Objective To establish the distribution of serum human epididymis protein 4 by gender and menopausal status in apparently healthy individuals. Methods The chemiluminecence immunoassay technology was used to determine the levels of serum HE4 in 500 apparently healthy individuals which included 200 males, 150 premenopausal females and 150 postmenopausal females. Results The scatter plots showed a weak correlation between age and HE4 level in male (Pearson coefficient 0.228) and a moderate correlation in female (Pearson coefficient 0.472). The serum HE4 levels were 46.2±10.7 pmol/L in male, 45.9±10.3 pmol/L in premenopausal female and 51.7±11.7 pmol/L in postmenopausal female respectively. Conclusion Gender and menopausal status influence on serum HE4 level was demonstrated. The reference range of serum HE4 in Beijing population are <63.7 pmol/L for male, <62.8 pmol/L for premenopausal female and <70.9 pmol/L for postmenopausal female.