Development and Validation of a HPLC Method for 6-Gingerol and 6-Shogaol in Joint Pain Relief Gel Containing Ginger (Zingiber officinale)

Authors: Tanwarat Kajsongkram, Saowalux Rotamporn, Sirinat Limbunruang, Sirinan Thubthimthed

Abstract:

High Performance Liquid Chromatography (HPLC) method was developed and validated for simultaneous estimation of 6-Gingerol(6G) and 6-Shogaol(6S) in joint pain relief gel containing ginger extract. The chromatographic separation was achieved by using C18 column, 150 x 4.6mm i.d., 5μ Luna, mobile phase containing acetonitrile and water (gradient elution). The flow rate was 1.0 ml/min and the absorbance was monitored at 282 nm. The proposed method was validated in terms of the analytical parameters such as specificity, accuracy, precision, linearity, range, limit of detection (LOD), limit of quantification (LOQ), and determined based on the International Conference on Harmonization (ICH) guidelines. The linearity ranges of 6G and 6S were obtained over 20- 60 and 6-18 μg/ml respectively. Good linearity was observed over the above-mentioned range with linear regression equation Y= 11016x- 23778 for 6G and Y = 19276x-19604 for 6S (x is concentration of analytes in μg/ml and Y is peak area). The value of correlation coefficient was found to be 0.9994 for both markers. The limit of detection (LOD) and limit of quantification (LOQ) for 6G were 0.8567 and 2.8555 μg/ml and for 6S were 0.3672 and 1.2238 μg/ml respectively. The recovery range for 6G and 6S were found to be 91.57 to 102.36 % and 84.73 to 92.85 % for all three spiked levels. The RSD values from repeated extractions for 6G and 6S were 3.43 and 3.09% respectively. The validation of developed method on precision, accuracy, specificity, linearity, and range were also performed with well-accepted results.

Keywords: Ginger, 6-gingerol, HPLC, 6-shogaol.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1338522

References:

[1] Ajinkya R. Nikam,Lohidasan Sathiyanarayanan, Kakasaher R. Mahadik. 2013. Validation of reversed-phase high pressure liquid chromatography method for simultaneous determination of 6-, 8-, and 10-shogaol from ginger preparations. Int J Pharm Pharm Sci, 5(1) : 432-437.
[2] Balladin, D. A., O. Headley, I. Chang-yen and D. Mc.Gaw. 1997. Extraction and evaluation of the main pungent principles of solar dried west indian ginger (Zingiber officinale Roscoe) rhizome. Renewable Energy. 12(2) : 125-130.
[3] Balladin, D. A., O. Headley, I. Chang-yen and D. Mc.Gaw. 1998. High pressure liquid chromatography of the main pungent principles of solar dried west indian ginger (Zingiber officinale Roscoe). Renewable Energy. 13(4) : 531-536.
[4] He, X., Bernart, M., Lian, L., and Lin, L. 1998. High-perfomance liquid chromatography – electrospray mass spectromertric analysis of pungent constituents of ginger. J.Chromatography A, 796 : 327-334.
[5] Ghasemzadeh Ali, Hawa Z. E. Jaafar and Asmah Rahmat 2015. Optimization protocol for the extraction of 6-gingerol and 6-shogaol from Zingiber officinale var. rubrum Theilade and improving antioxidant and anticancer activity using response surface methodology. BMC Complementary and Alternative Medicine.15:1-10.
[6] Jolad S. D., Lantz R. C., Solyom A. M., Chen G. J. 2004. Fresh organically grown ginger (Zingiber officinale) : composition and effects on LPS induced PGE2 production. Phytochemistry. 65: 1937-1954.
[7] Kamal Yoonus, Mhaveer Singh, Sayeed Ahmad, Prawez Alam and Shahana Salam. Stability - indicating RP-HPLC method for the determination of 6-gingerol in polyherbal formulations Journal of Analytical Science and Technology (2015) 6 :1-7.
[8] Rafael Zarate, Michael M. Yeoman 1996 Change in the amount of 6- gingerol and derivatives during a culture cycle of ginger, Zingiber officinale. Plant Science. 121, 115-122.
[9] Ranjit Singh.2013. HPLC method development and validation- an overviewJ Pharm Educ Res. 4(1) : 26-33.
[10] Sushila Bhattarai, Van Hoan Tran, Colin Charl Duke. 2001.The stability of gingerol and shogaol in aqueos solution .J.of pharmaceutical science. 90(10) 1658-1663
[11] Suzanna M. Zick, Mack T. Ruffin, Zora Djuric, Daniel Normolle, Dean E. Brenner. 2010. Quantitation of 6-, 8- and 10-Gingerols and 6-Shogaol in Human Plasma by High-Performance Liquid Chromatography with Electrochemical Detection. Int J Biomed Sci. 6(3) : 233-240
[12] USP, 24th Rev, 2000 Rockrille.MD : The United States Pharmacopeia Covertion, Inc.2005 : p. 2149-2152: 2254-2692.