Biological Verification and Quality Assessment of a Natural Hepatoprotective Recipe

Objective: Biological verification of an eight-ingredient hepatoprotective recipe available in the Egyptian market was carried out followed by biologically guided reduction of non-essential ingredients aiming determination of the most potent formula containing the least number of ingredients to facilitate quality assessment procedure. Methods: Investigation of hepatoprotective effect afforded by the marketed and the other reduced four recipes using carbon tetrachloride as hepatic toxicant in rats with reference to Silymarin as standard where values of certain hepatic and antioxidant enzymes were estimated. The chosen recipe was subjected to quality assessment procedure including microscopical ident-ification of diagnostic elements; determination of pharmacopoeial constants; detection of heavy metals, mycotoxinsʹ and microbial contamination, detection of pesticidesʹ and radioactive residues and quantitative HPLC estimation of certain biochemical marker compounds. Results: The most potent hepatoprotective recipe formed of three ingredients namely, Milkthistle or Silybum (Silybum marianum Gaertn. fruits), Curcuma (Curcuma longa L. rhizomes) and Liquorice (Glycyrrhiza glabra L. roots and rhizomes) in its ethanol and aqueous extracts which reduced the elevated levels of liver and antioxidant enzymes to 69.43±2.26, 163.26±2.16, 40.85±2.18 and 280.39±8.45 & 70.50±2.40, 164.05±2.20, 40.65±1.80 and 279.03±7.90 for ALT, AST, GSH and SOD respectively while had non-significant effect on AP levels. Quality assessment procedure revealed that it is formed of genuine medicinal plants, devoid of hazardous heavy metals, mycotoxins, pathogenic organisms and radioactive residues while screening of pesticide residues revealed the existence of acetamiprid, carbendazim, chlorpyrifos, cyper-methrin, lambda-cyhalothrin and malathion at levels of 0.01, 0.09, 0.04, 0.01, 0.01 and 0.02 mg kg-1 respectively which considered to be harmless. Quantitative HPLC investigation revealed that ethanol and aqueous extracts contain Silymarin, Curcumin and Glycyrrhetinic acid at levels of (75, 120 and 40 mg ml-1) and (100, 80 and 45 mg ml-1 Conclusion: The gained results proved that the three-ingredient recipe is the best among the screened ones as both ethanol and aqueous extracts, in addition, it meets the quality standards required by WHO, moreover, quantitative HPLC investigation revealed that both extracts contain comparable quantities of biochemical marker compounds leading to adequate hepatoprotective and so, the recipe fitted the basic requirements for developing natural hepatoprotective agents. ) respectively.