Ko R J, Li W Y, Koda R T
School of Pharmacy, University of Southern California, Los Angeles 90033.
J Chromatogr. 1990 Feb 23;525(2):411-8. doi: 10.1016/s0378-4347(00)83417-9.
In an effort to characterize the pharmacokinetic behavior of the antimitotic agent N-desacetylcolchicine a selective, sensitive high-performance liquid chromatographic method was developed for the determination of N-desacetylcolchicine, demecolcine and colchicine in serum or urine. To 0.5 ml of serum or 0.1 ml of urine diluted to 0.5 ml were added 50 microliters demecolcine (2 micrograms/ml) which serves as the internal standard. The sample was extracted using a C2 reversed-phase solid extraction column. N-Desacetyl-colchicine, colchicine and the internal standard were eluted from the column with methanol. The combined eluates were evaporated to dryness and the residue was reconstituted with water. The reconstituted sample was injected into a C18 reversed-phase column and eluted using a mobile phase consisting of 0.1 M potassium dihydrogenphosphate, 5 mM 1-pentanesulfonic acid in methanol and acetonitrile with a final pH of 6.0, at a flow rate of 1.5 ml/min. N-Desacetylcolchicine, colchicine and the internal standard were detected using a variable-wavelength ultraviolet detector at 254 nm. The limit of detection was 0.4 ng/ml for desacetylcolchicine and 4.0 ng/ml for colchicine. The method is linear over a concentration range of 1.0-200 ng/ml. The method has been shown to be a rapid, reliable method to monitor N-desacetylcolchicine levels in clinical trials in cancer patients.
为了表征抗有丝分裂剂N-去乙酰基秋水仙碱的药代动力学行为，开发了一种选择性、灵敏的高效液相色谱法，用于测定血清或尿液中的N-去乙酰基秋水仙碱、去甲秋水仙碱和秋水仙碱。向0.5 ml血清或稀释至0.5 ml的0.1 ml尿液中加入50微升去甲秋水仙碱（2微克/毫升）作为内标。使用C2反相固相萃取柱萃取样品。N-去乙酰基秋水仙碱、秋水仙碱和内标用甲醇从柱上洗脱。合并的洗脱液蒸发至干，残留物用水复溶。将复溶后的样品注入C18反相柱，使用由0.1 M磷酸二氢钾、5 mM 1-戊烷磺酸的甲醇和乙腈组成的流动相洗脱，最终pH为6.0，流速为1.5 ml/min。使用可变波长紫外检测器在254 nm处检测N-去乙酰基秋水仙碱、秋水仙碱和内标。去乙酰基秋水仙碱的检测限为0.4 ng/ml，秋水仙碱的检测限为4.0 ng/ml。该方法在1.0 - 200 ng/ml的浓度范围内呈线性。在癌症患者的临床试验中，该方法已被证明是一种监测N-去乙酰基秋水仙碱水平的快速、可靠的方法。
