A Review on Reported Analytical Method Development by UPLC & HPLC of Selected Anti-Diabetic Drugs

Sayyad Simran Shabbir *, Shaikh Sadiya Mehmood, Ansari Shaheenanjum Babuddin, Pathan Najiya Shahnoor, Dr. Aejaz Ahmed

J.I.I.U' S Ali Allana College of Pharmacy Akkalkuwa, Dist. Nandurbar (425415), Maharashtra, India

INTRODUCTION

A contemporary method called UPLC offers liquid chromatography a new path. Ultra performance liquid chromatography, or UPLC, is characterised by improvements in three key areas: sensitivity, speed, and resolution. High-performance liquid chromatography (HPLC) lacks the resolution, speed, and sensitivity of ultra performance liquid chromatography (UPLC), which is applicable to particles smaller than 2 μm in diameter. Pharmaceutical companies are looking for innovative ways to boost economy and speed up drug development in the twenty-first century. Analytical laboratories are not an exception to this trend, as UPLC analysis currently yields better product quality. At pressures as high as 100 million Pa, UPLC uses extreme pressures to perform separation and quantification. In contrast to HPLC, it has been observed that high pressure has no detrimental effects on the analytical column. Additionally, UPLC uses less time and solvent overall.

An important qualitative and quantitative method for estimating biological and pharmaceutical samples is high performance liquid chromatography (HPLC). For the purpose of ensuring the quality control of drug components, this chromatographic technique is the most reliable, fast, safe, and adaptable.

The word "diabetes mellitus" refers to a multifactorial metabolic illness marked by persistent hyperglycaemia and abnormalities in the metabolism of proteins, fats, and carbohydrates brought on by deficiencies in either or both of the actions or secretion of insulin. Diabetes mellitus causes long-term harm as well as organ failure and dysfunction. Diabetes mellitus can cause thirst, polyuria, blurred vision, and weight loss, among other symptoms. In its most severe forms, ketoacidosis or a non– ketotic hyperosmolar state may develop and lead to stupor, command, in absence of effective treatment, death1,2. Frequently, there may be no symptoms at all or only mild ones, which means that hyperglycaemia severe enough to induce pathological and functional alterations may exist for a long time before a diagnosis is made. Diabetes mellitus causes long-term consequences that include retinopathy, which can result in blindness, nephropathy, which can cause renal failure, and/or neuropathy, which can cause foot ulcers, amputations, Charcot joints, and symptoms of autonomic dysfunction, such as sexual dysfunction. Diabetes increases a person's risk of peripheral vascular, cerebral, and cardiovascular disease.

v  Types of Diabetes Mellitus Classification

Earlier classifications

The World Health Organisation (WHO) published the first widely recognised classification of diabetes mellitus in 1980 (1) and again in 1985 (3). In 1980 and 1985, there were two statistical risk classes in addition to clinical classes for diabetes mellitus and related categories for glucose intolerance. IDDM, or Type 1, and NIDDM, or Type 2, are the two main classes of diabetes mellitus that the 1980 Expert Committee proposed. The terms Type 1 and Type 2 were dropped from the 1985 Study Group Report, but the classes IDDM and NIDDM were kept, and a new class called Malnutrition-related Diabetes Mellitus (MRDM) was added. In addition to Gestational Diabetes Mellitus (GDM), other classes of diabetes included in the 1980 and 1985 reports were Other Types and Impaired Glucose Tolerance (IGT). The tenth revision of the International Classification of Diseases (ICD–10) in 1992 and the International Nomenclature of Diseases (IND) in 1991 both took these into account. The 1985 classification is still in use today and was widely accepted. It allowed for the classification of individual subjects and patients in a way that was clinically useful even in situations where the precise cause or aetiology was unknown. It represented a compromise between clinical and aetiological classification. The suggested classification consists of a corresponding aetiological classification as well as diabetes mellitus staging based on clinical descriptive criteria.

Revised classification

As proposed by Kuzuya and Matsuda, the classification includes both clinical stages and aetiological types of diabetes mellitus as well as other categories of hyperglycemia. Regardless of its aetiology, diabetes progresses through a number of clinical stages over the course of its natural history, as shown by the clinical staging. Individual subjects are also free to go in either direction between stages. Individuals with diabetes mellitus, or those at risk of developing it, can be grouped based on their clinical features into stages even if the underlying cause is unknown. The increased knowledge of the etiological types of diabetes mellitus leads to the classification of the disease.

Antidiabetic drugs are used to lower the concentration of glucose in the blood of people with diabetes mellitus. By keeping the blood sugar at or close to the normal range, these medicines reduce some of the risks associated with diabetes. Antidiabetic drugs exert their useful effects through: (1) increasing insulin levels in the body or (2) increasing the body's sensitivity (or decreasing its resistance) to insulin, or (3) decreasing glucose absorption in the intestines.

Anti-diabetic medications Insulin.

The pancreatic β cells naturally secrete the hormone insulin. Individuals suffering from type 1 diabetes mellitus exhibit a complete lack of insulin, while those with type 2 diabetes may also have reduced levels of endogenous insulin production. All individuals with type 1 diabetes need to take insulin for the rest of their lives. Insulin is frequently used as monotherapy as the disease progresses or as an adjunct therapy to oral antidiabetic medications in patients with type 2 diabetes.Various substitutions on insulin molecule and other modification led to multiple types of insulin. These characterized and administered based on their pharmacodynamic and pharmacokinetic characteristics such as onset, peak, duration of action. Most significantly they are classified as rapid-acting, short- acting, intermediate-acting or long-acting types of insulin.

Classification of anti-diabetic drug

Different Types of Validation characteristics using in HPLC & UPLC

Ø  Precision.

Ø  Accuracy.

Ø  Specificity.

Ø  Linearity.

Ø  Range.

Ø  Detection Limit.

Ø  Quantitation Limit.

Ø  Ruggedness.

Ø  Robustness.

Precision:

      The degree of agreement between several measurements made at different times from the same homogeneous sample under specified circumstances. The standard deviation or relative standard deviation of a set of measurements is typically used to express the precision of a test method. Three levels of precision can be distinguished: reproducibility, intermediate precision, and repeatability.

Accuracy:

It is the degree to which the measured value and the actual value agree. The percentage of recovery by assaying the known additional amount of the analyte in the sample or the difference between the mean and accepted true value along with confidence intervals are used to calculate accuracy. According to the ICH guidelines, three replicates of each concentration should be analysed, with a minimum of three concentration levels covering the designated range (totaling three * three = nine determinations).

Specificity:

The capacity to definitively evaluate the analyte in the presence of elements that might be anticipated to be present, such as matrix elements, degradation products, and impurities.

Linearity:

    The ability of an analytical method to produce test results that are exactly proportionate to the concentration (amount) of analyte in the sample, within a specified range, is known as linearity.

Range:

    The interval between the upper and lower concentrations of analyte in an analytical procedure with an appropriate degree of linearity, accuracy, and precision is known as the range of the analytical procedure.

Detection Limit:

     It is the smallest concentration of analyte in a sample that, in the given experimental circumstances, can be detected but not necessarily quantified.

Quantitation Limit:

      It is the smallest concentration of analyte in a sample that can be accurately and precisely quantified.

Ruggedness:

The degree to which test results obtained by analysing the same samples under various test conditions—such as different laboratories, analysis, instruments, reagent lots, elapsed assay times, temperature, days, etc.—can be repeated is known as ruggedness. It can be described as an absence of environmental and operational variable influence on the analytical method test results.

Robustness:

     It indicates the method's dependability under typical operating conditions and measures how well it can withstand slight but intentional changes in method parameters. Should measurements be subject to fluctuations in analytical conditions, either appropriately controlled analytical conditions should be used, or a precautionary note should be incorporated into the process.

REPORTED UPLC METHODS:

1. LIRAGLUTIDE:

"UPLC-MS/MS Determination of GLP-1 Analogue, Liraglutide A Bioactive Peptide in Human Plasma" was published in a report by Taposh Gorella et al. in 2019. With a linear gradient and 0.3% formic acid in water and acetonitrile: methanol (50:50) mobile phases, chromatographic separation was accomplished using an ACQUITY UPLC Peptide BEH C18, 300 Å, 1.7 µm, 2.1 × 150 mm Column at a flow rate of 0.3 mL/min. The cycle took eight minutes in total. 4.77 minutes of retention.

2. GLIBENCLAMIDE:

"Rapid, Validated UPLC-MS/MS Method for Determination of Glibenclamide in Rat Plasma" was published in 2018 by Mohd Aftab Alam et al. Glibenclamide was eluted using a 2.1 x 50 mm Acquity UPLC®BEH C18 1.7 μm column. Acetonitrile (0.1% formic acid) and water (0.1% formic acid) made up component (A) and component (B) of the mobile phase. In gradient mode, the mobile phase was pumped at 150 μl/min. The sample ran for a total of two minutes. After injecting the 10 μl sample, the autosampler's temperature was maintained at 20 ± 3°C. At m/z 516.11, the parent sodium ion [Na+] adduct of glibenclamide was detected. At m/z 513.19, the parent sodium ion [Na+] adduct of glimepiride was detected. The daughter fragments of the sodium ion glibenclamide adduct (m/z 516.11) had molecular masses of 417 and 391.The retention period is 80 minutes.

3. TOLBUTAMIDE:

"UPLC– MS-MS Method for Simultaneous Determination of Caffeine, Tolbutamide, Metoprolol, and Dapsone in Rat Plasma and its Application to Cytochrome P450 Activity Study in Rats" was published in 2013 by Yan Liu et al. Utilising an Acquity UPLC–MS–MS, the chromatographic separation was executed on a Waters Acquity UPLC BEH HILIC C18 column (2.1 × 50 mm, 1.7 µm). At 40°C, the column temperature was kept constant. The auto sampler’s chamber temperature was maintained at 10°C. The mobile phase was 15:85, v/v, acetonitrile and water with 0.1% formic acid at a flow rate of 0.25 mL/min. Each injection took 5 minutes to complete. 3.10 is the retention time.

4. NATEGLINIDE:

"RP-UPLC Method Development and Validation for The Determination of Nateglinide in Bulk Drug and Pharmaceutical Formulations: A Quality by Design Approach" was published in 2012 by Basavaiah Kanakapura et al. The Acquity UPLC BEH C-18 (100 × 2.1) mm chromatographic column, featuring a particle size of 1.7 μm, was utilised. The isocratic elution process was used for the entire examination. The buffer used in the mobile phase was acetonitrile: potassium dihydrogen orthophosphate at a pH of 2.8:40:60. The mobile phase's isocratic flow rate was kept constant at 0.40 mL min–1. The temperature of the column was set to 35°C. 2 μL was the injection volume. The eluted sample ran for 6.0 minutes under 210 nm monitoring. The sample was retained for approximately 2.8 minutes.

5. SITAGLIPTIN:

"Simultaneous Determination of Sitagliptin Phosphate Monohydrate and Metformin Hydrochloride in Tablets by a Validated UPLC Method" was published in 2012 by Chellu S. N. Malleswararao et al. As the stationary phase, Acquity UPLC BEH C8 (100 × 2.1 mm, 1.7 μm) was employed. The buffer 10 mM potassium dihydrogen phosphate, 2 mM sodium salt of hexane-1-sulfonic acid (pH adjusted to 5.50 with diluted phosphoric acid), and acetonitrile with gradient programme [Time(min)/% acetonitrile): 0/8, 2/8, 4/45, 6/45, 8/8, 10/8] made up the mobile phase composition. The mobile phase was filtered using a 0.2 μm filter before use. Water was utilised as the sample diluent, and the mobile phase flow rate was kept constant at 0.2 mL min−1. Eluents were observed at 210 nm, and the column temperature was 25°C. For both standards and samples, the injection volume was 0.5 μL. Ten minutes were spent on the analysis in total. MH and SP were found to have retention periods of two and seven minutes, respectively.

REPORTED HPLC METHODS:

1. NATEGLINIDE:

Development and Validation of RP-HPLC Method for Metformin Hydrochloride and Nateglinide in Bulk and Combined Dosage Form was published by Prasanthi Chengalva et al. (2016). Utilising a reverse phase C18 column, Waters Inertsil ODS 3V (250x4.6 mm, 5μ), a mobile phase consisting of phosphate buffer (pH 4.0): acetonitrile: methanol (30:60:10), a flow rate of 1.0 ml/min, and a UV detector, the developed method detected wavelengths of 221 nm. It lasts 20 minutes in total.Elution of metformin hydrochloride took 2.45 minutes and nateglinide 4.21 minutes using the developed method.

2. SITAGLIPTIN:

"Method development and validation of sitagliptin and metformin using reverse phase HPLC method in bulk and tablet dosage form" was published in 2013 by Vasanth P M et al. Potassium dihydrogen orthophosphate and methanol (50:50 v/v) make up the mobile phase. Ophosphoric acid was used to adjust the pH to 8.5, and the flow rate is 1.0 ml/min. A PDA detector is used to observe the elution at 215 nm, and 10 µL of injection volume was used. Waters model 2695 HPLC device with Empower 2.0 software. 2996 PDA Detector is watered by the detector. Sartorius Digital Balance with Elico Ph Metre (0.1 mg to 205 gm). On a Hypesil BDS C18 Column (100 x 4.6 mm, 5µm particle size), separation was accomplished. One hour is the total run time. Metformin and Sitaglipitin had respective retention periods of 17.113 and 2.3 minutes.

3. TOLBUTAMIDE:

"Development and Validation of RP - HPLC Method for Quantitative Analysis Tolbutamide in Pure and Pharmaceutical Formulations" was published in 2013 by D. Madhu Latha et al. The Zodiac C18 column (250 mm × 4.6 mm × 5 µ particle size) was used for the analysis, and the mobile phase consisted of methanol, 0.1% orthophosphoric acid, and acetonitrile (10: 30: 60). UV was used for detection at 231 nm. 1.0 ml/min flow rate and a 10-minute run time overall. Time of retention: 4.60 min.

4. LIRAGLUTIDE:

"Validated RP - HPLC Method For The Estimation Of Liraglutide In Tablet Dosage" was published in 2012 by P.V.V. Satyanarayana et al. Temporary phase Methane: 0.1% OPA in acetonitrile (35:60:5) pH 5.1, UV detection range of 245 nm, analytical column C18, flow rate of 1.0 ml/min, ambient temperature, injection volume of 20µl, duration of run (10 min), and retention period (6.11 min).

5. GLIBENCLAMIDE:

The article "RP - HPLC method for the estimation of glibenclamide in human serum" was published in 2007 by S. D. Rajendran et al. Glibenclamide and the internal standard, glimepride, were separated chromatographically using a 250 mm × 4.6 mm ID Luna phenomenex 5u C18 analytical column (Phenomenex, USA). Just prior to the analytical column, a Guard-Pak precolumn module (Phenomenex, USA) with an ODS cartridge insert was positioned serially. Acetonitrile and 25 mM phosphate buffer (pH: 3.5) in a 60:40 v/v ratio made up the mobile phase. At room temperature, the mobile phase was pumped at an isocratic flow rate of 1 ml/min. At 253 nm, the UV detection wave length was chosen. The ultraviolet maximum of glibenclamide in acetonitrile:water at a 1:1 ratio was measured at a wavelength of 253 nm. The retention period was 8.12 minutes, and the analytical run time was less than 12 minutes.

Better resolution and separation are found in UPLC chromatograms than in HPLC chromatograms, which also perform more sensitive analyses, use less solvent, and analyse data more quickly. Advanced technology has led to the development of a brand-new system design known as Ultra High-Performance Liquid Chromatography (UPLC). The use of LC techniques for different groups of drug active compounds is justified by the benefits of short turnaround time, method reliability, method sensitivity, and drug specificity. Some of the fundamentals of UPLC and HPLC, method validation, system suitability testing, and method application to pharmaceutical analysis are covered in this review.

ü   When it comes to LIRAGLUTIDE, the UPLC method offers a lower flow rate (0.3 ml/min), runtime (8min), and retention time (4.77min) in comparison to the HPLC method.

ü   In GLIBENCLAMIDE, the UPLC method offers a lower flow rate (150µl/min), run time (2min), and retention time (0.80min) in comparison to the HPLC method.

ü   When it comes to TOLBUTAMIDE, the UPLC method offers a lower flow rate (0.25 ml/min), run time (2 min), and retention time (3.10 min) in comparison to the HPLC method.

ü   In NATEGLINIDE, the UPLC method offers a lower flow rate (0.40 ml/min), run time (6min), and retention time (2.8min) in comparison to the HPLC method.

ü   In SITAGLIPTIN, the UPLC method offers a lower flow rate (0.2 ml/min), a shorter run time (10min), and a shorter retention time (7min) than the HPLC method.

CONCLUSION:

When traditional HPLC has nearly reached its separation barriers, UPLC has demonstrated its ability to broaden the application of separation science. A quicker and better UPLC separation can save time and other resources, as most pharmaceutical companies aim to cut costs and R&D timelines. Higher resolution methods with shorter columns, smaller packing particle sizes, and higher flow rates under high pressure can be performed with the UPLC technique. When working with UPLC systems, a notable reduction in solvent consumption and column equilibration time is also accomplished. Compared to HPLC, the injection volume is significantly lower. The primary drawback of UPLC is the instrument's high cost. Additionally, the column life is shortened by the increased back pressure. However, UPLC's benefits—low solvent consumption, quicker separation times, and improved selectivity and sensitivity—outweigh its drawbacks. We can infer from this review that UPLC offers more benefits than HPLC.

ACKNOWLEDGMENT

Authors are thankful to Principal and Management J.I.I.U' S Ali Allana College of Pharmacy Akkalkuwa, Dist. Nandurbar for providing moral support and necessary facilities for completion of this work.

REFERENCES

1.        RPUPLC method development and validation for the determination of Nateglinide in bulk drug and pharmaceutical formulations: a quality by design approach," research gate2012, 10(1),23-44, Basavaiah Kadakapura, Madathil Cijo, Cijo Xavier, and Jaganathamurthi, Ramesh.

2.        Development and validation of RP-HPLC method for quantitative analysis of tolbutamide in pure and pharmaceutical formulations, D. Madhu Latha, K. Ammani, P. Jitendra Kumar, tsijournals-2013,11(4),1607-1614.

3.        UPLC: a contemporary chromatographic technique, Kinjal Shanshyambhai Sheliya and Ketan Shah, research gate.net-2013,4(3);78.

4.        HPLC: a brief review, Malviya R, Bansal V, Pal O.P., and Sharma P.K. Journal of Global Pharma Technology, 2010; 2(5): 22–26.

5.         Mazhar Abbas, Ultra Performance Liquid Chromatography, SlideServe, 2020;9.

6.        Rapid, validated UPLC-MS/MS method for determination of Glibenclamide in rat plasma," hindawi (international journal of analytical chemistry) 2018;1–9, Mohd Aftab Alam, Fahad Ibrahim al Jeonoobi, and Abdullah Mohammed alMohizea.

7.        Development and validation of RP-HPLC method for metformin hydrochloride and nateglinide in bulk and combined dosage form, Prasanthi Chengalva, Angala Parameswar S, and Aruna G, International Journal of Pharmacy and Pharmaceutical Sciences, 2016; 8(4):267-271.

8.        The article "Validated RP-HPLC method for the estimation of Liraglutide in tablet dosage" was published in the International Journal of Science Invention Today in 2012 and was written by P.V.V. Satyanarayana and Alavala Siva Madhavi.

9.        Chellu S. "Simultaneous determination of Sitagliptin phosphate monohydrate and metformin hydrochloride in tablets by a validated UPLC method," S.N. Malleswararao, Mulwkutta V Suryanarayana, and Khagga Mukkanti, National Library of Medicine-2012; 80(1), 139-152.

10.    "RP-HPLC method for the estimation of Glibenclamide in human serum," S.D. Rajendran, B.K. Philip, R. Gopinath, and B. Suresh, Indian Journal of Pharmaceutical Science-2007, 69(6), 796-799.

11.    Taposh Gorilla, DR. Padmakar Wah, and P.M.M. Rajesh. "Determination of glp-1 analogue, Liraglutide, by UPLC-MS/MS

12.    A. Agalya, E. Vikram, and Dr. G Abirami Review of Selected Anti-Diabetic Drugs' Reported Analytical Method Development by UPLC and HPLC published in International Journal of Pharmaceutical Research and Applications, Volume 6, Issue 1, January–February 2021.

13.    WHO Diabetes Mellitus Expert Committee. Report No. 2. WHO, Geneva, 1980. Report No. 646.

14.    The Expert Committee on Diabetes Mellitus Diagnosis and Classification. The expert committee's report on the diagnosis and categorization of type 2 diabetes. Diabetes Care 20:0183–97, 1997.

15.    National Diabetes Information Centre. Diabetes mellitus classification and diagnosis, as well as other forms of glucose intolerance. Diabetes 1979;28(10)1039–1059.

16.    In Harrison's Principles of Internal Medicine, 14th edition (Isselbacher, K.J., Braunwald, E., Wilson, J.D., Martin, J.B., Fauci, A.S., and Kasper, D.L., eds), McGraw-Hill, Inc. (Health Professions Division), 1998;2060-2080, Foster, D.W. Diabetes Mellitus.

17.    Pancreatic Hormones and Antidiabetic Drugs, Karam JH, Basic and Clinical Pharmacology, Appleton-Lange, 1998; 684-703 (Katzung, B. G., ed.).

18.    Fantus IG and Cheng AY. Type 2 diabetes mellitus treated with oral antihyperglycemic medication. CMAJ 172(2): 213-26; 2005.

INTRODUCTION

A contemporary method called UPLC offers liquid chromatography a new path. Ultra performance liquid chromatography, or UPLC, is characterised by improvements in three key areas: sensitivity, speed, and resolution. High-performance liquid chromatography (HPLC) lacks the resolution, speed, and sensitivity of ultra performance liquid chromatography (UPLC), which is applicable to particles smaller than 2 μm in diameter. Pharmaceutical companies are looking for innovative ways to boost economy and speed up drug development in the twenty-first century. Analytical laboratories are not an exception to this trend, as UPLC analysis currently yields better product quality. At pressures as high as 100 million Pa, UPLC uses extreme pressures to perform separation and quantification. In contrast to HPLC, it has been observed that high pressure has no detrimental effects on the analytical column. Additionally, UPLC uses less time and solvent overall.

An important qualitative and quantitative method for estimating biological and pharmaceutical samples is high performance liquid chromatography (HPLC). For the purpose of ensuring the quality control of drug components, this chromatographic technique is the most reliable, fast, safe, and adaptable.

The word "diabetes mellitus" refers to a multifactorial metabolic illness marked by persistent hyperglycaemia and abnormalities in the metabolism of proteins, fats, and carbohydrates brought on by deficiencies in either or both of the actions or secretion of insulin. Diabetes mellitus causes long-term harm as well as organ failure and dysfunction. Diabetes mellitus can cause thirst, polyuria, blurred vision, and weight loss, among other symptoms. In its most severe forms, ketoacidosis or a non– ketotic hyperosmolar state may develop and lead to stupor, command, in absence of effective treatment, death1,2. Frequently, there may be no symptoms at all or only mild ones, which means that hyperglycaemia severe enough to induce pathological and functional alterations may exist for a long time before a diagnosis is made. Diabetes mellitus causes long-term consequences that include retinopathy, which can result in blindness, nephropathy, which can cause renal failure, and/or neuropathy, which can cause foot ulcers, amputations, Charcot joints, and symptoms of autonomic dysfunction, such as sexual dysfunction. Diabetes increases a person's risk of peripheral vascular, cerebral, and cardiovascular disease.

v  Types of Diabetes Mellitus Classification

Earlier classifications

The World Health Organisation (WHO) published the first widely recognised classification of diabetes mellitus in 1980 (1) and again in 1985 (3). In 1980 and 1985, there were two statistical risk classes in addition to clinical classes for diabetes mellitus and related categories for glucose intolerance. IDDM, or Type 1, and NIDDM, or Type 2, are the two main classes of diabetes mellitus that the 1980 Expert Committee proposed. The terms Type 1 and Type 2 were dropped from the 1985 Study Group Report, but the classes IDDM and NIDDM were kept, and a new class called Malnutrition-related Diabetes Mellitus (MRDM) was added. In addition to Gestational Diabetes Mellitus (GDM), other classes of diabetes included in the 1980 and 1985 reports were Other Types and Impaired Glucose Tolerance (IGT). The tenth revision of the International Classification of Diseases (ICD–10) in 1992 and the International Nomenclature of Diseases (IND) in 1991 both took these into account. The 1985 classification is still in use today and was widely accepted. It allowed for the classification of individual subjects and patients in a way that was clinically useful even in situations where the precise cause or aetiology was unknown. It represented a compromise between clinical and aetiological classification. The suggested classification consists of a corresponding aetiological classification as well as diabetes mellitus staging based on clinical descriptive criteria.

Revised classification

As proposed by Kuzuya and Matsuda, the classification includes both clinical stages and aetiological types of diabetes mellitus as well as other categories of hyperglycemia. Regardless of its aetiology, diabetes progresses through a number of clinical stages over the course of its natural history, as shown by the clinical staging. Individual subjects are also free to go in either direction between stages. Individuals with diabetes mellitus, or those at risk of developing it, can be grouped based on their clinical features into stages even if the underlying cause is unknown. The increased knowledge of the etiological types of diabetes mellitus leads to the classification of the disease.

Antidiabetic drugs are used to lower the concentration of glucose in the blood of people with diabetes mellitus. By keeping the blood sugar at or close to the normal range, these medicines reduce some of the risks associated with diabetes. Antidiabetic drugs exert their useful effects through: (1) increasing insulin levels in the body or (2) increasing the body's sensitivity (or decreasing its resistance) to insulin, or (3) decreasing glucose absorption in the intestines.

Anti-diabetic medications Insulin.

The pancreatic β cells naturally secrete the hormone insulin. Individuals suffering from type 1 diabetes mellitus exhibit a complete lack of insulin, while those with type 2 diabetes may also have reduced levels of endogenous insulin production. All individuals with type 1 diabetes need to take insulin for the rest of their lives. Insulin is frequently used as monotherapy as the disease progresses or as an adjunct therapy to oral antidiabetic medications in patients with type 2 diabetes.Various substitutions on insulin molecule and other modification led to multiple types of insulin. These characterized and administered based on their pharmacodynamic and pharmacokinetic characteristics such as onset, peak, duration of action. Most significantly they are classified as rapid-acting, short- acting, intermediate-acting or long-acting types of insulin.

Classification of anti-diabetic drug

Different Types of Validation characteristics using in HPLC & UPLC

Ø  Precision.

Ø  Accuracy.

Ø  Specificity.

Ø  Linearity.

Ø  Range.

Ø  Detection Limit.

Ø  Quantitation Limit.

Ø  Ruggedness.

Ø  Robustness.

Precision:

      The degree of agreement between several measurements made at different times from the same homogeneous sample under specified circumstances. The standard deviation or relative standard deviation of a set of measurements is typically used to express the precision of a test method. Three levels of precision can be distinguished: reproducibility, intermediate precision, and repeatability.

Accuracy:

It is the degree to which the measured value and the actual value agree. The percentage of recovery by assaying the known additional amount of the analyte in the sample or the difference between the mean and accepted true value along with confidence intervals are used to calculate accuracy. According to the ICH guidelines, three replicates of each concentration should be analysed, with a minimum of three concentration levels covering the designated range (totaling three * three = nine determinations).

Specificity:

The capacity to definitively evaluate the analyte in the presence of elements that might be anticipated to be present, such as matrix elements, degradation products, and impurities.

Linearity:

    The ability of an analytical method to produce test results that are exactly proportionate to the concentration (amount) of analyte in the sample, within a specified range, is known as linearity.

Range:

    The interval between the upper and lower concentrations of analyte in an analytical procedure with an appropriate degree of linearity, accuracy, and precision is known as the range of the analytical procedure.

Detection Limit:

     It is the smallest concentration of analyte in a sample that, in the given experimental circumstances, can be detected but not necessarily quantified.

Quantitation Limit:

      It is the smallest concentration of analyte in a sample that can be accurately and precisely quantified.

Ruggedness:

The degree to which test results obtained by analysing the same samples under various test conditions—such as different laboratories, analysis, instruments, reagent lots, elapsed assay times, temperature, days, etc.—can be repeated is known as ruggedness. It can be described as an absence of environmental and operational variable influence on the analytical method test results.

Robustness:

     It indicates the method's dependability under typical operating conditions and measures how well it can withstand slight but intentional changes in method parameters. Should measurements be subject to fluctuations in analytical conditions, either appropriately controlled analytical conditions should be used, or a precautionary note should be incorporated into the process.

REPORTED UPLC METHODS:

1. LIRAGLUTIDE:

"UPLC-MS/MS Determination of GLP-1 Analogue, Liraglutide A Bioactive Peptide in Human Plasma" was published in a report by Taposh Gorella et al. in 2019. With a linear gradient and 0.3% formic acid in water and acetonitrile: methanol (50:50) mobile phases, chromatographic separation was accomplished using an ACQUITY UPLC Peptide BEH C18, 300 Å, 1.7 µm, 2.1 × 150 mm Column at a flow rate of 0.3 mL/min. The cycle took eight minutes in total. 4.77 minutes of retention.

2. GLIBENCLAMIDE:

"Rapid, Validated UPLC-MS/MS Method for Determination of Glibenclamide in Rat Plasma" was published in 2018 by Mohd Aftab Alam et al. Glibenclamide was eluted using a 2.1 x 50 mm Acquity UPLC®BEH C18 1.7 μm column. Acetonitrile (0.1% formic acid) and water (0.1% formic acid) made up component (A) and component (B) of the mobile phase. In gradient mode, the mobile phase was pumped at 150 μl/min. The sample ran for a total of two minutes. After injecting the 10 μl sample, the autosampler's temperature was maintained at 20 ± 3°C. At m/z 516.11, the parent sodium ion [Na+] adduct of glibenclamide was detected. At m/z 513.19, the parent sodium ion [Na+] adduct of glimepiride was detected. The daughter fragments of the sodium ion glibenclamide adduct (m/z 516.11) had molecular masses of 417 and 391.The retention period is 80 minutes.

3. TOLBUTAMIDE:

"UPLC– MS-MS Method for Simultaneous Determination of Caffeine, Tolbutamide, Metoprolol, and Dapsone in Rat Plasma and its Application to Cytochrome P450 Activity Study in Rats" was published in 2013 by Yan Liu et al. Utilising an Acquity UPLC–MS–MS, the chromatographic separation was executed on a Waters Acquity UPLC BEH HILIC C18 column (2.1 × 50 mm, 1.7 µm). At 40°C, the column temperature was kept constant. The auto sampler’s chamber temperature was maintained at 10°C. The mobile phase was 15:85, v/v, acetonitrile and water with 0.1% formic acid at a flow rate of 0.25 mL/min. Each injection took 5 minutes to complete. 3.10 is the retention time.

4. NATEGLINIDE:

"RP-UPLC Method Development and Validation for The Determination of Nateglinide in Bulk Drug and Pharmaceutical Formulations: A Quality by Design Approach" was published in 2012 by Basavaiah Kanakapura et al. The Acquity UPLC BEH C-18 (100 × 2.1) mm chromatographic column, featuring a particle size of 1.7 μm, was utilised. The isocratic elution process was used for the entire examination. The buffer used in the mobile phase was acetonitrile: potassium dihydrogen orthophosphate at a pH of 2.8:40:60. The mobile phase's isocratic flow rate was kept constant at 0.40 mL min–1. The temperature of the column was set to 35°C. 2 μL was the injection volume. The eluted sample ran for 6.0 minutes under 210 nm monitoring. The sample was retained for approximately 2.8 minutes.

5. SITAGLIPTIN:

"Simultaneous Determination of Sitagliptin Phosphate Monohydrate and Metformin Hydrochloride in Tablets by a Validated UPLC Method" was published in 2012 by Chellu S. N. Malleswararao et al. As the stationary phase, Acquity UPLC BEH C8 (100 × 2.1 mm, 1.7 μm) was employed. The buffer 10 mM potassium dihydrogen phosphate, 2 mM sodium salt of hexane-1-sulfonic acid (pH adjusted to 5.50 with diluted phosphoric acid), and acetonitrile with gradient programme [Time(min)/% acetonitrile): 0/8, 2/8, 4/45, 6/45, 8/8, 10/8] made up the mobile phase composition. The mobile phase was filtered using a 0.2 μm filter before use. Water was utilised as the sample diluent, and the mobile phase flow rate was kept constant at 0.2 mL min−1. Eluents were observed at 210 nm, and the column temperature was 25°C. For both standards and samples, the injection volume was 0.5 μL. Ten minutes were spent on the analysis in total. MH and SP were found to have retention periods of two and seven minutes, respectively.

REPORTED HPLC METHODS:

1. NATEGLINIDE:

Development and Validation of RP-HPLC Method for Metformin Hydrochloride and Nateglinide in Bulk and Combined Dosage Form was published by Prasanthi Chengalva et al. (2016). Utilising a reverse phase C18 column, Waters Inertsil ODS 3V (250x4.6 mm, 5μ), a mobile phase consisting of phosphate buffer (pH 4.0): acetonitrile: methanol (30:60:10), a flow rate of 1.0 ml/min, and a UV detector, the developed method detected wavelengths of 221 nm. It lasts 20 minutes in total.Elution of metformin hydrochloride took 2.45 minutes and nateglinide 4.21 minutes using the developed method.

2. SITAGLIPTIN:

"Method development and validation of sitagliptin and metformin using reverse phase HPLC method in bulk and tablet dosage form" was published in 2013 by Vasanth P M et al. Potassium dihydrogen orthophosphate and methanol (50:50 v/v) make up the mobile phase. Ophosphoric acid was used to adjust the pH to 8.5, and the flow rate is 1.0 ml/min. A PDA detector is used to observe the elution at 215 nm, and 10 µL of injection volume was used. Waters model 2695 HPLC device with Empower 2.0 software. 2996 PDA Detector is watered by the detector. Sartorius Digital Balance with Elico Ph Metre (0.1 mg to 205 gm). On a Hypesil BDS C18 Column (100 x 4.6 mm, 5µm particle size), separation was accomplished. One hour is the total run time. Metformin and Sitaglipitin had respective retention periods of 17.113 and 2.3 minutes.

3. TOLBUTAMIDE:

"Development and Validation of RP - HPLC Method for Quantitative Analysis Tolbutamide in Pure and Pharmaceutical Formulations" was published in 2013 by D. Madhu Latha et al. The Zodiac C18 column (250 mm × 4.6 mm × 5 µ particle size) was used for the analysis, and the mobile phase consisted of methanol, 0.1% orthophosphoric acid, and acetonitrile (10: 30: 60). UV was used for detection at 231 nm. 1.0 ml/min flow rate and a 10-minute run time overall. Time of retention: 4.60 min.

4. LIRAGLUTIDE:

"Validated RP - HPLC Method For The Estimation Of Liraglutide In Tablet Dosage" was published in 2012 by P.V.V. Satyanarayana et al. Temporary phase Methane: 0.1% OPA in acetonitrile (35:60:5) pH 5.1, UV detection range of 245 nm, analytical column C18, flow rate of 1.0 ml/min, ambient temperature, injection volume of 20µl, duration of run (10 min), and retention period (6.11 min).

5. GLIBENCLAMIDE:

The article "RP - HPLC method for the estimation of glibenclamide in human serum" was published in 2007 by S. D. Rajendran et al. Glibenclamide and the internal standard, glimepride, were separated chromatographically using a 250 mm × 4.6 mm ID Luna phenomenex 5u C18 analytical column (Phenomenex, USA). Just prior to the analytical column, a Guard-Pak precolumn module (Phenomenex, USA) with an ODS cartridge insert was positioned serially. Acetonitrile and 25 mM phosphate buffer (pH: 3.5) in a 60:40 v/v ratio made up the mobile phase. At room temperature, the mobile phase was pumped at an isocratic flow rate of 1 ml/min. At 253 nm, the UV detection wave length was chosen. The ultraviolet maximum of glibenclamide in acetonitrile:water at a 1:1 ratio was measured at a wavelength of 253 nm. The retention period was 8.12 minutes, and the analytical run time was less than 12 minutes.

Better resolution and separation are found in UPLC chromatograms than in HPLC chromatograms, which also perform more sensitive analyses, use less solvent, and analyse data more quickly. Advanced technology has led to the development of a brand-new system design known as Ultra High-Performance Liquid Chromatography (UPLC). The use of LC techniques for different groups of drug active compounds is justified by the benefits of short turnaround time, method reliability, method sensitivity, and drug specificity. Some of the fundamentals of UPLC and HPLC, method validation, system suitability testing, and method application to pharmaceutical analysis are covered in this review.

ü   When it comes to LIRAGLUTIDE, the UPLC method offers a lower flow rate (0.3 ml/min), runtime (8min), and retention time (4.77min) in comparison to the HPLC method.

ü   In GLIBENCLAMIDE, the UPLC method offers a lower flow rate (150µl/min), run time (2min), and retention time (0.80min) in comparison to the HPLC method.

ü   When it comes to TOLBUTAMIDE, the UPLC method offers a lower flow rate (0.25 ml/min), run time (2 min), and retention time (3.10 min) in comparison to the HPLC method.

ü   In NATEGLINIDE, the UPLC method offers a lower flow rate (0.40 ml/min), run time (6min), and retention time (2.8min) in comparison to the HPLC method.

ü   In SITAGLIPTIN, the UPLC method offers a lower flow rate (0.2 ml/min), a shorter run time (10min), and a shorter retention time (7min) than the HPLC method.

CONCLUSION:

When traditional HPLC has nearly reached its separation barriers, UPLC has demonstrated its ability to broaden the application of separation science. A quicker and better UPLC separation can save time and other resources, as most pharmaceutical companies aim to cut costs and R&D timelines. Higher resolution methods with shorter columns, smaller packing particle sizes, and higher flow rates under high pressure can be performed with the UPLC technique. When working with UPLC systems, a notable reduction in solvent consumption and column equilibration time is also accomplished. Compared to HPLC, the injection volume is significantly lower. The primary drawback of UPLC is the instrument's high cost. Additionally, the column life is shortened by the increased back pressure. However, UPLC's benefits—low solvent consumption, quicker separation times, and improved selectivity and sensitivity—outweigh its drawbacks. We can infer from this review that UPLC offers more benefits than HPLC.

ACKNOWLEDGMENT

Authors are thankful to Principal and Management J.I.I.U' S Ali Allana College of Pharmacy Akkalkuwa, Dist. Nandurbar for providing moral support and necessary facilities for completion of this work.

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