Determination of medroxyprogesterone acetate in serum by HPLC with peroxyoxalate chemiluminescence detection using a fluorogenic reagent, 4-(N,N-dimethylaminosulphonyl)-7-hydrazino-2,1,3-benzoxadiazole.
The high-performance liquid chromatographic determination of medroxyprogesterone acetate (MPA) with peroxyoxalate chemiluminescence (PO-CL) detection is described. The spiked serum containing MPA was extracted on Bond-Elut C18 columns and derivatized with 4-(N,N-dimethylaminosulphonyl)-7-hydrazino-2,1,3-benzoxadiazole (DBD-H). The hydrazone of MPA with DBD-H was confirmed to be a mono-DBD-derivative. The reaction mixture was separated by direct injection onto a C18 analytical column, and quantified by PO-CL detection. The linear range of the standard curve, in serum, was 15.6-96.6 ng ml-1 with a detection limit of 9 ng ml-1 using only 100 microliters of serum, while the detection limit of standard MPA derivatized with DBD-H was 8.7 fmol per injection. The relative standard deviation of the method was 7.4% at 19.3 ng and 1.7% at 77.3 ng ml-1.
Uzu
S
S
Branch Hospital Pharmacy, University of Tokyo, Japan.
Imai
K
K
Nakashima
K
K
Akiyama
S
S
D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes; D006801:Humans; D008163:Luminescent Measurements; D017258:Medroxyprogesterone Acetate / Q000097:blood*; D010069:Oxadiazoles; D010070:Oxalates / Q000737:chemistry; D011485:Protein Binding; D013449:Sulfonamides
D005456:Fluorescent Dyes; D010069:Oxadiazoles; D010070:Oxalates; D013449:Sulfonamides; C080612:4-(N,N-dimethylaminosulfonyl)-7-hydrazino-2,1,3-benzoxadiazole; C044622:peroxyoxalate; D017258:Medroxyprogesterone Acetate