*3.4. Extraction and Isolation*

After 48 days, the fermented rice substrate was mechanically fragmented and then extracted three times with 300 mL EtOAc every flask. All of the EtOAc extracts were filtered and evaporated under reduced pressure to yield a crude extract (52.3 g).

The crude extract was subjected to a silica gel vacuum liquid chromatography (VLC), eluting with different solvents of increasing polarity from petroleum ether (PE) to MeOH to yield ten fractions (Frs. 1–10) based on TLC and HPLC analysis. Fr. 5 (2.1 g) was further purified by reversed-phase column chromatography (CC) over Lobar LiChroprep RP-18 with a MeOH-H2O gradient (from 10: 90 to 100: 0) to afford four subfractions (Frs. 5.1–5.4). Fr. 5.2 was further purified by CC on Sephadex LH–20 (MeOH) and then by preparative TLC (plate: 20 × 20 cm, developing solvents: CH2Cl2/MeOH, 30: 1) to obtain **5** (2.6 mg). Fr. 5.3 was subjected to CC on silica gel eluted with CH2Cl2-MeOH (100:1 to 5:1) to obtain **1** (4.1 mg) and **2** (3.0 mg). Fr. 6 (1.7 g) was further fractionated by CC over Lobar LiChroprep RP-18 with a MeOH/H2O gradient (from 10:90 to 100:0) to yield six subfractions (Frs. 6.1–6.6). Fr. 6.1 (112.4 mg) was further purified by prep. TLC (plate: 20 × 20 cm, developing solvents: petroleum ether/acetone, 2:1) and then on Sephadex LH–20 (MeOH) to obtain **7** (3.2 mg). Fr. 6.5 was subjected to CC on silica gel eluted with CH2Cl2-MeOH (150:1 to 70:1) to obtain **4** (5.7 mg). Further purification of Fr. 7 (3.6 g) by CC over Lobar LiChroprep RP-18 with a MeOH/H2O gradient (from 10:90 to 100:0) yielded seven subfractions (Frs. 7.1–7.7). Fr. 7.1 (736.2 mg) was purified by CC on silica gel eluting with a petroleum ether-acetone gradient (from 10:1 to 2:1), and further fractionated by Sephadex LH-20 (MeOH) to afford **3** (91.6 mg). Fr. 7.2 (960.7 mg) was further separated by CC on silica gel eluting with a petroleum ether-acetone gradient (from 10:1 to 1:1) purification, to afford **6** (23.4 mg).

5*R*-Hydroxyrecifeiolide (**1**): Colourless oil; [α] 25 <sup>D</sup> +33.33 (*c* 0.09, MeOH); UV (MeOH) λmax (log ε) 205 (3.06), 220 (3.01); ECD (7.55 mM, MeOH) <sup>λ</sup>max (Δε) 233 (−0.01) nm; 1H and 13C NMR data, see Table 1; ESIMS *m*/*z* 235 [M + Na]+; (+)-HRESIMS at *m*/*z* 235.1298 [M + Na]<sup>+</sup> (calcd for C12H20O3Na, 235.1305).

5*S*-hydroxyrecifeiolide (**2**): Colourless oil; [α] 25 <sup>D</sup> +23.07 (*c* 0.13, MeOH); UV (MeOH) λmax (log ε) 205 (3.01), 220 (3.06); ECD (8.49 mM, MeOH) <sup>λ</sup>max (Δε) 230 (−0.03) nm; 1H and 13C NMR data, see Table 1; ESIMS *m*/*z* 235 [M + Na]+; (+)-HRESIMS at *m*/*z* 235.1299 [M + Na]<sup>+</sup> (calcd for C12H20O3Na, 235.1305).

*ent*-Cladospolide F (**3**): Colorless crystal (MeOH); mp 59–62 ◦C; [α] 25 <sup>D</sup> −29.41 (*c* 0.17, MeOH); UV (MeOH) λmax (log ε) 206 (3.42); ECD (7.82 mM, MeOH) λmax (Δε) 210 (+0.28) nm, 267 (+0.03) nm; 1H and 13C NMR data, see Table 2; ESIMS *m*/*z* 231 [M + H]+, *m*/*z* 253 [M + Na]+; (+)-HRESIMS at *m*/*z* 231.1589 [M + H]<sup>+</sup> (calcd for C12H23O4, *m*/*z* 231.1591), at *m*/*z* 253.1407 [M + Na]<sup>+</sup> (calcd for C12H22O4Na, *m*/*z* 253.1410).

Cladospolide G (**4**): Yellow oil; [α] 25 <sup>D</sup> –24.56 (*c* 0.57, MeOH); UV (MeOH) λmax (log ε) 206 (3.49), 220 (3.27), 275 (2.62); ECD (4.04 mM, MeOH) <sup>λ</sup>max (Δε) 207 (+0.80) nm, 323 (−0.10) nm; 1H and 13C NMR data, see Table 2; ESIMS *m*/*z* 273 [M + H]+, *m*/*z* 295 [M + Na]+; (+)-HRESIMS at *m*/*z* 273.1700 [M + H]<sup>+</sup> (calcd for C14H25O5, *m*/*z* 273.1697), at *m*/*z* 290.1970 [M + NH4] <sup>+</sup> (calcd for C14H28O5N, *m*/*z* 290.1962), at *m*/*z* 295.1515 [M + Na]<sup>+</sup> (calcd for C14H24O5Na, *m*/*z* 295.1516).

Cladospolide H (**5**): pale yellow oil; 1H and 13C NMR data, see Table 2; ESIMS *m*/*z* 233 [M + Na]+; (+)-HRESIMS at *m*/*z* 233.1151 [M + Na]<sup>+</sup> (calcd for C12H18O3Na, *m*/*z* 233.1148). (The optical rotation and ECD of **5** could not be detected due to the limited quantity).

*Iso*-cladospolide B (**6**): colorless crystal (MeOH); mp 105–112 ◦C; [α] 25 <sup>D</sup> −90.91 (*c* 0.11, MeOH); UV (MeOH) <sup>λ</sup>max (log <sup>ε</sup>); ECD (9.21 mM, MeOH) <sup>λ</sup>max (Δε) 213 (−5.69) nm; 1H and 13C NMR data, see Table 2; (+)-HRESIMS at *m*/*z* 229.1432 [M + H]<sup>+</sup> (calcd for C12H21O4, 229.1434), at *m*/*z* 246.1699 [M + NH4] <sup>+</sup> (calcd for C12H24O4N, 246.1700).
