Concurrent analysis of bioactive triterpenes oleanolic acid and β-amyrin in antioxidant active fractions of Hibiscus calyphyllus , Hibiscus deflersii and Hibiscus micranthus grown in Saudi Arabia by applying validated HPTLC method

Abstract

In this study, we developed a validated HPTLC method for concurrent analysis of two natural antioxidant triterpenes, oleanolic acid (OA) and β-amyrin (BA) in the biologically active fractions (petroleum ether, toluene, chloroform, ethyl acetate and -butanol) of aerial parts of three species (, and). The chromatography was conducted on normal HPTLC (ready to use glass-plate coated with silica gel 60 F254) plate with chloroform and methanol (97:3, V/V) used as mobile phase. The derivatization of the developed plate was done with p-anisaldehyde and scanned at λ = 575 nm. Well resolved and intense peaks of OA and BA were obtained at Rf = 0.36 and 0.57, respectively. The linear regression equation/correlation coefficient (r) for OA and BA were Y = 6.65x + 553.35/0.994 and Y = 9.177x + 637.23/0.998, respectively in the linearity range of 100-1200 ng/spot indicated good linear relationship. The low values of %RSD for intra-day/inter-day precision of OA (1.45-1.61/1.38-1.59) and BA (1.52-1.57/1.50-1.53) suggested that the method was precise. The recovery/RSD (%) values for OA and BA were found to be 99.21-99.62/1.39-1.95 and 98.75-99.70/1.56-1.80, respectively assures the reasonably good accuracy of the proposed method. Fifteen samples were analyzed to check the content of OA and BA by using the developed HPTLC methods. The content of OA in different samples were found to be 3.87 (HmP) > 1.212 (HcP) > 0.673 (HdC) > 0.493 (HdP) > 0.168 (HdE) > 0.059 (HcC) > 0.015 (HcE) > 0.008 (HmT) µg/mg of the dried weight of extract. However the content of BA was found as: 2.293 (HmP) > 1.852 (HdT) > 0.345 (HdC) > 0.172 (HmT) > 0.041 (HdE) > 0.008 (HcC) µg/mg of the dried weight of extract. Some species fractions exhibited good antioxidant potential like: HcE (IC50 = 17.6 ± 1.8)> HdB (IC50 = 32.16 ± 0.9) > HmP (IC50 = 80.4 ± 4.5) > HmT (IC50 = 99.7 ± 8.2) when compared with ascorbic acid (IC50 = 14.2 ± 0.5), while other fractions exhibited only mild antioxidant capability. The developed HPTLC method can be further exploited for analysis of these markers in the quality assessment of raw material as well as herbal formulations available in the market.

Biological Potential and Chemical Profile of European Varieties of Ilex

“…The biological activity of polyphenol fractions should be noted at this point. It has been shown that the presence of polyphenolic compounds in the ingested diet reduces the risk of obesity, cardiovascular disease, and type 2 diabetes [ 48 , 49 , 50 ].…”

Antidiabetic, antioxidant, molecular docking and HPTLC analysis of miquelianin isolated from Euphorbia schimperi C. Presl

“…All the compounds along with ESME were applied on the plate by ATS-4 (Automatic TLC sampler-4; 160 nL/s) and the plate development was accomplished in Automatic Development Chamber at controlled temperature (25 ± 2 °C) and humidity (60 ± 5%). The plate was dried and analyzed quantitatively (at λ max = 350 nm) in absorbance mode (Siddiqui et al; 2018).…”

Anticancer activity and concurrent analysis of ursolic acid, β-sitosterol and lupeol in three different Hibiscus species (aerial parts) by validated HPTLC method

“…The extraction of powder (400 g) of all three Hibiscus species was carried out in 95% ethanol according to the Siddiqui et al (2018). The obtained extracts were fractionated using various solvents (petroleum ether, toluene, chloroform, ethyl acetate, and n-butanol), yield was calculated and stored at 4 °C in refrigerator as H. calyphyllus [HcP (petroleum ether fraction, 2.25 g), HcT (toluene fraction, 2.21 g), HcC (chloroform fraction, 3.32 g), HcE (ethyl acetate fraction, 2.3 g) and HcB (n-butanol fraction, 2.01 g)]; H. deflersii [HdP (petroleum ether fraction, 1.19 g), HdT (toluene fraction, 2.10 g), HdC (chloroform fraction, 2.46 g), HdE (ethyl acetate fraction, 2.37 g) and HdB (n-butanol fraction, 1.98 g)] and H. micranthus [HmP (petroleum ether fraction, 1.96 g), HmT (toluene fraction, 2.36 g), HmC (chloroform fraction, 2.55 g), HmE (ethyl acetate fraction, 2.86 g) and HmB (n-butanol fraction, 2.01 g)] until the time of use.…”

“…It is more precise and calibrated, and has several advantages over other analytical technique like HPLC (high performance liquid chromatography) in quantification of different markers (both UV active or inactive). The broad dimensions of stationary phases has increased the utilization of HPTLC for a wide range of samples (Siddiqui et al, 2018, Alam et al, 2017, Alam et al, 2015a, Alam et al, 2015b, Alam et al, 2015c, Siddiqui et al, 2015, Alajmi et al, 2015, Alam et al, 2014).…”