Analytical Method Development & Estimation of Salbutamol Sulphate in Inhaler Dosage from by Using UV, FTIR And HPLC

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:

Mode of instrument

1. Spectrum Mode: obtains sample spectra using wavelength scanning.
2. Photometric Mode: measures the absorbance or transmittance at a single wavelength or at multiple wavelengths.

3.3 Procedure

1. Switch on the FTIR spectrophotometer & start Spectra Manager Software.
2. Weigh Kbr and triturate it in the mortar & pestle.
3. Prepare the pellet of Kbr with the help of hydraulic press.
4. Transfer it in pellet holder & place it in sample compartment.
5. Select "Background" from the "measure" menu of the spectra manager program. The "Background" window opens.
6. Click on the background scan and Background spectrum will be displayed. Now click on 'OK' so that it will scan background spectra.
7. Weigh KBr and Salbutmol sulphate, accurately transfer them into a mortar and pestle & triturate it.
8. Transfer this mixture into the disc plate to prepare pellet of sample using hydraulic press, ie, pressed pellet technique
9. 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
10. 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.
11. 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.
12. For printout click "print" and select "Display Current".

Analysis of Salbutmol Sulphate Using Ft-Ir Spectrophotometer

Procedure:

1. Switch on the FTTR spectrophotometer & start Spectra Manager Software
2. Weigh Kfr and triturate it in the mortar & pestle.
3. Prepare the pellet of KBe with the help of frydraulic press.
4. Transfer it in pellet holder & place it in sample compartment.
5. Select "Background from the measure" mems of the spectra manager program The Background window opens.
6. Click on the background scan and Background spectrum will be displayed. Now click on "OK" so that it will scan background spectra.
7. Weigh KBr and Salbutamol sulphate, accurately transfer them into a mortar and pestle & triturate it.
8. Transfer this mixture into the disc plate to prepare pellet of sample using hydraulic press, i.e. pressed pellet technique.
9. 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.
10. 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.
11. 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.
12. For printout click "print" and select "Display Current".

Analysis Of Salbutmol Sulphate Using HPLC chromatography:

Column use for analysis

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:

Mode – Photometric

• Observations at λmax 208nm

Observation of FTIR:

Observation HPLC:

Practical using UV spectroscopy

5.2 UV spectrum of salbutamol sulphate solution in distilled water:

Practical using FTIR spectrometer: