Development and Validation of a Green Stability-Indicating HPTLC Method for Estimation of Curcumin, Gallic Acid, and Ursolic Acid From Polyherbal Formulation Jatyadi Taila

Abstract

Background Jatyadi Taila (JT) is a well-known Ayurvedic wound healing product, comprising 16 different medicinally important plants including; Curcuma longa, Terminalia chebula, and Jasminum officinale. Objective The proposed work discusses the development and validation of the green and economic stability-indicating HPTLC method for quantification of the key marker phytoconstituents; curcumin (CUR), gallic acid (GA), and ursolic acid (UA) from Jatyadi Taila (JT). Method Quality standard parameters for JT were determined following standard procedures. The marker constituents CUR, GA, and UA were resolved from JT using toluene: ethyl acetate: formic acid (6: 2: 1, %v/v/v) as mobile phase and subsequently derivatized to estimate UA. The developed plates were subjected to HPTLC-MS analysis. All constituents were subjected to forced degradation to determine the proposed technique's stability-indicating property and the accelerated stability studies of marketed formulation and marker constituents. Greenness evaluation of the method was aided by the AGREE methodology. Results The Rf values of CUR, GA, and UA were found to be 0.60 and 0.60; 0.27 and 0.28; and 0.74 and 0.77 from reference standard and oil samples respectively, when analyzed at 366 nm, 290 nm, and 366 nm respectively. HPTLC-MS was carried out to verify the active constituents present in JT. The constituents followed first-order degradation kinetics. The quantity of CUR, GA, and UA in JT was reduced at the end of accelerated stability studies. The developed approach was validated in compliance with the ICH Q2 (R2) guideline. Conclusion Amongst the chosen key markers, GA was highly unstable during forced degradation. JT should be stored at a controlled temperature using more protective packaging material to ensure its quality and efficacy. Highlights The developed method can be used as a quality control tool for JT as it can be used to determine the stability of the key marker compounds the herbal formulation.

Formulation and validation of analytical methods for ursolic acid in Plantago major gel preparations

“…Second, after plotting the peak areas (y-axis) against UA concentrations (x-axis), a good linear correlation (r  0.998) was formed between the two for the UA in the range of 20-110 ng/spot (Figure 8). Previous research showed that UA has a good linear relationship (r=0.9986) over the concentration range 200-600 ng/spot with respect to peak area [21]. Our findings indicate that the developed TLCdensitometric method is capable of analyzing low UA concentrations in the gel dosage forms.…”

“…The lowest UA level at which the method could detect (LOD) and quantify (LOQ) was, respectively, 4.55 ng/spot (LOD) and 15.17 ng/spot (LOQ). These LOD and LOQ values are lower than that found by Patel and Vyas [21] which is 10.02 ng/spot and 30.36 ng/spot, respectively and even much lower than that found by Jamal et al [30] i.e. 561.85 ng/spot and 1702.58 ng/spot, respectively.…”

“…The visible spectra of the UA standard and sample showed a similar pattern, each at a maximum wavelength of 536 nm. This maximum wavelength corresponds to the one used in a previous study, i.e., 530 nm [21] and 540 nm [30]. Second, after plotting the peak areas (y-axis) against UA concentrations (x-axis), a good linear correlation (r  0.998) was formed between the two for the UA in the range of 20-110 ng/spot (Figure 8).…”

“…The RSD values of the intraday precision (1.24-1.96%) and interday precision ( 6.29%) indicated a precise method. With a different concentration of UA, 200-600 ng/spot [21] and 1000-3000 ng/spot [30], the HPTLC-densitometer in previous studies also had good intraday and interday precision (RSD ≤ 2%). Fifth, the last parameters of method validation are LOD and LOQ.…”

“…Various analytical techniques such as GC-MS and HPLC were conducted to measure the UA content [18,19]. However, because these methods are costly and require specialized expertise, the TLC-densitometric method was used in the UA analysis [3,20,21]. Moreover, this method was able to separate UA from its isomer commonly found in herbs, oleanolic acid, after simple pre-derivatization with iodine [22].…”