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A Validated HPLC Method for Simultaneous Determination of 2-Hydroxy-4-methoxybenzaldehyde and 2-Hydroxy-4-methoxybenzoic Acid in Root Organs of Hemidesmus indicus

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Abstract

A reverse phase HPLC method was developed and validated for the simultaneous determination of 2-hydroxy-4-methoxybenzaldehyde and 2-hydroxy-4-methoxybenzoic acid in the root extracts of Hemidesmus indicus. A comprehensive validation of the method including sensitivity, linearity, reproducibility, accuracy, limit of detection (LOD) and limit of quantification (LOQ) was conducted using the optimized chromatographic conditions. The method was found to be linear (r > 0.998) in the range of 5–350 μg mL−1 for 2-hydroxy-4-methoxybenzaldehyde and for 2-hydroxy-4-methoxybenzoic acid (r > 0.999) in the range 10–300 μg mL−1. The method was found to be precise with inter-day precision values (% RSD) in the ranges of 0.61–1.76% for 2-hydroxy-4-methoxybenzaldehyde and 1.3–2.8% for 2-hydroxy-4-ethoxybenzoic acid while intra-day precisions (% RSD) of two analytes were in the range of 0.41–1.07 and 0.95–2.5%. The limits of detection (LODs) for 2-hydroxy-4-methoxybenzaldehyde and 2-hydroxy-4-methoxybenzoic acid were 0.84 and 2.34 μg mL−1. The described method was fast, sensitive and reproducible, and thus well suited for routine analysis of these two compounds from root extracts of H. indicus and other plants.

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Acknowledgment

This work was supported by a research grant (No.BT/PR4731/PBD/17/306/2004 to A. Mitra) from the Department of Biotechnology (DBT), Government of India. D. Sircar and G. Dey acknowledge the Council of Scientific and Industrial Research (CSIR) for the awards of ‘junior research fellowship’ and ‘research associateship’, respectively. A Mitra thanks DBT and the Indian Institute of Technology Kharagpur for concurrently awarding a long-term ‘biotechnology overseas associateship’ and a ‘sabbatical leave’.

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Authors and Affiliations

  1. Natural Product Biotechnology Group, Agricultural and Food Engineering Department, Indian Institute of Technology Kharagpur, Kharagpur, 721 302, India

    D. Sircar, G. Dey & A. Mitra

  2. Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Wakneghat, Solan, 173 215, India

    G. Dey

  3. Institut für Pharmazeutische Biologie, Technische Universität Braunschweig, Mendelssohnstrasse 1, 38106, Braunschweig, Germany

    A. Mitra

Authors
  1. D. Sircar
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  2. G. Dey
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  3. A. Mitra
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Correspondence to A. Mitra.

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Sircar, D., Dey, G. & Mitra, A. A Validated HPLC Method for Simultaneous Determination of 2-Hydroxy-4-methoxybenzaldehyde and 2-Hydroxy-4-methoxybenzoic Acid in Root Organs of Hemidesmus indicus . Chroma 65, 349–353 (2007). https://doi.org/10.1365/s10337-006-0146-x

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  • DOI: https://doi.org/10.1365/s10337-006-0146-x

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