Brenner D E, Galloway S, Cooper J, Noone R, Hande K R
Cancer Chemother Pharmacol. 1985;14(2):139-45. doi: 10.1007/BF00434353.
We compared doxorubicin and metabolite pharmacokinetic data obtained from thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) assay of plasma samples from six patients who had been treated with doxorubicin. Duplicate 1-ml samples were extracted with chloroform: isopropanol (1:1) and assayed using a sensitive HPLC system incorporating a dual pump gradient with tetrahydrofuran as the mobile phase and fluorescence detection. Duplicate 1-ml samples from the same specimens were assayed using a modification of a previously described TLC assay. Areas under the curve for doxorubicin by HPLC (3.36 +/- 2.30 microM X h) and TLC (4.16 +/- 2.50 microM X h) were not significantly different (P = 0.5). Terminal half-life of doxorubicin by HPLC (28.0 +/- 6.98 h) and TLC (23.2 +/- 7.8) (P = 0.29) and the calculated total-body clearances by HPLC (0.55 +/- 0.29 l/min) and TLC (0.45 +/- 0.23) (P = 0.55) were not significantly different. Areas under the curve for doxorubicinol by HPLC (2.75 +/- 1.4 microM X h) and TLC (2.53 +/- 7.1 microM X h) (P = 0.73) showed no significant differences. HPLC detected a mixed 7-deoxydoxorubicinol aglycone-doxorubicin aglycone peak, 7-deoxydoxorubicin aglycone, and two nonpolar, unidentified metabolites. TLC detected the following aglycone metabolites: doxorubicin aglycone, doxorubicinol aglycone, 7-deoxydoxorubicinol aglycone, an unidentified polar metabolite, and several unidentified nonpolar metabolites. From these data we conclude that HPLC and TLC detect concentrations of doxorubicin and doxorubicinol from human plasma equally well to concentrations of 7.0 nM (4 pmol injected doxorubicin). Aglycones do circulate in human plasma at concentrations above the detection limits of both assays. Doxorubicinol aglycone, which is detected by TLC but not by HPLC, may be formed from artifactual breakdown of doxorubicinol during TLC development. Unidentified nonpolar compounds seen on HPLC and TLC may represent further doxorubicin metabolism than previously described.
我们比较了通过薄层色谱法（TLC）和高效液相色谱法（HPLC）对6例接受阿霉素治疗患者的血浆样本进行检测所获得的阿霉素及其代谢产物的药代动力学数据。取1 ml样本重复两份，用氯仿：异丙醇（1:1）萃取，然后使用一个灵敏的HPLC系统进行检测，该系统采用双泵梯度，以四氢呋喃作为流动相并进行荧光检测。对相同样本的1 ml重复样本采用之前描述的TLC检测法的改良方法进行检测。HPLC法测得阿霉素的曲线下面积为（3.36±2.30微摩尔×小时），TLC法测得为（4.16±2.50微摩尔×小时），两者无显著差异（P = 0.5）。HPLC法测得阿霉素的终末半衰期为（28.0±6.98小时），TLC法测得为（23.2±7.8）（P = 0.29）；HPLC法计算的全身清除率为（0.55±0.29升/分钟），TLC法为（0.45±0.23）（P = 0.55），均无显著差异。HPLC法测得阿霉素醇的曲线下面积为（2.75±1.4微摩尔×小时），TLC法测得为（2.53±7.1微摩尔×小时）（P = 0.73），无显著差异。HPLC检测到一个混合的7-脱氧阿霉素醇苷元 - 阿霉素苷元峰、7-脱氧阿霉素苷元以及两种非极性、未鉴定的代谢产物。TLC检测到以下苷元代谢产物：阿霉素苷元、阿霉素醇苷元、7-脱氧阿霉素醇苷元、一种未鉴定的极性代谢产物以及几种未鉴定的非极性代谢产物。从这些数据我们得出结论，对于人血浆中阿霉素和阿霉素醇浓度的检测，HPLC和TLC在检测浓度达7.0 nM（注射4皮摩尔阿霉素）时效果相当。苷元在人血浆中的循环浓度高于两种检测方法的检测限。TLC能检测到而HPLC检测不到的阿霉素醇苷元，可能是在TLC展开过程中由阿霉素醇的人为分解形成的。HPLC和TLC上看到的未鉴定的非极性化合物可能代表了比之前描述的更深入的阿霉素代谢情况。
