How to Perform Reverse Engineering in Pharmaceuticals?

Reverse engineering/deformulation is a complex process consisting of several phases, such as the identification and quantification of formulation components. It is used at the beginning of the development of a generic drug product. 

It is generally performed on an innovator’s formulation. This is performed at the laboratory level for the identification, quantification and characterization of API and excipients used.

This can be performed by using various analytical techniques. From the information obtained after doing analysis, one will get a fair idea about how to formulate a product.

Reverse engineering is a method that is used to reproduce and redesign an existing product. By using different methods, the design of the product can be studied and the product can be improved.

How HPTLC is useful for Reverse Engineering?

Camag HPTLC in pharmaceutical analysis is one of the effective techniques for various pharma-related analysis. High-performance Thin Layer chromatography is a modern version of TLC wherein the principles are the same but the practice is fully automated and GLP/USP/EP compliant.

HPTLC has the latest generation equipment with USP-compliant software. It is 21CFR Rule 11 compliant and designed for rapid analysis, and more output at a reduced cost.

Unique features of HPTLC are that it is the fastest, simplest, most economical and flexible, “visible” technique which can analyse in parallel more than 100 samples. It is risk-free and multiple detections can be made without repeating chromatography.

HPTLC is used for the analysis of non-volatile organics such as pharmaceuticals, APIs, and botanicals including AYUSH samples, forensics, foods, speciality chemicals, etc. for establishing purity, impurities, fingerprint, identification, quantification and reverse engineering.

After doing a single analysis, the analyst gets many data points. Data is obtained at 254nm, 366nm, in white light and after derivatization in white light or at 366nm with a suitable TLC photo documentation unit. Chromatograms can be derived along with spectral evaluation by the scanner. The scanner is used for qualitative and quantitative analysis.

Applications of HPTLC in drug analysis:

• Identification, quantification and validation studies of API, bulk drug and formulations.
• Related substance analysis, impurity studies
• Process monitoring of synthetic reaction mixtures
• Cleaning validation
• Identification of known/unknown impurities
• Content uniformity test
• Forced degradation/stability studies

Thus, HPTLC plays a crucial role in identifying potential molecules for drug discovery, development and further steps of identification, quantification, in-process monitoring, impurity profiling, stability testing, post-degradation studies, assessing biological activity etc. for both natural and synthetic compounds. 

Reference:

1. Warad T. et al., Recent trends in pharmaceutical reverse engineering, Indo American

Journal of Pharmaceutical research, 8(9):1670-1675, 2018.

Anchrom Enterprises Pvt. Ltd is one of the leaders in HPTLC in drug analysis. Please contact us at lab@anchrom.in for HPTLC analysis of plant extracts, drugs, ingredients in cosmetics, and forensic science.