Screening and identifying the myocardial-injury protective ingredients from Sheng-Mai-San

Figures & data

Table 1. Identification of main components in SMS-Bu by UPLC–MS/MS.

Peak no.	Retention time (min)	M	Negative ions (m/z)	Positive ions (m/z)	Identification
1	8.065	800	799[M − H]^− 845[M + HCOO]^−	603[M + H − Glc − 2H2O]^+ 441[Aglycone + H − 3H2O]^+ 423[Aglycone + H − 4H2O]^+ 405[Aglycone + H − 5H2O]^+	Ginsenoside Rf*
2	9.301	784	783[M − H]^− 829[M + HCOO]^−	749[M + H − 2H2O]^+ 441[Aglycone + H − 3H2O]^+ 423[Aglycone + H − 4H2O]^+ 405[Aglycone + H − 5H2O]^+	20(S)-Ginsenoside Rg2
3	9.495	638	637[M − H]^− 683[M + HCOO]^−	621[M + H − H2O]^+ 603[M + H − 2H2O]^+ 441[Aglycone + H − 3H2O]^+ 423[Aglycone + H − 4H2O]^+ 405[Aglycone + H − 5H2O]^+	20(S)-Ginsenoside Rh1
4	9.838	638	683[M + HCOO]^−	621[M + H − H2O]^+ 603[M + H − 2H2O]^+ 441[Aglycone + H − 3H2O]^+ 423[Aglycone + H − 4H2O]^+ 405[Aglycone + H − 5H2O]^+	20(R)-Ginsenoside Rh1
5	11.959	432		433[M + H]^+ 415[M + H − H2O]^+ 384[M + H − H2O − CH3O]^+	Schizandrol A*
6	12.880	530		531[M + H]^+ 485[M + H − CH2O2]^+ 401[M + H − C6H10O3]^+ 383[M + H − C6H10O3 − H2O]^+	Gomisin D
7	13.098	388		389 [M + H]^+ 358 [M + H − CH3O]^+	Gomisin J
8	13.315	416		417[M + H]^+ 399[M + H − H2O]^+	Schizandrol B
9	13.741	620	665[M + HCOO]^−	621[M + H]^+ 441[Aglycone + H − 3H2O]^+ 423[Aglycone + H − 4H2O]^+	Ginsenoside Rh4/Rk3
10	14.977	500		501[M + H]^+ 483[M + H − H2O]^+ 431[M + H − C5H10]^+ 401[M + H − C5H8O2]^+	Trigloylgomisin H
11	15.219	784	783[M − H]^− 829[M + HCOO]^−	785[M + H]^+ 767[M + H − H2O]^+ 749[M + H − 2H2O]^+ 587[M + H − Glc − 2H2O]^+ 443[Aglycone + H − 2H2O]^+ 425[Aglycone + H − 3H2O]^+ 407[Aglycone + H − 4H2O]^+	20(S)-Ginsenoside Rg3
12	15.434	784	783[M − H]^− 829[M + HCOO]^−	785[M + H]^+ 767[M + H − H2O]^+ 749[M + H − 2H2O]^+ 605[M + H − Glc − H2O]^+ 587[M + H − Glc − 2H2O]^+ 443[Aglycone + H − 2H2O]^+ 425[Aglycone + H − 3H2O]^+ 407[Aglycone + H − 4H2O]^+	20(R)-Ginsenoside Rg3
13	15.782	500		501[M + H]^+ 483[M + H − H2O]^+ 431[M + H − C5H10]^+ 401[M + H − C5H8O2]^+	Angeloylgomisin H
14	16.192	522		523[M + H]^+ 505[M + H − H2O]^+ 401[M + H − C7H6O2]^+	Benzoylgomisin H
15	16.345	552		553[M + H]^+ 431[M + H − C7H6O2]^+ 415[M + H − C7H6O3]^+	Benzoylgomisin Q
16	16.692	514		532 [M + NH4]^+ 487 [M + H − CO]^+ 415 [M + H−C5H8O2]^+ 371 [M + H − C5H8O2 − C2H4O]^+	Tigloylgomisin P
17	17.119	536		537 [M + H]^+ 415 [M + H − C7H6O2]^+ 371 [M + H − C7H6O2 − C2H4O]^+	Gomisin G
18	17.431	536		559 [M + Na]^+ 415 [M + H − C7H6O2]^+ 371 [M + H − C7H6O2 − C2H4O]^+ 340 [M + H − C7H6O2 − C2H4O − CH3O]^+	Schisantherin A
19	17.386	766	765[M − H]^− 811[M + HCOO]^−	767[M + H]^+ 749[M + H − H2O]^+ 731[M + H − 2H2O]^+587[M + H − Glc − H2O]^+ 443[Aglycone + H − 2H2O]^+ 425[Aglycone + H − 3H2O]^+ 407[Aglycone + H − 4H2O]^+	Ginsenoside Rk1/Rg5
20	18.035	514		515[M + H]^+ 415[M + H − C5H8O2]^+	Gomisin B
21	18.134	514		515[M + H]^+ 415[M + H − C5H8O2]^+	Gomisin E
22	18.290	514		515[M + H]^+ 415[M + H − C5H8O2]^+	Gomisin F
23	18.615	416		417[M + H]^+ 347[M + H − C5H10]^+	Schizandrin A*
24	19.197	400		401[M + H]^+ 371[M + H − CH3O]^+ 331[M + H − C5H10]^+	Schizandrin B*

Glc, glucose; M, molecular mass; SMS, Sheng-Mai-San; SMS-Bu, n-butanol extraction of SMS; UPLC-MS/MS, ultra-performance liquid chromatography tandem mass spectrometry.

*Compared with authentic compounds.

Figure 1. The influence of SMS and SMS-Bu on T-wave amplitude AUC of the myocardial injury mice. Data were expressed as mean ± SD, n = 8 in each group. *p < 0.05 versus ISO group **p < 0.01 versus ISO group ##p < 0.01 versus control group. Abbreviations: PRO, propranolol group; ISO, isoproterenol group; SMS, Sheng-Mai-San group; SMS-Bu, n-butanol extraction of SMS group; AUC, area under curve.

Figure 2. Effects of SMS and SMS-Bu on histopathological changes. (A–E) Representative light microscopic appearance of mice myocardial histopathological morphology (hematoxylin staining; original magnification ×200) for control group (A), ISO group (B), propranolol group (C), SMS group (D), SMS-Bu group (E). Abbreviations: ISO, isoproterenol; SMS, Sheng-Mai-San; SMS-Bu, n-butanol extraction of SMS.

Figure 3. Effects of SMS and SMS-Bu on serum LDH activities in ISO-induced myocardial injury mice. Data were expressed as mean ± SD, n = 8 in each group. *p < 0.05 versus ISO group **p < 0.01 versus ISO group ##p < 0.01 versus control group. Abbreviations: SMS, Sheng-Mai-San group; SMS-Bu, n-butanol extraction of SMS group; LDH, lactate dehydrogenase; ISO, isoproterenol group; PRO, propranolol group; L, low dose; M, medium dose; H, high dose.

Figure 4. Effects of SMS and SMS-Bu on serum CK activities in ISO-induced myocardial injury mice. Data were expressed as mean ± SD, n = 8 in each group. **p < 0.01 versus ISO group ##p < 0.01 versus control group. Abbreviations: SMS, Sheng-Mai-San group; SMS-Bu, n-butanol extraction of SMS group; CK, creatine kinase; ISO, isoproterenol group; PRO, propranolol group; L, low dose; M, medium dose; H, high dose.

Figure 5. Effects of SMS and SMS-Bu on the MPO activities in ISO-induced myocardial injury mice. Data were expressed as mean ± SD, n = 8 in each group. **p < 0.01 versus ISO group ##p < 0.01 versus control group. Abbreviations: SMS, Sheng-Mai-San group; SMS-Bu, n-butanol extraction of SMS group; MPO, myeloperoxidase; ISO, isoproterenol group; PRO, propranolol group; L, low dose; M, medium dose; H, high dose.

Figure 6. Effects of SMS and SMS-Bu on the MDA levels in ISO-induced myocardial injury mice. Data were expressed as mean ± SD, n = 8 in each group. **p < 0.01 versus ISO group ##p < 0.01 versus control group. Abbreviations: SMS, Sheng-Mai-San group; SMS-Bu, n-butanol extraction of SMS group; MDA, malondialdehyde; ISO, isoproterenol group; PRO, propranolol group; L, low dose; M, medium dose; H, high dose.

Figure 7. Effects of SMS and SMS-Bu on SOD activities in ISO-induced myocardial injury mice. Data were expressed as mean ± SD, n = 8 in each group. *p < 0.05 versus ISO group **p < 0.01 versus ISO group ##p < 0.01 versus control group. Abbreviations: SMS, Sheng-Mai-San group; SMS-Bu, n-butanol extraction of SMS group; SOD, superoxide dismutase; ISO, isoproterenol group; PRO, propranolol group; L, low dose; M, medium dose; H, high dose.

Figure 8. UPLC-MS/MS analysis of SMS-Bu. (A) Chromatogram at 203 nm; (B) negative ion mode MS spectra; (C) positive ion mode MS spectra. Abbreviations: SMS, Sheng-Mai-San; SMS-Bu, n-butanol extraction of SMS; UPLC-MS/MS, ultra-performance liquid chromatography tandem mass spectrometry.