[A novel fluorescence reagent for the analysis of trace free oestradiol and oestriol in urine by reversed-phase high performance liquid chromatography with fluorescence detection and mass spectrometry identification].
A new fluorescent derivatization reagent, 10-ethyl-9-oxo-9, 10-dihydroacridine-2-sulfonyl chloride (EASC), was synthesized. A pre-column derivatization with EASC and reversed-phase high performance liquid chromatographic (RP-HPLC) method with fluorescence (FL) detection and mass spectrometry (MS) identification was performed for the trace analysis of oestradiol (E2) and oestriol (E3) in urine. This reagent shows higher sensitivities in ultraviolet (UV), FL and MS detection than those of dansyl chloride (DNS-Cl), and the fluorescence intensity of EASC is 1000 times higher than that of DNS-Cl. The results of derivatization indic ted that the derivatives can be obtained by the labeling reaction of EASC with estradiol (E2 and estriol (E3) in the presence of NaHCO3 buffer (pH 10.5) at 60 degrees C for 3 min. The excitation wavelength (lamda ex) and emission wavelength (lamda em) were 270 nm and 430 nm, respectively. The established method exhibited excellent reproducibility and recovery. The calibration curve were linear with regression coefficients over 0.9990, and the detection limits (S/N = 3) w 40, 31 fmol for the studied compounds. The practical applicability of the method was demonstrated by analyzing trace of free oestradiol and oestriol in the urine of root voles.
Zhao
Huaixin
H
Key Laboratory of Ldfe-Organic Analysis of Shandong Province, College of Chemistry Science, Qufu Normal University, Qufu 273165, China.
Sun
Zhiwei
Z
Xia
Lian
L
Sun
Xuejun
X
Suo
Yourui
Y
Li
Yulin
Y
You
Jinmao
J
D000166:Acridines / Q000138:chemical synthesis; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004958:Estradiol / Q000652:urine*; D004964:Estriol / Q000652:urine*; D005456:Fluorescent Dyes / Q000138:chemical synthesis; D013058:Mass Spectrometry / Q000379:methods*; D051381:Rats; D013050:Spectrometry, Fluorescence / Q000379:methods; D013450:Sulfones / Q000138:chemical synthesis
C552900:10-ethyl-9-oxo-9,10-dihydroacridine-2-sulfonyl chloride; D000166:Acridines; D005456:Fluorescent Dyes; D013450:Sulfones; D004958:Estradiol; D004964:Estriol