Jullien Vincent, Tréluyer Jean-Marc, Pons Gérard, Rey Elisabeth
Service de Pharmacologie Clinique, Université René Descartes, Groupe Hospitalier Cochin-Saint-Vincent-de-Paul, Assistance Publique-Hôpitaux de Paris, 82 avenue Denfert-Rochereau, 75674 Paris Cedex 14, France.
J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Mar 5;785(2):377-81. doi: 10.1016/s1570-0232(02)00933-9.
A sensitive high-performance liquid chromatography method was developed with spectrofluorimetric detection for the determination of tenofovir, a new HIV reverse transcriptase inhibitor, in human plasma. After precipitation of 200 microl of plasma samples by methanol and evaporation of the supernatant, fluorescent derivatized compounds were obtained by a 40-min incubation at 80 degrees C with chloroacetaldehyde 0.34% at pH 4.5. The assay was performed isocratically using 5 mM Na(2)HPO(4) (pH 6), containing tetrabutylammonium (TBA) chloride 5 mM, and acetonitrile (85:15, v/v) as mobile phase, and a Cluzeau C(8) plus satisfaction column maintained at 35 degrees C. Detection was performed at excitation and emission wavelengths set at 236 and 420 nm, respectively. In these conditions, tenofovir can be separated from adefovir, the internal standard, and endogenous substances. The method was found to be linear and has been validated over a concentration range of 5-1000 microg/l. The average coefficient of the limit of quantification (5 microg/l) was 5.38% and at this concentration, a signal-to-noise ratio of 500 was measured
建立了一种灵敏的高效液相色谱法，采用荧光分光光度检测法测定人血浆中新型HIV逆转录酶抑制剂替诺福韦。取200微升血浆样品，用甲醇沉淀，上清液蒸发后，在80℃下与0.34%氯乙醛在pH 4.5条件下孵育40分钟，得到荧光衍生化化合物。采用等度洗脱，以5 mM Na2HPO4（pH 6）、含5 mM四丁基氯化铵（TBA）和乙腈（85:15，v/v）作为流动相，在35℃下使用Cluzeau C8加满意柱进行测定。分别在激发波长236 nm和发射波长420 nm处进行检测。在此条件下，替诺福韦可与内标阿德福韦及内源性物质分离。该方法呈线性，在5-1000微克/升的浓度范围内得到验证。定量限（5微克/升）的平均系数为5.38%，在此浓度下，测得信噪比为500
