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OBJECTIVE: To establish a method for simultaneous detection of 5 anticoagulant rodenticides including brodifacoum, bromadiolone, flocoumafen, warfarin and difenacoum in whole blood by high-performance liquid chromatogarphy. METHODS: The 0. 5 m L of blood sample was extracted by 2. 0 m L ethyl acetate,then separated by Diamonsil C18 column( 250. 0 mm × 4. 6 mm × 5. 0 μm) using acid-ammonium acetate( 20. 0 mmol / L,p H = 5. 5) /methyl alcohol( 2∶ 8,V / V) as a mobile phase and detected by diode-array detector under the ultraviolet spectrum of 310 nm through high-performance liquid chromatogarphy. RESULTS: The good linear range of the 5 anticoagulant rodenticides was0. 50-10. 00 mg / L,and the correlation coefficients were > 0. 999 00. The detection limits of brodifacoum,bromadiolone,flocoumafen,warfarin and difenacoum were 0. 08,0. 06,0. 09,0. 04 and 0. 10 mg / L and the lower limits of quantitation were 0. 79,0. 58,0. 92,0. 45 and 0. 96 mg / L,respectively. The recovery rate was 98. 40%-104. 00%. The within-run relative standard deviation( RSD) was 0. 61%-7. 84%,and the between-run RSD was 1. 10%-9. 62%. The samples can be stored for 14 days in the refrigerator at 4 ℃. CONCLUSION: The method has advantages of simple operation,good separating effect,high sensitivity,precision and accuracy,which is suitable for detection of whole blood samples in rodenticide poisoning patients.
RESUMO
Objective To explore the acquired deficiencies of vitamin K-dependent coagulation factors in etiology, clinical characteristics and treatment. Methods Retrospective analysis was performed on the data of etiology, clinical manifestations of 45 patients with acquired deficiencies of vitamin K-dependent coagulation factor. All patients were treated with Vitamin K1 10 -40 mg/d, i. v. , for three months. Some patients with severe blooding were additionally treated with fresh freezing plasma or prothromibin complex. Prothrombin time(PT) and activated partial thromboplastic time(APTT) were measured using Stago automatic blood coagulation analyzer before and after treatment. Ⅱ , Ⅶ, Ⅸ and Ⅹ were measured in some patients. Results Among the 45 cases, no certain cause was found in 19 cases (42.2%), anticoagulant rodenticides poison was a common cause ( 11 cases,42.3% ). The main presentations was hemorrhage, the most common bleeding sites were mucosa (77.8%) (35/45)and hematuria (46.7%) ( 21/45 ). After vitamin K1 treatment, PT and APTT had shortened remarkably from ( 110.35 ± 35.36 ) s,(98.91 ±48.98)s to (13.48 ±2. 17)s,(33.25 ±6.95)s,respectively(t=19.10 and 6.19,Ps ＜0.01)and the activities of factor Ⅱ、Ⅶ、Ⅸ、Ⅹ had rapidly increased from ( 17.48 ± 10.93 ) %, ( 10.23 ± 5.68 )%, ( 11.98 ±4.69)%,(12.93±7.48)% to (70. 12 ±21.31)%,(92.76 ±29. 15)%,(88.64 ±40. 21)%,(63.97 ±20.11)%(t=12.13,14.43,13.27and9. 74,respectively,Ps＜0. 01).Conclusions The histories of patients with acquired deficiencies of vitamin K-dependent coagulation factors are usually hiding, therefore it is easily misdiagnosed. It is worth of detecting PT and APTT in diagnosis and monitoring. Using vitamin K1 10 -40 mg/d is effective and safety.