5β-Pregnane-3α,20α-diol glucuronide is a progesterone metabolite predominant in urine, during pregnancy. Presence of 5β-Pregnane-3α,20α-diol glucuronide in the urine indicates pregnancy, but it might also account for pseudopregnancy. It is useful in predicting the delivery time/end of pseudopregnancy.

Urinary pregnanediol glucuronide (PdG) seems to serve as an effective clinical marker for monitoring progesterone supplementation. PdG, along with other urinary metabolites such as luteinizing hormone (LH), follicle stimulating hormone (FSH), and estrone glucuronide (ElG), exhibit a strong association with ovarian excretion rates and demonstrate a high correlation with serum reproductive hormones. As a result, they can be used interchangeably to effectively monitor ovarian steroidogenesis.

5β-Pregnane-3α,20α-diol glucuronide has been used in a cross-sectional analysis of hormone levels. It has also been used as a standard to measure estradiol-17β, pregnanediol-glucuronide (PdG) by a double-antibody enzyme immunoassay.

5β-Pregnane-3α,20α-diol glucuronide has been used in steroid enzyme immunoassay.