Journal of Population Therapeutics

& Clinical Pharmacology

RESEARCH ARTICLE
DOI: 10.47750/jptcp.2023.30.13.026

Development of an RP-HPLC Method to estimation Glimepiride in Bulk and

Solid Dosage Form
Khaldoon S. Alhadad1, H. N. K. AL-Salman*2
1,2
Department of Pharmaceutical Chemistry, College of Pharmacy, University of Basrah, Iraq.
*Corresponding Author: H. N. K. AL-Salman, Department of Pharmaceutical Chemistry,
College of Pharmacy, University of Basrah, Iraq. Email: [email protected]

Submitted: 24 March 2023; Accepted: 10 April 2023; Published: 13 May 2023

ABSTRACT
AIM- Two straightforward and touchy RP-HPLC techniques for the quantitative estimation of
Glimepiride in mass and drug portion structures have been contrived in this review. MATERIAL
& METHODS- Glimepiride was resolved utilizing a RP-HPLC technique using a C-18 evenness
section and a portable period of methanol: phosphate support pH 4.0 (50:50 V/V). The versatile
stage was siphoned at a pace of 0.5 ml/min, and the location was done at a frequency of 239 nm.
RESULTS- 2.470 minutes was discovered to be the retention time. Linearity, Accuracy,
Precision, System Suitability, LOD, LOQ, Ruggedness, and Robustness are all validated for this
approach. The proposed method is an excellent way to get accurate results and has been
demonstrated to be suitable for regular Glimepiride analysis in bulk and dose forms.
CONCLUSION- The method was used to find out how much of a compound was included in
commercial pharmaceutical dosage forms. The approach is straightforward, repeatable, and
accurate, and it is a better option for routine quality control than other chromatographic
techniques.

Keywords: RP-HPLC Method, Estimation of Glimepiride, Bulk & Solid Dosage Form, Linearity,
Accuracy, Precision, System suitability

INTRODUCTION is the most broadly utilized insightful

HPLC is the most adaptable and generally innovation [1, 2]. HPLC works by
utilized chromatographic procedure. It is an constraining dissolvable through shut sections
actual division procedure in the fluid stage containing tiny particles under high tension,
where an example is isolated into its bringing about high goal detachments. In a
constituent parts (or analytes) by conveying scope of natural, inorganic, and organic
between the portable stage (a streaming fluid) materials, the system is utilized to isolate and
and the fixed stage (a fixed strong) (sorbents decide species. The motivation behind this
pressed inside a segment). A web-based study is to make an UV/noticeable
identifier creates a chromatogram by checking Spectrophotometric and RP-HPLC technique
the centralization of each isolated part in the for assessing Glimepiride in mass and drug
segment gushing. For the quantitative definitions [3-6].
examination of drugs, biomolecules,
polymers, and other natural substances, HPLC
J Popul Ther Clin Pharmacol Vol 30(13):e265–e270; 13 May 2023.
This article is distributed under the terms of the Creative Commons Attribution-Non
Commercial 4.0 International License. ©2021 Muslim OT et al.

e265
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Development of an RP-HPLC Method to estimation Glimepiride in Bulk and Solid Dosage Form

MATERAIL & METHODS channel paper was utilized to channel the

We concocted a straightforward and delicate RP- resultant arrangement [10].
HPLC technique for quantitative estimation of Columns must be cleaned
Glimepiride in mass medications and drug plans Molding of the segments was performed before
in the ongoing review. to the following HPLC show to streaming HPLC
Chromatographic system and conditions grade methanol through them at a stream pace of
The proposed procedure was carried out under 1 ml/min for 30 minutes. In order to erase any
the aforementioned chromatographic conditions. remainders of the earlier run that might be
Mobile phase selection available in the segment.
Glimepiride was put into the HPLC system as a Mobile phase loading
pure drug and ran in various solvent systems. In The pipe was filled with filtered and degassed
order to discover the optimal conditions for the mobile phase. Each channel was primed
separation of Glimepiride, different portable separately with newly prepared mobile phase.
stages, for example, methanol and water, Validated RP-HPLC method
methanol and different phosphate cushions with An insightful technique's approval is the most
pH of 6, 5.5, and 3.5 were investigated. In common way of laying out through research
comparison to other mobile phases, methanol and facility examinations that the strategy's
potassium dihydrogen orthophosphate buffer exhibition trademark meets the prerequisites for
(pH: 4) yielded good results. At long last, the expected logical application. Analytical
methanol and phosphate cushion were parameters are used to express performance
demonstrated to be the best versatile stage parts characteristics [11, 12].
(pH: 4). The medication had high resolution, a Linearity
moderate retention period, and adequate peak Proper aliquots of standard Glimepiride stock
symmetry in this mobile phase [7]. arrangements (100 g/ml) were set in different 10
Preparation of phosphate buffer ml volumetric flagons, and the subsequent
7.0 gm KH2PO4 was weighed into a 1000 ml arrangement was weakened sufficient with
volumetric flask, dissolved, and diluted with diluent to get a last centralization of 10-50 g/l.
HPLC water to 1000 ml. Ortho phosphoric acid The chromatographic apparatus was injected
was used to get the pH to 4.0 [8]. with these solutions. The Glimepiride calibration
Preparation of mobile phase curve was created by graphing peak area vs.
Blend 500 mL of the previously mentioned Glimepiride applied concentration [13].
cushion (half) with 500 mL of methanol HPLC Precision
(half) and degas in a ultrasonic water shower for Between day and intra-day difference tests
5 minutes. Under vacuum filtration, channel demonstrated the technique's accuracy. Six
through a 0.45 channel [9]. rehashed infusions of standard arrangement were
Preparation of working standard stock solution done in the intra-day tests, and the reaction
Around 10 mg of Glimepiride was painstakingly component of medication pinnacle and percent
gauged and disintegrated in 50 ml of methanol in RSD were determined. A chromatogram was
a 100 ml volumetric jar, then, at that point, shown. Six rehashed infusions of standard
weakened to the ideal focus with methanol. arrangement were finished six successive days in
Whatman channel paper No. 41 was utilized to the between day variety preliminaries, and the
channel the subsequent arrangement. reaction component of meds pinnacle and
Sample Solution Preparation percent RSD were figured. The made technique
Twenty tablets were painstakingly pummeled was viewed as exact in light of the information
and unequivocally weighted. In a 100 ml got [14].
volumetric flagon containing methanol (around Accuracy
50 ml), tablet powder comparable to 10 mg of The method's accuracy refers to how near the test
Glimepiride was broken down by sonication and findings are to the genuine value. Twenty tablets
separated utilizing Whatman channel paper (No. of each formulation were weighed and
41). The filtrate was gathered by washing the pulverised, and the results were analysed to see
channel paper with extra dissolvable. To how accurate they were. Recuperation
accomplish a centralization of 100 g/ml, the examinations were led out utilizing the standard
filtrate volume was changed in accordance with option strategy, which included adding a known
the imprint with a similar dissolvable. Whatman measure of standard medication answer for the
J Popul Ther Clin Pharmacol Vol 30(13):e265–e270; 13 May 2023.
This article is distributed under the terms of the Creative Commons Attribution-Non
Commercial 4.0 International License. ©2021 Muslim OT et al.

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Development of an RP-HPLC Method to estimation Glimepiride in Bulk and Solid Dosage Form

example arrangement (50, 100, and 150 percent).

Limit of detection (LOD)
Utilizing the made RP-HPLC technique, the
fostered strategy's Limit of Detection not entirely
set in stone by infusing progressively lower
measures of standard arrangements. The LOD is
the analyte focus at which a perceptible reaction
can be gotten (sign to commotion proportion of
3:1) [15, 16].
Limit of quantitation (LOQ) FIGURE 2: RP-HPLC Calibration curve of
The LOQ values were derived using a three-fold Glimepiride
multiplication method based on the LOD
strength.
Ruggedness
Glimepiride tests weighing 10 mg were gauged
and broken down in a 100 ml volumetric cup
containing portable stage (50 ml), then sonicated
for 30 minutes prior to making the last volume
with versatile stage. 3 ml of the standard stock
arrangement was pipette out and saved into a 10
ml volumetric cup, which was then loaded up
with versatile stage to the ideal volume. FIGURE 3: Overlain chromatogram of
Infusions of the examples were made into the Glimepiride by RP-HPLC method
segment [15].
Robustness
Various technique boundaries, for example, pH,
stream rate, segment temperature, infusion
volume, and versatile stage creation, are changed
inside a practical reach to test a strategy's vigor,
and the quantitative impact of the not set in
stone. There were no massive changes in the
chromatograms, exhibiting the heartiness of the
RP-HPLC strategy laid out [16]. FIGURE 4: Chromatogram of precision

RESULTS

FIGURE 5: Chromatogram of standard drug i.e.

Glimepride solution

TABLE 1: Adjustment information of

FIGURE 1: Standard HPLC Chromatogram of
Glimepiride by RP-HPLC strategy
Glimepiride at 239 nm S. No. Concentration (µg/ml) Area
1 0 0
2 10 10734
3 20 21576
4 30 32967
5 40 43029
6 50 54803

J Popul Ther Clin Pharmacol Vol 30(13):e265–e270; 13 May 2023.

This article is distributed under the terms of the Creative Commons Attribution-Non
Commercial 4.0 International License. ©2021 Muslim OT et al.

e267
Development of an RP-HPLC Method to estimation Glimepiride in Bulk and Solid Dosage Form

TABLE 2: Glimepiride characterization parameters

Parameter RP-HPLC
Calibration range (µg/ml) 10-50
Detection wavelength (nm) 239
Mobile phase (methanol: 0.051 M) phosphate 50:50
buffer) (v/v; pH: 4.0)
Retention time (min) 2.470±0.035
Regression equation (Y*) Y=109290 X -17080
Slope (m) 109293
Intercept (c) -17082
Correlation coefficient (r2) 0.9961
Limit of detection (µg/ml) 0.02
Limit of quantitation (µg/ml) 0.07

TABLE 3: RP-HPLC technique precision results for Glimepiride

S. No. Concentration (µg/ml) Intraday Interday
1 30 3305558 3305441
2 30 3292266 3297255
3 30 3293512 3302419
4 30 3285834 3302324
5 30 3296448 3302637
6 30 3298422 3299541
Avg. - 3295355 3301622
SD* - 6606.34 2836.59
%RSD* - 0.21 0.084

TABLE 4: Glimepiride recovery data using the RP-HPLC method

Level of Amount of drug Amount of drug
recovery (%) added (µg) Recovered (µg)* % Recovery ± SD*
50 5 4.96 99.3±0.42
100 10 9.99 99.4±0.38
150 15 14.79 98.1±0.49

TABLE 5: Robustness studies of Glimepiride by changing the flow rate

Flow Rate System Suitability Results Retention Time (tR)
Sl. No (ml/min) Plate Count Tailing
1 0.4 2812 1.2 2.736
2 *0.5 2874 1.3 2.473
3 0.6 2798 1.2 2.224

TABLE 6: RP-HPLC technique was utilized to decide the roughness of Glimepiride

Analyst I Analyst II
Label Claim (mg) Amount found (mg) % Recovery Amount found (mg) % Recovery
Samples ± SD** ± SD**
Amryl 1 1.0037 100.38±0.0732 1.0027 100.27± 0.0916
Glemstar 1 1.0025 100.26±0.1146 1.0024 100.24±0.0794

DISCUSSION as the maintenance time. In the fixation scope of

Due to their significance in quality control of 10-50 g/ml, when the centralizations of
prescriptions and medication items, the Glimepiride and their particular pinnacle regions
improvement of a logical technique for deciding were exposed to relapse examination utilizing the
medications by RP-HPLC has drawn in a ton of least squares strategy, a decent direct relationship
consideration lately. The objective of this work (r2= 0.9997) was seen between the convergence
was to make a RP-HPLC technique for dissecting of Glimepiride and their separate pinnacle
Glimepiride in mass drug and drug portion regions. Glimepiride's relapse condition was
structure using the most normally utilized RP-C found to be Y= 109290 X-17080, where 'Y'
18 section with UV discovery [17]. addresses the pinnacle region and 'X' addresses
Each example was infused multiple times, with a the grouping of Glimepiride.
5-minute run span. 2.470 0.035 min was viewed In the focus scope of 10-50 g/ml, linearity was
J Popul Ther Clin Pharmacol Vol 30(13):e265–e270; 13 May 2023.
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Development of an RP-HPLC Method to estimation Glimepiride in Bulk and Solid Dosage Form

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J Popul Ther Clin Pharmacol Vol 30(13):e265–e270; 13 May 2023.

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Commercial 4.0 International License. ©2021 Muslim OT et al.

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