Volume : 13, Issue : 05, May – 2026
Title:
COMPARATIVE EXTRACTION AND QUANTITATIVE ESTIMATION OF KHELLIN AND FUMARIC ACID FROM AMMI MAJUS L. AND FUMARIA PARVIFLORA L. BY HPLC AND HPTLC METHODS
Authors :
Uma Shankar Joshi*,Dr. S. K. Gupta ,Dr. Vivek Gupta, Dr. Jitendra Kumar Malik
Abstract :
The present study was undertaken to evaluate different extraction techniques for the isolation and quantitative estimation of Khellin from Ammi majus L. and Fumaric acid from Fumaria parviflora L. using UV spectrophotometric, HPLC, and HPTLC methods. Various extraction methods including maceration, reflux extraction, Soxhlet extraction, and ultrasound-assisted extraction (UAE) were employed using solvents of different polarities such as methanol, ethanol, chloroform, acetone, petroleum ether, and n-hexane. Among all extraction techniques, ultrasound-assisted extraction demonstrated the highest extraction efficiency for both phytoconstituents. The methanolic UAE extract of Ammi majus L. showed the maximum Khellin content (6.22% w/w), whereas methanolic UAE extract of Fumaria parviflora L. exhibited the highest Fumaric acid content (6.6% w/w). Quantitative estimation by HPLC revealed that the methanolic extract of Ammi majus L. contained 6.01% w/w of Khellin at a retention time of 12.394 minutes, while the methanolic extract of Fumaria parviflora L. contained 6.35% w/w of Fumaric acid at a retention time of 4.45 minutes. Calibration curves of standard Khellin and Fumaric acid showed good linearity between concentration and peak area, confirming the suitability of the developed HPLC method. HPTLC fingerprinting profiles of both plant extracts revealed the presence of several phytoconstituents at different Rf values. Khellin was identified at an Rf value of 0.73 in Ammi majus L. extract, whereas Fumaric acid was identified at an Rf value of 0.06 in Fumaria parviflora L. extract. Quantitative HPTLC analysis showed 6.38% w/w Khellin and 6.69% w/w Fumaric acid in methanolic extracts, respectively. The HPTLC findings were found to be comparable with HPLC results, confirming the reliability and reproducibility of both analytical methods. The results of the present investigation indicate that ultrasound-assisted extraction using methanol is an effective extraction method for isolation of Khellin and Fumaric acid. Furthermore, HPLC and HPTLC methods were found to be accurate, sensitive, rapid, and reproducible techniques for qualitative and quantitative analysis of these phytoconstituents. The study may be useful for standardization, quality control, and further pharmacological evaluation of these medicinal plants and their bioactive compounds.
Keywords: Ammi majus L., Fumaria parviflora L., Khellin, Fumaric acid, Ultrasound-assisted extraction, HPLC, HPTLC, Phytochemical analysis, Quantitative estimation, Medicinal plants.
Cite This Article:
Please cite this article in press Uma Shankar Joshi et al., Comparative extraction and quantitative estimation of khellin and fumaric acid from ammi majus l. And fumaria parviflora l. By hplc and hptlc methods.., Indo Am. J. P. Sci, 2026; 13(05).
REFERENCES:
1. Shakya AK. Medicinal plants: Future source of new drugs. International journal of herbal medicine. 2016 Jun 4;4(4):59-64.
2. Dar RA, Shahnawaz M, Ahanger MA, Majid IU. Exploring the diverse bioactive compounds from medicinal plants: a review. J. Phytopharm. 2023 Jun 30;12(3):189-95.
3. Khalil N, Bishr M, Desouky S, Salama O. Ammi visnaga L., a potential medicinal plant: A review. Molecules. 2020 Jan 12;25(2):301.
4. El-Ahmady S, Ibrahim N, Farag N, Gabr S. Apiaceae plants growing in the East: Centuries of healing traditions and culture. InEthnopharmacology of Wild Plants 2021 Feb 14 (pp. 246-300). CRC Press.
5. Kumar S, Sharma AK, Kamboj A. Fumaria parviflora Lam.(Fumitory): A traditional herbal medicine with modern evidence. Asian j pharm pharmacol. 2017;3(6):200-7.
6. Kumar A, P N, Kumar M, Jose A, Tomer V, Oz E, Proestos C, Zeng M, Elobeid T, K S, Oz F. Major phytochemicals: recent advances in health benefits and extraction method. Molecules. 2023 Jan 16;28(2):887.
7. Rasul MG. Conventional extraction methods use in medicinal plants, their advantages and disadvantages. Int. J. Basic Sci. Appl. Comput. 2018 Dec;2(6):10-4.
8. Mutha RE, Kalaskar M, Khan ZG. Modern analytical techniques for quality control and chemical identification of phytochemicals. Pharmacognosy and phytochemistry: principles, techniques, and clinical applications. 2025 Mar 13:167-88.
9. Khalil N, Bishr M, El-Degwy M, Abdelhady M, Amin M, Salama O. Assessment of conventional solvent extraction vs. supercritical fluid extraction of khella (Ammi visnaga L.) furanochromones and their cytotoxicity. Molecules. 2021 Feb 27;26(5):1290.
10. Alqasoumi SI, Alam P, Anwer MK, Abdel-Kader MS. Qualitative and quantitative analysis of khellin in Ammi visnaga fruits and pharmaceutical preparations using HPTLC and HPLC. Journal of Liquid Chromatography & Related Technologies. 2014 Jan 2;37(1):61-72.
11. Hooli V, Malviya R. HPLC Estimation of Khellin in Ammi majus L. Extracts and Fumaric acid in Fumaria parviflora L. Extracts. International Journal of Pharmacy & Life Sciences. 2025 Jan 1;16(1).
12. Bhargava A, Shrivastava P, Tilwari A. HPTLC analysis of Fumaria parviflora (Lam.) methanolic extract of whole plant. Future Journal of Pharmaceutical Sciences. 2021 Jan 4;7(1):1.