# Analytical method development for simultaneous estimation of kojic acid, ascorbic acid and niacinamide in cosmetics and its validation

**Authors:** Vishnu Venkatesan, Gayatri Sukumaran, R. Lakshmi Sundaram, Chetan Ashok, S. Devaraja Dravida Pandiyan, Srikanth Jeyabalan, Mahendran Sekar, Ling Shing Wong, Vetriselvan Subramaniyan

PMC · DOI: 10.1186/s13065-026-01741-0 · 2026-02-21

## TL;DR

A new fast and accurate HPLC method was developed to measure three common cosmetic ingredients in one analysis, improving efficiency for quality control.

## Contribution

A validated rapid HPLC method for simultaneous quantification of kojic acid, ascorbic acid, and niacinamide in cosmetics.

## Key findings

- The method achieves separation and quantification of three analytes within 15 minutes using a C18 column and gradient elution.
- Detection limits were as low as 0.06 µg/mL for kojic acid, with recoveries between 98% and 102%.
- Precision (%RSD < 2%) and robustness met ICH standards, ensuring reliability for routine quality control.

## Abstract

A novel HPLC method was developed and validated for the concurrent measurement of kojic acid, ascorbic acid, and niacinamide in cosmetic formulations. Using methanol and 0.1% acetic acid gradient elution on a C18 column, the method enables efficient separation and quantification of all three active components within 15 min, markedly improving analytical speed and solvent economy compared to previous approaches. Rigorous method validation demonstrated outstanding precision, specificity, linearity (r2 > 0.99), and robustness were developed in accordance with ICH criteria with the system appropriateness factors, like the tailing factor and theoretical plate count, consistently meeting acceptance criteria. The limits of detection were as low as kojic acid (0.06 µg/mL), niacinamide (0.10 µg/mL), and ascorbic acid (0.15 µg/mL), while percent recoveries for all analytes ranged between 98% and 102% at multiple concentration levels, confirming accuracy. Repeatability and intermediate precision (%RSD < 2%) were upheld across replicate assays, underlining the method’s reliability for routine quality control. The unique advantage of this method lies in its simultaneous, rapid assessment of multi-functional vitamins and antioxidants common to cosmetics, streamlining regulatory compliance and product development. This robust protocol provides a practical solution for manufacturers and research laboratories seeking fast, precise, and economical analysis of active ingredients in commercial skincare products.

## Linked entities

- **Chemicals:** kojic acid (PubChem CID 3840), ascorbic acid (PubChem CID 9888239), niacinamide (PubChem CID 936), methanol (PubChem CID 887), acetic acid (PubChem CID 176)

## Full-text entities

- **Chemicals:** methanol (MESH:D000432), kojic acid (MESH:C011890), niacinamide (MESH:D009536), acetic acid (MESH:D019342), ascorbic acid (MESH:D001205)

## Figures

6 figures with captions in the complete paper: https://tomesphere.com/paper/PMC13032711/full.md

---
Source: https://tomesphere.com/paper/PMC13032711