Author refers to test article as HCFO-1224yd(Z).

"The test substance supplied from the supplier was used as is. The test substance was gasified by passing through pressurized air at a specified flow rate into the test substance in a stainless-steel cylinder held into a constant temperature water bath set at 15°C. The generated gas was passed through a heat exchanger set at 15°C. Then, the test atmosphere at the intended concentration was prepared by mixing the test substance gas with pressurized air at specified flow rate. The test atmosphere was exposed to the rats by continuously supplying it to the chamber. The air from the chamber was exhausted to outside air after passing through a filter and scrubber. The rats held in the restraint tubes (Muenster Ltd.) were connected to the chamber to start exposure, when hydrocarbon concentration in the test atmosphere continuously monitored by a hydrocarbon monitor reached temporally equilibrium after the start of generating the test atmosphere containing the test substance. The rats were disconnected from the chamber to terminate exposure 6 hours after the start of exposure. In the control group, the rats held in the restraint tubes were connected to the chamber supplied air at a specified flow rate to start exposure. The rats were disconnected from the chamber to terminate exposure 6 hours after the start of exposure. Hydrocarbon concentration in the test atmosphere was continuously monitored with the hydrocarbon monitor (HCM-1B: Shimadzu Corp., Kyoto, Japan) every exposure. Temporal variability of the concentration was confirmed by recording the output voltage of the hydrocarbon monitor. The amount of test substance used during the inhalation exposure was calculated from the weight difference of the cylinder containing the test substance between before and after the test atmosphere generation. The total air volume supplied to the chamber was obtained by multiplying the elapsed time of the test atmosphere generation to the air flow rate of the test atmosphere supply (minimum unit: 0.1 L/min). The nominal concentration (minimum unit: 10 ppm) was calculated with the following equation from the amount of the test substance used and the total air volume supplied to the chamber. The test substance in the test atmosphere was quantified by a gas chromatography (hereinafter abbreviated as “GC”) according to the method validated in the acute inhalation toxicity study of the test substance (study No. B130803), then, the exposure concentration was calculated.

The target exposure concentrations of this study were set at 0 (control), 30,364, 60,728 and 121,456 mg/m (5000, 10000, and 20000 ppm respectively). The rats On GD 20, the animals were subjected to necropsy and cesarean section, and the embryo-fetus and placenta were examined. The rats (20 animals per group) were exposed to each test atmosphere from gestation day (GD) 6 to GD 19 for 6 hours a day. The actual exposure concentrations were 31,639, 62,489 and 124553 mg/m (5210, 10290, and 20510 ppm, respectively)."

F1 Male and Female Sprague-Dawley Rat