Rajagopalan P, Gao Z, Chu C K, Schinazi R F, McClure H M, Boudinot F D
Department of Pharmaceutics, College of Pharmacy, University of Georgia, Athens 30602-2353, USA.
J Chromatogr B Biomed Appl. 1995 Oct 6;672(1):119-24. doi: 10.1016/0378-4347(95)00197-q.
(-)-beta-D-2,6-Diaminopurine dioxolane (DAPD) and its metabolite dioxolane guanosine (DXG) have potent activity against hepatitis B virus and HIV, in vitro. A reversed-phase HPLC analytical method using UV and on-line radiochemical detection for the determination of DAPD and DXG in monkey serum and urine is described in this report. Retention times for DXG, DAPD and internal standard (2',3'-didehydro-2'deoxythymidine, D4T) were 5.0, 6.0 and 13.0 min, respectively. The extraction recovery was greater than 97% for DAPD and 94% for DXG. The limit of quantitation for UV detection was 100 ng/ml and 125 ng/ml for DXG and DAPD in monkey serum. The standard curves were linear from 0.1 microgram/ml to 5 micrograms/ml for DXG and 0.125 microgram/ml to 5 micrograms/ml for DAPD. For radiochemical detection, calibration curves of standard solutions of DAPD and DXG were linear in the range of 3500 Bq to 32,000 Bq and 7500 Bq to 60,000 Bq. The intra- and inter-day relative standard deviations were less than 7.2% using UV and less than 8.6% using on-line radiochemical detection. The HPLC method was applied to serum and urine samples collected from a male rhesus monkey that was administered 33.3 mg/kg DAPD with 200 microCi of [3H]DAPD intravenously.
（-）-β-D-2,6-二氨基嘌呤二氧戊环（DAPD）及其代谢产物二氧戊环鸟苷（DXG）在体外对乙型肝炎病毒和艾滋病毒具有强大的活性。本报告描述了一种使用紫外和在线放射化学检测的反相高效液相色谱分析方法，用于测定猴血清和尿液中的DAPD和DXG。DXG、DAPD和内标（2'，3'-二脱氢-2'-脱氧胸苷，D4T）的保留时间分别为5.0、6.0和13.0分钟。DAPD的提取回收率大于97%，DXG的提取回收率大于94%。猴血清中DXG和DAPD的紫外检测定量限分别为100 ng/ml和125 ng/ml。DXG的标准曲线在0.1微克/毫升至5微克/毫升范围内呈线性，DAPD的标准曲线在0.125微克/毫升至5微克/毫升范围内呈线性。对于放射化学检测，DAPD和DXG标准溶液的校准曲线在3500 Bq至32000 Bq以及7500 Bq至60000 Bq范围内呈线性。使用紫外检测时，日内和日间相对标准偏差小于7.2%，使用在线放射化学检测时小于8.6%。该高效液相色谱方法应用于从一只雄性恒河猴采集的血清和尿液样本，该猴静脉注射了33.3 mg/kg DAPD和200微居里的[3H]DAPD。
