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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.
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, Au
This article was published in the following journal.
Name: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
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A family of the order DIPTERA that includes the TSETSE FLIES. These flies occur only in Africa.
Bloodsucking flies of the genus Glossina, found primarily in equatorial Africa. Several species are intermediate hosts of trypanosomes.
A genus of gram-negative, rod-shaped bacteria in the family ENTEROBACTERIACEAE. They exist only as primary endosymbionts of five species of TSETSE FLIES, found in specialized organelles called mycetomes. The bacteria supply crucial B vitamins (VITAMIN B COMPLEX) which the flies require for fertility.
A species of Trypanosome hemoflagellates that is carried by tsetse flies and causes severe anemia in cattle. These parasites are also found in horses, sheep, goats, and camels.
A hemoflagellate subspecies of parasitic protozoa that causes Rhodesian sleeping sickness in humans. It is carried by Glossina pallidipes, G. morsitans and occasionally other species of game-attacking tsetse flies.
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