Alameen R, Amirtha R, Anand S, N. Santhi, S.S. Rajendran*
*Corresponding Author E-mail: rajendran@rvsgroup.com
ABSTRACT:
A Simple, sensitive, precise, rapid and economical UV Spectrophotometric method for determining Teneligliptin Hydrobromide Hydrate (THH) in both bulk and various marketed tablet dosage forms was developed and validated. The standard solutions of Teneligliptin Hydrobromide Hydrate (THH) were prepared using Distilled water. The standard solution of THH showed maximum absorption (λ max) at wavelength 244nm. This method obeys Beer’s law in the concentration range from 10-50μg/ml. The regression of the curve was found Y=0.00182x+0.0096 and the correlation coefficient was found to be 0.9994. The developed method was applied to marketed brand and generic tablet dosage forms and the % amount of drug estimated was 101.33% and 102% respectively. It was in accordance with the label claim. The Accuracy was found to be 97.7%. The limit of detection and limit of quantification was found to be 0.5439µg/ml and 1.648µg/ml respectively. Precision studies were performed as intraday and interday variations %RSD value within the limit of ≤ 2. The ICH guidelines were followed in the development and validation of the method. This method was simple, rapid and economical for the routine analysis of Teneligliptin Hydrobromide Hydrate (THH) in bulk and pharmaceutical dosage forms.
KEYWORDS: Teneligliptin Hydrobromide Hydrate (THH), UV Spectrophotometric, Distilled Water, Method Validation, ICH Guidelines.
INTRODUCTION:
Teneligliptin Hydrobromide Hydrate (INN; trade name Tenelia) is a pharmaceutical drug for treating type 2 diabetes mellitus. It belongs to the anti-diabetic drug class known as dipeptidyl peptidase-4 inhibitors, also known as "gliptins"1.
Teneligliptin Hydrobromide Hydrate is a Dipeptidyl peptidase-4 (DPP-4) inhibitor that has lately appeared as a new antidiabetic class that shows positive outcomes in improved glycemic control. By inhibiting DPP-4 enzymatic activity, DPP-4 inhibitors increase effective GLP-1 (Glucagon-like Peptide 1 Hormone) and GIP (Glucose-dependent insulin tropic peptide) concentrations, improving glucose-dependent hyperglycemia in diabetic patients by increasing serum insulin levels and decreasing serum glucagon levels2. DPP-4 inhibitors are promising drugs that can reduce glucose fluctuations in diabetic patients and have emerged as a new class of anti-diabetic agents3,4.
The IUPAC name of TeneligliptinHydrobromide Hydrate, [(2S,4S)-4-[4-(5-methyl-2-phenylpyrazol-3-yl) piperazin-1-yl] pyrrolidin-2-yl] -(1,3-thiazolidin-3-yl) methanone; hydrate; pentahydrobromide5.
Figure 1: Chemical structure of Teneligliptin Hydrobromide Hydrate (THH)6
Literature survey showed that various methods have been reported for the estimation of Teneligliptin hydrobromide hydrate usinganalytical methods like HPTLC7, RP-HPLC8-11, RP-UFLC12 and UVSpectrophotometry13-17. The objective of this Research is to develop and validate a simple, rapid, and economical UV spectrophotometric method for the determination of Teneligliptin Hydrobromide Hydrate in its bulk and in marketed brand and generic tablet dosage forms using distilled water as the solvent. Unlike some existing methods that use different solvents, our approach utilises distilled water as the solvent, aiming for simplicity and cost-effectiveness.
MATERIALS AND METHODS:
MATERIALS:
Instruments:
A double-beamUV-visible spectrophotometer (Lab India UV 3000+) configured with UV Win Software.
Cuvettes 1cm quartz cell was used to measure the absorbance of solutions.
Chemicals:
Brand and generic tablets of Teneligliptin were purchased from a local pharmacy in Sulur, Coimbatore and Each tablet is labelled to contain 20mg of Teneligliptin Hydrobromide Hydrate.
Pure drug of Teneligliptin Hydrobromide Hydrate.
Selection of solvent:
THH is freely soluble in Dimethyl Sulphoxide (DMSO), methanol, and water18. Hence Distilled water was selected as the solvent for dissolving Teneligliptin Hydrobromide Hydrate (THH).
Preparation of Standard Stock Solution:
Accurately weighed 100mg of Teneligliptin hydrobromide hydrate (THH) was transferred to a 100ml volumetric flask with 10ml distilled waterand dissolved. The volume was adjusted to 100ml with distilled water to obtain a standard solution having a concentration of 1000μg/ml.
Selection of wavelength of maximum absorbance (λmax):
1ml of the standard stock solution was transferred to a 100ml volumetric flask, and the volume was adjusted to 100ml with distilled water to obtain a Teneligliptin Hydrobromide Hydrate (THH) solution with a concentration of 10μg/ml. The solution was then scanned between 230 and 300nm against distilled water as a blank. The absorption maxima of THH were found to be at 244nm. (Figure 2)
Figure 2: Wavelength of maximum absorption of THH(λmax)
Preparation of calibration curve for Teneligliptin hydrobromide Hydrate:
1, 2, 3, 4, and 5 ml of the standard stock solution (1000 μg/ml) were pipetted into a series of 100 ml volumetric flasks. The volumes were then adjusted to 100 ml with distilled water to obtain solutions with concentrations of 10, 20, 30, 40, and 50 μg/ml of the drug. The absorbance of these resulting solutions was measured at 244 nm against distilled water as a blank. (Figure 3) (Table 1) A graph was plotted correlating the absorbance values with the concentrations of the solutions. (Figure 4)
Figure 3: Combined Spectrumof THH (10-50µg/ml)
Table 1: Linearity table of Teneligliptin Hydrobromide Hydrate.
S. No |
Concentration (μg/ml) |
Absorbance |
1. |
10 |
0.197 |
2. |
20 |
0.362 |
3. |
30 |
0.559 |
4. |
40 |
0.740 |
5. |
50 |
0.917 |
Figure 4: Linearity Graph of Teneligliptin Hydrobromide Hydrate.
Assay of Teneligliptin Hydrobromide Hydrate in Marketed Dosage form:
(Branded and Generic tablet dosage form):
Twenty tablets were weighed their average weight was calculated. The tablets were then finely powdered. Powder equivalent to 100mg of Teneligliptin Hydrobromide Hydrate (THH) was accurately weighed and transferred to a 100ml volumetric flask. The volume was adjusted to 100ml with distilled water and it was filtered through a Whatman filter paper grade 41.
Pipette out 3ml from this solution and transfer into a separate 100ml volumetric flask. The volumes were then adjusted to 100ml with distilled water to obtain a solution with a concentration of 30μg/ml of the drug. The absorbance of this solution was measured at 244nm against distilled water as a blank. The concentration of THH in the tablet formulation was estimated by the developed method. (Table 2)
Table 2: Assay of THH in tablet dosage form.
Tablet Form |
Label Claim |
Amount added |
Amount Found |
Assay % |
Branded Tablet |
20mg |
30μg/ml |
30.4μg/ml |
101.33% |
Generic Tablet |
20mg |
30µg/ml |
30.6µg/ml |
102% |
Method Validation:
The developed UV Spectrophotometric method was validated according to ICH19 to study different parameters.
Linearity:
Linearity was observed between absorbance and concentration in the working range of 10-50μg/ml THH.
Accuracy:
To ensure the accuracy of the method, recovery studies were carried out by adding standard solutions of THH to the pre-analyzed sample solution at 80%, 100%, and 120% of the assay concentration.
Precision:
Precision was determined by measuring the drug at a concentration (10μg/ml) 6 times on the same day is Intraday precision and the same concentration (10μg/ml) was determined for consecutive days is interday precision. RSD% values were then calculated.
Repeatability:
Repeatability, referred to as intra-assay precision, was determined by analyzing six samples of the same concentrations of drug (10μg/mL) under the same operating conditions over a short period.
Limit of Detection:
The detection limit was calculated using the standard deviation of absorbance of the same concentration, which is a standard solution of THH (10g/ml) prepared six times and LOD calculated by LOD = 3.3 (SD/S)
Where SD - is the standard deviation; S - is the slope of the curve.
Limit of Quantification:
The quantification limit was calculated using the standard deviation of the peak area at the same concentration, which was standard solution THH (10g/ml) prepared six times and LOQ calculated by LOD = 10 (SD/S)
Where SD - is the standard deviation; S - is the slope of the curve.
RESULT AND DISCUSSION:
Linearity:
The linearity of the proposed method was determined at ranging from 10-50μg/ml for THHand was found to be linear. The obtained regression analysis data is summarized. (Table 3)
Accuracy:
The accuracy of the developed method for THH was determined at 80%, 100% and 120% levels of standard solution. (Table 4)
Table 3: Linearity data of the developed method
Parameters observed |
Values |
Wavelength (nm) |
244nm |
Linearity Range (µg/ml) |
10-50 µg/ml |
Regression Equation (y = mx +c) |
y = 0.0182x + 0.0096 |
Correlation coefficient (r2) |
0.9994 |
Slope (m) |
0.0182 |
Intercept (c) |
0.0096 |
Table 4: Accuracy study of THH.
S. No |
Level of % recovery |
Amount of standard-stock solution added (ml) |
Amount added (µg) |
Amount found (µg) |
% Recovery |
1. |
80 |
0.8 |
8 |
7.87 |
78.7% |
2. |
100 |
1 |
10 |
9.68 |
96.8% |
3. |
120 |
1.2 |
12 |
11.76 |
117.6% |
Precision:
Precision was determined by measuring the drug a concentration (10μg/ml) for 6 times both intra day and interday (table 5). Percentage relative standard deviation was within the limit of ±2%. This indicated that the method was highly precise.
Table 5: Intraday and Interday Precision study of THH.
Intraday Precision. |
||||
S.No |
Concentration (µg/ml) |
Absorbance |
SD |
%RSD |
1. |
10 |
0.168 |
0.003061
|
1.8091%
|
2. |
10 |
0.173 |
||
3. |
10 |
0.170 |
||
4. |
10 |
0.171 |
||
5. |
10 |
0.164 |
||
6. |
10 |
0.169 |
||
Interday Precision. |
||||
S. No |
Concentration (µg/ml) |
Absorbance |
SD |
%RSD |
1. |
10 |
0.169 |
0.001789
|
1.0522%
|
2. |
10 |
0.169 |
||
3. |
10 |
0.173 |
||
4. |
10 |
0.171 |
||
5. |
10 |
0.168 |
||
6. |
10 |
0.170 |
Repeatability:
Table 6: Repeatability studyof THH.
S. No |
Concentration (µg/ml) |
Absorbance |
SD |
%RSD |
1. |
10 |
0.197 |
0.003033
|
1.5396%
|
2. |
10 |
0.200 |
||
3. |
10 |
0.193 |
||
4. |
10 |
0.201 |
||
5. |
10 |
0.195 |
||
6. |
10 |
0.196 |
Limit of detection (LOD) and limit of quantification (LOQ):
Table 7: Limit of Detection and Limit of Quantification of THH.
LOD (µg/ml) |
0.5439 µg/ml |
LOQ (µg/ml) |
1.648 µg/ml |
Summary:
Table 8: Summary of Developed method for THH.
S. No |
Parameters |
Values |
1. |
Linearity studies |
|
Wavelength (nm) |
244nm |
|
Linearity Range (µg/ml) |
10-50 µg/ml |
|
Regression Equation (y = mx +c) |
y = 0.0182x + 0.0096 |
|
Correlation coefficient (r2) |
0.9994 |
|
Slope (m) |
0.0182 |
|
Intercept (c) |
0.0096 |
|
2. |
Accuracy |
97.7% |
3. |
Precision (%RSD) |
|
Intraday Precision (%RSD) |
1.8091% |
|
Interday Precision (%RSD) |
1.0522% |
|
4. |
Repeatability (%RSD) |
1.5396% |
5. |
LOD (µg/ml) |
0.5439µg/ml |
6. |
LOQ (µg/ml) |
1.648µg/ml |
8. |
Assay % |
|
Branded tablet |
101.33% |
|
Generic tablet |
102% |
CONCLUSION:
A simple, precise, rapid and economical UV spectrophotometric method was developed and was found suitable for the determination of Teneligliptin Hydrobromide Hydrate (THH) in both bulk and marketed Brand and Generic Tablet Dosage forms. The percentage assay was in good agreement with the label claim and all results were validated according to the parameters of ICH guidelines and the results are within the limits. This method is applicable for the routine analysis of Teneligliptin Hydrobromide Hydrate (THH) in bulk and pharmaceutical dosage forms.
CONFLICT OF INTEREST:
The authors have no conflicts of interest regarding this investigation.
ACKNOWLEDGMENTS:
The authors are thankful to the Principal, Management and Department of Pharmaceutical Chemistry of RVS College of Pharmaceutical Sciences, Sulur, Coimbatore, Tamilnadu, India, for providing all the facilities throughout the work.
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Received on 12.12.2023 Revised on 08.04.2024 Accepted on 20.07.2024 Published on 10.12.2024 Available online on December 30, 2024 Asian Journal of Pharmaceutical Analysis. 2024; 14(4):217-221. DOI: 10.52711/2231-5675.2024.00039 ©Asian Pharma Press All Right Reserved
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