HPLC法测定血浆和肺部组织、体液中左氧氟沙星浓度

Determination of the concentration of levofloxacin in plasma and tissues and body fluids of lung by high performance liquid chromatography

目的建立测定血浆、肺组织、支气管黏膜、肺泡灌洗液和痰液中左氧氟沙星浓度的方法。方法固定相为TSK-gel ODS-80TM C18柱（150mm×4．6mm，5μm）；流动相为50mmol·L^-1 KH2PO4（pH2．O）四氢呋喃-1mol·L^-1醋酸铵（92／7／1，V／V／V）；荧光检测波长Ex／Em为296／504mm，流速1．0mL·min^-1。结果各质控浓度的血浆、肺组织、支气管黏膜、肺泡灌洗液和痰液中左氧氟沙星的萃取回收率分别为94．2％-98．4％、100．2％～103．7％、99．3％～100．5％、99．9％～102．2％和100．6％-100．9％。各质控浓度的血浆、肺泡灌洗液、痰液和缓冲液测定结果日内RSD分别≤3．6％、≤5．3％、≤5．2％和≤1.7％，方法回收率分别为91．5％～110％、96．7％～100．5％、99．5％～103．8％和96．3％～103％；日间RSD分别≤3．5％、≤6．3％、≤6．3％和≤4．1％。血浆、肺泡灌洗液、痰液和缓冲液中左氧氟沙星质量浓度为0．01～5．00mg·L^-1时具有良好的线性关系（r=0．9996～0．9999）。左氧氟沙星在上述组织体液中最低检测限浓度为0．01mg·L^-1。结论本试验建立的HPLC方法灵敏准确、特异性高，可用于测定血浆、肺组织、支气管黏膜、肺泡灌洗液和痰液中左氧氟沙星的浓度。

AIM To establish the high performance liquid chromatography （HPLC） method for detmination of the concentration of levofloxacin in plasma, lung, bronchus membrane, bronchoalveolar lavage fluid （BALF） and sputum. METHODS The HPLC was peffonned on TSK-gel ODS-80TM Cl8 （ 150 mm× 4.6 mm, 5μm） columm. The mobile phase consisted of 50 mmol· L^-1 KH2PO4 （pH 2.0）-tetrahydrofuran-1 mol ·L^-1 CH3COONH4（92/7/1, V/V/V）. The detection wavelength（Ex/Em）wassetat296nm/504nm.The flow rate was 1.0 mL·min^-1. RESULTS The extraction recovery of each quality control concentration of plasma, lung, bronchus membrane, BALF and sputum was 94.2% - 98.4%, 100.2% - 103.7%, 99.3% - 100.5%, 99.9% - 102.2% and 100.6% - 100.9%, respectively. The inter-day coefficient variation of each quality control concentration of plasma, BALF, sputum and buffer was less than 3.6%,5.3%, 5.2% and 1.7%, respectively. The method recovery was 91.5% - 110%, 96.7% - 100.5%, 99.5% - 103.8% and 96.3% - 103% ,respectively. The intra-day coefficient variation was less than 3.5%, 6.3%, 6.3% and 4.1% ,respectively. A good linearity was shown in the concentration range 0.01 - 5.00 mg·L^-1 for all constituents （ r = 0.999 6 - 0.999 9）. The minimum detection limit concerttration of levofloxacin in above tissues and body fluids was 0.01 mg· L^-1. CONCLUSION The method of establishment is sensitive, accurate and has a high specificity.It is a suitable method for determining the drug concentration of levofloxacin in plasma , lung, bronchus membrane, BALF and sputum.