The invention discloses anti-dandruff shampoo with an effect of improving the microbial flora on the scalp skin. The anti-dandruff shampoo is prepared from the following raw materials: water, polydimethylsiloxane, glycol stearate, sodium lauroyl sarcosine, polyquaternium-10, cetostearyl alcohol, ammonium chloride, vaccinium uliginosum berry extract, celery seed extract, silk hydrolysate, zinc pyrithione, piroctone olamine, EDTA disodium, surfactant, preservatie, artemisinin and rosemary water. The invention further discloses a preparation process of the shampoo. The shampoo can be used for improving the microbial environment of the scalp skin and improving the resistance of the scalp skin and can achieve the effect of effectively removing dandruff and preventing dandruff reoccurrence.