In the present study, the characterization and comparison of the pungent components in commercial Z. bungeanum oils and Z. schinifolium oils were investigated. By high-performance liquid chromatography (HPLC)-mass spectrometry (MS/MS) analysis, the major alkylamides in Z. bungeanum and Z. schinifolium oils were identified as hydroxy-ε-sanshool, hydroxy-α-sanshool, hydroxy-β-sanshool, hydroxy-γ-sanshool, hydroxy-γ-isosanshool, bungeanool, isobungeanool, and tetrahydrobungeanool, respectively. Hydroxy-α-sanshool was found to be the most abundant alkylamide in all oils. The levels of hydroxy-ε-sanshool and hydroxy-β-sanshool in Z. bungeanum oils were comparable to that in Z. schinifolium oils, whereas Z. bungeanum oils contained significantly (P less than 0.05) higher levels of hydroxy-γ-isosanshool, bungeanool, isobungeanool, and tetrahydrobungeanool, compared with Z. schinifolium oils. Furthermore, principal component analysis (PCA) indicated that Z. bungeanum oil and Z. schinifolium oil were clearly classified by HPLC fingerprinting profiles and concentrations of alkylamides. In addition, the results of PCA suggested that alkylamides, such as hydroxy-γ-sanshool and bungeanool, could be potential markers to distinguish Z. bungeanum oil and Z. schinifolium oil. The results from this study could be used to discriminate the different flavor characterization and control the quality of commercial Z. bungeanum oil and Z. schinifolium oil.

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Current tools for detecting transgenic crops, such as polymerase chain reaction (PCR), require professional equipment and complex operation. Herein, we introduce a clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system to analyze transgenes by designing an isothermal amplification to serve as the amplified reporter, allowing an isothermal and label-free detection of transgenic crops. The use of Cas12a allowed direct and specific recognition of transgenes. To enhance the sensitivity of the assay, we used rolling circle amplification (RCA) to monitor the recognition of transgenes by designing the RCA primer as the cleavage substrate of Cas12a. The presence of transgenes can be detected by monitoring the G-quadruplex in RCA amplicon using a G-quadruplex binding dye, N-methyl mesoporphyrin IX (NMM). We termed the assay as isoCRISPR and showed that the assay allowed distinguishing transgenic corn cultivars (“Bt11” and “MON89034”) from nontransgenic corn cultivars (“yellow”, “shenyu”, “xianyu”, and “jingke”). The isoCRISPR assay will enrich the toolbox for transgenic crop identification and broaden the application of CRISPR/Cas in food authenticity and safety.

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