Comparative Quality Assessment of Branded and Generic Metformin Prolonged-Release Tablets and Glimepiride Tablets using Assay, In-vitro Dissolution Testing and Statistical Approaches (Model-Dependent and Model-Independent Approaches)
Deepak Mandal1,2, Manisha Trivedi2, Meenakshi Dahiya2, Robin Kumar2, Dharam Pal Pathak1, Nupur Tomar2, Rajan Choudhary2
1,2Quality Assurance, Deepak Mandal, Delhi Pharmaceutical Science and Research University, New Delhi, India.
2Reference Standard Division, Dr. Manisha Trivedi, Indian Pharmacopoeia Commission, Ghaziabad, India.
2Quality Assurance Head, Dr. Meenakshi Dahiya, Indian Pharmacopoeia Commission, Ghaziabad, India.
2Reference Standard Division, Dr. Robin Kumar, Indian Pharmacopoeia Commission, Ghaziabad, India.
1Pharmaceutical Chemistry, Dr. Dharam pal Pathak, Delhi Pharmaceutical Science and Research University, New Delhi, India.
2Reference Standard Division, Dr. Nupur Tomar, Indian Pharmacopoeia Commission, Ghaziabad, India.
2Reference Standard Division, Mr. Rajan Choudhary, Indian Pharmacopoeia Commission, Ghaziabad, India.
*Corresponding Author E-mail: rudrasanty2001@gmail.com, ipcmanisha@gmail.com, meenakshi.ipc@gov.in, robinkumar21@gmail.com, drdppathak@gmail.com, nupurtomar85@gmail.com, rajanchoudhary413@gmail.com
ABSTRACT:
Background: This study explores the pharmacological features of metformin hydrochloride and glimepiride, two widely recommended medicines for controlling type 2 diabetes mellitus. Objective: The primary objectives were to assess the dissolving profiles, confirm the uniformity of the active pharmaceutical components, and evaluate conformity to pharmacopoeial standards in branded and generic formulations. Methods: We employed UV spectroscopy and high-performance liquid chromatography (HPLC) to conduct extensive evaluations. The absorption profile of UV spectroscopy revealed the measurement of the concentration of the medication in different formulations. HPLC had provided precise separation and quantitation and proved the purity and concentration of various samples. The dissolving studies were performed to detect the differences in the release of branded and generic preparations. Univariate ANOVA, and t-tests were applied to determine consistency and reproducibility of data through statistical analysis. The use of Graphical representation and interpretation, non mathematical models (f1, f2, and the Rescigno ratio), mathematical models (zero order, first order, Higuchi, Pappas, Sahlin, Hixson-Crowell, Baker-Lader, Kos Meyer-Pappas) used to describe the dissolution profiles. Results: The dissolution studies showed there were major variations between the brand and generic formulations concerning the rate of release which could affect the bioavailability and efficacy of a drug. Consistency and trustworthiness were supported with the help of statistical study. In conclusion, our results emphasize the high importance of standard analytical measures in the process of ensuring drug quality and drug efficacy. The given work contributes insightful experience to the pharmaceutical analysis of the anti-diabetic medications, which benefit establishing the reliable formulations of generics and highlighting the importance of quality control in the pharmaceutical industry.
KEYWORDS: Metformin hydrochloride, glimepiride, UV spectroscopy, high-performance liquid chromatography, Pharmacopoeial standards, bioavailability, therapeutic efficacy, quality control, generic formulations.
1. INTRODUCTION:
The assurance of the efficacy and safety of pharmaceuticals is essential in the sphere of pharmaceutical sciences1. The given research takes a new step to inquire into the quality of metformin prolonged-release formulations and their generic substitutes through extended assay and in-vitro dissolution testing.2 In the study, the researchers explain the importance of such tests as essential quality control methods that are used to ensure that both branded and generic copies of drugs have high pharmaceutical standards.3,4 Diabetes mellitus (DM) is a prolonged condition of high sugar blood levels, which requires effective pharmaceutical remedies to control.5 Glimepiride and metformin hydrochloride are two of the key medicines used in treating type two diabetic mellitus (T2DM).6 Metformin is a biguanide that enhances insulin sensitivity and reduces glucose production in the liver, glimepiride, a member of a sulfonylurea, increases insulin secretion by the pancreatic beta cells.7 Since these pharmaceuticals are used widely, it is crucial to ensure that their extent of performance in branded and generic form is assured in terms of quality, efficacy and safety.8,9 This work utilises high performance liquid chromatography (HPLC) and ultraviolet-visible (UV-Vis) spectroscopy to analyse the assay of metformin and glimepiride formulations.10,11,12 This HPLC is a strong analysis tool that allows an accurate quantification of a product and also guarantees the detection of contaminants that may be present in the given product.13,14,15,16 This makes this technique more valid assessing the purity and the potency in the given drug.17,18 The overall research utilizes current spectroscopic tools like the in the analytical process that was used here is quite proficient as it not only strengthens this study in terms of its findings due to its reliability but also marks the beginning of future computations involving quality assurance of drugs.19 Through the advanced form of statistical testing, such as the use of univariate ANOVA and the t-tests, the study confirms the reliability and recyclability of its conclusions, which ensures that the findings are scientifically sound and clinically applicable.20,21,22
1.1. Graphical Representation Of Dissolution Profile Data: Metformin Samples:
Fig. 1. % Release vs. Time
Initial time point (0.25hours):
Ref.1 and Ref.2 showed close values (17.78, 18.98). Test samples varied, highest in Test.1 (21.51) and lowest in Test.6 (15.17).
Early time point (1hour):
Ref.1 and Ref.2 remained similar (36.13, 35.67). Test.2 and Test.3 were close to reference (36.51, 36.45), while Test.5 and Test.6 showed lower values (31.07, 32.75).
Mid time point (3 hours):
Ref.1 and Ref.2 were similar (48.32, 47.81). Test.1 matched Ref.1 (48.22), while Test.2 - 5 showed higher values (59.79 - 58.89). Test.6 was slightly lower (55.15)
Later time point (5 hours):
Ref.1 and Ref.2 were close (66.63, 69.19). All tests showed higher values, with Test.4 highest (73.67) and others slightly above references (68.64 -72.95).
Final time point (8 hours):
Ref.1 and Ref.2 were similar (93.52, 92.96). Test samples aligned closely, except T3 (88.63). Reference profiles were uniform; tests varied at 3 h but matched by 8 h.
1.2. Symbols:
Symbols:
1. K0, K1, KH, KKP, KHC, KBL, K1, K2 - Rate constants for various drug release models (Zero-order, First-order, Higuchi, Korsmeyer-Peppas, Hixson-Crowell, Baker-Lonsdale, Peppas-Sahlin).
2. RSQR_ADJ - Adjusted R-square, MSE_ROOT - Root Mean Square Error
3. AIC - Akaike Information Criterion, MSC - Model Selection Criterion.
4. n - Exponent, f1 - Difference factor ,f2 - Similarity factor
5. ζ1, ζ2 - Rescigno indices
Table1. List of instruments
|
Instrument |
Brand/Model |
Manufacturer |
|
HPLC System |
Agilent 1200 |
Agilent Technologies |
|
Uv-Visible Spectrophotometer |
Lambda 35 |
Perkin Elmer |
|
Dissolution Apparatus |
Ds 8000 |
Lab India |
|
Sonicator |
Branson Sonicator |
Mettler Toledo |
|
Water Purification System |
Integral 3 Q-Pod |
Millipore Co. Bedford USA |
|
Ph Meter |
Seven Compact Ph/Ion S220 |
Mettler Toledo |
|
Syringe Filter |
RanDisk |
Rankem |
|
Digital Weighing Balance |
- |
Mettler Toledo |
|
Analytical Column |
Inertsil (C18 150*4.6mm 5µm) |
- |
|
Membrane Filters |
Millipore |
Axiva |
|
Hot Air Oven |
- |
- |
|
Glassware |
- |
Borosil |
Table2. List of the Chemicals and Reagents
|
Chemicals |
Manufacturer/Supplier |
|
Metformin (Working Standard) |
Torrent pharmaceuticals |
|
Glimepiride (IPRS) |
Indian pharmacopoeia commission Ghaziabad |
|
Potassium dihydrogen phosphate |
Finar |
|
Sodium hydroxide |
Sigma-Aldrich |
|
Sodium dihydrogen phosphate |
Finar |
|
Methanol (HPLC grade) |
Rankem |
|
Acetonitrile (HPLC grade) |
Finar |
|
Distilled water |
Millipore |
Table 3. Metformin Hydrochloride Prolonged-Release 500 mg Tablets
|
Sample Type |
Sample |
Assay Results (%) |
Time (Hr.) |
Dissolution Profile (%) |
|
Branded |
R1 (EXERMET 500 SR) |
100.25 |
0.25 |
17.78 |
|
1 |
35.49 |
|||
|
3 |
48.32 |
|||
|
5 |
66.63 |
|||
|
8 |
93.52 |
|||
|
Branded |
R2 (GLUCOFORMIN XL 500) |
99.85 |
0.25 |
18.98 |
|
1 |
35.67 |
|||
|
3 |
47.81 |
|||
|
5 |
69.19 |
|||
|
8 |
92.96 |
|||
|
Generic |
T1 (METFORNIL SR 500) |
100.27 |
0.25 |
21.50 |
|
1 |
34.60 |
|||
|
3 |
48.22 |
|||
|
5 |
68.64 |
|||
|
8 |
94.55 |
|||
|
Generic |
T2 (METFORM SR 500) |
101.36 |
0.25 |
15.17 |
|
1 |
32.75 |
|||
|
3 |
55.15 |
|||
|
5 |
72.18 |
|||
|
8 |
93.15 |
|||
|
Generic |
T3 (DAILYGLIM 500 SR) |
101.08 |
0.25 |
15.39 |
|
1 |
31.07 |
|||
|
3 |
58.89 |
|||
|
5 |
72.95 |
|||
|
8 |
90.75 |
|||
|
Generic |
T4 (METROSE 500 SR) |
99.88 |
0.25 |
15.49 |
|
1 |
36.51 |
|||
|
3 |
59.79 |
|||
|
5 |
69.81 |
|||
|
8 |
90.15 |
|||
|
Generic |
T5 (ELCEPHASE 500 SR) |
101.48 |
0.25 |
16.61 |
|
1 |
36.14 |
|||
|
3 |
58.66 |
|||
|
5 |
73.67 |
|||
|
8 |
92.29 |
|||
|
Generic |
T6 (METFORMIN HCL (J.A)) |
100.92 |
0.25 |
18.38 |
|
1 |
36.45 |
|||
|
3 |
60.90 |
|||
|
5 |
69.94 |
|||
|
8 |
88.63 |
All samples met pharmacopoeial standards (API 90 -110%). Both branded and generic forms released >85% within 8 hours, confirming consistent efficacy.
Table 4. Glimepiride 1 mg Tablets
|
Sample Type |
Samples |
Assay (%) |
Dissolution Profile at 30 mins (%) |
|
Branded |
R3 (Glimda-1) |
92.79 |
96 |
|
|
R4 (Glimestar-1) |
102.67 |
99 |
|
Generic |
T7 Glimepiride HCl (Jan Aushadhi) |
99.38 |
91 |
|
|
T8 (Glidum-1) |
96.29 |
89 |
|
|
T9 (Glyzee-1) |
101.08 |
94 |
|
|
T10 (Glyfic-1) |
99.88 |
83 |
All samples met IP 2022 standards (API 90 -110%). Both branded and generic forms released >75% in 30 minutes, confirming consistent efficacy.
Non-Mathematical Models (Model-Independent Approach):
Table 5. f1, f2, and Rescigno Index (ζ1, ζ2) for Test Products
|
Test Product |
f1 (Ref.1) |
f1 (Ref.2) |
f2 (Ref.1) |
f2 (Ref.2) |
ζ1 (Ref.1) |
ζ1 (Ref.2) |
ζ2 (Ref.1) |
ζ2 (Ref.2) |
|
Test.1 |
3.20 |
2.32 |
81.98 |
87.72 |
0.0115 |
0.0078 |
0.0132 |
0.0087 |
|
Test.2 |
7.89 |
7.46 |
62.14 |
61.73 |
0.0396 |
0.0342 |
0.0490 |
0.0487 |
|
Test.3 |
8.00 |
7.12 |
61.02 |
61.18 |
0.0422 |
0.0362 |
0.0520 |
0.0511 |
|
Test.4 |
7.54 |
7.12 |
62.08 |
63.14 |
0.0424 |
0.0371 |
0.0509 |
0.0467 |
|
Test.5 |
10.33 |
9.54 |
60.19 |
61.04 |
0.0519 |
0.0451 |
0.0537 |
0.0498 |
|
Test.6 |
7.14 |
6.52 |
67.39 |
68.51 |
0.0360 |
0.0299 |
0.0384 |
0.0339 |
|
Overall Statistics |
Mean |
SE |
Mean |
SE |
Mean |
SE |
Mean |
SE |
|
Mean_R Vs Individual |
6.92 |
0.99 |
66.78 |
3.94 |
0.0341 |
0.0056 |
0.0408 |
0.0067 |
|
Mean_R Vs Mean_T |
6.18 |
|
67.64 |
|
0.0323 |
|
0.0381 |
|
The mean f1 (6.18) and f2 (67.64) values fall within acceptable limits, confirming similarity between test and reference profiles. Low Rescigno Index (ζ₁) and minimal errors further support their close equivalence in dissolution behaviour.
UNIVARIATE ANOVA OUTPUT
Table 6. Anova Results
|
Source of Variation |
Sum of Squares (SS) |
Degrees of Freedom (DF) |
Mean Square (MS) |
F-Value |
p-Value |
F Critical (F Crit) |
|
Between Groups |
50,836.47 |
1 |
50,836.47 |
141.68 |
3.23E-19 |
3.96 |
|
Within Groups |
27,987.87 |
78 |
358.82 |
|
|
|
|
Total |
78,824.33 |
79 |
|
|
|
|
The F-value (141.68 > 3.96) and low P-value (<0.05) indicate significant differences in release over time, confirming a sustained-release profile.
T-Test Results
Table 7. p-Values for Pair-wise Comparisons
|
Comparison |
P-Value |
|
Ref.1 vs. Test.1 |
0.956574307 |
|
Ref.1 vs. Test.2 |
0.921271729 |
|
Ref.1 vs. Test.3 |
0.903645267 |
|
Ref.1 vs. Test.4 |
0.876254211 |
|
Ref.1 vs. Test.5 |
0.945382522 |
|
Ref.1 vs. Test.6 |
0.95094842 |
|
Ref.2 vs. Test.1 |
0.975350028 |
|
Ref.2 vs. Test.2 |
0.939827323 |
|
Ref.2 vs. Test.3 |
0.922554102 |
|
Ref.2 vs. Test.4 |
0.894298915 |
|
Ref.2 vs. Test.5 |
0.963547946 |
|
Ref.2 vs. Test.6 |
0.969040683 |
All p-values >0.05, indicating no significant difference and confirming similarity between test and reference dissolution profiles.
MATHEMATICAL MODELS:
(Model-dependent Approach):Table 8. Interpretation Of Data for Different Models
|
Time (hour) |
Ref.1 |
Ref.2 |
Test.1 |
Test.2 |
Test.3 |
Test.4 |
Test.5 |
Test.6 |
|
0.25 |
17.78 |
18.98 |
21.51 |
15.49 |
18.38 |
16.61 |
15.39 |
15.17 |
|
1 |
36.13 |
35.67 |
34.6 |
36.51 |
36.45 |
36.14 |
31.07 |
32.75 |
|
3 |
48.32 |
47.81 |
48.22 |
59.79 |
60.19 |
58.66 |
58.89 |
55.15 |
|
5 |
66.63 |
69.19 |
68.64 |
69.81 |
69.94 |
73.67 |
72.95 |
72.18 |
|
8 |
93.52 |
92.96 |
94.55 |
90.15 |
88.63 |
92.29 |
90.75 |
93.15 |
Table 9. Parameters of the mathematical models and descriptive statistics for the dissolution data
|
Metric |
NO.1 |
NO.2 |
NO.3 |
NO.4 |
NO.5 |
NO.6 |
NO.7 |
NO.8 |
|
|
Zero Order |
K0 |
10.203 |
10.112 |
9.795 |
10.150 |
10.039 |
10.188 |
10.198 |
10.177 |
|
RSQR_ADJ |
0.5563 |
0.4377 |
0.2569 |
0.3848 |
0.4717 |
0.5418 |
0.6133 |
0.5808 |
|
|
MSE_ROOT |
21.2688 |
23.5171 |
25.0017 |
24.5673 |
23.3288 |
22.2707 |
20.3130 |
20.861 |
|
|
AIC |
48.3435 |
49.5494 |
50.2840 |
50.0736 |
49.4529 |
48.8959 |
47.7918 |
48.1113 |
|
|
MSC |
0.4792 |
0.2424 |
-0.0364 |
0.1525 |
0.3047 |
0.4472 |
0.6167 |
0.5362 |
|
|
First Order |
K1 |
0.275 |
0.303 |
0.298 |
0.314 |
0.297 |
0.293 |
0.270 |
0.273 |
|
RSQR_ADJ |
0.9068 |
0.9506 |
0.9210 |
0.9575 |
0.9764 |
0.9703 |
0.9177 |
0.9195 |
|
|
MSE_ROOT |
9.7483 |
6.9724 |
8.1497 |
6.4551 |
4.9317 |
5.6693 |
9.3688 |
9.1398 |
|
|
AIC |
38.9818 |
34.9602 |
36.8325 |
34.0350 |
30.8047 |
32.4775 |
38.5052 |
38.2083 |
|
|
MSC |
2.0395 |
2.6740 |
2.2056 |
2.8256 |
3.4128 |
3.1836 |
2.1644 |
2.1867 |
|
|
Higuchi |
KH |
30.596 |
30.634 |
29.903 |
30.840 |
30.393 |
30.699 |
30.498 |
30.498 |
|
RSQR_ADJ |
0.9643 |
0.9652 |
0.9306 |
0.9517 |
0.9617 |
0.9739 |
0.9746 |
0.9724 |
|
|
MSE_ROOT |
6.0301 |
5.8498 |
7.6427 |
6.8831 |
6.2776 |
5.3196 |
5.2010 |
5.3565 |
|
|
AIC |
33.2179 |
32.8535 |
36.0616 |
34.8055 |
33.7004 |
31.7134 |
31.4427 |
31.7964 |
|
|
MSC |
3.0002 |
3.0251 |
2.3340 |
2.6972 |
2.9302 |
3.3109 |
3.3415 |
3.2553 |
|
|
Korsmeyer-Peppas |
KKP |
33.780 |
36.080 |
37.661 |
36.985 |
34.929 |
34.091 |
32.578 |
33.250 |
|
n |
0.449 |
0.416 |
0.381 |
0.407 |
0.429 |
0.446 |
0.466 |
0.456 |
|
|
RSQR_ADJ |
0.9633 |
0.9827 |
0.9752 |
0.9734 |
0.9700 |
0.9767 |
0.9716 |
0.9714 |
|
|
MSE_ROOT |
6.1142 |
4.1253 |
4.5684 |
5.1039 |
5.5567 |
5.0270 |
5.5042 |
5.4524 |
|
|
AIC |
34.0452 |
29.3234 |
30.5477 |
31.8777 |
32.8978 |
31.6957 |
32.7838 |
32.6704 |
|
|
MSC |
2.8623 |
3.6134 |
3.2530 |
3.1851 |
3.0639 |
3.3139 |
3.1180 |
3.1097 |
|
|
Hixson-Crowell |
KHC |
0.073 |
0.080 |
0.080 |
0.084 |
0.080 |
0.078 |
0.071 |
0.072 |
|
RSQR_ADJ |
0.8986 |
0.9145 |
0.8661 |
0.9258 |
0.9504 |
0.9541 |
0.9102 |
0.9099 |
|
|
MSE_ROOT |
10.1652 |
9.1700 |
10.6123 |
8.5300 |
7.1510 |
7.0465 |
9.7867 |
9.6695 |
|
|
AIC |
39.4843 |
38.2479 |
40.0008 |
37.3797 |
35.2637 |
35.0869 |
39.0289 |
38.8844 |
|
|
MSC |
1.9558 |
2.1260 |
1.6775 |
2.2681 |
2.6696 |
2.7487 |
2.0771 |
2.0740 |
|
|
Baker-Lonsdale |
KBL |
0.028 |
0.029 |
0.028 |
0.030 |
0.028 |
0.029 |
0.027 |
0.027 |
|
RSQR_ADJ |
0.9457 |
0.9880 |
0.9927 |
0.9881 |
0.9821 |
0.9746 |
0.9472 |
0.9521 |
|
|
MSE_ROOT |
7.4384 |
3.4360 |
2.4705 |
3.4227 |
4.2948 |
5.2413 |
7.5070 |
7.0511 |
|
|
AIC |
35.7365 |
26.4682 |
22.5097 |
26.4219 |
29.1455 |
31.5354 |
35.8466 |
35.0949 |
|
|
MSC |
2.5804 |
4.0893 |
4.5927 |
4.0944 |
3.6893 |
3.3406 |
2.6075 |
2.7056 |
|
|
Peppas-Sahlin |
K1 |
19.688 |
38.230 |
40.504 |
38.607 |
35.138 |
34.434 |
16.099 |
13.475 |
|
K2 |
14.281 |
-3.458 |
-4.302 |
-3.800 |
-3.219 |
-2.920 |
16.660 |
19.899 |
|
|
M |
0.297 |
0.562 |
0.553 |
0.603 |
0.660 |
0.644 |
0.292 |
0.273 |
|
|
RSQR_ADJ |
0.9532 |
0.9908 |
0.9901 |
0.9918 |
0.9959 |
0.9893 |
0.9631 |
0.9625 |
|
|
MSE_ROOT |
6.9102 |
3.0013 |
2.8887 |
2.8354 |
2.0623 |
3.4060 |
6.2773 |
6.2390 |
|
|
AIC |
35.7876 |
25.7803 |
25.3213 |
25.0979 |
21.2777 |
27.2981 |
34.6349 |
34.5615 |
|
|
MSC |
2.5719 |
4.2039 |
4.1241 |
4.3151 |
5.0006 |
4.0468 |
2.8095 |
2.7945 |
K0: Zero-order rate constant; K1: First-order rate constant; KH: Higuchi constant; KKP: Korsmeyer-Peppas constant;
n: Release exponent; KHC: Hixson-Crowell constant; KBL: Baker-Lonsdale constant; K1 and K2: Release exponents;
RSQR_ADJ: Adjusted R2; MSE_ROOT: Root mean square error; AIC: Akaike Information Criterion; MSC: Model selection criterion.
2. MATERIALS AND METHOD:
2.1. Materials:
Metformin Hydrochloride Prolonged-Release 500 mg Tablets:
Reference (Branded):
Metformin Hydrochloride Prolonged-Release 500 mg Tablets (Reference No. R1, R2)
Test (Generic):
Metformin Hydrochloride Prolonged-Release 500 mg Tablets (Test No. T1, T2, T3, T4, T5, T6)
Glimepiride 1 mg Tablets:
Reference (Branded):
Glimepiride 1 mg Tablets (Reference No. R3, R4)
Test (Generic):
Glimepiride 1 mg Tablets (Test No. T7, T8, T9, T10)
2.2. Methods:
Metformin Hydrochloride (UV Spectrophotometry)
Diluent: Water
The assay of Metformin Hydrochloride, A 20 mg working standard was dissolved in water, diluted, and filtered (0.45 µm PVDF). For the sample, 20 tablets were powdered, and an equivalent of 100 mg was dissolved, diluted, and filtered. Absorbance was measured at 232 nm with water as blank.
For dissolution testing, Conducted in 1000 mL phosphate buffer (pH 6.8) at 37 ± 0.5 °C, 100 rpm (paddle). Samples were collected at 15 min, 1 h, 3 h, 5 h, and 8 h, diluted, filtered, and analyzed at 232 nm. The standard solution was prepared as per assay.
Glimepiride (HPLC):
Chromatographic Conditions:
· Column: Inertsil ODS, 150 x 4.6 mm, 5 µm
· Wavelength: 228 nm
· Injection Volume: 10 µl
· Flow Rate: 1.0 ml/min
· Mobile Phase: 50:50 buffer: acetonitrile
· Diluent: 90:10 acetonitrile: water
The assay for Glimepiride - A 10 mg IPRS standard was dissolved in 70 mL diluent, sonicated, diluted to 100 mL, and filtered (0.45 µm PVDF). For the sample, 20 tablets were powdered, and an amount equivalent to 10 mg was dispersed in solvent to prepare a 0.01% w/v solution, then filtered. Injections were run in the sequence: blank → standard → sample → bracketing standard. System suitability criteria: theoretical plates ≥2000, tailing factor ≤2.0, RSD ≤2.0%.
For dissolution testing, conducted in 900 mL phosphate buffer (pH 7.8) using a paddle at 75 rpm, 37 ± 0.5 °C. A 125 mg IPRS standard was prepared in solvent mixture (90:10 acetonitrile: water), diluted, and filtered. Samples were taken at 30 min, diluted, filtered, and analyzed, with blank, standard, and sample injections used to quantify dissolved drug.
Applied methods to compare dissolution profiles:
Statistical Analysis (ANOVA and t-tests)
Analysis of Variance (ANOVA):
· ANOVA was used to test whether mean differences among groups were statistically significant.
t-tests:
· These were utilised to compare the means between two groups. Both ANOVA and t-tests were done using [MS Excel 2019], with significance levels set at p < 0.05.
Model Independent Methods (Non-Mathematical Models: f1, f2, Rescigno ratio).
f1 (Difference Factor):
f2 (Similarity Factor):
f2=50×log {[1+ (Rt−Tt)2] −0.5×100}
Rescigno Ratio ζ1
Rescigno Ratio ζ2
where:
· Di,T, and Di,R are the Dissolution values at time i for test and reference products.
· dDi,T/dt and dDi,R/dt are the Dissolution values at time i for test and reference products.
· A ratio close to 1 indicates that the test and reference products have similar release profiles.
EXPERIMENTAL:
Metformin hydrochloride and glimepiride tablets were analyzed using UV spectroscopy and HPLC to compare branded and generic formulations.
· Metformin: Assayed at 232 nm by UV; dissolution tested in 1000 mL phosphate buffer (pH 6.8) at 37 °C, 100 rpm, with samples taken up to 8 h.
· Glimepiride: Quantified by HPLC (Inertsil ODS column, 50:50 buffer–acetonitrile); dissolution in 900 mL phosphate buffer (pH 7.8) at 37 °C, 75 rpm, sampled at 30 min.
3. RESULTS and DISCUSSION:
The study compared branded and generic Metformin HCl prolonged-release and Glimepiride tablets using UV spectroscopy, HPLC, and in-vitro dissolution tests, providing insights into quality, dissolution, and bioequivalence across formulations
3.1. Metformin Hydrochloride Prolonged Release 500 mg:
At 0.25 h, references were similar (R1: 17.78%, R2: 18.98), while generics varied (T1 highest 21.51%, T6 lowest 15.17%).
Early time point (1 hour): The dissolution values for R1 and R2 remained close (36.13% and 35.67%, respectively), with T2 and T3 nearly matching the reference profiles, whereas T5 and T6 showed lower dissolution values.
· Mid time point (3 hours): At this point, branded samples were similar (R1: 48.32%, R2: 47.81), while generics, especially T2 (59.79%) and T3 (60.19%), showed higher dissolution, indicating faster release.
· Later time point (5 hours): Branded samples remained consistent, while generics showed higher solubility, with T4 highest (73.67%).
· Final time point (8 hours): Dissolution of branded and generics converged, with all nearing reference levels except T3, which stayed slightly lower.
All samples met pharmacopoeial standards (API 90 -110%). Both branded and generic forms released >85% within 8 h, confirming consistent efficacy.
3.2. Glimepiride 1 mg:
Statistical Analysis:
· Model-independent techniques: f1 and f2 confirmed similarity between generic and branded profiles, while Rescigno ratios supported the closeness of their release characteristics.
· Model-based approaches: Dissolution data were fitted to various models; First Order, Higuchi, and Korsmeyer-Peppas gave the best fit, indicating a complex release mechanism involving diffusion and polymer relaxation.
ANOVA and t Tests:
ANOVA showed significant changes in release over time, confirming sustained release. Pairwise t-tests found no significant differences between test and reference profiles, supporting therapeutic similarity.
4. CONCLUSION:
This study highlights the importance of rigorous analysis to ensure the quality and efficacy of anti-diabetic drugs, focusing on metformin prolonged-release and glimepiride. Branded and generic formulations were compared using UV spectroscopy, HPLC, and in-vitro dissolution testing. Drug release kinetics were evaluated with model-dependent (Zero Order, First Order, Higuchi, Korsmeyer-Peppas, Hixson-Crowell, Baker-Lonsdale) and model-independent (f₁, f₂, Rescigno ratio) approaches. Results showed time-dependent differences in dissolution affecting bioavailability, validated through ANOVA and t-tests. The findings support the reliability of generic formulations and underline the need for thorough evaluation to strengthen diabetes management.
5. LIST OF ABBREVIATIONS:
1. T2DM - Type 2 Diabetes Mellitus
2. HPLC - High-Performance Liquid Chromatography
3. API - Active Pharmaceutical Ingredient
4. SR - Sustained Release
5. XR - Extended Release
6. USP - United States Pharmacopeia
7. IPRS - Indian Pharmacopoeia Reference Standard
8. PVDF - Polyvinylidene Fluoride
9. NMR - Nuclear Magnetic Resonance
6. CONFLICT OF INTEREST:
The authors declare no financial or personal conflicts of interest. The study was conducted without external funding, and the authors take full responsibility for the results.
7. ACKNOWLEDGEMENTS:
My special thanks to the Indian Pharmacopoeia Commission, which has generously assisted me both with resources and moral support, thus performing a great role in the successful completion of this work.
1. Ghante M, Dargude S, Patil A, Bhusari V. Introduction: Quality Assurance from Perspective of Pharmaceutical Industry. In: Modern Aspects of Pharmaceutical Quality Assurance. Singapore: Springer Nature Singapore. 2024. p. 1–7.
2. Soumya K, Susanna S, Murthy TEGK. In vitro dissolution studies on commercial brands containing immediate release pioglitazone and metformin hydrochloride extended release tablets . Research Journal of Pharmaceutical Dosage Forms and Technology. 2017; 9(1).
3. Gomeni R, Bressolle-Gomeni F. Deconvolution Analysis by Non-linear Regression Using a Convolution-Based Model: Comparison of Nonparametric and Parametric Approaches. AAPS J. 2020 Jan 9;22(1):9.
4. Sun ML, Liu HJ, Luo XD, Wang Y, Zhang W, Liu C, et al. Bioequivalence and Safety Assessment of Two Formulations of Metformin Hydrochloride Sustained-Release Tablets (Yuantang® SR and Glucophage® XR) Under Fed Conditions in Healthy Chinese Adult Subjects: An Open-Label, Two-Way Crossover, Sequence Randomized Phase I Clinical Trial. Drugs R D. 2022 Mar 21;22(1):51–60.
5. Mamari R, Ibrahim R. The effect of Chronic treatments of Type 2-diabetes mellitus on COVID-19 Morbidity and Symptoms Severity. Res J Pharm Technol. 2023;16(11).
6. Manikandan M, Kannan K, Selvamuthukumar S, Manavalan R. Design, development and evaluation of Metformin Hydrochloride and Glimepiride immediate release tablets. Res J Pharm Technol. 2012; 5(4).
7. Proks P, Reimann F, Green N, Gribble F, Ashcroft F. Sulfonylurea Stimulation of Insulin Secretion. Diabetes. 2002 Dec 1;51(suppl_3):S368–76.
8. Jain A, Chaudhary J, Saini A, Mehan N. Quality assessment and comparative study of different marketed brands of metformin. Res J Pharm Technol. 2019; 12(3).
9. Gupta VK, Singh A, Gupta AK, Trigunayat A, Kumar B. A Pilot Study on Knowledge and Experience on use of Generic Medicines among Healthcare Professionals. Res J Pharm Technol. 2022; 15(12).
10. Parthiban C, Bhagavan Raju M, Sudhakar M. A novel simultaneous estimation of metformin, glimepiride and rosiglitazone in tablet dosage form by RP-HPLC method. Res J Pharm Technol. 2011; 4(10).
11. Sandhya SM, Shiji Kumar PS. Simple and rapid simultaneous rp-hplc method for determination of glimepiride and metformin in tablet dosage form. Res J Pharm Technol. 2014; 7(8).
12. Gholve R, Pekamwar S, Wadher S, Kalyankar T. Stability-indicating RP-HPLC method development and validation for simultaneous estimation of telmisartan and rosuvastatin calcium in bulk and in tablet dosage form. Futur J Pharm Sci. 2021 Dec 4; 7(1): 224.
13. Bayas JP, Sumithra M. Analytical method development and validation of dasatinib in bulk and pharmaceutical formulation using quality by design. Res J Pharm Technol. 2021; 14(3).
14. Mohd AB, Sanka K, Gullapelly R, Diwan P V, Shastri N. Development and validation of RP-HPLC method for glimepiride and its application for a novel self-nanoemulsifying powder (SNEP) formulation analysis and dissolution study. J Anal Sci Technol. 2014 Dec 2; 5(1): 27.
15. Neelima K, Prasad YR. Analytical Method Development and Validation of Metformin, Voglibose, Glimepiride in Bulk and Combined Tablet Dosage Form by Gradient RP-HPLC. Pharm Methods. 2014 Jul 18; 5(1): 27–33.
16. Xiong K, Ma X, Cao N, Liu L, Sun L, Zou Q, et al. Identification, characterization and HPLC quantification of impurities in apremilast. Analytical Methods. 2016; 8(8): 1889–97.
17. Desai RJ, Sarpatwari A, Dejene S, Khan NF, Lii J, Rogers JR, et al. Comparative effectiveness of generic and brand-name medication use: A database study of US health insurance claims. PLoS Med. 2019 Mar 13; 16(3): e1002763.
18. Cotia A, Oliveira Junior HA, Matuoka JY, Boszczowski Í. Clinical Equivalence between Generic Versus Branded Antibiotics: Systematic Review and Meta-Analysis. Antibiotics. 2023 May 21; 12(5): 935.
19. Baghel US, Sharma A, Gautam H, Mukim M, Singh D, Dave S, et al. Spectrophotometric Multi-Component Quantitation of Rosiglitazone, Glibenclamide and Metformin HCl in Pharmaceutical Dosages Form by using Cramer’s Matrix. Res J Pharm Technol. 2023; 16(12).
20. Yunita EP, Asriyanti NLPG, Gunawan A. Evaluation of Drug Combinations’ Effectiveness in Hyperkalemia Management of Chronic Kidney Disease Patients. Res J Pharm Technol. 2022; 15(1).
21. Karun KM, Deepthy MS. Generalized Estimating Equations in Longitudinal Studies: A Non-Parametric Alternative for Two-Way Repeated Measures Mixed ANOVA. Res J Pharm Technol. 2023; 16(5).
22. Zumbo BD. Univariate Tests. In: Encyclopedia of Quality of Life and Well-Being Research. Dordrecht: Springer Netherlands; 2014. p. 6819–20.
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Received on 03.08.2025 Revised on 05.12.2025 Accepted on 02.03.2026 Published on 01.07.2026 Available online from July 04, 2026 Research J. Pharmacy and Technology. 2026;19(7):2926-2932. DOI: 10.52711/0974-360X.2026.00417 © RJPT All right reserved
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