JPC - Journal of Planar Chromatography - Modern TLC
Volume/Issue:
Volume 30: Issue 3
High-performance thin-layer chromatography and indirect TLC—HRMS-based determination of 20-hydroxyecdysone in Sesuvium portulacastrum
Authors:
Niramaya S. Muchate Savitribai Phule Pune University, Pune, India

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Nitin S. Kadam Savitribai Phule Pune University, Pune, India
Savitribai Phule Pune University, Pune, India

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Nilima S. Rajurkar Savitribai Phule Pune University, Pune, India

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Tukaram D. Nikam Savitribai Phule Pune University, Pune, India

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Pages:
193–198
Online Publication Date:
Jun 2017
Publication Date:
01 Jun 2017
Article Category:
Research Article
DOI:
https://doi.org/10.1556/1006.2017.30.3.7
Keywords:
20-Hydroxyecdysone; High-performance thin-layer chromatography; Indirect TLC—high-resolution mass spectrometry; Sesuvium portulacastrum
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20-Hydroxyecdysone (20E), an imperative phytoecdysteroid, regulates several biochemical and physiological processes during the different developmental stages in insects. The aim of the present work was to validate an analytical method for the detection and identification of the 20E content in Sesuvium portulacastrum using high-performance thin-layer chromatography (HPTLC) combined with image analysis and using high-resolution mass spectrometry (HRMS). Using HPTLC, the better separation of 20E was achieved on TLC plate prewashed with ethanol—ethyl acetate—water (8:2:0.5) and using the solvent system consisting of chloroform—methanol—benzene (12.5:2.5:1.5) at RF value of 0.30. The bands were documented using a TLC visualizer and scanned at 254 nm in absorption mode. The method was validated for specificity, linearity, precision, robustness, stability, accuracy, limit of detection (LOD), and limit of quantification (LOQ) as per the International Conference on Harmonization (ICH) guidelines. The linear calibration curve was achieved in the range of 50–500 ng band−1 with the most significant correlation coefficient (r = 0.997). The relative standard deviations for intra-day and inter-day precisions were found to be 1.897 and 3.125 at 300 ng band−1, and the limit of detection and limit of quantification were 4.67 and 14.16 ng band−1, respectively. Furthermore, 20-hydroxyecdysone isolated from standard and plant sample bands was identified using indirect TLC—HRMS analysis. The HPTLC method and the indirect TLC—HRMS analysis were observed to be simple, precise, accurate, and suitable for the determination of 20E in S. portulacastrum. In the phytopharmaceutical industry, the present method can be applied for the screening and routine quality control of 20E in plant materials and formulations.

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Cover JPC - Journal of Planar Chromatography - Modern TLC
JPC - Journal of Planar Chromatography - Modern TLC
Print ISSN:
0933-4173
Online ISSN:
1789-0993

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Journal of Planar Chromatography - Modern TLC
Language English
Size A4
Year of
Foundation		 1988
Volumes
per Year		 1
Issues
per Year		 6
Founder Akadémiai Kiadó
Founder's
Address		 H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.
Publisher Akadémiai Kiadó
Springer Nature Switzerland AG
Publisher's
Address		 H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.
CH-6330 Cham, Switzerland Gewerbestrasse 11.
Responsible
Publisher		 Chief Executive Officer, Akadémiai Kiadó
ISSN 0933-4173 (Print)
ISSN 1789-0993 (Online)

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