Quantitative Analysis Of Volatile Bio-markers In Urine Of B Cell Non-hodgkin’s Lymphoma Patients And The Establishment Of The Diagnostic Function Models

Objective: To establish the technology platform of urine VOCs(volatile organic compounds), select volatile markers for B cell non-Hodgkin’s lymphoma(B-NHL) patients and assess the clinical value of the markers. Finally, to establish the diagnostic function models of B-NHL.Methods: We established a technology platform of urine VOCs test by testing the samples of a healthy person using headspace gas solid phase micro-extraction(SPME)-gas chromatography(GC)- mass spectrometry(MS). We collected urine samples from 35 patients with aggressive B-NHL, samples from 33 patients with indolent B-NHL, samples from 33 patients with benign lymphoma enlargement patients and 30 healthy volunteers were collected as controls. The samples were detected with the technology platform and the potential volatile markers were selected by using(partial least squares discriminant analysis, PLS-DA). Standard curves were used to quantitate the levels of urine VOCs. Wilcoxon/Kruskal–Wallis Test was used to compare the differences between groups. The volatile markers which were usesd to diagnose B-NHL and monitor the transformation of indolent B-NHL were assessed with receiver operator characteristic(ROC) curves. The diagnostic functions were established with Fisher discriminant analysis, and their diagnostic values of the functions were assessed with a leave-one-out classification procedure.Results: The technology of SPEM-GC/MS equal to the specification of breath test. There are 3 VOCs which have significant difference among the groups. They are hexane, 1-butanone and 4-heptanone. The levels of hexane in B-NHL patients’ urine were lower than non-lymphoma persons’. On the contrary, the levels of the latter 2 VOCs werehigher in B-NHL patients’ urine. Comparing the urine of B-NHL patients with indolent patients, the levels of 4-heptanone and 3-Nonen-2-one were significantly different between aggressive B-NHL patients and indolent B-NHL patients and they were higher in the former patients. The best volatile bio-marker of B-NHL was 4-heptanone, with a sensitivity of 91.8%and a specificity of82.3% between B-NHL patients and non-lymphoma persons and with a sensitivity of 92.7% and a specificity of 85.0% between aggressive B-NHL patients and indolent B-NHL patients. There had no significant difference between patients at stage I-II or stage III-IV(P > 0.05). The diagnostic function model established with hexane, 1-butanone and 4-heptanone had a sensitivity of 86.8% and a specificity of 92.1%. The leave-one-out classification procedure of this model had a sensitivity of 83.8% and a specificity of 88.9%. 4-heptanone and 3-Nonen-2-one were used to establish the second diagnostic function model for the transformation of indolent B-NHL, which had a sensitivity of 85.7% and a specificity of 93.9% and its leave-one-out classification procedure had a sensitivity of 82.9% and a specificity of 84.8%.Conclusions: We established the technology platform for urine VOCs testing by SPEM-GC/MS. hexane, 1-butanone, 4-heptanone and 3-Nonen-2-one selected as volatile markers of B-NHL patients could be used to early diagnosis for B-NHL and monitor the transformation of indolent B-NHL. The diagnostic function models had high sensitivity and specificity for B-NHL diagnosis and monitoring the transformation of indolent B-NHL which deserve further study.