【摘要】 目的:研究血浆中华法林对映体的浓度对华法林抗凝疗效的影响。方法:建立血浆中华法林对映体浓度的立体选择性高效液相色谱法,采用手性-AGP色谱柱(ULTRON ES-OVM 5μm,150 mm×4.6 mm),保护柱(ULTRON ES-OVM.G 10mm×4.6 mm),以乙腈-20 mmol·L-1磷酸盐缓冲液(14∶86,p H 5)为流动相,流速1.0 m L·min-1,检测波长308 nm,柱温30℃,内标峰面积法定量计算。对146名心脏瓣膜置换术后使用华法林进行抗凝治疗的患者,服药后11~13 h按上述方法进行华法林对映体血浆浓度的检测。结果:1对映体标准曲线范围为75~2500 ng·m L-1,最低检出限为25 ng·m L-1,批内、批间RSD均在10%以下,回收率>90%,血浆样品稳定性良好。2R(+)-华法林平均血浆浓度为(635.0±380.7)ng·m L-1,S(-)-华法林平均血浆浓度为(334.8±237.7)ng·m L-1;S(-)-华法林血浆浓度与国际标准化比值(INR)值呈显著正相关(r=0.252,P<0.05),随着S(-)-华法林血浆浓度的升高,华法林的抗凝疗效增强,R(+)-华法林血浆浓度与INR值无显著相关性(r=0.104,P>0.05)。INR值在安全范围(1.8~3.0)的125例患者中,有92例(73.6%)S(-)-华法林血浆浓度在(334.8±237.7)ng·m L-1。结论:本研究建立的华法林对映体血浆浓度的检测方法灵敏而可靠,可作为华法林抗凝疗效的监测指标,联合INR值有助于提高抗凝监测的准确性。更多还原

【Abstract】 Objective: To study the influence of concentration of warfarin enantiomers in human plasma on warfarin anticoagulation. Methods: A stereo selective HPLC method was established to determine warfarin enantiomers in human plasma. The HPLC method employed the chiral- AGP column( 150 mm × 4. 6 mm,5 μm) with a mobile phase of acetonitrile- 20 mmol·L- 1phosphate buffer( 14∶ 86,p H 5) at a flow rate of 1. 0 m L·min- 1. The UV detection wavelength was 308 nm and the column temperature was 30 ℃. The internal standard peak area method for quantitative calculation was used. One hundred and forty- six Chinese patients after heart valve replacement( HVR)receiving a long- term warfarin maintenance therapy participated in the study. Plasma concentration of warfarin enantiomers at post- dosing 11 to 13 hours were measured by means of the chiral HPLC. Results: 1 The range of standard curve of each enantiomer was 75- 2500 ng·m L- 1,with the lowest limit of detection of 25 ng·m L- 1. RSDs of within or between- assay were less than 10% and the average recovery rate was > 90%. Plasma samples has good stability. 2The mean concentration of R( +)- and S(-)- warfarin in plasma were( 635. 0 ± 380. 7) ng·m L- 1and( 334. 8 ± 237. 7) ng·m L- 1,respectively,indicating that a significant positive relationship between the international normalized ratio( INR) and S(-)- warfarin plasma concentration( r = 0. 252,P < 0. 05). With the increase of S(-)- warfarin plasma concentration,the anticoagulant effect was enhanced. There was no significant correlation between R( +)- warfarin plasma concentration with INR values( r = 0. 104,P > 0. 05). The results showed that the 125 cases had their INR within the safety range( 1. 8- 3. 0),of which 92( 73. 6%) cases had a S(-)- warfarin level at( 334. 8 ± 237. 7) ng·m L- 1. Conclusion: The method is sensitive and reliable. The plasma concentration of warfarin can serve as a sensitive index in monitoring anticoagulant effect,the combination of warfarin plasma concentration with INR would improve the precision of anticoagulant monitoring.更多还原