Akademik Veri Yönetim Sistemi
Kromatografi XX. Uluslararası Katılımlı, Ankara, Türkiye, 24 - 25 Şubat 2022
One of the widely known side effects of nifedipine and
phenytoin is gingival overgrowth. The aim of this study is method development
for determination of nifedipine and phenytoin concentration in gingival
crevicular fluid (GCF) and plasma by HPLC. And to develop analysis methods to
determine whether there is a relationship between plasma/GCF levels and
nifedipine, phenytoin levels. Separation
of nifedipine from GCF was performed by Microsphere, C18 (100 x 4.6 mm, particle size 3 mm) analytical column and methanol, sodium acetate
(pH=4.0, 10 mM) (60:40, v/v) containing mobile phase at 0.8 ml/min. Detection
of nifedipine and nitrendipine (internal standard, 0.5 mg/ml) was performed by UV/Vis detector at 235 nm. GCF
samples were extracted by using a mixture of methanol and water (50:50, v/v).
Plasma nifedipine content were extracted by using a mixture of hexane and
dichloromethane. The calibration curve for nifedipine was linear over
the concentration range of 0.01-0.5 mg/ml. The mean
recovery (±SD) from GCF was
99.05±3.72 % for nifedipine at a concentration of 0.1 mg/ml (n=6). The mean recovery (±SD) from plasma was
102.03±5.62 % for nifedipine at a concentration of 0.1 mg/ml (n=6).
Separation of phenytoin from GCF was performed by
XTerra, C18 (250 x 4.6 mm, particle size 5 mm) analytical column and acetonitrile, K2HPO4 (pH=5.0, 20 mM) (30:70, v/v) containing mobile
phase at 1.2 ml/min. Phenytoin and mephenytoin (as an internal standard, 5 mg/ml) were detected by UV/Vis detector at 240 nm. GCF samples were extracted by using a mixture of
acetonitrile and water (50:50, v/v). Plasma samples were extracted by using
acetonitrile. The calibration curve for phenytoin was linear over
the concentration range of 0.02-5 mg/ml. The mean
recovery (±SD) from GCF was
101.15±4.12 % for phenytoin at a concentration of 1 mg/ml (n=6). The mean recovery (±SD) from plasma was
105.22±3.17 % for phenytoin at a concentration of 1 mg/ml (n=6).
Consequently, this study describes a simple, sensitive, and practical
HPLC-UV/Vis method which permits determination of nifedipine and phenytoin in human gingival
crevicular fluid and plasma samples.