遗传性嗜铬细胞瘤家系血浆儿茶酚胺及其代谢物的含量测定
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摘要
目的建立高效液相色谱-电化学检测器(HPLC-ECD)法测定遗传性嗜铬细胞瘤家系血浆儿茶酚胺及其代谢产物的含量的方法。方法工作电极为玻璃电极,参比电极为Ag-AgCl电极,电压0.6 V,温度30℃。色谱柱:Ultimate XB-C18(4.6 mm×150 mm,5μm);以甲醇为流动相A,0.1%磷酸二氢钠(含EDTA-Na和辛烷磺酸)水溶液为流动相B,梯度洗脱,流速为0.8 mL/min;柱温为室温;检测波长:280 nm;进样量:20μL。结果正常健康人中肾上腺素(E)、去甲肾上腺素(NE)、甲氧基去甲肾上腺素(NMN)、甲氧基肾上腺素(MN)分别为(56.71±25.67)、(98.62±36.90)、(78.36±22.36)、(33.27±14.62) ng/L。嗜铬细胞瘤家系中确诊的2例嗜铬细胞瘤患者的NE分别为353.36、563.87 ng/L和NMN分别为425.71、527.65 ng/L,其余18位家属的相关检测值在正常健康成人的范围内,其他2位家属的相关检测值比正常值高,建议去医院进一步检查。结论该方法简便,灵敏、准确,可用于临床嗜铬细胞瘤患者的检测。
Objective To establish a method for the determination of catecholamines and their metabolites in plasma of hereditary pheochromocytoma families by high performance liquid chromatography-electrochemical detector(HPLC-ECD).Methods The working electrode was a glass electrode and the reference electrode was an Ag-AgCl electrode with a voltage of 0.6 V and a temperature of 30℃. Using Ultimate XB-C18(4.6 mm × 150 mm, 5 μm); using methanol as mobile phase A, 0.1% sodium dihydrogen phosphate(containing EDTA-Na and octanesulfonic acid) aqueous solution as mobile phase B, gradient elution, flow rate 0.8 mL/min; column temperature was room temperature; detection wavelength was280 nm; injection volume was 20 μL. Results In normal healthy people, the contents of epinephrine(E), norepinephrine(NE), methoxy norepinephrine(NMN), methoxy-adrenalin(MN) were(56.71±25.67),(98.62±36.90),(78.36±22.36),(33.27±14.62) ng/L. The NEs of 2 patients with pheochromocytoma diagnosed in pheochromocytoma were 353.36, 563.87 ng/L and NMN were 425.71 and 527.65 ng/L, respectively. The correlation detection values of the other 18 family members were within the range of normal healthy adults. The correlation detection values of the other two family members were higher than the normal values. it is recommended to go to the hospital for further examination. Conclusion This method is simple, sensitive and accurate and can be used for the detection of patients with clinical pheochromocytoma.
Objective To establish a method for the determination of catecholamines and their metabolites in plasma of hereditary pheochromocytoma families by high performance liquid chromatography-electrochemical detector(HPLC-ECD).Methods The working electrode was a glass electrode and the reference electrode was an Ag-AgCl electrode with a voltage of 0.6 V and a temperature of 30℃. Using Ultimate XB-C18(4.6 mm × 150 mm, 5 μm); using methanol as mobile phase A, 0.1% sodium dihydrogen phosphate(containing EDTA-Na and octanesulfonic acid) aqueous solution as mobile phase B, gradient elution, flow rate 0.8 mL/min; column temperature was room temperature; detection wavelength was280 nm; injection volume was 20 μL. Results In normal healthy people, the contents of epinephrine(E), norepinephrine(NE), methoxy norepinephrine(NMN), methoxy-adrenalin(MN) were(56.71±25.67),(98.62±36.90),(78.36±22.36),(33.27±14.62) ng/L. The NEs of 2 patients with pheochromocytoma diagnosed in pheochromocytoma were 353.36, 563.87 ng/L and NMN were 425.71 and 527.65 ng/L, respectively. The correlation detection values of the other 18 family members were within the range of normal healthy adults. The correlation detection values of the other two family members were higher than the normal values. it is recommended to go to the hospital for further examination. Conclusion This method is simple, sensitive and accurate and can be used for the detection of patients with clinical pheochromocytoma.
引文
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[5]张华,李永兴.高效液相电化学法检测尿儿茶酚胺和VMA诊断嗜铬细胞瘤[J].现代检验医学杂志,2003,18(3):8-9.
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[13]Eisenhofer G,Keiser H,Friberg P,et al.Plasma metanephrines are markers of pheochromocytoma produced by catecholO-methyItransferase within tumors[J].J Clin Endocrinol Metab,2008,83(6):2175-2185.
[14]黄航,王峰,张方毅,等.酶联免疫法检测间甲肾上腺素类物质在嗜铬细胞瘤诊断中的应用[J].浙江临床医学,23(12):1779-1781.
[15]Melmed S,Polonsky KS,Larsen PR,et al.Williams Textbook of Endocrinology[M].12th ed.Philadelphia:Saunders/Elservier,2011,69(8):56-60.
[16]朱正,赵心明,戴景蕊,等.腹膜后异位嗜铬细胞瘤的CT和MR表现[J].中华肿瘤杂志,2015,37(3):181-185.
[17]李西,刘燕娜,章春泉,等.嗜铬细胞瘤致Takotsubo型心肌病超声表现1例[J].中国医学影像技术,2017(9):1443.
[18]文进,李汉忠,纪志刚,等.35例肾上腺嗜铬细胞瘤/副神经节瘤3D腹腔镜手术分析[J].基础医学与临床,2016,36(7):1014-1016.
[19]Neary NM,King KS,Pacak K.Drugs and pheochromocytoma-don′t be fooled by every elevated metanephrine[J].N Engl J Med,2011,364(23):2268-2270.
[20]Bouhanick B,Fauvel J,Pont F.Biochemical misdiagnosis of pheochromocytoma in patients treated with sulfasalazine[J].JAMA,2010,304(17):1898-1901.
[21]刘洪旭,邓思珊,马丽红,等.1例嗜铬细胞瘤继发高血压脑出血患者的药学监护[J].中国现代医生,2018,56(3):137-139.
[2]Yu R,Nissen NN,Bannykh SI.Cardiac Complications as Initial Manifestation of Pheochromocytoma:Frequency,Outcome,and Predictors[J].Endocr Pract,20l2,18(4):483-492.
[3]Eisenhofer G Editorial:biochemical diagnosis of pheochromocytoma is it time to switch to plasma-free metanephrines[J].J Clin Endocrinol Metab,2003,88(2):550-552.
[4]Grouzmann E,Drouard-Troalen L,Baudin E,et al.Diagnostic accuracy of free and total metanephrines in plasma and fractionated metanephrines in urine of patients with pheochromocytoma[J].Eur J Endocrino1,2010,162(5):951-960.
[5]张华,李永兴.高效液相电化学法检测尿儿茶酚胺和VMA诊断嗜铬细胞瘤[J].现代检验医学杂志,2003,18(3):8-9.
[6]Ghnguly S,LeBeau S,Pierce K,et al.Multiple paragangliomas in a pregnant patient with a succinate dehydrogenaLse B mutation[J].Postgrad Med,2010,122(6):46-50.
[7]姜蕾,王卫庆,张军妮,等.高效液相色谱电化学法测定血间甲肾上腺素类物质方法的建立及临床应用[J].中华内分泌代谢杂志,2005,20(6):561-563.
[8]陈静,丘宏强.高效液相色谱-电化学法测定人尿液中儿茶酚胺的浓度[J].中国药房,2016,27(26):3627-3629.
[9]Foti A,Kimura S,DeQuattro V,et al.Liquid-chomatographic measurement of catecho1amines and metabolites in plasma and urine[J].Clin Chem,2007,33(12):2209-2213.
[10]Lagerstedt SA,O′Kane DJ,Singh RJ.Measurement of plasma free metanephrine and normetanephrine by liquid chromatography-tandem mass spectrometry for diagnosis of pheochromocytoma[J].Clin Chem,2004,50(3):603-611.
[11]Lenders JW,Pacak K,Walther MM,et al.Biochemical diagnosis of pheochromocytoma:which test is best[J].JAMA,2002,287(11):1427-1434.
[12]Campbell KA,Joseph SP,Whiting MJ,et al.The Author′s Reply:The Halflives of Plasma Free Metanephrines[J].Clin Endocrinol,2012,77(3):485-486.
[13]Eisenhofer G,Keiser H,Friberg P,et al.Plasma metanephrines are markers of pheochromocytoma produced by catecholO-methyItransferase within tumors[J].J Clin Endocrinol Metab,2008,83(6):2175-2185.
[14]黄航,王峰,张方毅,等.酶联免疫法检测间甲肾上腺素类物质在嗜铬细胞瘤诊断中的应用[J].浙江临床医学,23(12):1779-1781.
[15]Melmed S,Polonsky KS,Larsen PR,et al.Williams Textbook of Endocrinology[M].12th ed.Philadelphia:Saunders/Elservier,2011,69(8):56-60.
[16]朱正,赵心明,戴景蕊,等.腹膜后异位嗜铬细胞瘤的CT和MR表现[J].中华肿瘤杂志,2015,37(3):181-185.
[17]李西,刘燕娜,章春泉,等.嗜铬细胞瘤致Takotsubo型心肌病超声表现1例[J].中国医学影像技术,2017(9):1443.
[18]文进,李汉忠,纪志刚,等.35例肾上腺嗜铬细胞瘤/副神经节瘤3D腹腔镜手术分析[J].基础医学与临床,2016,36(7):1014-1016.
[19]Neary NM,King KS,Pacak K.Drugs and pheochromocytoma-don′t be fooled by every elevated metanephrine[J].N Engl J Med,2011,364(23):2268-2270.
[20]Bouhanick B,Fauvel J,Pont F.Biochemical misdiagnosis of pheochromocytoma in patients treated with sulfasalazine[J].JAMA,2010,304(17):1898-1901.
[21]刘洪旭,邓思珊,马丽红,等.1例嗜铬细胞瘤继发高血压脑出血患者的药学监护[J].中国现代医生,2018,56(3):137-139.