Wright A W, Watt J A, Kennedy M, Cramond T, Smith M T
Department of Pharmacy, University of Queensland, Royal Brisbane Hospital, Australia.
Ther Drug Monit. 1994 Apr;16(2):200-8. doi: 10.1097/00007691-199404000-00016.
An original, sensitive, and specific high-performance liquid chromatographic (HPLC) assay was developed for the quantitation of morphine and its two major metabolites, morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G), in human plasma and cerebrospinal fluid (CSF) and in rat plasma, using hydromorphone as the internal standard. Solid-phase extraction was used to separate morphine and its glucuronide metabolites from plasma constituents. Extraction efficiencies of morphine, M3G, and M6G from human plasma samples (0.5 ml) were 84, 87, and 88%, respectively. Extraction efficiencies of morphine, M3G, and M6G did not differ significantly (p > 0.05) between human plasma and CSF or rat plasma. Morphine, M3G, M6G, and hydromorphone were separated on a 10 mu C8 Resolve radially compressed cartridge using a mobile phase comprising methanol:acetonitrile:phosphate buffer, (0.0125M pH 7.5; 10:10:80), in which 11 mg/L of cetyltrimethylammonium bromide (cetrimide) was dissolved. Quantitation was achieved using a single electrochemical detector at ambient temperature (23 degrees C). Standard curves were linear over the ranges 0.020-2.190, 0.027-2.709, and 0.027-0.542 microM for morphine, M3G, and M6G, respectively. Lower limits of detection for morphine, M3G, and M6G in human plasma and CSF samples (0.5 ml) were 0.020, 0.027, and 0.027 microM, respectively. Corresponding lower limits of detection in rat plasma (0.1 ml) were 0.102, 0.135, and 0.135 microM, respectively. Intraassay precision for low and high concentrations of morphine, M3G, and M6G were < 23 and < 8% respectively. Similarly, interassay accuracy for low and medium concentrations of morphine, M3G, and M6G were < 17% and were < 9% for high concentrations.
建立了一种原始、灵敏且特异的高效液相色谱（HPLC）分析法，用于定量测定人血浆、脑脊液（CSF）以及大鼠血浆中的吗啡及其两种主要代谢物，即吗啡 - 3 - 葡萄糖醛酸苷（M3G）和吗啡 - 6 - 葡萄糖醛酸苷（M6G），采用氢吗啡酮作为内标。使用固相萃取法从血浆成分中分离吗啡及其葡萄糖醛酸代谢物。从人血浆样品（0.5 ml）中提取吗啡﹑M3G和M6G的效率分别为84%、87%和88%。人血浆与脑脊液或大鼠血浆之间，吗啡、M3G和M6G的提取效率无显著差异（p > 0.05）。吗啡、M3G、M6G和氢吗啡酮在10μm C8径向压缩柱上进行分离，流动相为甲醇∶乙腈∶磷酸盐缓冲液（0.0125M，pH 7.5；10∶10∶80），其中溶解有11 mg/L的十六烷基三甲基溴化铵（西曲溴铵）。在环境温度（23℃）下使用单通道电化学检测器进行定量分析。吗啡、M3G和M6G的标准曲线分别在0.020 - 2.190、0.027 - 2.709和0.027 - 0.542μM范围内呈线性。人血浆和脑脊液样品（0.5 ml）中吗啡、M3G和M6G的检测下限分别为0.020、0.027和0.027μM。大鼠血浆（0.1 ml）中相应的检测下限分别为0.102、0.135和0.135μM。低浓度和高浓度的吗啡、M3G和M6G的批内精密度分别< 23%和< 8%。同样，低浓度和中等浓度的吗啡、M3G和M6G的批间准确度< 17%，高浓度时< 9%。
