Quantitative Analysis Of Volatile Markers In Breath Of Hepatocellular Carcinoma Patients And The Establishment Of The Breath Diagnostic Function Models Of The Cancer

Objective To establish the methodology of breath test, select volatile markers for hepatocellular carcinoma (HCC) patients and assess the clinical value of the markers. Thereby, to investigate the levels of the markers in patients with different clinical stages and different levels of AFP. Finally, to establish the breath diagnostic function models of HCC.Methods We established the methodology of breath test by testing the samples of a healthy person using solid phase microextraction (SPME) /gas chromatography (GC) /mas spectrometry (MS). We collected breath samples from 30 patients with HCC using Tedlar? gas bags, samples from 27 patients with liver cirrhosis comorbid with hepatitis B and 36 healthy people were collected as controls, in the meantime, we collected ambient air for reference. The samples were detected with the technology and the volatile markers of HCC were selected by comparing the levels of VOCs in the breath of healthy people and HCC patients. Standard curvers were used to quantitate the levels of VOCs. Mann-Whitney U Test was used to compare between groups. The markers which were usesd to diagnose HCC were assessed with receiver operator characteristic (ROC) curves. The diagnostic functions of HCC were established with Fisher discriminant analysis, and the diagnostic abilities of the functions were assessed with a leave-one-out classification procedure or by testing the training and validation sets.Results The technology of SPEM/GC/MS was suitable for breath test. The levels of six VOCs in HCC patients'breath, including 3-hydroxy-2-butanone, toluene, ethylbenzene, xylene, styrene and decane, were of significant difference (P<0.05) compared to healthy persons'. 3-Hydroxy-2-butanone, styrene and decane might be the markers of HCC, whose levels in HCC patients'breath were higher than those in healthy persons'and ambient air. The optimal volatile marker of HCC was 3-hydroxy-2-butanone, with a sensitivity of 83.3% and a specificity of 91.7% between HCC patients and nomal controls and with a sensitivity of 70.0% and a specificity of 70.4% between HCC patients and liver cirrhosis patients, whose level was gradually escalating among healthy people, cirrhosis patients and HCC patients. It had no significant differences neither between the levels of styrene in the breath of healthy people or liver cirrhosis patients (P>0.05) nor between the levels of decane in the breath of liver cirrhosis or HCC patients (P>0.05). It showed no statistical significance (P>0.05) for the concentration differences of these markers neither between HCC patients with AFP>400μg/L or AFP<400μg/L nor between patients at stageⅠ~Ⅱor stageⅢ~Ⅳ. The two-classification diagnostic function models established with these markers could have a sensitivity of 86.7% and a specificity of 91.7% for HCC diagnosis.Conclusions We established the methodology for breath testing with SPEM/GC/MS. 3-Hydroxy-2-butanone, styrene and decane selected as volatile markers of HCC patients can be used to early diagnosis, for whose levels have no correlation between different clinical stages or different levels of AFP in HCC patients. The two-classification diagnostic function models have high sensitivity and specificity for HCC diagnosis.