The concentration of monosodium glutamate (MSG) was evaluated in ten samples of formulated spices in Pakistan. The samples were analyzed by modified HPLC method after water extraction and derivatization of MSG by dinitrofluorobenzene (DNFB), using reversed phase C18 column, mobile phase was consisted of methanol: water (1:1) followed by UV detection at 254 nm. The standard curve of derivatized MSG of 500, 250 and 125 mg/mL was plotted to determine the quantity of MSG in spices samples. The peak of MSG was identified by comparing it with retention time of MSG standards, that is, 8.6 min. MSG concentration in different samples was in the range of 2.6-7.7%, showing a wide range of added MSG. Glutamate contents in the samples were in the permissible limit established by the European Directive, 95/2/CE. Furthermore, MSG levels in food prepared by the formulated Pakistani spices samples are also less than the largest palatable dose of MSG for humans.