Research On New Analysis Method Of Amifostine And Its Intermediates

Amifostine is a broad spectrum cytoprotective agent, which was first approved by US FDA in1995and was approved by CFDA in2001. Amifostine is widely used in the prevention of the damage to normal tissue by cytotoxic reagents, without affecting the efficacy of the cancer treatment. Compared with other cytoprotective agent amifostine can protect almost all normal tissue. Therefore, it is of great significance for patient medication safety to improve quality standards.A reverse-phase HPLC method determinining N-(2-bromoethyl)-1,3-propanediamine dihydrobromide (bromine salt) was first established. The optimum separation condition was column Phenomenex Luna C18, mobile phase methanol-0.94g/L sodium1-hexanesulfonate(adjusted with phosphoric acid to a pH of3.0)=40-60, the flow rate1.0mL/min, the UV detector220nm, and injection volume10μL. Bromine salt demonstrated good linearity (r=0.9998), LOD0.74μg/mL, average recovery99.48%.In this paper, we have studied the stability of amifostines which were synthesized using DMSO and DMF as accelerator respectively. In the condition of acceleration test and long-time test, which temperature was40℃、25℃and2-8℃、-20℃, No significant change in physical and chemical properties(include clarity, pH and moisture) was observed.It turned out that the stability of the two kinds of amifostine had no significant difference.An efficient and reliable RP-HPLC method was developed for determinining amifostine and amifostine thiol. The optimum separation was carried out on Phenomenex Luna C8using methanol-0.5g/L nonfluorobutane sulfonate potassium in water (with pH adjusted to2.5by trifluoroacetic acid)=38-62as mobile phase. The flow rate was1.0mL/min. The UV detection was at220nm and injection volume was10μL. Amifotine demonstrated good linearity (r=0.9999) in the range from0.029to2.417mg/mL, LOD2.05μg/mL, average recovery99.71%. Amifotine thiol demonstrated good linearity (r=0.9999) in the range of32.490-126.360μg/mL, LOD3.25μg/mL, average recovery100.90%.The optimum conditions of preparing amifostine oxidative degradation products were explored by the methods of3factors and3levels orthogonal test. The process parameters were optimized with30%H2O2as oxidant, the molar ratio of the reactant1:1,and ultrasound for10minutes (0℃), the conversion rate was up to30.29%.