A small‐scale neutral alumina column chromatography method for carbon isotope determination of hopanes in crude oils or rock extracts.

• Using neutral alumina as a stationary phase and disposable glassware such as Pasteur pipettes can separate hopanes successfully. • The sterane content of the collected hopanes decreased from the original 12% to 0.84%, compared to urea complexation. • Neutral alumina column chromatography meets the needs of experimenters for smaller and simpler devices. • The separated hopanes can meet the requirements of stable carbon isotopes of monomer hydrocarbons detection. This paper presents a small-scale column chromatography method for separating hopanes in crude oil or rock extracts using neutral alumina as a solid phase adsorbent and a Pasteur pipette as a separation device. Three oil samples were selected to study the effect of solid phase adsorbent type and column length on the separation of hopanes. The oil samples were eluted with mixed reagents (V hexane : V petroleum ether = 8:2) and collected at intervals of 0.5 ml. Ten Fractions were collected and tested for the compounds using GC–MS. A quantitative approach was used to reveal distribution characteristics of compounds in each eluted Fraction. Experimental results showed that 100–200 um neutral alumina exhibited significant differences in the adsorptive capacity of biomarkers from oil samples and rock extracts. The elution order of the biomarkers in the chromatographic column (the length is 180 mm) was n-alkanes, steranes and hopanes. The separation of hopanes could be realized by collecting the eluted Fractions 4 and 5. Compared with the urea complexation, the purity of hopanes separated by column chromatography was higher. The concentration of n-alkanes (nC 16 - nC 34) could be reduced from 1.99 to 4.83 mg/ml to 0.79–0.94 mg/ml, and the content of steranes can be reduced from the original 12% to 0.45%. Residual n-alkanes and steranes were not visible in the GC–MS detection. The Total Ion Chromatography (TIC) of hopanes was consistent with the distribution characteristics of the m/z 191 mass chromatogram. The isolated hopanes could meet the detection requirements of isotope ratio mass spectrometry. The C 29 Ts / C 29 αβ ratio of hopanes decreased gradually from 1.63 to 0.73 as the column length increased. It is speculated that the variation of the C 29 Ts/C 29 αβ ratio is not only affected by maturity but also by the oil and gas migration. This method is a new attempt in the field of compound purification and can be widely used in the study of stable carbon isotopes of hopanes monomeric hydrocarbons. [ABSTRACT FROM AUTHOR]