Food and Environmental Protection Laboratory, FAO/IAEA Agriculture and Biotechnology Laboratories, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture, International Atomic Energy Agency, Wagramer Strasse 5, PO Box 100, A-1400 Vienna, Austria.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Sep 15;878(26):2384-90. doi: 10.1016/j.jchromb.2010.07.008. Epub 2010 Jul 22.
A new simple, sensitive and precise liquid chromatography-tandem mass spectrometry method has been developed and validated for the determination of valacyclovir-HCl and acyclovir in tsetse flies (Glossina pallipides). Tsetse flies were extracted by ultrasonication with acidified methanol/acetonitrile, centrifuged and cleaned up by solid phase dispersion using MgSO(4) and MSPD C(18) material. Samples were analysed using a Waters Alliance 2695 series HPLC with a C(18) Gemini analytical column (150 mm x 4.6 mm x 5 microm) and a guard cartridge column connected to a Waters Quattro-Micro triple-quadrupole mass spectrometer. The isocratic mobile phase consisted of methanol:acetonitrile:water (60:30:10, v/v/v) plus formic acid (0.1%) at a flow rate of 0.25 ml/min. The precursor>product ion transition for valacyclovir (m/z 325.1>152) and acyclovir (m/z 226.1>151.9) were monitored in positive electrospray multiple reaction monitoring mode. The method was validated at fortification levels of 0.5, 1 and 2 microg/g. The range of calibration for both drugs was 0.45-4.5 microg/g. The overall accuracy of the method was 92% for valacyclovir and 95% for acyclovir with corresponding within-laboratory reproducibilities of 4.4 and 3.4%, respectively. Mean recoveries were above 80% for both drugs and repeatability ranged from 0.7 to 6.1%. For both drugs the limits of detection and quantification were 0.0625 and 0.2 microg/g, respectively. The method was applied in experiments on the mass rearing of tsetse flies for sterile insect technique (SIT) applications, in which the flies were fed with blood meals containing acyclovir or valcyclovir-HCl prior to analysis to assess effects on Glossina pallidipes Salivary Gland Hypertrophy syndrome.
建立并验证了一种新的灵敏、准确、简便的液相色谱-串联质谱法，用于检测采采蝇（Glossina pallipides）中盐酸伐昔洛韦和阿昔洛韦的含量。采用酸化甲醇/乙腈对采采蝇进行超声提取，离心后通过固相分散法（MgSO4 和 MSPD C18 材料）进行净化。采用 Waters Alliance 2695 系列 HPLC 系统，使用 C18 Gemini 分析柱（150mm×4.6mm×5μm）和保护柱连接 Waters Quattro-Micro 三重四极杆质谱仪进行样品分析。等度洗脱流动相由甲醇/乙腈/水（60：30：10，v/v/v）加甲酸（0.1%）组成，流速为 0.25ml/min。采用正电喷雾多反应监测模式监测伐昔洛韦（m/z 325.1→152）和阿昔洛韦（m/z 226.1→151.9）的前体/产物离子跃迁。该方法在 0.5、1 和 2μg/g 加标水平下进行验证。两种药物的校准范围均为 0.45-4.5μg/g。该方法对伐昔洛韦的总准确度为 92%，对阿昔洛韦的总准确度为 95%，相应的实验室内重现性分别为 4.4%和 3.4%。两种药物的平均回收率均高于 80%，重复性范围为 0.7-6.1%。两种药物的检测限和定量限分别为 0.0625 和 0.2μg/g。该方法应用于采采蝇大规模饲养的无菌昆虫技术（SIT）应用实验中，在分析前，让采采蝇吸食含有阿昔洛韦或盐酸伐昔洛韦的血液餐，以评估对 Glossina pallidipes 唾液腺增生综合征的影响。
