Tlc reprostar 3

Compounds 1, 2 and 3 (standards) were isolated from E. schimperi (Ahmed et al., 2017 ). Analytical grade reagents such as ABTS, DPPH, p-anisaldehyde, acarbose, α‐Amylase, α‐glucosidae and BHA were purchased from Sigma Aldrich. Other chemicals like dinitrosalicylic acid, sodium carbonate, p-nitrophenyl glucopyranoside and potassium persulphate are procured from different sources. Glass-backed silica gel 60F₂₅₄ HPTLC plates (10 × 20 cm) were purchased from Merck (Darmstadt, Germany). CAMAG automatic TLC sampler-4 (CAMAG, Muttenz, Switzerland) was used to apply the standards and samples band wise. Development was done in ADC2 (automatic development chamber), scanning in CATS 4 (CAMAG) and documentation by CAMAG TLC Reprostar 3.

For the application of different tracks of standards and GSLE, glass-backed silica gel 60F₂₅₄ HPTLC plates (Merck, Germany) were used. CAMAG automatic TLC sampler-4 was used for the application of the standards and GSLE, band-wise to the plates and developed in automatic development chamber (ADC2) (CAMAG, Muttenz, Switzerland). The developed HPTLC Plates were documented by CAMAG TLC Reprostar3 and scanned in CATS 4 (CAMAG).

The three anticancer biomarkers, ursolic acid, β-sitosterol and lupeol were procured from Sigma Aldrich (USA). The solvents used (petroleum ether, ethyl acetate, toluene, chloroform, ethyl acetate, methanol and n-butanol; AR grade) were purchased from BDH (UK). The HPTLC plates (glass-backed silica gel 60F₂₅₄) were purchased from Merck (Germany). The application of biomarkers and extracts (band wise) to the plate was carried out by Automatic TLC Sampler-4 (CAMAG, Switzerland) while the development took place in automatic development chamber (ADC2, Switzerland). The developed plate was derivatized with p-anisaldehyde reagent [the reagent was prepared according to the Wagner and Bladt (2004) by using anisaldehyde, glacial acetic acid, methanol and concentrated sulphuric acid. Initially anisaldehyde (0.5 mL) was mixed with glacial acetic acid (10 mL) followed by addition of methanol (85 mL) and then sulphuric acid (5 mL)]. The scanning and documentation of developed HPTLC plate was carried out by CATS 4 (CAMAG, Switzerland) and TLC Reprostar 3 (CAMAG, Switzerland), respectively.