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Planta Med 2010; 76(5): 481-488
DOI: 10.1055/s-0029-1186220
Analytical Studies
Original Papers
© Georg Thieme Verlag KG Stuttgart · New York

Physicochemical Stability and Biological Activity of Withania somnifera Extract under Real-Time and Accelerated Storage Conditions

Dada Patil1 , Manish Gautam3 , Umesh Jadhav2 , Sanjay Mishra1 , Suresh Karupothula3 , Sunil Gairola3 , Suresh Jadhav3 , Bhushan Patwardhan1
  • 1Bioprospecting Laboratory, Interdisciplinary School of Health Sciences, University of Pune, Pune, India
  • 2Poona College of Pharmacy and Research Center, Bharati Vidyapeeth University, Pune, India
  • 3Serum Institute of India Limited, Pune, India
Further Information

Publication History

received May 11, 2009 revised August 28, 2009

accepted Sept. 16, 2009

Publication Date:
20 October 2009 (online)

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Abstract

Stability testing at preformulation stages is a crucial part of drug development. We studied physicochemical stability and biological activity of Withania somnifera (ashwagandha) dried root aqueous extract during six months real-time and under accelerated storage conditions. The characteristic constituents of ashwagandha roots include withanolides such as withaferin A and withanolide A. We modified and validated the HPLC‐DAD method for quantitative measurement of withanolides and fingerprint analysis. The results suggest a significant decline in withaferin A and withanolide A content under real and accelerated conditions. The HPLC fingerprint analysis showed significant changes in some peaks during real and accelerated storage (> 20 %). We also observed incidences of clump formation and moisture sensitivity (> 10 %) under real-time and accelerated storage conditions. These changes were concurrent with a significant decline in immunomodulatory activity (p < 0.01) during the third month of the accelerated storage. Thus, adequate control of temperature and humidity is important for WSE containing formulations. This study may help in proposing suitable guidance for storage conditions and shelf life of ashwagandha formulations.

Key words

Withania somnifera - Solanaceae - Ayurveda - ashwagandha - stability - withaferin A - withanolide A

References

  • 1 EMEA .Reflection paper on stability testing of herbal medicinal products and traditional herbal medicinal products. EMEA/HMPC/3626/2009. 2009: 1-3
    PubMedGoogle Scholar
  • 2 Goppel M, Franz G. Stability control of senna leaves and senna extracts.  Planta Med. 2004;  70 432-436
    Article in Thieme ConnectPubMedGoogle Scholar
  • 3 Anuchit P, Sirirat P, Suthimaln I, Tanomjit S. Stability of andrographolide in powdered Andrographis herb under accelerated conditions.  Planta Med. 2006;  72 954-956
    Article in Thieme ConnectPubMedGoogle Scholar
  • 4 Kopleman S H, Nguyen P A, Zito W S, Muller F X, Augsburger L L. Selected physical and chemical properties of commercial Hypericum perforatum extracts relevant for formulated product quality and performance.  AAPS Pharm Sci. 2001;  3 1-18
    PubMedGoogle Scholar
  • 5 Sahoo N K, Sarkar S, Patwardhan B. Stability study on Boswellia serrata (hydro-alcoholic) extract.  Asian J Chem. 2009;  21 3529-3534
    PubMedGoogle Scholar
  • 6 [no authors listed.]. Ayurvedic pharmacopoeia of India, Vol. 1. Delhi; The Controller of Publications Government of India 2001: 15-16
    Google Scholar
  • 7 Gautam M, Diwanay S S, Gairola S, Shinde Y S, Jadhav S S, Patwardhan B K. Immune response modulation to DPT vaccine by aqueous extract of Withania somnifera in experimental system.  Int Immunopharmacol. 2004;  4 841-849
    CrossrefPubMedGoogle Scholar
  • 8 Ziauddin M, Phansalkar N, Patki P, Diwanay S, Patwardhan B. Studies on the immunomodulatory effects of Ashwagandha.  J Ethnopharmacol. 1996;  50 69-76
    CrossrefPubMedGoogle Scholar
  • 9 Patwardhan B, Mashelkar R A. Traditional medicine-inspired approaches to drug discovery: can Ayurveda show the way forward?.  Drug Discov Today. 2009;  DOI: 10.1016/j.drudis.2009.05.009
    PubMedGoogle Scholar
  • 10 Sumantran V N, Kulkarni A, Boddul S, Chinchwade T, Koppikar S J, Harsulkar A, Patwardhan B, Chopra A, Wagh U V. Chondroprotective potential of root extracts of Withania somnifera in osteoarthritis.  J Biosci. 2007;  32 299-307
    CrossrefPubMedGoogle Scholar
  • 11 Misra L, Mishra P, Pandey A, Sangwan R S, Sangwan N S, Tuli R. Withanolides from Withania somnifera roots.  Phytochemistry. 2008;  69 1000-1004
    CrossrefPubMedGoogle Scholar
  • 12 Pramanick S, Roy A, Ghosh S, Majumder H K, Mukhopadhyay S. Withanolide Z, a new chlorinated withanolide from Withania somnifera.  Planta Med. 2008;  74 1745-1748
    Article in Thieme ConnectPubMedGoogle Scholar
  • 13 Kaileh M, Vanden Berghe W, Heyerick A, Horion J, Piette J, Libert C, De Keukeleire D, Essawi T, Haegeman G. Withaferin A strongly elicits IkappaB kinase beta hyperphosphorylation concomitant with potent inhibition of its kinase activity.  J Biol Chem. 2007;  282 4253-4264
    PubMedGoogle Scholar
  • 14 ICH . Guidance for industry on stability data package for registration applications in climatic zones III and IV.  Q1F. 2002;  1-8
    PubMedGoogle Scholar
  • 15 Taylor M, Ginsburg J, Hickey A, Gheyas F. Composite method to quantify powder flow as a screening method in early tablet or capsule formulation development.  AAPS Pharm Sci Technol. 2000;  1 18E
    PubMedGoogle Scholar
  • 16 Ganzera M, Choudhary M I, Khan I A. Quantitative HPLC analysis of withanolides in Withania somnifera.  Fitoterapia. 2003;  74 68-76
    CrossrefPubMedGoogle Scholar
  • 17 Yuhua C, Wang L, Yu X, Ye J. Development of the chromatographic fingerprint of herbal preparations Shuang–Huang–Lian oral liquid.  J Pharm Biomed Anal. 2006;  41 845-856
    CrossrefPubMedGoogle Scholar
  • 18 Kai-Tai F, Yi-Zeng L, Xiao-lin Y, Kelvin C, Guang-Hua Lu. Critical value determination on similarity of fingerprints.  Chemometric Intelligent Lab Syst. 2006;  82 236-240
    PubMedGoogle Scholar
  • 19 Nelson D S, Mildenhall P. Studies on cytophillic antibodies. The production by mice of macrophage cytophillic antibodies to sheep erthrocytes; relationship to the production of other antibodies and development of delayed type hypersensitivity.  Aust J Exp Biol Med Sci. 1967;  45 113-130
    CrossrefPubMedGoogle Scholar
  • 20 Kaul A, Bani S, Zutshi U, Suri K, Satti N K, Suri O P. Immunopotentiating properties of Cryptolepis buchanani root extract.  Phytother Res. 2003;  17 1140-1144
    CrossrefPubMedGoogle Scholar
  • 21 Abdullah E C, Geldart D. The use of bulk density measurements as flowability indicators.  Powder Technol. 1999;  102 151-165
    PubMedGoogle Scholar
  • 22 Xie P, Chen S, Yi-Zeng L, Wang X, Tian R, Roy U, Van Beek T A, Schripsema J. Chromatographic fingerprint analysis: a rational approach for quality assessment of traditional Chinese herbal medicine.  J Chromatogr A. 2006;  1112 171-180
    CrossrefPubMedGoogle Scholar
  • 23 Xie Y, Jiang Z H, Zhoua H, Cai X, Wong Y F, Liu Z Q, Bian Z X, Xu H X, Liu L. Combinative method using HPLC quantitative and qualitative analyses for quality consistency assessment of a herbal medicinal preparation.  J Pharm Biomed Anal. 2007;  43 204-212
    CrossrefPubMedGoogle Scholar
  • 24 Enrique Ortega R. Bulk properties of food particulate materials: an appraisal of their characterization and relevance in processing.  Food Bioprocess Technol. 2009;  2 28-44
    CrossrefPubMedGoogle Scholar
  • 25 Kopelman S H, Augsburger L L. Some physical properties of Ginkgo biloba extracts important for tableting and encapsulation.  JANA. 2000;  3 32-37
    PubMedGoogle Scholar
  • 26 Chaurasiya N D, Uniyal G C, Lal P, Misra L, Sangwan N S, Tuli R, Sangwan R S. Analysis of withanolides in root and leaf of Withania somnifera by HPLC with photodiode array and evaporative light scattering detection.  Phytochem Anal. 2008;  19 148-154
    CrossrefPubMedGoogle Scholar
  • 27 Srivastava P, Tiwari N, Yadav A K, Kumar V, Shanker K, Verma R K, Gupta M M, Gupta A K, Khanuja S P. Simultaneous quantification of withanolides in Withania somnifera by a validated high-performance thin-layer chromatographic method.  J AOAC Int. 2008;  91 1154-1161
    PubMedGoogle Scholar
  • 28 Malik F, Kumar A, Bhushan S, Mondhe D M, Pal H C, Sharma R, Khajuria A, Singh S, Singh G, Saxena A K, Suri K A, Qazi G N, Singh J. Immune modulation and apoptosis induction: two sides of antitumoral activity of a standardized herbal formulation of Withania somnifera.  Eur J Cancer. 2009;  45 1494-1509
    CrossrefPubMedGoogle Scholar
  • 29 Stan S D, Hahm E R, Warin R, Singh S V. Withaferin A causes FOXO3a- and Bim-dependent apoptosis and inhibits growth of human breast cancer cells in vivo.  Cancer Res. 2008;  68 7661-7669
    CrossrefPubMedGoogle Scholar
  • 30 Bani S, Gautam M, Sheikh F A, Khan B, Satti N K, Suri K A, Qazi G N, Patwardhan B. Selective Th1 up-regulating activity of Withania somnifera aqueous extract in an experimental system using flow cytometry.  J Ethnopharmacol. 2006;  107 107-115
    CrossrefPubMedGoogle Scholar

Prof. Bhushan Patwardhan

Manipal Education
Chief Academics

14 HAL Airport Road

560 008 Bangalore

India

Phone: + 91 80 40 78 91 49

Fax: + 91 80 40 78 94 34

Email: bhushan.patwardhan@manipalu.com

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