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. 2023 Mar 15;9(11):eadf4166. doi: 10.1126/sciadv.adf4166

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Fig. 1. — (A) Purification scheme of hatching factors (HF). (B) Separation of HFs using preparative ODS-HPLC. Fractions were collected at 30-s intervals and tested for hatching against G. rostochiensis eggs. Fractions were first tested in groups; then, each of the fractions collected at 22 to 24 and 28 to 30 min was tested alone for HS activity. Hatching rate (%) is shown at two dilutions. (C) Solanoeclepin A (SEA) detection from fraction Fr-B1, which was obtained by further purification of Fr-B based on HS activity. A multiple reaction monitoring (MRM) transition of m/z 499 > 399 was selected to detect SEA. (D) Ultraperformance liquid chromatography (UPLC) fractionation of Fr-A1, which was obtained by further purification of Fr-A based on HS activity. The HS activity of each fraction on G. rostochiensis eggs was investigated at two different dilutions. The values indicate the means of two biological replicates; white dots are individual measurements. (E) Chemical structures of SEA and solanoeclepin B (SEB).