HPLC法测定儿童血浆中白消安的浓度
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摘要
目的建立灵敏、特异、准确、可靠的HPLC方法测定儿童血浆中白消安的浓度,监测造血干细胞移植(HSCT)前预处理方案中患儿静脉滴注白消安后血浆中药物的浓度,为分析白消安的药代动力学/药效学(PK/PD)及群体药代动力学(PPK)之间的关系奠定基础。方法取血浆样品,以甲醇400μL沉淀蛋白处理,待测物及内标经Diamonsil C_(18)色谱柱(150 mm×4. 6 mm,5μm)分离,以0. 1%甲酸水溶液-甲醇为流动相,色谱运行时间19 min。流速:1. 2m L·min~(-1),检测波长:280 nm,柱温:30℃,进样量:40μL。结果本研究所建立的HPLC-UV法测定患儿血浆中白消安的线性范围为0. 05～10μg·m L~(-1)。批内精密度为3. 22%～4. 42%,批间精密度为2. 60%～7. 19%。结论本研究所建立的检测方法有良好的特异性、灵敏度、精密度和准确度,适用于临床HSCT患儿血浆中白消安的治疗药物监测。
Objective To establish a method for determination of busulfan concentration in infants' plasma by high performance liquid chromatography detector method( HPLC). Methods Busulfan and the internal standard( IS) metronidazole were extracted from human plasma after protein precipitation by methanol( 400 μL),then detected with detection wavelength set at 280 nm after separation by Diamonsil C_(18)( 150 mm ×4. 6 mm,5 μm). The mobile phase consisted of methanol-0. 1%formic acid aqueous solution with a flow rate of 1. 2 m L·min~(-1). The method was validated for specificity,precision and accuracy,as well as stability. Results Linear calibration curves were obtained in the range of0. 05-10 μg·mL~(-1). RSD were in the ranges of 3. 22%-4. 42% and inter-day RSD were in the ranges of 2. 60%-7. 19%. Conclusion This validated method was successfully applied to quantify the concentrations of busulfan in the children undergoing hematopoietic stem cell transplantation.
Objective To establish a method for determination of busulfan concentration in infants' plasma by high performance liquid chromatography detector method( HPLC). Methods Busulfan and the internal standard( IS) metronidazole were extracted from human plasma after protein precipitation by methanol( 400 μL),then detected with detection wavelength set at 280 nm after separation by Diamonsil C_(18)( 150 mm ×4. 6 mm,5 μm). The mobile phase consisted of methanol-0. 1%formic acid aqueous solution with a flow rate of 1. 2 m L·min~(-1). The method was validated for specificity,precision and accuracy,as well as stability. Results Linear calibration curves were obtained in the range of0. 05-10 μg·mL~(-1). RSD were in the ranges of 3. 22%-4. 42% and inter-day RSD were in the ranges of 2. 60%-7. 19%. Conclusion This validated method was successfully applied to quantify the concentrations of busulfan in the children undergoing hematopoietic stem cell transplantation.
引文
[1]ANDERSSON B S,THALL P F,MADDEN T,et al.Busulfan systemic exposure relative to regimen-related toxicity and acute graft-versus-host disease:defining a therapeutic window for i.v.BuCy2 in chronic myelogenous leukemia[J].Biol Blood Marrow Transplant,2002,8(9):477-485.
[2]DIX S P,WINGARD J R,MULLINS R E,et al.Association of busulfan area under the curve with veno-occlusive disease following BMT[J].Bone Marrow Transplant,1996,17(2):225-230.
[3]GROCHOW L B,JONES R J,BRUNDRETT R B,et al.Pharmacokinetics of busulfan:correlation with veno-occlusive disease in patients undergoing bone marrow transplantation[J].Cancer Chemoth Pharm,1989,25(1):55-61.
[4]SLATTERY J T,CLIFT R A,BUCKNER C D,et al.Marrow transplantation for chronic myeloid leukemia:the influence of plasma busulfan levels on the outcome of transplantation[J].Blood,1997,89(8):3055-3060.
[5]EUROPEAN MEDICINES AGENCY(EMA).Guideline on bioanalytical method validation[EB/OL].London:EMA,2011-07-21[2016-05-12].http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2011/08/WC500109686.pdf.
[6]黄菁菁,陈冰,杨婉花.HPLC法测定人血浆中白消安的浓度[J].中国药师,2013,16(3):395-397.
[7]张善堂,方焱,屈建,等.柱前衍生高效液相色谱法测定人血浆中白消安的浓度[J].中国医院药学杂志,2007,27(4):458-461.
[8]PHILIPPE M,GOUTELLE S,GUITTON J,et al.Should busulfan therapeutic range be narrowed in pediatrics?Experience from a large cohort of hematopoietic stem cell transplant children[J].Bone Marrow Transplant,2016,51(1):72-78.
[9]MICHEL G,VALTEAU-COUANET D,GENTET J C,et al.Weight-based strategy of dose administration in children using intravenous busulfan:clinical and pharmacokinetic results[J].Pediatr Blood Cancer,2012,58(1):90-97.
[10]BOOTH B P,RAHMAN A,DAGHER R,et al.Population pharmacokinetic-based dosing of intravenous busulfan in pediatric patients[J].J Clin Pharmacol,2007,47(1):101-111.
[2]DIX S P,WINGARD J R,MULLINS R E,et al.Association of busulfan area under the curve with veno-occlusive disease following BMT[J].Bone Marrow Transplant,1996,17(2):225-230.
[3]GROCHOW L B,JONES R J,BRUNDRETT R B,et al.Pharmacokinetics of busulfan:correlation with veno-occlusive disease in patients undergoing bone marrow transplantation[J].Cancer Chemoth Pharm,1989,25(1):55-61.
[4]SLATTERY J T,CLIFT R A,BUCKNER C D,et al.Marrow transplantation for chronic myeloid leukemia:the influence of plasma busulfan levels on the outcome of transplantation[J].Blood,1997,89(8):3055-3060.
[5]EUROPEAN MEDICINES AGENCY(EMA).Guideline on bioanalytical method validation[EB/OL].London:EMA,2011-07-21[2016-05-12].http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2011/08/WC500109686.pdf.
[6]黄菁菁,陈冰,杨婉花.HPLC法测定人血浆中白消安的浓度[J].中国药师,2013,16(3):395-397.
[7]张善堂,方焱,屈建,等.柱前衍生高效液相色谱法测定人血浆中白消安的浓度[J].中国医院药学杂志,2007,27(4):458-461.
[8]PHILIPPE M,GOUTELLE S,GUITTON J,et al.Should busulfan therapeutic range be narrowed in pediatrics?Experience from a large cohort of hematopoietic stem cell transplant children[J].Bone Marrow Transplant,2016,51(1):72-78.
[9]MICHEL G,VALTEAU-COUANET D,GENTET J C,et al.Weight-based strategy of dose administration in children using intravenous busulfan:clinical and pharmacokinetic results[J].Pediatr Blood Cancer,2012,58(1):90-97.
[10]BOOTH B P,RAHMAN A,DAGHER R,et al.Population pharmacokinetic-based dosing of intravenous busulfan in pediatric patients[J].J Clin Pharmacol,2007,47(1):101-111.