Analysis of the antiviral drugs acyclovir and valacyclovir-hydrochloride in tsetse flies (Glossina pallidipes) using LC-MSMS.
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.
Sasanya
J J
JJ
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.
Abd-Alla
A M M
AM
Parker
A G
AG
Cannavan
A
A
D000212:Acyclovir / Q000031:analogs & derivatives* / Q000032:analysis*; D000824:Animal Nutritional Physiological Phenomena; D000818:Animals; D000998:Antiviral Agents / Q000032:analysis*; D002853:Chromatography, Liquid / Q000379:methods*; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014370:Tsetse Flies / Q000737:chemistry*; D000077483:Valacyclovir; D014633:Valine / Q000031:analogs & derivatives* / Q000032:analysis
D000998:Antiviral Agents; D014633:Valine; D000077483:Valacyclovir; D000212:Acyclovir