[Determination of nitrite and nitrate in rat nervi ischiadicus by high performance liquid chromatography]

To study the effect of nitric oxide on the diabetic neuropathy, a quantitative method for the determination of nitrite(NO2-) and nitrate(NO3-) in rat nervi ischiadicus by high performance liquid chromatography(HPLC) with UV-VIS detection was developed. In the range of 1 mumol/L-25 mumol/L, correlation coefficient between concentration and peak area for NO2- and NO3- was more than 0.991. The minimum detection limits for both ions were 0.2 mumol/L and 0.5 mumol/L, respectively. Relative standard deviation(RSD) of inter-day and intra-day was less than 14%. The effects of different acetonitrile concentration and pH of mobile phase on separation as well as the stability of both salty ions were studied. In comparison with normal group, the levels of NO2- and NO3- for diabetic groups were lower. Measured mean values were, in turn, as follows: normal rats, (2.62 +/- 0.38) mumol/L and (24.43 +/- 5.61) mumol/L (n = 8); diabetic-controlled rats (saline-given only), (2.24 +/- 0.20) mumol/L and (17.15 +/- 7.79) mumol/L (n = 6); diabetic rats treated with bovine colostrum insulin-like growth factor(BctIGF-1) or with recombinant human insulin-like growth factor (rhIGF-1), (2.55 +/- 0.36) mumol/L and (6.12 +/- 2.14) mumol/L (n = 5) and (1.46 +/- 0.08) mumol/L and (9.95 +/- 3.34) mumol/L (n = 7), respectively.