The invention discloses a preparation method and a preparation control method of a mulberry leaf extract. The preparation method comprises the steps of cutting mulberry leaves into shreds, adding water, decocting and extracting for two times, combining two decoction-filtering liquors, filtering through a sieve, enabling a primary filtrate to pass through a microfiltration membrane with the bore diameter of 0.01 to 0.02 micron, enabling a refined filtrate to pass through a reverse osmosis membrane with the bore diameter of 0.0001 to 0.001 micron, concentrating until a solid content is 15 to 18 percent, performing vacuum concentration on a concentrated initial solution under 60 DEG C, to form an extractum with the relative density of 1.10 to 1.13, and spray-drying the extractum to obtain the mulberry leaf extract. A quality control method is characterized by adopting high efficiency liquid chromatography to measure the content of rutin in the mulberry leaf extract. The preparation method can avoid rutin content loss caused by washing a mulberry leaf medicinal material and performing high temperature concentration, the rutin in the mulberry leaves is well extracted and reserved, microorganisms such as bacteria can be well removed, and a microorganism indicator is enabled to meet national health food material regulations; the quality control method is accurate, reliable and good in repeatability, and can be taken as a method for measuring the content of the mulberry leaf extract rutin.