Development of a HPLC Method for the Estimation of Metronidazole in Human Plasma

Abstract

Varying amount of metronidazole (200 to 5000 ng) and fixed
quantity (120 ng) of nevirapine (internal standard) was added to the
blank human plasma (0.5 mL), and extracted with diethyl ether. The
mixture was centrifuged and diethyl ether layer separated, dried and
reconstituted with mobile phase. Fifty microlitres of this solution
were injected into a reverse phase C-18 column using a mobile
phase consisting of acetonitrile and 0.01 M potassium dihydrogen
phosphate buffer (pH adjusted to 3.0 with 5% orthophosphoric acid)
in the ratio of 20 : 80% v/v and the eluents were monitored at
318 nm. The method was validated for its linearity, precision and
accuracy. The method is simple, precise, specific, less time consuming
and accurate for the estimation of metronidazole in plasma
samples.