Determination of N-ethylglycine in urine of cancer patients with metastatic bone disease by HPLC using 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsulfonyl chloride as a fluorescent labeling reagent.
A simple and highly sensitive HPLC for the determination of N-ethylglycine in urine was developed. The labeling reaction of N-ethylglycine with 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsulfonyl chloride was carried out at 70 degrees C for 15 min at pH 9.0. The fluorescent derivative was separated on a reversed-phase column and detected at excitation and emission wavelengths of 320 and 400 nm, respectively. The detection limit of N-ethylglycine was 15 fmol (S/N = 3). The recovery of N-ethylglycine added to urine was 101.9%. The concentration of N-ethylglycine in urine of cancer patients with metastatic bone disease was 11.3 +/- 22.0 nmol/mg creatinine, and that of normal subject was 0.4 +/- 0.4 nmol/mg creatinine.
Tsuruta
Yasuto
Y
Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, Hiroshima 729-0292, Japan. tsuruta@fupharm.fukuyama-u.ac.jp
Matsumoto
Maya
M
Inoue
Hirofumi
H
Munemura
Sayaka
S
Yamano
Shigeru
S
Iguchi
Haruo
H
D000328:Adult; D001859:Bone Neoplasms / Q000556:secondary* / Q000652:urine*; D016022:Case-Control Studies; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008875:Middle Aged; D034442:N-substituted Glycines / Q000737:chemistry / Q000652:urine*; D010797:Phthalimides / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D055815:Young Adult; D009764:o-Phthalaldehyde / Q000737:chemistry
C117624:4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsulfonyl chloride; D005456:Fluorescent Dyes; C518321:N-ethylglycine; D034442:N-substituted Glycines; D010797:Phthalimides; D009764:o-Phthalaldehyde