Fig. 1. Structures of some triterpene glycosides and aromatic compounds of Cimicifuga species. 1–17 reference standards. 18–26 compounds identified by MS analysis.

Fig. 2. Typical UV spectra of aromatic Cimicifuga compounds, obtained on-line in mobile phase as part of the HPLC fingerprint of Cimicifuga species. (A) Isoferulic acid (17); (B) Cimicifugic acid B (11); and (C) Cimifugin (12).

Fig. 3. Comparison of HPLC–UV chromatograms of aromatic constituents (solvent system I). Because all major aromatic compounds elute before 25 min, only the 0–25 min windows are shown: (A) C. racemosa; (B) C. americana; (C) C. rubifolia; (D) C. acerina; (E) C. biternat; (F) C. dahurica; (G) C. foetida-1; (H) C. foetida-2; (I) C. heracleifolia; (J) C. japonica; (K) C. simplex; (L) Remifemin; (M) CimiPure; (N) Product A; and (O) Product B.

Fig. 4. TIC and mass spectrum of reference standards obtained by hyphenated LC–MS separation: (A) TIC; (B) 1 Actein; (C) 2 23-epi-26-Deoxyactein; (D) 3 Cimigenol3-O-arabinoside; (E) 5 25-O-Acetylcimigenol xyloside; (F) 6 Acetylshengmanol3-O-xyloside; (G) 7 24-O-Acetyl hydroshengmanol3-O-xyloside; (H) 8 Cimiracemoside A; and (I) 9 Cimicifugoside H-1. The mass spectrum of 4 is not displayed as it is almost identical to that of 3; xyl: xylose, ara: arabinose.

Fig. 5. Comparison of the total ion current (TIC) traces of Cimicifuga species (solvent system II): (A) C. racemosa; (B) C. americana; (C) C. rubifolia; (D) C. acerina; (E) C. biternat; (F) C. dahurica; (G) C. foetida-1; (H) C. foetida-2; (I) C. heracleifolia; (J) C. japonica; (K) C. simplex-1; and (L) C. simplex-2 (see Section 2 for botanical details).

Fig. 6. Comparison of the TIC traces of C. racemosa fingerprints demonstrating the conversion of triterpene glycosides under heat treatment: (A) fresh 75% ethanol extract; (B) 75% ethanol extract (liquid) settled for more than 24 months; (C) extract heated to 90–95 °C for 3 h; and (D) extract heated to 90–95 °C for 48 h.

Fig. 7. Hypothetic conversion of triterpenes: acetyl-shengmanol type triterpenes are first converted into hydroshengmanol and subsequently into cimigenol type triterpenes [3] and [37].

Fig. 8. Comparison of selected ion chromatograms (SIC) at m/z 307 for cimifugin (12) and aglycone fragment of cimifugin glucoside (18): (A) C. racemosa; (B) C. americana; (C) C. rubifolia; (D) C. acerina; (E) C. biternat; (F) C. dahurica; (G) C. foetida; (H) C. heracleifolia; (I) C. japonica; (J) C. simplex; (K) Remifemin; (L) CimiPure; and (M) Product B.

Fig. 9. Comparison of selected ion chromatograms (SIC) at m/z 621 for cimigenol arabinoside (3): (A) fresh extract of C. racemosa; (B) heat treated C. racemosa; (C) C. americana; (D) C. rubifolia; (E) C. acerina; (F) C. biternat; (G) C. dahurica; (H) C. foetida; (I) C. heracleifolia; (J) C. japonica; (K) C. simplex; (L) Remifemin; (M) CimiPure; and (N) Product B.

Fig. 10. Comparison TIC fingerprints of commercial black cohosh product: (A) Remifemin; (B) CimiPure; (C) Product A; and (D) Product B.

Fig. 11. HPLC–ELSD fingerprint chromatograms of the Cimicifuga triterpene glycosides (INA method, solvent system II): (A) C. racemosa; (B) C. americana; (C) C. rubifolia; (D) C. acerina; (E) C. biternat; (F) C. dahurica; (G) C. foetida-1; (H) C. foetida-2; (I) C. heracleifolia; (J) C. japonica; (K) C. simplex-1; (L) C. simplex-2; (M) Remifemin; (N) CimiPure; and (O) Product B.

Summary of the phenolic and triterpene glycoside constituents identified from the fingerprints of the 10 different Cimicifuga species