OBJECTIVETo screen the factors that affect indirubin-generated significantly in the process of preparing indigo naturalis, optimize level combination and determine the optimum technology for indirubin-generated.

METHODUsing concentration of indirubin (mg x g(-1)) that generated by fresh leaf as an index, Plackett-Burman design, Box-Behnken design response surface analysis as the statistical method, we screened the significantly influencing factors and the optimal level combination.

RESULTThe soaking and making indirubin process in preparing indigo naturalis was identified as the wax is not removed before immersion with immersion pH 7, solvent volume-leaf weight (mL: g)15, soaked not avoided light, soaking 48 h, temperature 60 degrees C, ventilation time of 180 min, and added ammonia water to adjust pH to 10.5.

CONCLUSIONThe soaking and making indirubin process in preparing indigo naturalis is optimized systematically. It clarify the various factors on the impact of the active ingredient indirubin which controlled by industrialized production become reality in the process of preparing indigo naturalis, at the same time, it lay the foundation for processing principle of indigo naturalis.