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F. C. CHOW, D. W. HAMAR and R. H. UDALL

Chow, Hamar & Udall (1972) reported that a water-soluble (`aqueous') fraction extracted from the needles of yellow pine (Pinus ponderosa) could disrupt fetal development of mice and some constituent of the pine needles was believed to be responsible for the activity. However, the aqueous fractions of pine needles harvested in the following 2 years induced little reproductive failure in mice. This inconsistency in activity raised doubts concerning the toxicity of the pine needles.

A preliminary study was undertaken to determine whether metabolites produced by fungi which we had observed on the pine needles could be the cause of the reproductive failure. Several pine needles were placed in the aqueous fraction. Fungi started to grow within 2 days of incubation at room temperature. After 2 weeks of growth, the

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FU-HO C. CHOW, KATHLEEN J. HANSON, D. W. HAMAR and R. H. UDALL

Colorado residents of long standing can recall the legend that Indian women used to take an aqueous extract of pine needles to induce abortion. The effects of pine needle ingestion on pregnancy of cattle and mice have been reported by Bruce (1927), MacDonald (1952), Deem, Osborn & Maag (1959, unpublished data reported by Faulkner, 1968), Allen & Kitts (1961), and Cook & Kitts (1964), but the active agent and the effect on reproduction have not been defined. The object of this investigation was to isolate the agent which affects reproduction.

Three fractions (volatile, water-soluble and acetone-soluble) were prepared from needles of yellow pine (Pinus ponderosa) collected from Rist Canyon, west of Fort Collins, Colorado, and stored in plastic bags at −20° C. Fractions were prepared as follows.