Iloprost solution was carried out using HPLC apparatus (Varian Inc., Palo Alto, USA) equipped with 200-2031 Metachem online degasser, a M210 binary pump, a ProStart 410 auto sampler, a G1316A thermostatic column, a 25 μl CSL20 Cheminert Sample Loop injector. Data was acquired and processed using Galaxie chromatography manager software (Varian Inc., Palo Alto, USA). A GraceSmart RP C18 column (4.6 × 250 mm, 5 μm, Alltech Grom GmbH, Rottenburg-Hailfingen, Germany) was used as a stationary phase during analysis. Deionized water (HPLC grade) acidified with 0.1% v/v trifluoroacetic acid (TFA, HPLC grade) and acetonitrile (HPLC grade) 60:40% v/v was used as the mobile phase. The flow rate was 1 ml/min; while, Iloprost solution was acquired at a maximum wavelength of 205 nm. The drug was separated as an enantiomer at the retention times of 20.5 and 22 min, respectively. Enantiomers were widely separated during analysis and no interference peaks or overlapping of compounds were observed (Supplementary Fig. 1
AUC = 0.5899x + 0.0234
where, x is the Iloprost concentration (μg/ml) and AUC (mAU × min) is the area under the curve. A linear correlation with R2 = 0.9998 was carried out in the range from 0.01 to 10 μg/ml.
AUC = 0.5899x + 0.0234
where, x is the Iloprost concentration (μg/ml) and AUC (mAU × min) is the area under the curve. A linear correlation with R2 = 0.9998 was carried out in the range from 0.01 to 10 μg/ml.