鸡血浆中磺胺氯吡嗪钠和甲氧苄啶含量HPLC检测方法的建立

Establishment of a HPLC method for the detection of sulfachlorpyrazine sodium and trimethoprim in broiler chicken plasma

关键词:  磺胺氯吡嗪钠  甲氧苄啶  肉鸡  HPLC  血浆

Key words:  sulfachloropyrazine sodium  trimethoprim  broiler chicken  HPLC  plasma

摘要:

为了建立鸡血浆中磺胺氯吡嗪钠和甲氧苄啶含量测定的HPLC方法。本研究对鸡血浆中磺胺氯吡嗪和甲氧苄啶采用乙腈提取和净化，梯度洗脱，高效液相色谱紫外检测法测定，外标法定量。流动相为甲醇-0.02 mol/L磷酸二氢钾水溶液。紫外测定时采用双波长检测：磺胺氯吡嗪检测波长为268 nm，甲氧苄啶检测波长为240 nm。空白血浆添加磺胺氯吡嗪和甲氧苄啶分别在0.1~70.0 μg/mL和0.05~5.00 μg/mL范围内线性关系良好（R2均大于0.9999），磺胺氯吡嗪和甲氧苄啶的检测限分别为0.05 μg/mL和0.025 μg/mL，定量限分别为0.1 μg/mL和0.05 μg/mL。空白血浆添加磺胺氯吡嗪和甲氧苄啶在各自线性范围内，批内变异系数分别小于6.84%和7.55%，批间系数分别小于5.23%和6.81%。本研究建立的鸡血浆中磺胺氯吡嗪和甲氧苄啶的提取和净化方法，适用于鸡血浆中磺胺氯吡嗪和甲氧苄啶含量的单个或同时测定。

Abstract:

Abstract: In order to establish a high performance liquid chromatographic ( HPLC) method for the detection of the amounts of sulfachloropyrazine sodium and trimethoprim in broiler’s plasma. The plasma sample containing sulfachloropyrazine sodium and trimethoprim was firstly extracted and purified by acetonitrile, and then underwent the procedure of gradient elution，before being measured using high performance liquid chromatography ultraviolet detection method，lastly quantified through external standard method. The isocratic mobile phase was a aqueous solution made of a mixture of methyl alcohol and 0.02 mol/L potassium dihydrogen phosphate. Dual-wavelength detection was used for ultraviolet detection: the detective UV wavelength of sulfachlorpyrazine was 268 nm, and that of trimethoprim was 240 nm. The linear relationship was good when blank plasma containing the amounts of sulfachlorpyrazine and trimethoprim in the range of 0.1-70.0 ug/mL and 0.05-5.00 ug/mL respectively(R2 > 0.9999).The detection limits of sulfachlorpyrazine and trimethoprim were 0.05 ug/mL and 0.025 ug/mL, and the quantitative limits were 0.1 ug/mL and 0.05 ug/mL respectively. In the blank plasma, sulfamlopyrazine and trimethoprim were added within their linear ranges.,the intra-batch variation coefficients were less than 6.84% and 7.54% respectively, and the inter-batch coefficients were less than 5.27% and 6.80% respectively.This research provides a method for the extraction and purification of sulfachlorpyrazine and trimethoprim in broiler chicken plasma，and it is applicable for both single and simultaneous detection of sulfachlorpyrazine and trimethoprim in broiler chicken plasma. .Key words:sulfachloropyrazine sodium; trimethoprim; broiler chicken; HPLC; plasma