start
Click on 'Spectral Analysis in the spectrum manager software then click of Processing" then click on "correction' then 'smoothing and finally click on "Apply" and then 'OK' this is done to obtain the spectrum of the actual sample Click on "processing" the click on "Peak find then adjust Y axis parameters and click on "Apply" and "OK" this is done to obtain the peaks in the spectrum.
Then save the observations and results obtained by clicking "save as" save the data by the name of the Drug and Initials of the person performing the procedure.
For printout click "print" and select "Display Current".
Analysis of Salbutmol Sulphate Using Ft-Ir Spectrophotometer
Name of Instrument –
FT-IR Spectrophotometer
Software – Spectra Manager
Make – Jasco
SOP No.- SA/LAB/035
Balance Id. No. SA/LB/AB-01
Procedure:
Switch on the FTTR spectrophotometer & start Spectra Manager Software
Weigh Kfr and triturate it in the mortar & pestle.
Prepare the pellet of KBe with the help of frydraulic press.
Transfer it in pellet holder & place it in sample compartment.
Select "Background from the measure" mems of the spectra manager program The Background window opens.
Click on the background scan and Background spectrum will be displayed. Now click on "OK" so that it will scan background spectra.
Weigh KBr and Salbutamol sulphate, accurately transfer them into a mortar and pestle & triturate it.
Transfer this mixture into the disc plate to prepare pellet of sample using hydraulic press, i.e. pressed pellet technique.
Remove the previous KBr pellet from the sample compartment and place sample pellet disc plate into the sample compartment of FT-IR spectrophotometer. Click on Sample measurement. Enter sample details and click OK, Scan will start.
Click on 'Spectral Analysis' in the spectrum manager software then click of "Processing" then click on 'correction' then 'smoothing' and finally click on Apply' and then 'OK' this is done to obtain the spectrum of the actual sample Click on "processing" the click on "Peak find" then adjust Y axis parameters and click on "Apply" and "OK" this is done to obtain the peaks in the spectrum.
Then save the observations and results obtained by clicking "save as save the data by the name of the Drug and Initials of the person performing the procedure.
For printout click "print" and select "Display Current".
Analysis Of Salbutmol Sulphate Using HPLC chromatography:
Name of Instrument –
HPLC Chromatography
Software – Lab solution
Make – Shimadzu
Id. No. SA/LB/HPLC/01
Column use for analysis
Column use for analysis
C 18, 250 mm x 4.6 mm, 5 µ
Make
Shimadzu, software- lab solution
Mobile phase
Acetonitrile and 0.1%
Orthophosphoric acid (70:30)
Sample concentration
100 µg/ml
Run time
6 min
Injection value
20 µl
Flow
0.70 ml/min
Wavelength
224 nm
Procedure:
Switch on the computer.
Switch on the UV detector and HPLC pump.
Double click on the LC solution icon on the desktop, followed by the HPLC icon.
Check there is sufficient elution solvents: Please top-up if there is insufficient. Use only HPLC grade solvents to prevent clogging of the system.
If HPLC has not been in use for a while, air bubbles in the system are needed to purge out. Turn the knob on HPLC a quarter and press Purge. The purging process will take 10 minutes.
Once the purging is finished, turn the kriob back.
Attach the column in the correct flow direction.
Open the required method file or build a new method file. Ensure that the wavelength, flow rate, elution time and elution proportion are correct.
Monitor the baseline by clicking the Baseline icon in the program. Check tubing connection; ensure that there is no leakage.
Once the baseline is fiat, the system is ready for sample injection.
Prepare the sample in the appropriate solvent. Ensure there is no particulate by filtering the sample solution through a membrane. The sample must be purified by column chromatography.
OBSERVATION:
Observation of UV spectroscopy:
Mode of spectrum:
Sample Name
Wavelength(λmax)
Absorbance
Salbutamol sulphate
224.00 nm
0.375
Mode – Photometric
Observations at λmax 208nm
1.
1-1
Abs
0.375
2.
2-1
Abs
0.374
3.
3-1
Abs
0.373
Observation of FTIR:
Sr. No.
Functional groups
Observed wave number (cm-1)
Standard wave number (cm-1)
Compou nd class
1.
-OH bonding
3474.13 cm- 1
3650 – 3200 cm- 1
Alcohol
2.
=CH-H
3158.83 cm- 1
3100 – 3070 cm- 1
Hydrocarbon,
Alkane
3.
C=C aromatic
1614.13 cm- 1
1600-1500 cm- 1
Benzene
4.
C-M
1387.53 cm- 1
1350 cm- 1
Amines
Observation HPLC:
Peak
Name
RET. Time
Area
Area%
1
Salbutamol sulphate
3.263
6018391
98.477
2
3.995
30474
0.499
3
4.780
62610
1.024
Total
6111474
100.000
Theoretical plates / meter (USP)
Tailing factor
16314
1.148
54270
1.284
67154
1.127
REPORTS:
Practical using UV spectroscopy
5.2 UV spectrum of salbutamol sulphate solution in distilled water:
Practical using FTIR spectrometer:
Practical using HPLC:
RESULT AND CONCLUSION:
CONCLUSION:
I have learnt and completed hands-on experimental work on UV, FTIR Spectrophotometer and HPLC.
I have determined max and absorbance by using UV Spectrophotometer and Functional groups by using FTIR-Spectrophotometer of salbutamol sulphate.
I have determined the percentage purity of salbutamol sulphate by using HPLC.
The sample solution was found to be stable 48hrs and mobile phase was found stable up to 7 days.
This method can be consider for its intended purpose study to established the quality of drug product during stability study analysis.
RESULT:
The analysis of salbutamol sulphate was found to be absorbance and λ max by using UV spectroscopy.
The analysis of salbutamol sulphate was found to be functional group by using FTIR spectrometer.
The analysis of salbutamol sulphate was found to be % purity (by area normalisation method) such as 98.47% by using HPLC.
Formulation And Evaluation of Floating Tablets of Salbutamol Sulphate, G. Mahendar*, S.Jaya, International Research Journal of Pharmaceutical and Applied Sciences (Irjpas). Formulation and evaluation of Salbutamol sulphate microspheres by solvent evaporation method, V.V Prasanth, Akashmoy Chakraborty, Sam T Mathew, Rinku Mathappan,and V. Kamalakkannan, Journal of Applied Pharmaceutical Science 01 (05); 2011:133-137.
Standard Operating Procedure (SOP) of UV Visible Spectroscopy.
Standard Operating Procedure (SOP) of FT-IR Spectroscopy.
Vishruti Maniar, Krishna Kalsara, Dr Umesh Upadhyay, A Review of Ftir – a Useful Instrument, International Journal of Pharmaceutical Research and Applications Volume 8, Issue 1 Jan-Feb 2023, pp: 2486-2490 www.ijprajournal.com.
Anjoo kamboj, Pawan sidana, Upendra k jain. Development and validation of UV spectroscopy method of stimulanious. ISSN- 0975-1491 Vol 9, Issue 6, 201 http://dx.doi.org/10.22159/ijpps.2017v9i6.14938
Deepak Sharma, Gurmeet Singh, Dinesh Kumar, Mankaran Singh. Formulation Development and Evaluation of Fast Disintegrating Tablets of Salbutamol l Sulphate, Cetirizine Hydrochloride in Combined Pharmaceutical Dosage Form: A New Era in Novel Drug Delivery for Pediatrics and Geriatrics. https://onlinelibrary.wiley.com/doi/full/10.1155/2015/640529#
G.R. Gadekar, S.S. Ptil, R.R. Shah, D.S. Ghodke. Development and validation of simple UV spectrometric method for the estimation of salbutamol sulphate from pharmaceuticals formulation. ol 11, Issue 5, 2019 https://core.ac.uk/reader/482244590
Ivan Bezruk, Anna Materiienko, Svitlana Gubar, Vera Bunyatyan, Sergiy M. Kovalenko, Victoriya Georgiyants, Liudas Ivanauska. Development and validation of a HPLC method for quantification of degradation impurities of salbutamol sulfate with following long-term stability studies in multicomponent cough syrup. 69, 211–217.
Formulation And Evaluation of Floating Tablets of Salbutamol Sulphate, G. Mahendar*, S.Jaya, International Research Journal of Pharmaceutical and Applied Sciences (Irjpas). Formulation and evaluation of Salbutamol sulphate microspheres by solvent evaporation method, V.V Prasanth, Akashmoy Chakraborty, Sam T Mathew, Rinku Mathappan,and V. Kamalakkannan, Journal of Applied Pharmaceutical Science 01 (05); 2011:133-137.
Standard Operating Procedure (SOP) of UV Visible Spectroscopy.
Standard Operating Procedure (SOP) of FT-IR Spectroscopy.
Vishruti Maniar, Krishna Kalsara, Dr Umesh Upadhyay, A Review of Ftir – a Useful Instrument, International Journal of Pharmaceutical Research and Applications Volume 8, Issue 1 Jan-Feb 2023, pp: 2486-2490 www.ijprajournal.com.
Anjoo kamboj, Pawan sidana, Upendra k jain. Development and validation of UV spectroscopy method of stimulanious. ISSN- 0975-1491 Vol 9, Issue 6, 201 http://dx.doi.org/10.22159/ijpps.2017v9i6.14938
Deepak Sharma, Gurmeet Singh, Dinesh Kumar, Mankaran Singh. Formulation Development and Evaluation of Fast Disintegrating Tablets of Salbutamol l Sulphate, Cetirizine Hydrochloride in Combined Pharmaceutical Dosage Form: A New Era in Novel Drug Delivery for Pediatrics and Geriatrics. https://onlinelibrary.wiley.com/doi/full/10.1155/2015/640529#
G.R. Gadekar, S.S. Ptil, R.R. Shah, D.S. Ghodke. Development and validation of simple UV spectrometric method for the estimation of salbutamol sulphate from pharmaceuticals formulation. ol 11, Issue 5, 2019 https://core.ac.uk/reader/482244590
Ivan Bezruk, Anna Materiienko, Svitlana Gubar, Vera Bunyatyan, Sergiy M. Kovalenko, Victoriya Georgiyants, Liudas Ivanauska. Development and validation of a HPLC method for quantification of degradation impurities of salbutamol sulfate with following long-term stability studies in multicomponent cough syrup. 69, 211–217.
Ghule Ujjawala
Corresponding author
Dr. Kolpe Institute of Pharmacy, Kolpewadi, Ahmednagar, Maharashtra- 423602
Ghule Ujjawala*, Shivam Gangurde, Pooja Gidhad, Smita Ghuge, Analytical Method Development & Estimation of Salbutamol Sulphate in Inhaler Dosage from by Using UV, FTIR And HPLC, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 5, 282-292. https://doi.org/10.5281/zenodo.15332560