Abu-Qare A W, Abou-Donia M B
Department of Pharmacology and Cancer Biology, Duke University Medical Center, PO Box 3813, Durham, NC 27710, USA.
J Pharm Biomed Anal. 2001 Dec;26(5-6):939-47. doi: 10.1016/s0731-7085(01)00448-4.
A method was developed for the separation and quantification of the anti-nerve agent pyridostigmine bromide (PB; 3-dimethylaminocarbonyloxy-N-methyl pyridinium bromide), the analgesic drugs acetaminophen and acetylsalicylic acid, and the stimulant caffeine (3,7-dihydro-1,3,7-trimethyl-1-H-purine-2,6-dione) in rat plasma and urine. The compounds were extracted using C(18) Sep-Pak(R) cartridges then analyzed by high performance liquid chromatography (HPLC) with reversed phase C18 column, and UV detection at 280 nm. The compounds were separated using gradient of 1-85% acetonitrile in water (pH 3.0) at a flow rate ranging between 1 and 1.5 ml/min in a period of 14 min. The retention times ranged from 8.8 to 11.5 min. The limits of detection were ranged between 100 and 200 ng/ml, while limits of quantitation were 150-200 ng/ml. Average percentage recovery of five spiked plasma samples were 70.9+/-9.5, 73.7+/-9.8, 88.6+/-9.3, 83.9+/-7.8, and from urine 69.1+/-8.5, 74.5+/-8.7, 85.9+/-9.8, 83.2+/-9.3, for pyridostigmine bromide, acetaminophen, acetylsalicylic acid and caffeine, respectively. The relationship between peak areas and concentration was linear over range between 100 and 1000 ng/ml. The resulting chromatograms showed no interfering peaks from endogenous plasma or urine components. This method was applied to analyze these compounds following oral administration in rats.
已开发出一种用于分离和定量大鼠血浆及尿液中抗神经毒剂溴化吡啶斯的明（PB；3 - 二甲基氨基甲酰氧基 - N - 甲基吡啶溴化物）、镇痛药对乙酰氨基酚和乙酰水杨酸以及兴奋剂咖啡因（3,7 - 二氢 - 1,3,7 - 三甲基 - 1 - H - 嘌呤 - 2,6 - 二酮）的方法。使用C(18) Sep - Pak(R) 柱进行化合物萃取，然后通过配备反相C18柱的高效液相色谱（HPLC）进行分析，并在280 nm处进行紫外检测。在14分钟内，以1至1.5 ml/分钟的流速，使用1 - 85%乙腈水溶液（pH 3.0）的梯度对化合物进行分离。保留时间在8.8至11.5分钟之间。检测限在100至200 ng/ml之间，定量限为150 - 200 ng/ml。五个加标血浆样本中溴化吡啶斯的明、对乙酰氨基酚、乙酰水杨酸和咖啡因的平均回收率分别为70.9±9.5、73.7±9.8、88.6±9.3、83.9±7.8，尿液中的平均回收率分别为69.1±8.5、74.5±8.7、85.9±9.8、83.2±9.3。峰面积与浓度之间的关系在100至1000 ng/ml范围内呈线性。所得色谱图未显示来自内源性血浆或尿液成分的干扰峰。该方法用于分析大鼠口服给药后的这些化合物。
