药学实践杂志
藥學實踐雜誌
약학실천잡지
THE JOURNAL OF PHARMACEUTICAL PRACTICE
2014年
4期
270-272,293
,共4页
吕磊%赵亮%兰红涛%金柔男%张国庆
呂磊%趙亮%蘭紅濤%金柔男%張國慶
려뢰%조량%란홍도%금유남%장국경
血浆%胆汁酸测定%质谱%肝癌
血漿%膽汁痠測定%質譜%肝癌
혈장%담즙산측정%질보%간암
plasma%bile acid determination%MS%liver cancer
目的:建立HPLC-MS方法同时检测健康人和原发性肝癌患者血浆中13种胆汁酸的含量。方法血浆样品前处理采用乙腈-蛋白沉淀法,色谱分离采用SHISEIDO MG C18(100 mm ×3.0 mm,3.0μm)柱,流动相:乙腈-水(含0.1%甲酸和0.5%氨水),梯度洗脱,流速0.4 ml/min,柱温25℃,进样量5μl。质谱监测采用ESI源负离子模式,选择离子(SIM)分段监测。结果13种胆汁酸在3.3~4400 ng/ml的浓度范围内线性关系良好;日内和日间精密度均<7%;平均回收率为82.67%~109.75%。样品测定结果显示,与健康对照组相比,原发性肝癌患者血浆中胆汁酸含量升高,其中以结合型胆汁酸TUDCA、TCDCA、TCA、GCA和GCDCA的升高最为明显。结论所建HPLC-MS方法灵敏度高、选择性好,可用于原发性肝癌患者与健康人体内血清胆汁酸的含量差异比较,该研究为肝癌患者体内胆汁酸的代谢和疾病的临床诊治奠定了基础。
目的:建立HPLC-MS方法同時檢測健康人和原髮性肝癌患者血漿中13種膽汁痠的含量。方法血漿樣品前處理採用乙腈-蛋白沉澱法,色譜分離採用SHISEIDO MG C18(100 mm ×3.0 mm,3.0μm)柱,流動相:乙腈-水(含0.1%甲痠和0.5%氨水),梯度洗脫,流速0.4 ml/min,柱溫25℃,進樣量5μl。質譜鑑測採用ESI源負離子模式,選擇離子(SIM)分段鑑測。結果13種膽汁痠在3.3~4400 ng/ml的濃度範圍內線性關繫良好;日內和日間精密度均<7%;平均迴收率為82.67%~109.75%。樣品測定結果顯示,與健康對照組相比,原髮性肝癌患者血漿中膽汁痠含量升高,其中以結閤型膽汁痠TUDCA、TCDCA、TCA、GCA和GCDCA的升高最為明顯。結論所建HPLC-MS方法靈敏度高、選擇性好,可用于原髮性肝癌患者與健康人體內血清膽汁痠的含量差異比較,該研究為肝癌患者體內膽汁痠的代謝和疾病的臨床診治奠定瞭基礎。
목적:건립HPLC-MS방법동시검측건강인화원발성간암환자혈장중13충담즙산적함량。방법혈장양품전처리채용을정-단백침정법,색보분리채용SHISEIDO MG C18(100 mm ×3.0 mm,3.0μm)주,류동상:을정-수(함0.1%갑산화0.5%안수),제도세탈,류속0.4 ml/min,주온25℃,진양량5μl。질보감측채용ESI원부리자모식,선택리자(SIM)분단감측。결과13충담즙산재3.3~4400 ng/ml적농도범위내선성관계량호;일내화일간정밀도균<7%;평균회수솔위82.67%~109.75%。양품측정결과현시,여건강대조조상비,원발성간암환자혈장중담즙산함량승고,기중이결합형담즙산TUDCA、TCDCA、TCA、GCA화GCDCA적승고최위명현。결론소건HPLC-MS방법령민도고、선택성호,가용우원발성간암환자여건강인체내혈청담즙산적함량차이비교,해연구위간암환자체내담즙산적대사화질병적림상진치전정료기출。
Objective To develop a method for determination of 13 kinds of bile acid in healthy people and patients with pri-mary liver cancer by HPLC-MS.Methods Acetonitrile was used to precipitate protein in plasma samples .The separation was per-formed on a SHISEIDO MG C18 reverse phase column (3.0 mm ×100 mm,3μm).The mobile phase were acetonitrile and water (con-taining 0.1%formic acid and 0.5%ammonia), gradient elution.The flow rate was 0.4 ml/min, the temperature of column was 25℃and the injection volume was 5 μl.Electrospray ion source (ESI) and selected ion mode (SIM) were applied for quantitative analysis under negative ion mode .Results The calibration curves of 13 kinds of bile acids were linear in the range of 3.3-4 400 ng/ml, intra-day and inter-day precision results were all less than 7%, the average recovery were 82.67%-109.75%.The result showed that the content of conjugated bile acid increased in the plasma of patients with primary liver cancer compared with the healthy control group , especially in TUDCA, TCDCA, TCA, GCA and GCDCA.Conclusion The LC -MS method was sensitive and selective which could be suitable for determination of bile acid in healthy people and patients with primary liver cancer .The study laid a foundation for the me-tabolism of bile acid in vivo and the clinical diagnosis and treatment of patients with hepatocellular carcinoma .