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Abstract

A methodical approach to development, Quality by Design (QbD) starts with predetermined goals for a product, process comprehension, and process control based on knowledge and quality risk management. When developing and validating a traditional approach, it is possible that it will not achieve the desired outcome. AQbD is the application of the QbD idea to the validation of analytical methods. A systematic and trustworthy strategy for developing analytical processes that cover all stages of a product's lifecycle is offered by AQbD. The AQbD technique for creating the HPTLC method is predicated on changing one parameter while maintaining the other parameters constant in order to obtain the desired result. The development of the conventional HPTLC method necessitates numerous tries and errors, which impacts the accuracy,precision and resilience of the process.Therefore, AQbD is used for analytical method validation in order to reduce time, complexity, and?most importantly?validation failure. Method intent definition, experimental design, experimental result evaluation, method condition selection, and risk assessment with varying analytical parameters and evaluation conditions are some of the phases that make up the QbD Approach to method development. AQbD in HPTLC is essential to understand different factors showing significant impact on method outcome. The HPTLC method should display robustness to facilitate use for a longer period along with very low potential of failure.

Keywords

Design in HPTLC, Approaches, analytical methods

Introduction

Introduction to HPTLC

HPTLC is a more complicated and automated version of thin-layer chromatography (TLC) that has better detection limits and separation efficiency. It is also known as flat-bed chromatography, planar chromatography, or high-pressure thin layer chromatography. It is a strong analytical instrument that works well for both qualitative and quantitative issues. [1,2] Depending on the kind of adsorbents used to the plates and the solvent system utilized during development, separation may arise from partitioning, adsorption, or both.  Phytochemical analysis, biomedical analysis, quantification of herbal medications, analytical analysis, fingerprint analysis, and the possibility of hyphenation (HPTLC-MS, HPTLC-FTIR, and HPTLC- Scanning Diode Laser) are among the applications cited. [3] High resolution sorbents with specific particle sizes or chemically altered surfaces, more effective elution methods, the capacity to integrate with other instrumental techniques, the development of computer programmes for method optimization, and the application of modern tools such as chromatographic chambers, densitometers, and video scanners. [4] HPLC, a very sophisticated chemical standardization technique, is far more reliable and reproducible when used to standardize herbal compositions, both single and compound. The separation method called high-pressure liquid chromatography uses a stationary phase and liquid mobile phase. The size of stationary phase utilized determines whether ion exchange, adsorption, or partitioning is employed for separations. [5].

Principle:

  1. Separation: Components are separated by HPTLC. based on affinity between stationary and mobile phases
  2. Adsorption: Components interact with the adsorbent surface, resulting in separation.
  3. Components split into mobile and stationary phases.

Fig No.1. Principles of HPTLC

Benefits and drawbacks of HPTLC in comparison to TLC: In recent years, HPTLC has become a reliable identification technique and has replaced traditional TLC.The device is managed by software. The most efficient silica gel hydrophilic phasethat meets the requirements of most pharmacopeias is being utilized for HPTLC pollutants identification. [6]

Table No. 1: Difference between TLC and HPTLC [6]

Sr.No.

Feature

TLC

HPLC

1

Technique

Manual

Instrumental

2

Plates

Lab-made

Pre-coated

3

Plate height

30 μm

12 μm

4

Layer of Sorbent

250 μm

100 μm

5

Stationary Phase

Silica gel, alumina, &Kiesulgur

Numerous stationary phase options, such as C8, C18 for reversed phase and silica gel for normal phase

6

Separations

10-15 cm

3-5 cm

7

Analysis time

20-200 min

1-3 min

8

Average Size of Particles

10-12 μm

5-6 μm

9

Efficiency

Less

High

10

Sample Holder

Capillary/Pipette

Syringe

11

Sample Spotting

Manual Spotting

Autosampler

12

Size of Sample

Uncontrolled

Controlled solvent Independence

13

Sample Shape

Circular

Rectangular

14

Sample tracks for each plate

<10

<36

15

Volume Range

1-10 μl

0.1 to 500μl

16

Development Chamber

More amount

Less amount

17

Wavelength range

254 or 366

190 or 180

18

Detection limit

1-5 pg

100-500 pg

19

Limit of detection (fluorescence)

50-100pg

5-10 pg

20

PC Connectivity

No

Yes

21

Quantitative Analysis

No

Yes

22

Scanning

No

UV/Visible/Fluorescence scanner

23

Analysis Judgement

By Analyst

By machine

Automation for HPTLC:

Modern TLC, often referred to as HPTLC, is primarily used for quantification, requires tools, and can only be carried out on precoated layers. Consequently, the terms TLC and HPTLC are used interchangeably. TLC is widely used around the world to teach the principle of chromatography. The sample's visibility during chromatography, the demonstration apparatus's extremely low cost, and its user-friendliness are the main justifications for this method. It employs a multifaceted technique to improve resolution under capillary flow-controlled circumstances. Planar chromatography can separate molecules in one or two dimensions. External control of mobile-phase velocity is also possible, as seen in forced-flow development. [7] HPTLC is the quickest chromatographic method. The samples are chromatographed in parallel. Each stage of the technique is completed individually, making In addition to being quicker, HPTLC is adaptable enough to evaluate several samples at once. The amount of stationary and mobile phase used depends on the number of samples being analyzed. [8]

Steps involved in HPTLC [9]:

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  54. By Design Approach Amruta, S., & Khurd, P. B. (n.d.). Sandip R. More1, Kajal V. Doshi1, Vandana Gawande1, Abhijeet S. Sutar2, Arun M. Kashid1, Shraddha V. Tathe1, Shubhangee S. Gaikwad1.

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L. P. Jain
Corresponding author

Department of Pharmaceutical Chemistry, Government College of Pharmacy, Karad

Photo
M. S. Charde
Co-author

Department of Pharmaceutical Chemistry, Government College of Pharmacy, Karad

Photo
S. J. Momin
Co-author

Department of Pharmaceutical Chemistry, Government College of Pharmacy, Karad

Photo
S. V. Potdar
Co-author

Department of Pharmaceutics, Government College of Pharmacy, Karad

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N. D. Kulkarni
Co-author

Department of Pharmaceutical Chemistry, Government College of Pharmacy, Karad

L. P. Jain*, M. S. Charde, S. J. Momin, S. V. Potdar, N. D. Kulkarni, Quality by Design in HPTLC: A Review of Method Development Approaches, Int. J. Sci. R. Tech., 2025, 2 (6), 179-197. https://doi.org/10.5281/zenodo.15585927

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