TY - JOUR
T1 - Rapid, quantitative high-performance liquid column chromatography of pseudouridine
AU - Kuo, Kenneth C.
AU - Gehrke, Charles W.
AU - McCune, Roy A.
AU - Waalkes, T. Phillip
AU - Borek, Ernest
N1 - Funding Information:
The work was supported in part by a contract fiorn the National Cancer Institute, Bethesda, Md. (Contract No. hi NO1 CM 12323).
PY - 1978/5/1
Y1 - 1978/5/1
N2 - A rapid, precise, and accurate chromatographic method for the determination of pseudo-uridine (ψ) in urine by high-performance liquid chromatography (HPLC) has been developed. The ribonucleosides were first isolated with an affinity gel containing immobilized phenylboronic acid. The response for ψ was linear well above and below the range necessary to determine urinary ψ. Good precision was obtained for both matrix-dependent and matrix-independent samples. Supporting experimental data are presented on precision, recovery, chromatographic methods, sample cleanup and application to the analysis of urine samples from normal males and females, and patients with advanced colon cancer. In a comparison of 40 normals with 10 colon cancer patients, 9 of the 10 patients had a ψ: creatinine (Cr) ratio greater than x̄ + 2σ for the normal population. This HPLC method is now being used extensively in our laboratory as a routine method for determination of ψ in urine from patients with various types of cancer and in chemotherapy response studies. Data are presented on the dynamics of ψ excretion by normal males and females. When the excretion of ψ was normalized with the excretion of creatinine, it was noted that samples collected at random have the same ψ: Cr ratio value as for the 24-h total collection, thus, allowing the use of random samples. The constancy of the ψ: Cr ratio implies that RNA turnover is constant and ψ excretion is independent of diet. Base values are presented for the ψ: Cr.
AB - A rapid, precise, and accurate chromatographic method for the determination of pseudo-uridine (ψ) in urine by high-performance liquid chromatography (HPLC) has been developed. The ribonucleosides were first isolated with an affinity gel containing immobilized phenylboronic acid. The response for ψ was linear well above and below the range necessary to determine urinary ψ. Good precision was obtained for both matrix-dependent and matrix-independent samples. Supporting experimental data are presented on precision, recovery, chromatographic methods, sample cleanup and application to the analysis of urine samples from normal males and females, and patients with advanced colon cancer. In a comparison of 40 normals with 10 colon cancer patients, 9 of the 10 patients had a ψ: creatinine (Cr) ratio greater than x̄ + 2σ for the normal population. This HPLC method is now being used extensively in our laboratory as a routine method for determination of ψ in urine from patients with various types of cancer and in chemotherapy response studies. Data are presented on the dynamics of ψ excretion by normal males and females. When the excretion of ψ was normalized with the excretion of creatinine, it was noted that samples collected at random have the same ψ: Cr ratio value as for the 24-h total collection, thus, allowing the use of random samples. The constancy of the ψ: Cr ratio implies that RNA turnover is constant and ψ excretion is independent of diet. Base values are presented for the ψ: Cr.
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U2 - 10.1016/S0378-4347(00)81367-5
DO - 10.1016/S0378-4347(00)81367-5
M3 - Article
C2 - 659524
AN - SCOPUS:0018193361
SN - 0378-4347
VL - 145
SP - 383
EP - 392
JO - Journal of Chromatography B: Biomedical Sciences and Applications
JF - Journal of Chromatography B: Biomedical Sciences and Applications
IS - 3
ER -