Sigma Institute of Pharmacy, Sigma University, Vadodara, Gujarat, India 390019
One second-generation sulfonylurea that is frequently given to treat type 2 diabetes mellitus is gliclazide. Reliable analytical techniques are crucial for guaranteeing the efficacy, safety, and purity of gliclazide in pharmaceutical formulations. For drug analysis, High-Performance Thin-Layer Chromatography (HPTLC) has become widely accepted as a quick, easy, accurate, and economical method. An overview of documented HPTLC techniques created and approved for the quantitative measurement of gliclazide, both alone and in conjunction with other antidiabetic medications, is provided in this paper. Along with validation parameters like accuracy, precision, linearity, specificity, robustness, and sensitivity in compliance with regulatory guidelines, important aspects of method development are covered, including the choice of stationary and mobile phases, detection wavelength, and chromatographic conditions. The review also emphasizes the useful benefits of HPTLC, including its minimal solvent consumption, capacity to analyze numerous samples at once, and suitability for regular quality control. All things considered, this review is a helpful resource for academics and analysts working on gliclazide analysis using HPTLC.
Introduction to Type-2 Diabetes Mellitus:[1]
Blood glucose levels rise in diabetes mellitus as a result of either inadequate insulin synthesis or poor insulin action. Type 2 diabetes is growing more widespread globally and is mostly caused by genetic factors and unhealthy lifestyle choices. This disorder causes a disruption in the usual balance between insulin and glucagon, which results in chronically elevated blood sugar levels and long-term problems that impact multiple organs.
Pathophysiology:[2]
This explanation explains both the anomalies associated with type 2 diabetes mellitus (T2DM) and the normal management of blood glucose. When glucose enters the bloodstream following a meal, the beta cells in the pancreas are stimulated to release insulin. Insulin makes it easier for bodily tissues like the muscles and liver to absorb glucose for storage or energy generation. Insulin production decreases when blood glucose levels drop.
Due to insufficient insulin release from beta cells or decreased body tissue responsiveness to insulin (insulin resistance), this regulatory system is compromised in type 2 diabetes. Consequently, there is a reduction in the uptake of glucose, which results in chronic hyperglycemia—a condition where blood glucose levels are consistently high.
Causes:[28]
Treatment:[29]
Introduction to Gliclazide:[3]
1-(3-azabicyclo[3.3.0]oct-3-yl)-3-(p-tolylsulfonyl) urea is the chemical name for gliclazide. Type 2 Diabetes Mellitus is treated with this oral sulfonylurea antidiabetic medication. Although it is not sold in the US, the medication was patented in 1966, received worldwide permission for medical use in 1972, and is sold under a number of brand names. By increasing the release of insulin from the pancreatic beta cells and strengthening overall glycemic control, it lowers blood glucose.
Mechanism of Action of Gliclazide:
Figure 1: Mechanism of Action of Gliclazide[7]
HPTLC (High-Performance Thin-Layer Chromatography) [4,5]
A sophisticated analytical method for both qualitative and quantitative analysis is called High-Performance Thin-Layer Chromatography (HPTLC). It improves resolution, accuracy, and precision by using pre-coated plates with thin layers and small particles.
Compounds migrate at varying rates according to their affinity for the stationary phase in HPTLC, and separation takes place while the mobile phase moves by capillary action. Densitometric scanning under UV or visible light, solvent migration, and precise sample application are all part of the procedure. HPTLC is more reproducible than traditional TLC and is appropriate for routine quality control and regulatory applications.
Steps in the HPTLC Process:
Figure 2: Camag Linomat Applicator[30] Figure 3: Developing Chamber[31]
Figure 4: TLC Scanner[32] Figure 5: Digital Camera[33]
INSTRUMENTATION OF HPTLC
DRUG PROFILE
Drug profile of Gliclazide: [6]
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Physicochemical Properties of Gliclazide |
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Name |
Gliclazide |
|
IUPAC Name |
3-[(3aR,6aS)-octahydrocyclopenta[c]pyrrol-2-yl]-1-(4-methylbenzenesulfonyl)urea |
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Category |
anti-diabetic drug |
|
Class |
Sulfonylurea (oral antihyperglycemic agent) |
|
CAS no. |
21187-98-4 |
|
Molecular Formula |
C15H21N3O3S |
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Structural Formula |
|
|
Molecular Weight |
323.41 g/mol |
|
Official Status |
Included in British Pharmacopoeia (BP), European Pharmacopoeia (EP) |
|
Appearance |
Solid crystalline powder |
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Solubility |
Approximately 0.19 mg/mL in water |
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pKa |
Around 4.07 (strongest acidic), 1.38 (strongest basic) |
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Melting point |
180-182 °C |
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Partition Co-efficient (log p) |
2.6 |
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Therapeutic Properties of Gliclazide |
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Uses |
Treatment of type 2 diabetes mellitus to lower blood glucose by stimulating insulin secretion from pancreatic beta cells and improving peripheral insulin sensitivity |
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Side Effects |
Common side effects: hypoglycemia, dizziness, headache, nauseaSevere effects may include allergic reactions and blood disorders. |
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Dose and Dosage form |
Available as oral tablets, usually in doses of 40-120 mg.Dose adjusted based on glucose control and patient response |
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Pharmacokinetic Properties of Gliclazide |
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Absorption |
After oral treatment, gliclazide is effectively absorbed and reaches peak plasma levels in two to eight hours. Food may reduce the peak concentration and somewhat postpone absorption. The drug's distribution is primarily limited to extracellular fluids due to its modest volume of distribution (13–24 L) and high plasma protein binding (85–97%). |
|
Distribution |
Gliclazide shows a low volume of distribution upon absorption, ranging from roughly 13 to 24 liters, suggesting a restricted distribution mostly to extracellular fluid. With protein binding typically ranging from 85% to 97%, it is strongly bound to plasma proteins. In addition to limiting quick clearance, this substantial protein binding limits the amount of free medication available in the bloodstream for activity. |
|
Metabolism |
CYP2C9 and CYP2C19 enzymes primarily hydroxylate and oxidize gliclazide in the liver. The parent medication is primarily in charge of controlling blood sugar levels because its metabolites have little or no hypoglycemic impact. Additionally, recent research suggests that gut bacteria may contribute to the formation of other hydroxylated metabolites with distinct pharmacokinetic characteristics. |
|
Excretion |
About 60–70% of gliclazide metabolites are excreted in the urine through the renal system. Ten to twenty percent of the medicine is eliminated in feces, mostly as metabolites rather than the drug itself. The medicine undergoes an effective metabolic transformation in the liver, as evidenced by the fact that less than 1% of it remains intact in urine. |
|
Half life |
About 60–70% of gliclazide metabolites are excreted in the urine through the renal system. Ten to twenty percent of the medicine is eliminated in feces, mostly as metabolites rather than the drug itself. The medicine undergoes an effective metabolic transformation in the liver, as evidenced by the fact that less than 1% of it remains intact in urine. |
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Toxicity |
Gliclazide's primary toxicity risk is hypoglycemia, which can result in headache, dizziness, sweating, confusion, and in extreme situations, coma. Due to delayed medication excretion, this risk rises with overdose or in individuals with liver and kidney disease. Studies on animals reveal a large oral LD50, indicating that when used properly, gliclazide has a wide safety margin. |
|
Protein binding |
94% |
LITERATURE REVIEW
A One-Time Literature Review on Analytical Methods of Gliclazide.
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Sr. No. |
Title |
Method |
Description |
Ref. No. |
|
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Gliclazide Prolonged-release Tablets – BP 2025 (HPLC (Reversed-phase method)) |
Official Method |
Column: Macherey-Nagel Nucleosil 100-5 C8 (250 mm × 4.0 mm, 5 µm) Mobile Phase: Triethylamine : Trifluoroacetic acid : Acetonitrile : Water (0.1 : 0.1 : 40 : 60 v/v/v/v) Diluent: Acetonitrile : Water (45 : 55 v/v) Flow Rate: 0.9 mL/min λ max: 235 nm |
8 |
|
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Method development and validation for estimation of Gliclazide in bulk and tablet form by UV Spectrophotometer |
UV |
Solvent: Methanol λ max: 232 nm Range: 2–20 µg/mL |
9 |
|
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Ultraviolet-visible Spectrophotometric Method for Estimation of Gliclazide in Presence of Excipients Interacting in UV-visible Region
|
UV |
λ max: 226 nm, 221 nm, 231 nm Range: 4–28 µg/mL |
10 |
|
|
Development and validation of UV spectrophotometric method for the determination of Gliclazide in tablet dosage form |
UV |
λ max: 229.5 nm Range: 7–27 µg/mL |
11 |
|
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UV Spectrophotometric Method Development and Validation for the Estimation of Gliclazide in Bulk and Pharmaceutical Dosage Form |
UV |
Solvent: methanol + distilled water (50:50) λ max: 224.05 nm Range: 2–20 µg/mL |
12 |
|
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Spectrophotometric Quantification of Gliclazide in Pharmaceutical Dosage Form |
UV |
Solvent: Methanol λ max: Method A (Gliclazide + Cresol Red): 510 nm Method B (Gliclazide + Bromophenol Blue): 445 nm Range: Method A: 30–200 µg/mL Method B: 70–230 µg/mL |
13 |
|
|
New Spectrophotometric Gliclazide Determination in Tablets Formulations by Charge Transfer Complexation |
UV |
Solvent: Toluene λ max: 415 nm Range: 4.85 – 113.19 µg/mL |
14 |
|
|
Analytical Method Development and Optimization on High Performance Liquid Chromatography for Related Substances Test of Gliclazide Drug as Active Pharmaceutical Ingredient |
HPLC |
Stationary Phase: Reverse Phase (C18, assumed standard RP-HPLC column) Mobile Phase: 0.1% Potassium Phosphate in Water : 0.1% Potassium Phosphate in Methanol (MeOH) Flow Rate: Optimized for precise separation λ max: ~226–230 nm for Gliclazide |
15 |
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UV and RP-HPLC Method Development of Gliclazide in Tablet Dosage Form and it’s Validation
|
RP-HPLC |
Stationary Phase: C18 column (250 mm × 4.6 mm, 5 µm particle size) Mobile Phase: Propylene Glycol : Water (pH 4.5, adjusted with Orthophosphoric Acid) : Methanol = 50:40:10 Flow Rate: 1.0 mL/min λ max: UV Detector at 215 nm Range: 10–50 µg/mL |
16 |
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A RP-HPLC Method Development and Validation for the Estimation of Gliclazide in bulk and Pharmaceutical Dosage Forms |
RP-HPLC |
Stationary Phase: Symmetry C18 column Mobile Phase: Methanol : Phosphate Buffer = 50:50 (v/v) Flow Rate: 1.2 mL/min λ max: UV Detector at 210 nm Range: 1–100 µg/mL |
17 |
|
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Analytical Method Development and Validation of Gliclazide using RP-HPLC from Pharmaceutical Dosage Form |
RP-HPLC |
Stationary Phase: Hypersil OSD C18 column (4.6 mm × 250 mm, 5 µm) Mobile Phase: Phosphate Buffer : Acetonitrile = 10:90 (v/v), pH 3 Flow Rate: 1.0 mL/min λ max: UV Detector at 228 nm |
18 |
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Development and validation of a new analytical HPLC method for simultaneous determination of the antidiabetic drugs, metformin and gliclazide
|
HPLC |
Stationary Phase: Alltima CN column (250 mm × 4.6 mm, 5 µm) Mobile Phase: 20 mM Ammonium Formate Buffer (pH 3.5) : Acetonitrile = 45:55 (v/v) λ max: UV Detector at 227 nm Range: Gliclazide: 1.25–150 µg/mL Metformin: 2.5–150 µg/mL |
19 |
|
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HPLC Method for Determination of Gliclazide in Human Serum |
HPLC |
Stationary Phase: C18 column Mobile Phase: Acetonitrile : Methanol : Water = 50:30:20 (v/v), pH 3 Flow Rate: Gliclazide: 4.85 min Internal Standard (Phenytoin): 3.8 min λ max: UV Detector at 230 nm Range: 50–10,000 ng/mL |
20 |
|
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Stability Indicating Analytical Method Development and Validation for Assay of Gliclazide in Tablet Dosage Form by using Reverse Phase High Performance Liquid Chromatography |
RP-HPLC |
Stationary Phase: C18 column (150 × 4.6 mm, 5 µm) Mobile Phase: Water : Acetonitrile : Methanol : TFA : TEA = 40:40:20:0.1:0.1 (v/v) Flow Rate: 1.5 mL/min λ max: UV Detector at 235 nm |
21 |
|
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Development and Validation of HPTLC Method for Determination of Gliclazide in API and Pharmaceutical Dosage Form |
HPTLC |
Stationary Phase: Silica Gel 60 F??? TLC plate Mobile Phase: Toluene: Chloroform: Methanol (4:4:2 v/v) λ max: 230 nm Range: 4–14 ng/spot |
22 |
|
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Simultaneous HPTLC analysis of Gliclazide and Metformin hydrochloride in bulk and tablet dosage form |
HPTLC |
Stationary Phase: Silica Gel 60 F??? TLC plate Mobile Phase: Toluene : Acetonitrile : Ethanol : Ammonium Sulphate (0.25%) (4:4:4:3 v/v/v/v) λ max: 228 nm Range: 200–1000 ng/spot for both GLZ and MET |
23 |
|
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Simultaneous HPTLC Determination of Gliclazide and Rosiglitazone in Tablets |
HPTLC |
Stationary Phase: Precoated Silica Gel 60 F??? TLC plate Mobile Phase: Toluene : Ethyl acetate : Methanol (85:5:10 v/v/v) λ max: 225 nm Range: Gliclazide: 1–3 µg/spot Rosiglitazone: 0.05–0.15 µg/spot |
24 |
|
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Simultaneous Determination of Metformin Hydrochloride in its Multicomponent Dosage forms with Sulfonyl Ureas like Gliclazide and Glimepiride using HPTLC |
HPTLC |
Stationary Phase: Precoated Silica Gel 60 F??? Mobile Phase: Ammonium sulfate (0.25%) : Methanol : Ethyl acetate (10.0 : 2.5 : 2.5 v/v/v) Range: Gliclazide: 100–500 ng/mL Metformin hydrochloride (Combination I): 1000–5000 ng/mL Glimepiride: 300–500 ng/mL Metformin (Combination II): 150000–250000 ng/mL |
25 |
|
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Liquid Chromatography – Mass Spectrometry Method for the Determination of Gliclazide in Human Plasma and Application to a Pharmacokinetic Study of Gliclazide Sustained Release Tablets |
LC-MS |
Column: C18 Mobile Phase: Acetonitrile–water containing 10 mmol/L ammonium acetate 75:25 (v/v) Flow Rate: 1.0 mL/min Range: 0.025–2.5 µg/mL |
26 |
|
|
Liquid Chromatography-Tandem Mass Spectrometry Method for the Estimation of Gliclazide in Human Plasma: Application to Bioequivalence Study |
LC-MS |
Column: C18 Mobile Phase: Methanol : Water : Formic acid (90 : 10 : 0.1, v/v/v) Flow Rate: Optimized for rapid separation Range: 20–9125 ng/mL |
27 |
CONCLUSION
The documented HPTLC techniques created and approved for the quantitative measurement of gliclazide are compiled in this review. According to the literature, HPTLC is a straightforward, precise, accurate, and affordable method that can be used to analyze gliclazide in pharmaceutical dosage forms and bulk medications. The examined techniques work well in normal quality control and meet regulatory validation requirements. All things considered, HPTLC is a dependable and valuable analytical instrument for gliclazide estimate and a helpful resource for upcoming analytical and research projects.
REFERENCES
Drashti Pandya, Dr. Mitali Dalwadi, Kajal Vable, Dr. Priyanka Patil, A Review on HPTLC Method Development and Validation for Quantitative Estimation of Gliclazide, Int. J. of Pharm. Sci., 2026, Vol 4, Issue 1, 885-895. https://doi.org/10.5281/zenodo.18200888
10.5281/zenodo.18200888
