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

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.





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