Comparative Evaluation of Spectrophotometric and HPLC-PDA Methods for Carotenoid Quantification in Cold Pressed Capsicum annuum Oil

Laksana Charoenchai; Tawanchai Phohom; Bancha Yingngam; Charuwan Thanawiroon

doi:10.53848/ssstj.v13i1.1306

Authors

Laksana Charoenchai Rangsit University Pathum
Tawanchai Phohom Ubon Ratchathani University
Bancha Yingngam Ubon Ratchathani University https://orcid.org/0000-0001-7215-9123
Charuwan Thanawiroon Ubon Ratchathani University

DOI:

https://doi.org/10.53848/ssstj.v13i1.1306

Keywords:

Capsicum annuum, Carotenoids , β-carotene, UV‒Vis spectrophotometry, HPLC-PDA, Quality control

Abstract

Accurate and validated analytical methods for the measurement of carotenoids in plant oils are needed for quality control, regulatory compliance and product development in the food, nutraceutical, and pharmaceutical industries. In this work, the methods of UV‒Vis spectrophotometry and high-performance liquid chromatography with photodiode array detection (HPLC‒PDA) were evaluated for the quantification of carotenoids in cold-pressed Capsicum annuum oil under harmonized validation conditions. Both techniques were validated according to ICH Q2 (R2) principles, which include predetermined analytical target profiles, risk-based validation criteria and measurement uncertainty estimation. The spectrophotometric approach exhibited good linearity (R²=0.9999),a recovery of 97.66–100.96% and an interday precision of ≤3.34% RSD, indicating its suitability for quick screening of apparent total carotenoids represented as β-carotene equivalents. Greater specificity for β-carotene was obtained with the developed HPLC-PDA method, yielding R² = 0.9992, with 82.75–95.14% recovery and LODs and LOQs of 5.41 and 9.00 µg/mL, respectively. The apparent total carotenoid content was estimated at 161.36 ± 0.83 mg/g oil by spectrophotometric analysis, and a β-carotene concentration of 8.5 mg/g oil was determined by HPLC-PDA measurements. This example demonstrates that the two methods do not measure the same endpoint but rather address different analytical concerns. Overall, the two approaches are complementary: spectrophotometry, as a high-throughput low-cost approach, is good for quality control, whereas HPLC-PDA is suitable when compound-specific identification and regulatory evidence are needed.

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