Optimization of Betulinic Acid Extraction from Tecomella undulata Bark Using a Box-Behnken Design and Its Densitometric Validation

Abstract

Betulinic acid (BA) is a pentacyclic triterpenoid acid obtained from the stem bark of Tecomella undulata Seem. (Bignoniaceae). Development of an efficient extraction method for the isolation of BA is important as it has a wide range of pharmacological activity. A Box-Behnken design (BBD) was used to investigate the effect of extraction variables such as temperature (30-60˝C), time (4-8 h) and solvent to drug ratio (300-500 mL/100 g) on the maximization of BA yield and its quantification using validated densitometric high performance thin layer chromatography coupled with ultraviolet detection (HPTLC-VIS). A quadratic polynomial model was found to best fit the model with R 2 = 0.99. The optimized Soxhlet extraction yielded 2.449% w/w of BA at a temperature 53.86˝C, time 6.38 h and solvent to drug ratio 371 mL/100 g. BA in Tecomella undulata bark was detected at Rf value of 0.65 at 510 nm using the solvent system toluene-ethyl acetate-glacial acetic acid (8.5:1.5:0.02 v/v/v). The analytical method was validated and the linear regression analysis reflects good linear relationship (R 2 = 0.9902). Lower %RSD and SEM suggested that the developed HPTLC-VIS method was precise, accurate and robust. Therefore, these economical techniques are very efficient and promising for the extraction and quantification of pharmaceutically important BA.

Optimization of Ultrasonicated Kaempferol Extraction from Ocimum basilicum Using a Box–Behnken Design and Its Densitometric Validation

“…Validation parameters such as the LOD and the LOQ were used to confirm the accuracy and precision of the proposed method with reproducible results. The validation parameters in the current study were large (LOD = 8.16 ng; LOQ = 18.142 ng), indicating that the method used here was more sensitive than the HPTLC methods used for previous KA analyses [ 33 ]. A recovery exceeding 98% was observed for the accuracy study ( Table 4 ).…”

Parameter Optimization and Potential Bioactivity Evaluation of a Betulin Extract from White Birch Bark

“…For single-factor experiments, five critical factors affecting the betulin yield were investigated, i.e., the solvent type (water, methanol, ethanol, butanol, and amyl alcohol), ethanol concentration (45%, 55%, 65%, 75%, and 85%), extraction time (20,25,30,35, and 40 min), extraction temperature (20,30,40,50, and 60 • C), and solid/solvent ratio (1:20, 1:25, 1:30, 1:35, and 1:40 mg/mL). In the single-factor experiment design, only one factor was varied at a time, while the other factors were kept constant in each experiment [13]. For each experimental test, 5 g of dried outer bark of Betula platyphylla Suk.…”

“…Tálos-Nebehaj and co-workers reported that the tree outer bark consists of more bioactive compounds than the inner bark, and that these bioactive compounds showed high antioxidant activities [18]. It was reported that these birch triterpenes showed antiviral, antimicrobial, and hepatoprotective pharmacological activities [12,13,15]. According to the standard curve of betulin ( Figure 3C), the betulin concentration in the extract sample was calculated to be 50%.…”

Identification of RoCYP01 (CYP716A155) enables construction of engineered yeast for high-yield production of betulinic acid

“…Unfortunately, the content of BA in the commercially used bark from birch (genus Betula) is quite low, 0.025% by weight, although this is alleviated to some degree by the alcoholic precursor betulin, which is found in higher amounts and can be converted to BA (Pezzuto and Kim 1998). Much effort has gone into increasing yields from phytochemical isolation of BA and betulin (Krasutsky et al 2003;Siddiqui and Aeri 2016;Šiman et al 2016), as well as conversion of betulin to BA (Pezzuto and Kim 1998;Csuk et al 2006;Tulisalo et al 2013;Ressmann et al 2017). However, all these methods are based on isolation of BA and/or betulin from plant materials, which consumes large amounts of plant biomass, and requires a tedious and laborious extraction and purification process that is environmental unfriendly.…”