The present study was designed to investigate can the amplitude modulation (AM) of Photoplethysmogram (PPG) be used as an indicator of blood loss and if so what is the best PPG probe site. PPG from ear, finger and forehead probe sites, standard ECG, and Finapres blood pressure waveforms were continuously recorded from 8 healthy volunteers during baseline, blood withdrawal of 900 ml followed by the blood reinfusion. The instantaneous amplitude modulations present in heart rate (AM(HR)) and breathing rate (AM(BR)) band frequencies of PPG were extracted from high-resolution time-frequency spectrum. HR and pulse pressure showed no significant changes during the protocol. The AM(HR) significantly (P<0.05) decreased at 100 ml through 900 ml blood loss from ear and finger probe sites. The mean percent decrease in AM(HR) at 900 ml blood loss compared to baseline value was 45.2%, 42.0%, and 42.3% for ear, finger and forehead PPG signals, respectively. In addition, significant increases in AM(BR) were found due to blood loss in ear and finger PPG signals. Even without baseline AM(HR) values, 900 ml blood loss detection was shown possible with specificity and sensitivity both 87.5% from ear PPG signals. The present technique has great potential to serve as a valuable tool in the intraoperative and trauma settings to detect hemorrhage.