QuECHERS-HPLC-MS测定人血浆中3种大环内酯类抗菌药物及其代谢物含量

Determination of 3 Macrolide Antibiotics and Their Metabolites in Plasma Using QuECHERS-HPLC-MS

关键词:  大环内酯类抗菌药物  代谢物  红霉素  地红霉素  螺旋霉素Ⅰ  高效液相色谱串联质谱  血浆

Key words:  macrolide antibiotics  metabolites  erythromycin A  dirithromycin  spiramycin Ⅰ  HPLC-MS  plasma

摘要:

目的 建立测定人血浆中3种大环内酯类抗菌药物（红霉素、地红霉素、螺旋霉素Ⅰ）及其代谢物（脱水红霉素、红霉素A烯醇醚、红霉胺、新螺旋霉素Ⅰ）的分析方法。方法 用乙腈溶液提取，QuECHERS方法净化，采用无水硫酸镁和无水乙酸钠沉淀蛋白和盐析分层，无水硫酸镁和石墨碳烯净化，上清液氮气吹干后甲醇复溶待测，用Eclipse Plus C₁₈色谱柱分离，电喷雾正离子模式，多反应离子监测扫描，内标法定量。结果 3种大环内酯类抗菌药物及其代谢物浓度在4~900 ng·mL^-1内的线性关系良好（r>0.999），定量限<10 ng·mL^-1；在3个添加水平下，3种大环内酯类抗菌药物及其代谢物的提取回收率可达81.45%~96.82%，日内精密度<5.00%，日间精密度<10.00%。结论 QuECHERS-液相色谱串联三重四级杆质谱测定血浆中3种大环内酯类抗菌药物及其代谢物的分析方法简单，快速，灵敏，特异性强，适用于血浆中大环内酯类抗菌药物及其代谢物的测定。

Abstract:

OBJECTIVE To establish a determination method of 3 macrolide antibiotics (erythromycin A, dirithromycin, spiramycin Ⅰ), and it's metabolite (anhydroerythromycin A, erythromycin A ene alcohol ether, erythromycylamine, neospiramycin Ⅰ) in plasma using QuECHERS followed by HPLC-MS. METHODS The macrolide antibiotics in samples were extracted with acetonitrile. The plasma was directly purified by QuECHERS pretreatment method. MgSO₄ and sodium acetate anhydrous were added into plasma sample for precipitating protein and salting out stratification. Then, MgSO₄ and PestiCarb were used for dehydration and removing impurity. The upper layer was evaporated to dryness under a stream of nitrogen and the residue obtained was redissolved in methanol. Three macrolide antibiotics were separated by Eclipse Plus C₁₈ chromatographic column. Electro spray ionization was used to analyze these macrolide antibiotics by using positive ion mode. Multiple reaction monitoring was used to monitor ions. Internal standard method was used to quantitative calculation. RESULTS Three macrolide antibiotics showed good linear relationship in the range of 4-900 ng·mL^-1(r>0.999). The quantitation limits was <10 ng·mL^-1. At three levels, the average extract recoveries were 81.45%-96.82%, and the intra-day and inter-day precisions were respectively lower than 5.00% and 10.00%, respectively. CONCLUSION The analytical method of 3 macrolide antibiotics in plasma using QuEChERS and HPLC-MS is simple, rapid, sensitive and specific. The analytical method is suitable to quantitate macrolide antibiotics and their metabolites in plasma.