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Development And Validation Of An Hptlc Method For The Quantitative Determination Of Rutin In Hemidesmus Indicus (L.) R. Br. Mother Tincture.
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| Author:
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, SUMAN SHRIVASTAVA, S.J.DAHARWAL
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| Abstract:
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Hemidesmus indicus (L.) R. Br. relate to the Asclepiadaceae family. The dried roots of plant introduced as Indian sarsaparilla or Anantmool, are always the most popular medicinal herbs employed to the treatment of syphilis, chronic rheumatism, urinary disorders and often include antibacterial effects. This study aimed to identify the content of rutin in H. indicus and to validate the high-performance thin-layer chromatography method. The plant was collected, and the chromatographic test established the phytochemical screening. The separation was operated on TLC aluminum plates precoated with silica gel 60 F254. Determination and quantitation were done in reflection/absorbance mode with densitometric scanning at 371 nm. TLC study of the compound ascertained a dark black spot at Rf 0.88 when the solvent system composed Acetonitrile: water (5:5% v/v). The developed technique has been validated according to ICH Q2 (R1) guideline with respect to linearity, accuracy, precision, robustness, limit of detection and quantitation. Rutin was verified and its content was determined using a validated HPTLC method with good linearity within the range of 200-1000 ng/spot with r2 = 0.9998, correlation coefficient with calibration curve equation y = 0.0297x + 0.0001. The average % recovery values varied from 99.03-101.15 % for in-house. The peak area determination at 3 different concentration levels shows low values of %RSD (< 2 %) for inter-day (0.04-0.06%) and intra-day (0.04-0.05%) rutin variation. The LOD and LOQ have proven to be 10 and 50 ng/spot, respectively. The average content of rutin in extract and marketed mother tincture was 0.23 ± 0.001 and 0.21 ± 0.0005 µg/g, respectively. The proposed method indicates that simple, precise and accurate for quantification of active constituents with frequent quality control assessment of H. indicus in homeopathic mother tincture.
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| Keyword:
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Hemidesmus indicus (L.) R. Br., Rutin, High Performance Thin Layer Chromatography, ICH Q2 (R1) guidelines
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2021.13.02.318
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| Download:
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