UV-Spectrophotometric Method Development and Determination of Suitable Medium for Lornoxicam by Comparing In-Between Three Body Fluids
Sourav Duari*, Ayan Das, R.N. Pal
Department of Pharmaceutics, Calcutta Institute of Pharmaceutical Technology and Allied Health Sciences, Maulana Abul Kalam Azad University of Technology, Uluberia, West Bengal, India.
*Corresponding Author E-mail: souravduari2017@gmail.com
ABSTRACT:
The study was aimed to create and verify a new, straight-forward, accurate, and specific UV-Spectrophotometric analysis method of Lornoxicam, by comparing in-between three different types of body fluids, having three different pH, like a) gastric fluid, b) physiological fluid c) intestinal fluid, and to determine the best site for administration also. UV spectrophotometric analysis was carried out by utilizing a Shimadzu UV-visible spectrophotometer, using three different mediums for the drug lornoxicam. The standard solution's spectrum was run from 200 to 400 nm range in order to figure out the absorption maximum (λmax) of lornoxicam, which was discovered at 353 nanometers. The standard solution’s absorbance of 0.3, 1, 3, 5, 7, 9, 10, 15, and 20 micrograms per milliliter was determined the solution of 353 nm against the blank. Then the graph was plotted by taking concentration on x-axis and absorbance on the y-axis. The same thing was performed for the identification of another two individual solutions in the UV spectrophotometry method. And then they were compared among themselves for suitability. Accordingly, the site of drug administration was determined. The linearity of the UV spectrophotometry method for three fluids were noted. Then it was noticed that the absorption of physiological fluid, having a pH value of 7.4, was found to be the best option for determining the UV spectrophotometry method. The drug lornoxicam was best soluble in pH 7.4 buffer as compared with pH 1.2 and 6.8 buffers. In the suggested UV visible spectrophotometry, ICH criteria were followed, and the findings and statistical parameters showed that the development technique was sensitive, exact, and easy to use for lornoxicam estimation in bulk form.
KEYWORDS: Lornoxicam, UV-spectroscopy, Nanometer, Concentration, Administration.
INTRODUCTION:
Lornoxicam is a NSAID type of drug, this is a member of the “oxicam class”. It has powerful anti-inflammatory and analgesic properties1. The Chemical name of this drug is “6-chloro-4-hydroxy-2-methyl-N-2-pyridyl-5H-thieno-[2,3-e]-1,2-thiazine-2-carboxamide-1,1-dioxide2.”
It additionally has antipyretic properties. The powder form of lornoxicam is yellow in color3. This drug is soluble in methanol and ethanol, isopropyl alcohol and slightly soluble in water. There are parenteral and oral preparations available4.
For the management of rheumatoid arthritis and osteoarthritis, lornoxicam’s oral dose is 12 milligrams daily, in two or three divided doses5. For the treatment of pain, Oral lornoxicam dosages range from 8 to 16mg per day. Similar dosages can be injected intramuscularly or intravenously, but the maximum dose can be increased up to 24mg daily, in some cases6,7.
The mechanism of action of Lornoxicam is to inhibit the enzyme cyclooxygenase non-selectively. It acts on both cox-1 and Cox- 2 isoenzymes8. Prostaglandin acts as a messenger molecule in the process of inflammation. Cyclooxygenase catalyze the conversion of arachidonic acid to thromboxane and prostaglandins9,10.
The primary goal of this study was to locate an appropriate method of analysis by UV-Spectrophotometer. The solubility of the drug Lornoxicam was studied and compared in-between three different types of body fluids with three different pHs those are physiological buffer, stomach buffer and intestinal buffer. The three different types of buffers represent the three different body solutions. The best suitable solution was found out and the best suitable site for administration was determined.
MATERIALS AND METHODS:
Materials:
Lornoxicam was procured from Mr. Arnab Saha. Sodium chloride, Sodium dihydrogen phosphate and di-sodium nitrogen phosphate and potassium chloride, hydrochloric acid, pepsin, sodium hydroxide, calcium chloride, magnesium chloride, and sodium bicarbonate were given by CIPT College. The grade of all other chemicals utilized was analytical grade.
Spectrophotometric analysis was done with a double beam UV visible Spectrophotometer with a 10 millimeters path length in quartz cell, for analytical purpose11.
Preparation of physiological simulator buffer solution (7.4 pH)
0.8g of NaCl was precisely weighed, then dissolved into 10ml distilled water. Sodium di-hydrogen phosphate, di-sodium hydrogen phosphate and KCl were accurately weighed and put into the solution12. And the volume of solution contained up-to 100 millilitres of distilled water. Then the pH 7.4 was adjusted. This 100ml solution was applied as the diluent13,14.
Preparation a standard curve of lornoxicam in physiological solution (pH 7.4)
10 milligrams of the drug was diluted into ten millilitres of the buffer solution to get 1mg per ml or 1000mcg/ml stock solution15. Then further serial dilutions were done with the previously made buffer, to get the concentration range of 0.3, 1, 3, 5, 9, 10, 15 and 20mcg/ml. And the absorption was determined using a UV spectrophotometer at 353nm16 against a blank of pH 7.4 phosphate buffer. The standard calibration curve was obtained by plotting the absorbance with concentration (mcg/ml).
Preparation of gastric simulator buffer solution (1.2 pH):
0.2gm of NaCl was weighed accurately and dissolved into 0.7ml of HCl. And 0.32g of pepsin was accurately weighed and dissolved in 10ml of distilled water17 and the volume was made up to 100ml by adding more distilled water. Then the PH was adjusted to 1.2 by adding HCl18. This 100ml solution was used as the diluent.
Preparation of standard curve of lornoxicam in gastric simulator buffer solution (pH 1.2):
Ten milligrams of the drug was diluted into 10ml of the buffer solution to get 1mg/ml or 1000mcg/ml stock solution19. Then further serial dilutions were done to get the concentration range of 0.3, 1, 3, 5, 9, 10, 15 and 20 mcg/ml. And the absorption was evaluated in comparison to pH 1.2 buffer as the blank at utilizing a UV spectrophotometer, 353nm20. The “standard calibration curve” was obtained by plotting the absorbance with concentration (mcg/ml).
Preparation of intestine simulator buffer solution (6.8 pH):
0.1gm of NaOH, 1.19gm NaCl, 0.1g of KCl, 0.04gm of CaCl2, 0.02gm MgCl, 0.2gm of sodium bicarbonate were weighed accurately, and dissolved into 10ml of water and then a 100ml amount was added which was distilled water21. Then the pH was changed to 6.8. This 100ml solution was applied as the diluent.
Preparation of standard curve of lornoxicam in intestinal solution (pH 6.8):
10mg of the drug was diluted into 10ml of the buffer solution to get 1mg/ml or 1000mcg/ml stock solution22. Then further serial dilutions were done to get the concentration range of 0.3, 1, 3, 5, 9, 10, 15 and 20mcg /ml. And the absorption was measured against pH 6.8 phosphate buffer as the blank at 353nm using a UV-spectrophotometer23. The “standard calibration curve” was created by plotting the absorbance versus the concentration (mcg/ml).
RESULT:
The purpose of this investigation was to determine the best appropriate solution for developing a simple and precise UV-spectrophotometric method and to select a suitable absorption site for lornoxicam. This investigation includes figuring out the concentration of the drug in three types of body fluids spectrophotometrically.
The all three concentrations against absorbance standard curve graphs, determined by UV spectrophotometer, of the drug lornoxicam, in three different buffer media was observed and comparison was done in-between them. The simulator fluid which could stabilize the region with the highest drug concentration was chosen to be the focus of the administration of adjusted dose forms of lornoxicam. Findings and discussions of the above study is presented below.
Table 1: Calibration curve readings of lornoxicam in pH 7.4 buffer at 353 nm
|
Concentration (mcg/ml) |
Absorbance at 353 nm |
|
0.3 |
0.13 |
|
1 |
0.17 |
|
3 |
0.253 |
|
5 |
0.353 |
|
7 |
0.449 |
|
9 |
0.542 |
|
10 |
0.591 |
|
15 |
0.811 |
|
20 |
0.999 |
Figure 1: Standard curve of lornoxicam in pH 7.4 buffer at 353 nm.
Table 2: Calibration curve readings of lornoxicam in pH 1.2 buffer at 353nm
|
Concentration (mcg/ml) |
Absorbance at 353 nm |
|
0.3 |
0.011 |
|
1 |
0.016 |
|
3 |
0.031 |
|
5 |
0.045 |
|
7 |
0.065 |
|
9 |
0.08 |
|
10 |
0.088 |
|
15 |
0.132 |
|
20 |
0.175 |
Table 3: Calibration curve readings of lornoxicam in pH 6.8 buffer at 353 nm
|
Concentration (mcg/ml) |
Absorbance at 353 nm |
|
0.3 |
0.099 |
|
1 |
0.105 |
|
3 |
0.127 |
|
5 |
0.148 |
|
7 |
0.168 |
|
9 |
0.189 |
|
10 |
0.198 |
|
15 |
0.25 |
|
20 |
0.311 |
Figure 2: Standard curve of lornoxicam in pH 1.2 buffer at 353 nm.
Figure 3: Standard curve of lornoxicam in pH 6.8 buffer at 353 nm.
Table 4: Comparison between calibration curve of lornoxicam, in three body fluids, at 353 nm
|
Concentration (mcg/ml) |
Physiological solution (7.4 pH) |
Gastric solution(1.2pH) |
Intestinal solution (6.8pH) |
|
0.3 |
0.13 |
0.011 |
0.099 |
|
1 |
0.17 |
0.016 |
0.105 |
|
3 |
0.253 |
0.031 |
0.127 |
|
5 |
0.353 |
0.045 |
0.148 |
|
7 |
0.449 |
0.065 |
0.168 |
|
9 |
0.542 |
0.08 |
0.189 |
|
10 |
0.591 |
0.088 |
0.198 |
|
15 |
0.811 |
0.132 |
0.25 |
|
20 |
0.999 |
0.175 |
0.311 |
Figure 4: Comparison between Standard curves of lornoxicam, in three body fluids, at 353 nm
From the above comparison between three different pH buffers, we can see that the lornoxicam drug is mostly soluble in 7.4 pH phosphate buffer solution.
Sensitivity:
The equations LOD = 3*σ/ S and LOQ= 10*σ/ S were utilized for calculating the limits of detection (LOD) and quantification (LOQ), respectively. sigma refers to the standard deviation of intercept and S is the slope. The LOD of gastric fluid and intestinal fluid and physiological It was discovered that fluid 4.34mcg per ml 3.75mcg per ml and 6.72mcg per ml respectively. The LOQ of gastric fluid intestinal fluid and physiological It was discovered that fluid was 14.55mcg per ml 12.30mcg per ml and 22.32mcg per ml respectively.
DISCUSSION:
According to figure 4, from the comparison between three different pH buffers, we can conclude that the lornoxicam drug is mostly soluble in 7.4 pH phosphate buffer solution.
Figure 4 displays the solubility of lornoxicam in different bodily fluids. It was discovered through solubility research that exhibited the maximum solubility. In the blood or physiological fluid. And in the intestinal and gastric fluids, solubility is intermediate and lowest respectively.
So, the most suitable buffer for UV Spectrophotometric analysis of the drug lornoxicam would be the phosphate buffer of pH 7.4.
CONCLUSION:
The primary goal of the research was to design the most suitable UV-spectrophotometric determination method and to ascertain the body's appropriate absorption site for the creation of an appropriate adjusted dosage form for the medication lornoxicam. The comparison of three different body fluids solubility of the drug was done. The drug in each body fluid was assessed using spectrophotometry. The results of UV Spectrophotometry analysis verified that there was a higher drug concentration in the physiological fluid compared to gastric and intestinal fluid. Therefore, the formulation would be changed in a manner that would dosage form will deliver the maximum drug into the physiological fluid or the blood.
CONFLICT OF INTEREST:
The authors have no conflicts of interest regarding this investigation.
ACKNOWLEDGMENTS:
The authors would like to thank CIPT and AHS college authority for their kind support during all lab studies.
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Received on 09.11.2024 Revised on 02.02.2025 Accepted on 07.04.2025 Published on 06.05.2025 Available online from May 10, 2025 Asian Journal of Pharmaceutical Analysis. 2025; 15(2):75-79. DOI: 10.52711/2231-5675.2025.00012 ©Asian Pharma Press All Right Reserved
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