INTRODUCTION:

In the course of the pharmaceutical expansion of a new drug, it is essential to choose as early as promising formulation which having finest stability features. Guidelines regards with stability testing are provided by International Commission for Harmonization (ICH), which highlights the stress testing environments with prime purpose of evaluating the consequence of severe circumstances on the drug. Such kind of outcomes are play an crucial part in the estimation of a drug product shelf life throughout initial periods of its pharmaceutical growth. These results not only important for estimation of shelf life of drug but may also help as monitors for improved drug design, drug formulation and drug analysis.^1-6

Teneligliptin is a new drug, which has been used for the management of type 2 diabetes mellitus. It is an antidiabetic drug which belongs to dipeptidyl peptidase-4 inhibitors.⁷ Chemically, it is {(2S, 4S)-4- [4-(3-methyl-1-phenyl-1H-pyrazol-5-yl)-1-piperazinyl]-2- pyrrolidinyl} (1, 3-thiazolidin-3-yl) methanone. Fig. 1 shows that structure of Teneligliptine. Teneligliptin is permitted for use in India, Japan, and Korea in 2012.⁸ This antidiabetic drug show its action for approximately 24 h, with raise of activated glucagon-like peptide 1 (GLP-1) levels by conquering postprandial hyperglycemia after the meals. This drug showed a hopeful result in stabilizing the glycemic variations during the day and suppressing the diabetic difficulties.^9-10

The literature review revealed a simple UV spectroscopic method development and validation of teneligliptin HBr hydrate in tablet dosage form¹¹, and a stability indicating RP-HPLC method for development and validation of teneligliptin HBr hydrate in pure and tablet dosage for analytical method development.^12-13

Uptill there are, no UV stability indicating method has been described for the analysis of this drug in its pharmaceutical formulations. Amongst the several approaches existing for the determination of drugs, but still spectrophotometry is actual popular, due to its simplicity, specificity and low cost.

In this current study a simple, sensitive, selective, sensitive, economical, accurate and reproducible analytical method with superior finding range for estimation of Teneligliptine in pure form as well from pharmaceutical dosage form was developed and validated. Based on forced degradation studies, the method was also tested for its stability indicating capability permitting to the ICH requirements which can be used for the routine and quality control analysis of Teneligliptine in bulk and pharmaceutical formulation.

Fig.1: Chemical structure of Teneligliptine

MATERIAL AND METHODS:

Chemicals and reagents:

Teneligliptine was achieved as a gift sample from Ajanta Pharma Limited, Mumbai, India. All solvents and other chemicals used for above said work, were of analytical reagent grade procured from Research lab, Mumbai. Double distilled water used throughout experiment.

Instrumentation:

A projected work was conceded out on Lab India UV/VIS double beam spectrophotometer (model 3000+) with 1 cm matched quartz cells was used for all spectral measurements. All weighing was done on electronic balance (Sansui-Vibra MK-1890S).

Selection of Solvents:

On the basis of solubility distilled water used during the experiment as a solvent to dissolve the drug.

Preparation of Standard Stock Solution of Teneligliptin:

10 mg of Tenelig liptine was precisely weighed and transmitted to 100 ml volumetric flask and dissolved in around 20 ml of distilled water. The volume was completed up to the mark along with distilled water to give 100 µg/ml stock solution.

Preparation of calibration curve for Teneligliptine:

By scanning a suitable standard solution in the UV-VIS spectrophotometer in the wavelength range of 200-400 nm, the λ max of the drug was determined (Fig. 2). Aliquots (1,2,5ml) from standard solution of Teneligliptine were pipetted out in to a sequence of five volumetric flasks and the volume was made up to 10 ml with double distilled water. The absorbance was measured at 243 nm contrary to blank. The calibration curve was constructed by plotting absorbance v/s concentration (µg/ml). Correlation coefficient has been measured. The summary of analytical parameters and calibration curve data are presented in Table 1 and Table 2 respectively.

Estimation of Teneligliptine:

20 tablets of Tenure^® (Glenmark pharmaceutical Ltd.) were weighed correctly and powdered. Powder equivalent to 50 mg teneligliptin HBr hydrat been weighed and moved to a 100 ml volumetric flask. It was dissolved in 100 ml distilled water and sonicate for 15 minutes to get a uniform solution. Firstly it was filtered by 0.45μm Whatman filter paper. At last concentration of 100 μg/ml of teneligliptin has been prepared. This solution again filtered by the filter paper to eliminate certain un-dissolved excipients. After filtration, from this 2 ml was taken and diluted to 10 ml with distilled water which provides 20 μg/ml solution whereas absorbance of this solution was measured at 243 nm.

Method Validation:

The method was validated according to ICH Q2B guidelines to determine the Linearity, sensitivity, precision, and accuracy of the analyte.¹³Linearity of the recommended method was determined by determining the absorbance of the standard solutions in the concentration range of 5-25μg/ml and execution smallest square regression analysis. In addition, the accuracy of the proposed method has been patterned by standard addition method and recovery studies were passed out at 80%, 100% and 120% of required concentration. The percentage analytical recovery was calculated by comparing the concentration resulted with the addition of spiked samples with actual expected theoretical increase in concentration. Intra-day precision was determined by carrying out the analysis for six concentrations at two different time interval in a day. Similarly interday precision was determined by performing analysis on two consecutive days. LOD and LOQ of the proposed methods were calculated.^14-17 Recovery of the analyte of interest from a given matrix can be used as a measure of the accuracy or the bias of the method.^18-21

Stability Studies of Teneligliptine:

Stability studies were accomplished by forced degradation study of Teneligliptine and it comprises the study of consequence of photolysis, oxidation, temperature and prone to hydrolysis through a wide range of pH values. For acidic hydrolysis 0.1, 1.0 N HCl, for basic hydrolysis 0.1, 1 N NaOH, for neutral hydrolysis distilled water, for oxidation study 0.1%, 1% and 3% H₂O₂ was used. For carrying out photolysis studies the drug was treated by sunlight for 3 days and thermal stress was applied by heating the drug at 60⁰C for 2 hrs.

RESULTS AND DISCUSSION:

The development of a simple, economic, sensitive, and accurate analytical method for the quantitative determination of samples has been decrease needless sample preparations and the charge of materials as well labor. The λ max of Teneligliptine was taken in double distilled water is shown in (Fig 2).

For the the analysis of Teneligliptine λ max was measured at (243 nm) by making scans of the Teneligliptin solutions in the whole UV region. Data for calibration curve was made in the series of concentrations of 5-25 μg/ml. Beer’s law was followed above this concentration range (Fig 3). Overlain spectra for 5-25 μg/ml concentrations shown in (Fig 4).

The regression equation was found to be Y=0.0257x+0.0281. The correlation coefficient (r) of the standard curve was established 0.9991. The characteristic of the calibration plot is shown in Table 1. Performing repeat examines of the standard solutions was used to evaluate the accuracy and precision of the recommended methods (Table 3 and 4). The LOD and LOQ were found to be 0.19µg/ml and 0.83 µg/ml respectively.

To examine the accuracy of the projected technique and to check the interference due to excipients used in dosage forms, standard addition method has been used for recovery experiments. 100.10-100.20 is the average of recovery result has been concluded. The recommended methods can be effectively useful for assay in tablet dosage forms without any interference (Table 3).

Double distilled water was used for the performing calibration range and analyzed with the relevant calibration curves to determine the intra- and inter-day variability.

To determine the precision of the method Teneligliptine solutions at concentration 05, 15, 25 µg/ml were analyzed each in triplicate. Solutions for the standard curves were prepared fresh daily. The method was found to be precise. The % RSD values for interday precision at concentration 05, 15, 25 µg/ml was found to be 0.995, 0.012, 0.991 respectively and for intraday precision it was 0.451, 0.320, 0.128 respectively. Results are shown in (Table 4).

The use of this technique is elucidated in the experimental segment. The achieved consequences establish the validity and accuracy of the recommended method for the determination of Teneligliptine in sachets. The stability studies indicates that considerable alterations were detected by treating the drug with sun light, thermal stress, oxidation, acid, basic hydrolysis and neutral hydrolysis, however there was substantial variation with all these stress conditions. The results are summarize (Table 5).

These results disclose that the developed method was simple, sensitive, sensitive, inexpensive, accurate and reproducible and subsequently, can be applied to the determination of Teneligliptine tablet in pharmaceuticals deprived of any interference from the excipients. Based on forced degradation studies according to the ICH requirements, this method can be used for the routine and quality control analysis of Teneligliptine in raw material and pharmaceutical formulations.

Fig.2: UV spectrum of Teneligliptine

Table 1: Optical characteristics of the proposed method

Parameters	Result
Measured wavelength (λ max)	243 nm
Beers law limit (µg/ml)	5-25 ppm
Regression equation (y = m x + c)	Y=0.0257x+0.0281
Slope	0.0257
Intercept	0.0281
Correlation coefficient ®	0.9991
LOD µg/ml	0.19
LOD µg/ml	0.83

Table 2: Calibration curve data for Teneligliptine

Sr. No.	Conc. (µg/ml)	Absorbance
1	5	0.153
2	10	0.283
3	15	0.419
4	20	0.548
5	25	0.662

Fig.3: Calibration curve of Teneligliptine at 243 nm

Table 3: Result of recovery studies

Level of % recovery	*% Mean recovery**	S.D.	R.S.D	SE
Level of % recovery	Teneligliptine
80	100.10	0.610	0.617	0.338
100	100.11	0.510	0.517	0.301
120	100.20	0.475	0.465	0.291

*Mean of three determinations at each, SE- Standard error

Table 4: Statistical validation for interday and intraday precision

Parameters

Concentrations (µg/ml)

Intraday*

% mean±S.D

%RSD

2.06±0.208

0.995

0.0352

50.22±0.602

0.012

0.0405

79.77±0.769

0.991

0.0437

Interday*

% mean±S.D

%RSD

20.08 ± 0.32

0.451

0.0299

50.77±0.16

0.320

0.0456

80.47±0.119

0.128

0.0424

*Denotes average of three determinations, SE-Standard error

CONCLUSION:

A simple, sensitive, accurate, precise, reproducible and cost effective stability indicating UV spectrophotometric method has been established for quantitative determination of Teneligliptine in bulk and pharmaceutical formulation. This method decreases general method development period and solvents. Forced degradation studies comprises the influence of temperature, oxidation, neutral, photolysis and susceptibility to hydrolysis through extensive range of pH values, were accepted out approving to the ICH necessities which can be used for the routine and quality control analysis of Teneligliptine bulk as well in pharmaceutical formulations.

ACKNOWLEDGEMENT:

Authors are thankful to the trustees Hon. Prasad Bapu Hire of KBHSSTrust’s Institute of Pharmacy, Malegaon, Nashik for providing essential facilities.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

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Received on 17.12.2018 Accepted on 18.02.2019

Asian J. Pharm. Ana. 2019; 9(3):128-132.

DOI: 10.5958/2231-5675.2019.00024.3

Sr. No.	Situations useful	Conc. Taken(µg/ml)	Average Conc. Found (µg/ml)	Observation
1	Acidic hydrolysis (0.1, 1 N HCl)	20 µg/ml	19.94 µg/ml	Stable
2	Basic hydrolysis (0.1, 1 N NaOH)	20 µg/ml	8.33 µg/ml	Degraded
3	Neutral Hydrolysis	20 µg/ml	14.15 µg/ml	Degraded
4	H₂O₂(0.1, 1, 3%)	20 µg/ml	Change in λ max	Stable
5	Thermal stress (60⁰ C, 2 hrs)	20 µg/ml	Change in λ max	Degraded
6	Sunlight treatment 1 Day	20 µg/ml	17.00 µg/ml	Degraded
	Sunlight treatment 2 Day	20 µg/ml	13.50 µg/ml	Degraded
	Sunlight treatment 3 Day	20 µg/ml	10.95 µg/ml	Degraded