Characterization and Estimation of Harpagoside in Dried Root Extract and Oral Powder Formulations of Harpagophytum Procumbens by Validated RP-HPLC-PDA Method
The present investigation was undertaken to perform structural, physicochemical characterization and to estimate the harpagoside in dried root extract and flavoured powder formulations of harpagophytum procumbens (devils claw). The extraction of harpagoside from the root extract was carried out using different solvents and the harpagoside content was estimated using an RP-HPLC-PDA method. The HPLC analysis was carried out using methanol: 0.02% formic acid (60:40 v/v) as the mobile phase, pumped at flow rate of 1mL/min through C18 column (Kinetex XB, 150x4.6mm, 5µ). The structural and physicochemical characterization of the root extract was performed using FT-IR, DSC and SEM analysis. Flavoured powder formulations (effervescent and non-effervescent) of the root extract were prepared and the harpagoside content was estimated by HPLC method. Higher percentage of harpagoside (1.6 %) was extracted with water as extraction solvent when compared to methanol and methanol: water (50:50 v/v). The developed RP-HPLC-PDA method resulted in shorter elution time (harpagoside was eluted at 5.3min) and the method was linear with good regression coefficient (R2>0.998) within the concentration range tested (0.2-4µg/mL). The FT-IR spectra of root extract showed the presence of characteristic peaks corresponding to the harpagoside and the results from photomicrographs and SEM analysis revealed spherical morphology of the particles in the extract. The prepared powdered formulations readily dispersed and dissolved in water and showed more than 98% of harpagoside content even after 6months time period indicating the stability of the formulations. Overall, the results from the present investigation can form basis to establish standardization parameters for formulations containing root extract of devil’s claw.
Keywords: Harpagophytum procumbens, Structural and physicochemical characterization, RP-HPLC-PDA method, FT-IR, DSC, SEM.
2. Ekor M. The growing use of herbal medicines: issues relating to adverse reactions and challenges in monitoring safety. Front Pharmacol 2013; 4:177-186.
3. Kasilo OMJ, Trapsida JM. Decade of African traditional medicine 2001–2010. Afr Health Mon (Special Issue) 2011; 14:25-31.
4. Rivera JO, Loya AM, Ceballos R. Use of herbal medicines and implications for conventional drug therapy medical sciences. Altem Integ Med 2013; 2:1-6.
5. Saper RB, Phillips RS, Sehgal A, Khouri N, Davis RB, Paguin J et.al. Lead, mercury and arsenic in US and Indian manufactured Ayurvedic medicines sold via the internet. JAMA 2008; 300:915-923.
6. Cohen PA. American Roulette-Contaminated dietary supplements. N Engl J Med 2009; 361:1526-1525.
7. Kumari R, Mita K. A review on the standardization of herbal medicines. Int J Pharm Sci Res 2016; 7:97-106.
8. Pravin HN, Kareparamban J, Auruna J, Vilasrao K. Future Trend in Standardization of Herbal Drugs. J of Applied Pharmaceutical Sci 2012; 02:38-44.
9. Yadav NP, Mayank T, Dixit VK. Recent approaches in herbal drug standardization. IJIB 2008; 2:195-203.
10. Beatrice B, Aurore F, Anne-sylvie F, Emmanuel P, Fathi M, Farid C et.al. Extraction by solvent using microwave and ultrasound-assisted techniques followed by HPLC analysis of Harpagoside from Harpagophytum procumbens and comparison with conventional solvent extraction methods. C R Chimie 2016; 19:692-698.
11. Anauate MC, Torres LM, De Mello SB. Effect of isolated fractions of Harpagophytum procumbens D.C. (devils claw) on COX-1, COX-2 activity and nitric oxide production on whole-blood assay. Phytother Res 2010; 24:1365-1369.
12. Inaba K, Murata K, Naruto S, Matsuda H. Inhibitory effects of devil’s claw (secondary root of Harpagophytum procumbens) extract and harpagoside on cytokine production in mouse macrophages. J Nat Med 2010; 64:219-222.
13. Kikuchi T, Matsuda S, Kubo Y, Namba T. New iridoids for Harpagophytum procumbens D.C. Chem Pharm Bull 1983; 31:2296-2301.
14. Alexanader HS. Fast HPLC for quality control of Harpagophytum procumbens using monolithic silica column: method transfer from conventional particle-based silica column. J Chromatogy A 2005; 1073:377-381.
15. Chrubasik S, Junck H, Breitschwerdt H, Conradt C, Zappe H. Effectiveness of Harpagophytum extract WS 1531 in the treatment of exacerbation of low back pain: a randomized placebo-controlled double-blind study. Eur J Anaesthesiol 1999; 16:118-129.
16. Chrubasik S, Zimpfer Ch, Schutt U, Ziegler R. Effectiveness of Harpagophytum procumbens in treatment of acute low back pain. Phytomedicine 1996; 3:1-10.
17. Devils claw dry extract monograph. Ph. Eur 2013; 8.0:1125-1126.
18. Bai Y, Pengfei Y, Qinghui W, Shaoqing C. Determination of harpagide and harpagoside in scrophulariae radix by HPLC-UV. China journal of Chinese material medica 2011; 36:2697-2702.
19. Guillerault L, Ollivier E, Elias R, Balansard G. Determination of harpagide, 8-para-coumaroyl harpagide and harpagoside by HPLC in Harpagophytum procumbens drugs and in commercial extract. J Liq Chromatogr Relat Technol 1994; 17:2951-2960.
20. International Conference on Harmonization of Technical Requirements for registration of Pharmaceuticals for human use. Code Q2B Validation of Analytical Procedures, 1994.
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. that allows others to share the work with an acknowledgment of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (SeeÂ The Effect of Open Access).