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Box–Behnken supported development and validation of robust HPTLC method: an application in estimation of punarnavine in leaf, stem, and their callus of Boerhavia diffusa Linn

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Abstract

Boerhavia diffusa (BD) Linn. (Nyctaginaceae) is one of the most commonly used herbs in the Indian traditional system of medicine for the urinary disorders. The aim of the current investigation was to carry out initiation, development, and maintenance of BD callus cultures and quantitative estimation of punarnavine in plant and callus extracts. Leaves and stem of BD were used as explant for the tissue culture studies using Murashige and Skoog (MS) basal medium. MS Media comprising 2,4-Dichlorophenoxy acetic acid (2,4-D) (1 ppm) and 2,4-D (1 ppm) + Indole-3-acetic acid (IAA) (1.0 ppm) were found to yield friable callus from leaf explant; similarly, 2,4-D (0.3 ppm) + IAA (0.75 ppm) + Kinetin (0.3 ppm) and 2,4-D (0.5 ppm) + Naphthalene acetic acid (NAA) (1.5 ppm) + Kinetin (0.3 ppm) were found to yield friable callus from the stem explant. High-performance thin-layer chromatography method was been developed for the quantitative estimation of punarnavine (Rf = 0.73) using mobile phase containing toluene: ethyl acetate: formic acid in the ratio (7.0:2.5:0.7, v/v/v) at 262 nm. The validated method was found linear (r2 = 0.9971) in a wide range (100–1000 ng spot−1), precise, accurate, and robust. The values of limit of detection, LOD = 30.3 ng spot−1, and limit of quantification, LOQ = 100.0 ng spot−1. The robustness of the method was proved by applying the Box–Behnken design (BBD). The developed method found appropriate for the quality control of medicinal plants containing punarnavine as a constituent.

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Abbreviations

LOD:

Limit of detection

LOQ:

Limit of quantification

BBD:

Box–Behenken design

ICH:

International council for harmonization

HPTLC:

High-performance thin-layer chromatography

BD:

Boerhavia diffusa

MS:

Murashige and Skoog

TLC:

Thin-layer chromatography

Rf:

Retention factor

UV:

Ultra-violet

2–4, D:

2–4, Dichlorophenoxy acetic acid

IAA:

Indole acetic acid

NAA:

Naphthalene acetic acid

PPM:

Parts per million

PNE:

Punarnavine

RSD:

Relative standard deviation

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Acknowledgements

The authors acknowledge Jamia Hamdard, New Delhi, India for providing facility to conducting this study.

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Authors and Affiliations

  1. Department of Pharmacy, Mohammad Al-Mana College for Medical Sciences, Al-Safa, Dammam, 34222, Kingdom of Saudi Arabia

    Wasim Ahmad

  2. Department of Pharmacology, Mesco Institute of Pharmacy, Uttar Pradesh, Amroha, India

    Ibraheem Husain

  3. Department of Pharmaceutics, College of Clinical Pharmacy Imam, Abdulrahman Bin Faisal University, Dammam, 31441, Saudi Arabia

    Niyaz Ahmad

  4. Department of Natural Product & Alternative Medicines College of Clinical Pharmacy, Imam Abdulrahman Bin Faisal University, Dammam, 31441, Saudi Arabia

    Mohd Amir

  5. Department of Pharmaceutical Chemistry College of Clinical Pharmacy Imam, Abdulrahman Bin Faisal University, Dammam, 31441, Saudi Arabia

    Md. Sarafroz

  6. Department of Epidemic Disease Research, Institutes for Research and Medical Consultations (IRMC), Imam Abdulrahman Bin Faisal University, Dammam, 31441, Saudi Arabia

    Mohammad Azam Ansari

  7. Department of Pharmaceutics, College of Pharmacy, Jouf University, Al-Jouf, Sakaka, Saudi Arabia

    Ameeduz Zafar

  8. Department of Pharmacognosy, College of Pharmacy, Taif University, Taif, Saudi Arabia

    Abuzer Ali

  9. Department of Pharmacognosy and Phytochemistry, Faculty of Pharmacy, Jamia Hamdard, New Delhi, 110062, India

    Rasheeduz Zafar

  10. Faculty of Pharmacy, University Teknologi MARA (UITM), Cawangan Selangor, 42300, Kampus Puncak Alam, Seleangor Darul Ehsan, Malaysia

    Kamran Ashraf

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  1. Wasim Ahmad
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  2. Ibraheem Husain
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  3. Niyaz Ahmad
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  7. Ameeduz Zafar
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  8. Abuzer Ali
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Contributions

WA and RZ conceived and designed research. IB, NA, MS, and AZ analyzed data. WA, AA, MAA, and MA conducted the experiment and wrote the manuscript.

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Correspondence to Wasim Ahmad.

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Ahmad, W., Husain, I., Ahmad, N. et al. Box–Behnken supported development and validation of robust HPTLC method: an application in estimation of punarnavine in leaf, stem, and their callus of Boerhavia diffusa Linn. 3 Biotech 10, 165 (2020). https://doi.org/10.1007/s13205-020-2154-1

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