药学学报

Component	Precursor ion (m/z)	Product ion (m/z)	Dwell time/s	Fragmentor	Collision energy/eV	Polarity	Retention time/min
Magnoflorine	342.1	297.1	30	166	20	[M]⁺	4.62
Epiberberine	336.1	320.0	30	166	36	[M]⁺	6.48
Coptisine	320.1	292.1	30	166	32	[M]⁺	6.52
Jatrorrhizine	338.1	322.1	30	166	36	[M]⁺	6.52
Berberrubine	322.1	307.1	30	166	32	[M+H]⁺	6.76
Palmatine	352.1	336.1	30	166	36	[M]⁺	7.03
Berberine	336.1	320.1	30	166	36	[M]⁺	7.12
Ononin	431.1	269.1	30	166	17	[M+H]⁺	6.80
Calycosin	285.0	270.0	30	166	24	[M+H]⁺	7.21
Calycosin-7-glucoside	477.0	285.0	30	166	20	[M+H]⁺	5.45
Formononetin	269.0	197.0	100	166	48	[M+H]⁺	8.57
Astragaloside Ⅳ	807.5	807.5	100	166	0	[M+Na]⁺	8.43
Chlorogenic acid	355.1	162.9	100	166	10	[M+H]⁺	3.19
Neochlorogenic acid	355.1	162.9	100	166	10	[M+H]⁺	2.34
Cryptochlorogenic acid	355.1	162.9	100	166	10	[M+H]⁺	3.44
Carbamazepine	237.0	194.0	30	166	21	[M+H]⁺	7.88

Component

Precursor ion (m/z)

Product ion (m/z)

Dwell time/s

Fragmentor

Collision energy/eV

Polarity

Retention time/min

Magnoflorine

342.1

297.1

166

[M]⁺

4.62

Epiberberine

336.1

320.0

166

[M]⁺

6.48

Coptisine

320.1

292.1

166

[M]⁺

6.52

Jatrorrhizine

338.1

322.1

166

[M]⁺

6.52

Berberrubine

322.1

307.1

166

[M+H]⁺

6.76

Palmatine

352.1

336.1

166

[M]⁺

7.03

Berberine

336.1

320.1

166

[M]⁺

7.12

Ononin

431.1

269.1

166

[M+H]⁺

6.80

Calycosin

285.0

270.0

166

[M+H]⁺

7.21

Calycosin-7-glucoside

477.0

285.0

166

[M+H]⁺

5.45

Formononetin

269.0

197.0

100

166

[M+H]⁺

8.57

Astragaloside Ⅳ

807.5

100

166

[M+Na]⁺

8.43

Chlorogenic acid

355.1

162.9

100

166

[M+H]⁺

3.19

Neochlorogenic acid

355.1

162.9

100

166

[M+H]⁺

2.34

Cryptochlorogenic acid

355.1

162.9

100

166

[M+H]⁺

3.44

Carbamazepine

237.0

194.0

166

[M+H]⁺

7.88

Component	Precursor ion (m/z)	Product ion (m/z)	Dwell time/s	Fragmentor	Collision energy/eV	Polarity	Retention time/min
Magnoflorine	342.1	297.1	30	166	20	[M]⁺	4.62
Epiberberine	336.1	320.0	30	166	36	[M]⁺	6.48
Coptisine	320.1	292.1	30	166	32	[M]⁺	6.52
Jatrorrhizine	338.1	322.1	30	166	36	[M]⁺	6.52
Berberrubine	322.1	307.1	30	166	32	[M+H]⁺	6.76
Palmatine	352.1	336.1	30	166	36	[M]⁺	7.03
Berberine	336.1	320.1	30	166	36	[M]⁺	7.12
Ononin	431.1	269.1	30	166	17	[M+H]⁺	6.80
Calycosin	285.0	270.0	30	166	24	[M+H]⁺	7.21
Calycosin-7-glucoside	477.0	285.0	30	166	20	[M+H]⁺	5.45
Formononetin	269.0	197.0	100	166	48	[M+H]⁺	8.57
Astragaloside Ⅳ	807.5	807.5	100	166	0	[M+Na]⁺	8.43
Chlorogenic acid	355.1	162.9	100	166	10	[M+H]⁺	3.19
Neochlorogenic acid	355.1	162.9	100	166	10	[M+H]⁺	2.34
Cryptochlorogenic acid	355.1	162.9	100	166	10	[M+H]⁺	3.44
Carbamazepine	237.0	194.0	30	166	21	[M+H]⁺	7.88

Component

Precursor ion (m/z)

Product ion (m/z)

Dwell time/s

Fragmentor

Collision energy/eV

Polarity

Retention time/min

Magnoflorine

342.1

297.1

166

[M]⁺

4.62

Epiberberine

336.1

320.0

166

[M]⁺

6.48

Coptisine

320.1

292.1

166

[M]⁺

6.52

Jatrorrhizine

338.1

322.1

166

[M]⁺

6.52

Berberrubine

322.1

307.1

166

[M+H]⁺

6.76

Palmatine

352.1

336.1

166

[M]⁺

7.03

Berberine

336.1

320.1

166

[M]⁺

7.12

Ononin

431.1

269.1

166

[M+H]⁺

6.80

Calycosin

285.0

270.0

166

[M+H]⁺

7.21

Calycosin-7-glucoside

477.0

285.0

166

[M+H]⁺

5.45

Formononetin

269.0

197.0

100

166

[M+H]⁺

8.57

Astragaloside Ⅳ

807.5

100

166

[M+Na]⁺

8.43

Chlorogenic acid

355.1

162.9

100

166

[M+H]⁺

3.19

Neochlorogenic acid

355.1

162.9

100

166

[M+H]⁺

2.34

Cryptochlorogenic acid

355.1

162.9

100

166

[M+H]⁺

3.44

Carbamazepine

237.0

194.0

166

[M+H]⁺

7.88

Component	Regression equation	r	Linearity range/ng·mL^-1	LLOQ/ng·mL^-1
Magnoflorine	Y = 0.02X + 0.000 66	0.998 8	0.8-80	0.8
Epiberberine	Y = 0.16X + 0.012	0.997 5	0.8-80	0.8
Coptisine	Y = 0.05X + 0.003 4	0.997 6	0.8-80	0.8
Jatrorrhizine	Y = 0.05X + 0.003 5	0.996 5	0.4-40	0.4
Berberrubine	Y = 0.05X + 0.000 15	0.998 7	0.8-80	0.8
Palmatine	Y = 0.11X + 0.006 6	0.995 7	0.8-80	0.8
Berberine	Y = 0.11X - 0.002 8	0.998 0	1.6-160	1.6
Ononin	Y = 0.22X - 0.000 89	0.999 8	0.5-50	0.5
Calycosin	Y = 0.10X - 0.000 70	0.999 8	0.5-50	0.5
Calycosin-7-glucoside	Y = 0.15X - 0.001 6	0.999 8	0.5-50	0.5
Formononetin	Y = 0.24X + 0.005 4	0.998 7	0.5-50	0.5
Astragaloside Ⅳ	Y = 0.003 3X + 0.000 016	0.998 6	10-1 000	10.0
Chlorogenic acid	Y = 0.003 1X + 0.000 095	0.998 3	4-400	4.0
Neochlorogenic acid	Y = 0.000 87X - 0.000 029	0.997 1	5-500	5.0
Cryptochlorogenic acid	Y = 0.000 49X + 0.000 000 65	0.995 5	5-500	5.0

Component

Regression equation

Linearity range/ng·mL^-1

LLOQ/ng·mL^-1

Magnoflorine

Y = 0.02X + 0.000 66

0.998 8

0.8-80

0.8

Epiberberine

Y = 0.16X + 0.012

0.997 5

0.8-80

0.8

Coptisine

Y = 0.05X + 0.003 4

0.997 6

0.8-80

0.8

Jatrorrhizine

Y = 0.05X + 0.003 5

0.996 5

0.4-40

0.4

Berberrubine

Y = 0.05X + 0.000 15

0.998 7

0.8-80

0.8

Palmatine

Y = 0.11X + 0.006 6

0.995 7

0.8-80

0.8

Berberine

Y = 0.11X - 0.002 8

0.998 0

1.6-160

1.6

Ononin

Y = 0.22X - 0.000 89

0.999 8

0.5-50

0.5

Calycosin

Y = 0.10X - 0.000 70

0.999 8

0.5-50

0.5

Calycosin-7-glucoside

Y = 0.15X - 0.001 6

0.999 8

0.5-50

0.5

Formononetin

Y = 0.24X + 0.005 4

0.998 7

0.5-50

0.5

Astragaloside Ⅳ

Y = 0.003 3X + 0.000 016

0.998 6

10-1 000

10.0

Chlorogenic acid

Y = 0.003 1X + 0.000 095

0.998 3

4-400

4.0

Neochlorogenic acid

Y = 0.000 87X - 0.000 029

0.997 1

5-500

5.0

Cryptochlorogenic acid

Y = 0.000 49X + 0.000 000 65

0.995 5

5-500

5.0

Component	Regression equation	r	Linearity range/ng·mL^-1	LLOQ/ng·mL^-1
Magnoflorine	Y = 0.02X + 0.000 66	0.998 8	0.8-80	0.8
Epiberberine	Y = 0.16X + 0.012	0.997 5	0.8-80	0.8
Coptisine	Y = 0.05X + 0.003 4	0.997 6	0.8-80	0.8
Jatrorrhizine	Y = 0.05X + 0.003 5	0.996 5	0.4-40	0.4
Berberrubine	Y = 0.05X + 0.000 15	0.998 7	0.8-80	0.8
Palmatine	Y = 0.11X + 0.006 6	0.995 7	0.8-80	0.8
Berberine	Y = 0.11X - 0.002 8	0.998 0	1.6-160	1.6
Ononin	Y = 0.22X - 0.000 89	0.999 8	0.5-50	0.5
Calycosin	Y = 0.10X - 0.000 70	0.999 8	0.5-50	0.5
Calycosin-7-glucoside	Y = 0.15X - 0.001 6	0.999 8	0.5-50	0.5
Formononetin	Y = 0.24X + 0.005 4	0.998 7	0.5-50	0.5
Astragaloside Ⅳ	Y = 0.003 3X + 0.000 016	0.998 6	10-1 000	10.0
Chlorogenic acid	Y = 0.003 1X + 0.000 095	0.998 3	4-400	4.0
Neochlorogenic acid	Y = 0.000 87X - 0.000 029	0.997 1	5-500	5.0
Cryptochlorogenic acid	Y = 0.000 49X + 0.000 000 65	0.995 5	5-500	5.0

Component

Regression equation

Linearity range/ng·mL^-1

LLOQ/ng·mL^-1

Magnoflorine

Y = 0.02X + 0.000 66

0.998 8

0.8-80

0.8

Epiberberine

Y = 0.16X + 0.012

0.997 5

0.8-80

0.8

Coptisine

Y = 0.05X + 0.003 4

0.997 6

0.8-80

0.8

Jatrorrhizine

Y = 0.05X + 0.003 5

0.996 5

0.4-40

0.4

Berberrubine

Y = 0.05X + 0.000 15

0.998 7

0.8-80

0.8

Palmatine

Y = 0.11X + 0.006 6

0.995 7

0.8-80

0.8

Berberine

Y = 0.11X - 0.002 8

0.998 0

1.6-160

1.6

Ononin

Y = 0.22X - 0.000 89

0.999 8

0.5-50

0.5

Calycosin

Y = 0.10X - 0.000 70

0.999 8

0.5-50

0.5

Calycosin-7-glucoside

Y = 0.15X - 0.001 6

0.999 8

0.5-50

0.5

Formononetin

Y = 0.24X + 0.005 4

0.998 7

0.5-50

0.5

Astragaloside Ⅳ

Y = 0.003 3X + 0.000 016

0.998 6

10-1 000

10.0

Chlorogenic acid

Y = 0.003 1X + 0.000 095

0.998 3

4-400

4.0

Neochlorogenic acid

Y = 0.000 87X - 0.000 029

0.997 1

5-500

5.0

Cryptochlorogenic acid

Y = 0.000 49X + 0.000 000 65

0.995 5

5-500

5.0

Component	Content/μg·mL^-1
Magnoflorine	1 166.1 ± 35.1	983.6 ± 22.8^**
Epiberberine	1 643.7 ± 7.8	1 185.6 ± 32.1^**
Coptisine	2 100.6 ± 20.1	1 863.9 ± 11.7^**
Jatrorrhizine	1 187.0 ± 20	944.8 ± 16.0^**
Berberrubine	685.4 ± 26.4	501.6 ± 15.0^**
Palmatine	3 028.0 ± 15.5	2 550.8 ± 13.5^**
Berberine	20 912.4 ± 299.7	18 809.1 ± 524.6^**
Chlorogenic acid	13 795.4 ± 503.5	11 947.8 ± 688.3^**
Neochlorogenic acid	1 151.7 ± 24.8	973.1 ± 14.3^**
Cryptochlorogenic acid	2 121.8 ± 36.9	1 858.4 ± 28.8^**

Component

Content/μg·mL^-1

With excipient

Without excipient

Magnoflorine

1 166.1 ± 35.1

983.6 ± 22.8^**

Epiberberine

1 643.7 ± 7.8

1 185.6 ± 32.1^**

Coptisine

2 100.6 ± 20.1

1 863.9 ± 11.7^**

Jatrorrhizine

1 187.0 ± 20

944.8 ± 16.0^**

Berberrubine

685.4 ± 26.4

501.6 ± 15.0^**

Palmatine

3 028.0 ± 15.5

2 550.8 ± 13.5^**

Berberine

20 912.4 ± 299.7

18 809.1 ± 524.6^**

Chlorogenic acid

13 795.4 ± 503.5

11 947.8 ± 688.3^**

Neochlorogenic acid

1 151.7 ± 24.8

973.1 ± 14.3^**

Cryptochlorogenic acid

2 121.8 ± 36.9

1 858.4 ± 28.8^**

Component	Content/μg·mL^-1
Magnoflorine	1 166.1 ± 35.1	983.6 ± 22.8^**
Epiberberine	1 643.7 ± 7.8	1 185.6 ± 32.1^**
Coptisine	2 100.6 ± 20.1	1 863.9 ± 11.7^**
Jatrorrhizine	1 187.0 ± 20	944.8 ± 16.0^**
Berberrubine	685.4 ± 26.4	501.6 ± 15.0^**
Palmatine	3 028.0 ± 15.5	2 550.8 ± 13.5^**
Berberine	20 912.4 ± 299.7	18 809.1 ± 524.6^**
Chlorogenic acid	13 795.4 ± 503.5	11 947.8 ± 688.3^**
Neochlorogenic acid	1 151.7 ± 24.8	973.1 ± 14.3^**
Cryptochlorogenic acid	2 121.8 ± 36.9	1 858.4 ± 28.8^**

Component

Content/μg·mL^-1

With excipient

Without excipient

Magnoflorine

1 166.1 ± 35.1

983.6 ± 22.8^**

Epiberberine

1 643.7 ± 7.8

1 185.6 ± 32.1^**

Coptisine

2 100.6 ± 20.1

1 863.9 ± 11.7^**

Jatrorrhizine

1 187.0 ± 20

944.8 ± 16.0^**

Berberrubine

685.4 ± 26.4

501.6 ± 15.0^**

Palmatine

3 028.0 ± 15.5

2 550.8 ± 13.5^**

Berberine

20 912.4 ± 299.7

18 809.1 ± 524.6^**

Chlorogenic acid

13 795.4 ± 503.5

11 947.8 ± 688.3^**

Neochlorogenic acid

1 151.7 ± 24.8

973.1 ± 14.3^**

Cryptochlorogenic acid

2 121.8 ± 36.9

1 858.4 ± 28.8^**

Component	Sample	Batch
Magnoflorine	Capsule	1 203.0 ± 16.8	1 178.0 ± 17.9	1 166.2 ± 17.6	1 155.4 ± 65.4	1 181.3 ± 21.9
Without excipient	968.6 ± 13.0^**	981.0 ± 24.0^**	971.1 ± 22.8^**	951.1 ± 33.4^**	976.0 ± 18.8^**
Epiberberine	Capsule	1 539.7 ± 41.8	1 562.0 ± 50.0	1 550.2 ± 39.0	1 545.1 ± 43.0	1 570.5 ± 57.7
Without excipient	1 250.0 ± 146.1^**	1 258.0 ± 103.2^**	1 253.1 ± 51.7^**	1 219.9 ± 73.1^**	1 225.3 ± 81.1^**
Coptisine	Capsule	2 097.7 ± 31.8	2 071.3 ± 60.8	2 096.5 ± 19.6	2 102.2 ± 13.7	2 094.3 ± 35.6
Without excipient	1 744.1 ± 87.0^**	1 790.5 ± 24.7^**	1 789.7 ± 40.0^**	1 745.6 ± 18.6^**	1 771.2 ± 22.1^**
Jatrorrhizine	Capsule	1 166.3 ± 25.1	1 163.9 ± 16.7	1 176.4 ± 11.1	1 160.9 ± 17.6	1 190.3 ± 15.4
Without excipient	914.6 ± 6.7^**	924.1 ± 25.1^**	914.6 ± 19.6^**	903.7 ± 16.4^**	913.8 ± 19.6^**
Berberrubine	Capsule	653.2 ± 17.8	666.9 ± 14.4	673.2 ± 25.3	661.6 ± 12.4	650.1 ± 15.7
Without excipient	513.2 ± 15.8^**	515.7 ± 15.5^**	518.0 ± 20.8^**	516.7 ± 7.2^**	517.8 ± 13.3^**
Palmatine	Capsule	3 006.3 ± 51.1	2 986.3 ± 91.6	2 964.0 ± 28.4	2 914.8 ± 19.8	2 936.8 ± 46.4
Without excipient	2 520.2 ± 52.2^**	2 607.5 ± 77.5^**	2 599.8 ± 57.2^**	2 564.7 ± 34.4^**	2 577.8 ± 51.7^**
Berberine	Capsule	21 251.7 ± 389.4	21 457.7 ± 337.8	21 304.2 ± 100.1	20 638.4 ± 186.3	21 129.0 ± 161.0
Without excipient	17 987.4 ± 259.4^**	17 958.2 ± 103.9^**	17 622.4 ± 695.9^**	17 912.3 ± 432.4^**	18 083.6 ± 310.2^**
Ononin	Capsule	60.0 ± 1.2	59.9 ± 0.7	60.1 ± 0.9	60.1 ± 0.9	59.7 ± 1.4
Without excipient	58.8 ± 1.1^**	57.7 ± 1.2^**	58.4 ± 1.1^**	58.8 ± 1.2^**	58.6 ± 0.7^**
Calycosin	Capsule	57.7 ± 1.2	58.3 ± 0.6	58.4 ± 0.5	58.0 ± 0.4	58.4 ± 1.2
Without excipient	56.4 ± 1.1^**	56.5 ± 1.2^**	56.2 ± 0.8^**	56.4 ± 0.8^**	56.5 ± 0.6^**
Calycosin-7-glucoside	Capsule	152.1 ± 1.9	151.6 ± 2.9	150.5 ± 2.8	151.7 ± 1.1	150.6 ± 2.5
Without excipient	149.9 ± 2.8^**	148.4 ± 4.8^**	146.2 ± 2.4^**	146.6 ± 1.0^**	147.4 ± 1.2^**
Formononetin	Capsule	28.3 ± 1.0	27.8 ± 0.4	28.3 ± 0.3	28.0 ± 0.2	27.6 ± 0.4
Without excipient	26.8 ± 0.4^**	26.9 ± 0.5^**	27.2 ± 0.3^**	27.0 ± 0.2^**	27.1± 0.3^**
Astragaloside Ⅳ	Capsule	103.1 ± 3.0	100.3 ± 2.3	101.9 ± 3.7	98.6 ± 0.7	99.7 ± 2.0
Without excipient	95.2 ± 3.6^**	96.9 ± 3.7^**	97.4 ± 3.9^**	96.7 ± 2.9^**	97.6 ± 3.1^**
Chlorogenic acid	Capsule	14 671.6 ± 170.3	14 643.0 ± 330.0	14 972.4 ± 381.2	15 057.1 ± 325.9	15 095.4 ± 252.4
Without excipient	11 811.3 ± 577.7^**	11 630.7 ± 242.4^**	11 845.0 ± 255.4^**	11 840.7 ± 324.1^**	12 032.4 ± 416.1^**
Neochlorogenic acid	Capsule	1 164.8 ± 24.6	1 185.9 ± 22.9	1 162.0 ± 15.8	1 160.0 ± 15.6	1 183.3 ± 21.9
Without excipient	992.7 ± 10.3^**	992.0 ± 13.1^**	976.2 ± 8.3^**	978.0 ± 18.4^**	970.4 ± 30.9^**
Cryptochlorogenic acid	Capsule	2 088.6 ± 48.7	2 140.1 ± 64.6	2 108.2 ± 49.6	2 119.8 ± 62.1	2 115.7 ± 57.4
Without excipient	1 815.8 ± 38.8^**	1 841.3 ± 45.9^**	1 824.2 ± 41.9^**	1 807.9 ± 28.0^**	1 775.1 ± 46.0^**

Component

Sample

Batch

2403001

2403002

2403003

2404001

2404002

Magnoflorine

Capsule

1 203.0 ± 16.8

1 178.0 ± 17.9

1 166.2 ± 17.6

1 155.4 ± 65.4

1 181.3 ± 21.9

Without excipient

968.6 ± 13.0^**

981.0 ± 24.0^**

971.1 ± 22.8^**

951.1 ± 33.4^**

976.0 ± 18.8^**

Epiberberine

Capsule

1 539.7 ± 41.8

1 562.0 ± 50.0

1 550.2 ± 39.0

1 545.1 ± 43.0

1 570.5 ± 57.7

Without excipient

1 250.0 ± 146.1^**

1 258.0 ± 103.2^**

1 253.1 ± 51.7^**

1 219.9 ± 73.1^**

1 225.3 ± 81.1^**

Coptisine

Capsule

2 097.7 ± 31.8

2 071.3 ± 60.8

2 096.5 ± 19.6

2 102.2 ± 13.7

2 094.3 ± 35.6

Without excipient

1 744.1 ± 87.0^**

1 790.5 ± 24.7^**

1 789.7 ± 40.0^**

1 745.6 ± 18.6^**

1 771.2 ± 22.1^**

Jatrorrhizine

Capsule

1 166.3 ± 25.1

1 163.9 ± 16.7

1 176.4 ± 11.1

1 160.9 ± 17.6

1 190.3 ± 15.4

Without excipient

914.6 ± 6.7^**

924.1 ± 25.1^**

914.6 ± 19.6^**

903.7 ± 16.4^**

913.8 ± 19.6^**

Berberrubine

Capsule

653.2 ± 17.8

666.9 ± 14.4

673.2 ± 25.3

661.6 ± 12.4

650.1 ± 15.7

Without excipient

513.2 ± 15.8^**

515.7 ± 15.5^**

518.0 ± 20.8^**

516.7 ± 7.2^**

517.8 ± 13.3^**

Palmatine

Capsule

3 006.3 ± 51.1

2 986.3 ± 91.6

2 964.0 ± 28.4

2 914.8 ± 19.8

2 936.8 ± 46.4

Without excipient

2 520.2 ± 52.2^**

2 607.5 ± 77.5^**

2 599.8 ± 57.2^**

2 564.7 ± 34.4^**

2 577.8 ± 51.7^**

Berberine

Capsule

21 251.7 ± 389.4

21 457.7 ± 337.8

21 304.2 ± 100.1

20 638.4 ± 186.3

21 129.0 ± 161.0

Without excipient

17 987.4 ± 259.4^**

17 958.2 ± 103.9^**

17 622.4 ± 695.9^**

17 912.3 ± 432.4^**

18 083.6 ± 310.2^**

Ononin

Capsule

60.0 ± 1.2

59.9 ± 0.7

60.1 ± 0.9

59.7 ± 1.4

Without excipient

58.8 ± 1.1^**

57.7 ± 1.2^**

58.4 ± 1.1^**

58.8 ± 1.2^**

58.6 ± 0.7^**

Calycosin

Capsule

57.7 ± 1.2

58.3 ± 0.6

58.4 ± 0.5

58.0 ± 0.4

58.4 ± 1.2

Without excipient

56.4 ± 1.1^**

56.5 ± 1.2^**

56.2 ± 0.8^**

56.4 ± 0.8^**

56.5 ± 0.6^**

Calycosin-7-glucoside

Capsule

152.1 ± 1.9

151.6 ± 2.9

150.5 ± 2.8

151.7 ± 1.1

150.6 ± 2.5

Without excipient

149.9 ± 2.8^**

148.4 ± 4.8^**

146.2 ± 2.4^**

146.6 ± 1.0^**

147.4 ± 1.2^**

Formononetin

Capsule

28.3 ± 1.0

27.8 ± 0.4

28.3 ± 0.3

28.0 ± 0.2

27.6 ± 0.4

Without excipient

26.8 ± 0.4^**

26.9 ± 0.5^**

27.2 ± 0.3^**

27.0 ± 0.2^**

27.1± 0.3^**

Astragaloside Ⅳ

Capsule

103.1 ± 3.0

100.3 ± 2.3

101.9 ± 3.7

98.6 ± 0.7

99.7 ± 2.0

Without excipient

95.2 ± 3.6^**

96.9 ± 3.7^**

97.4 ± 3.9^**

96.7 ± 2.9^**

97.6 ± 3.1^**

Chlorogenic acid

Capsule

14 671.6 ± 170.3

14 643.0 ± 330.0

14 972.4 ± 381.2

15 057.1 ± 325.9

15 095.4 ± 252.4

Without excipient

11 811.3 ± 577.7^**

11 630.7 ± 242.4^**

11 845.0 ± 255.4^**

11 840.7 ± 324.1^**

12 032.4 ± 416.1^**

Neochlorogenic acid

Capsule

1 164.8 ± 24.6

1 185.9 ± 22.9

1 162.0 ± 15.8

1 160.0 ± 15.6

1 183.3 ± 21.9

Without excipient

992.7 ± 10.3^**

992.0 ± 13.1^**

976.2 ± 8.3^**

978.0 ± 18.4^**

970.4 ± 30.9^**

Cryptochlorogenic acid

Capsule

2 088.6 ± 48.7

2 140.1 ± 64.6

2 108.2 ± 49.6

2 119.8 ± 62.1

2 115.7 ± 57.4

Without excipient

1 815.8 ± 38.8^**

1 841.3 ± 45.9^**

1 824.2 ± 41.9^**

1 807.9 ± 28.0^**

1 775.1 ± 46.0^**

Component	Sample	Batch
Magnoflorine	Capsule	1 203.0 ± 16.8	1 178.0 ± 17.9	1 166.2 ± 17.6	1 155.4 ± 65.4	1 181.3 ± 21.9
Without excipient	968.6 ± 13.0^**	981.0 ± 24.0^**	971.1 ± 22.8^**	951.1 ± 33.4^**	976.0 ± 18.8^**
Epiberberine	Capsule	1 539.7 ± 41.8	1 562.0 ± 50.0	1 550.2 ± 39.0	1 545.1 ± 43.0	1 570.5 ± 57.7
Without excipient	1 250.0 ± 146.1^**	1 258.0 ± 103.2^**	1 253.1 ± 51.7^**	1 219.9 ± 73.1^**	1 225.3 ± 81.1^**
Coptisine	Capsule	2 097.7 ± 31.8	2 071.3 ± 60.8	2 096.5 ± 19.6	2 102.2 ± 13.7	2 094.3 ± 35.6
Without excipient	1 744.1 ± 87.0^**	1 790.5 ± 24.7^**	1 789.7 ± 40.0^**	1 745.6 ± 18.6^**	1 771.2 ± 22.1^**
Jatrorrhizine	Capsule	1 166.3 ± 25.1	1 163.9 ± 16.7	1 176.4 ± 11.1	1 160.9 ± 17.6	1 190.3 ± 15.4
Without excipient	914.6 ± 6.7^**	924.1 ± 25.1^**	914.6 ± 19.6^**	903.7 ± 16.4^**	913.8 ± 19.6^**
Berberrubine	Capsule	653.2 ± 17.8	666.9 ± 14.4	673.2 ± 25.3	661.6 ± 12.4	650.1 ± 15.7
Without excipient	513.2 ± 15.8^**	515.7 ± 15.5^**	518.0 ± 20.8^**	516.7 ± 7.2^**	517.8 ± 13.3^**
Palmatine	Capsule	3 006.3 ± 51.1	2 986.3 ± 91.6	2 964.0 ± 28.4	2 914.8 ± 19.8	2 936.8 ± 46.4
Without excipient	2 520.2 ± 52.2^**	2 607.5 ± 77.5^**	2 599.8 ± 57.2^**	2 564.7 ± 34.4^**	2 577.8 ± 51.7^**
Berberine	Capsule	21 251.7 ± 389.4	21 457.7 ± 337.8	21 304.2 ± 100.1	20 638.4 ± 186.3	21 129.0 ± 161.0
Without excipient	17 987.4 ± 259.4^**	17 958.2 ± 103.9^**	17 622.4 ± 695.9^**	17 912.3 ± 432.4^**	18 083.6 ± 310.2^**
Ononin	Capsule	60.0 ± 1.2	59.9 ± 0.7	60.1 ± 0.9	60.1 ± 0.9	59.7 ± 1.4
Without excipient	58.8 ± 1.1^**	57.7 ± 1.2^**	58.4 ± 1.1^**	58.8 ± 1.2^**	58.6 ± 0.7^**
Calycosin	Capsule	57.7 ± 1.2	58.3 ± 0.6	58.4 ± 0.5	58.0 ± 0.4	58.4 ± 1.2
Without excipient	56.4 ± 1.1^**	56.5 ± 1.2^**	56.2 ± 0.8^**	56.4 ± 0.8^**	56.5 ± 0.6^**
Calycosin-7-glucoside	Capsule	152.1 ± 1.9	151.6 ± 2.9	150.5 ± 2.8	151.7 ± 1.1	150.6 ± 2.5
Without excipient	149.9 ± 2.8^**	148.4 ± 4.8^**	146.2 ± 2.4^**	146.6 ± 1.0^**	147.4 ± 1.2^**
Formononetin	Capsule	28.3 ± 1.0	27.8 ± 0.4	28.3 ± 0.3	28.0 ± 0.2	27.6 ± 0.4
Without excipient	26.8 ± 0.4^**	26.9 ± 0.5^**	27.2 ± 0.3^**	27.0 ± 0.2^**	27.1± 0.3^**
Astragaloside Ⅳ	Capsule	103.1 ± 3.0	100.3 ± 2.3	101.9 ± 3.7	98.6 ± 0.7	99.7 ± 2.0
Without excipient	95.2 ± 3.6^**	96.9 ± 3.7^**	97.4 ± 3.9^**	96.7 ± 2.9^**	97.6 ± 3.1^**
Chlorogenic acid	Capsule	14 671.6 ± 170.3	14 643.0 ± 330.0	14 972.4 ± 381.2	15 057.1 ± 325.9	15 095.4 ± 252.4
Without excipient	11 811.3 ± 577.7^**	11 630.7 ± 242.4^**	11 845.0 ± 255.4^**	11 840.7 ± 324.1^**	12 032.4 ± 416.1^**
Neochlorogenic acid	Capsule	1 164.8 ± 24.6	1 185.9 ± 22.9	1 162.0 ± 15.8	1 160.0 ± 15.6	1 183.3 ± 21.9
Without excipient	992.7 ± 10.3^**	992.0 ± 13.1^**	976.2 ± 8.3^**	978.0 ± 18.4^**	970.4 ± 30.9^**
Cryptochlorogenic acid	Capsule	2 088.6 ± 48.7	2 140.1 ± 64.6	2 108.2 ± 49.6	2 119.8 ± 62.1	2 115.7 ± 57.4
Without excipient	1 815.8 ± 38.8^**	1 841.3 ± 45.9^**	1 824.2 ± 41.9^**	1 807.9 ± 28.0^**	1 775.1 ± 46.0^**

Component

Sample

Batch

2403001

2403002

2403003

2404001

2404002

Magnoflorine

Capsule

1 203.0 ± 16.8

1 178.0 ± 17.9

1 166.2 ± 17.6

1 155.4 ± 65.4

1 181.3 ± 21.9

Without excipient

968.6 ± 13.0^**

981.0 ± 24.0^**

971.1 ± 22.8^**

951.1 ± 33.4^**

976.0 ± 18.8^**

Epiberberine

Capsule

1 539.7 ± 41.8

1 562.0 ± 50.0

1 550.2 ± 39.0

1 545.1 ± 43.0

1 570.5 ± 57.7

Without excipient

1 250.0 ± 146.1^**

1 258.0 ± 103.2^**

1 253.1 ± 51.7^**

1 219.9 ± 73.1^**

1 225.3 ± 81.1^**

Coptisine

Capsule

2 097.7 ± 31.8

2 071.3 ± 60.8

2 096.5 ± 19.6

2 102.2 ± 13.7

2 094.3 ± 35.6

Without excipient

1 744.1 ± 87.0^**

1 790.5 ± 24.7^**

1 789.7 ± 40.0^**

1 745.6 ± 18.6^**

1 771.2 ± 22.1^**

Jatrorrhizine

Capsule

1 166.3 ± 25.1

1 163.9 ± 16.7

1 176.4 ± 11.1

1 160.9 ± 17.6

1 190.3 ± 15.4

Without excipient

914.6 ± 6.7^**

924.1 ± 25.1^**

914.6 ± 19.6^**

903.7 ± 16.4^**

913.8 ± 19.6^**

Berberrubine

Capsule

653.2 ± 17.8

666.9 ± 14.4

673.2 ± 25.3

661.6 ± 12.4

650.1 ± 15.7

Without excipient

513.2 ± 15.8^**

515.7 ± 15.5^**

518.0 ± 20.8^**

516.7 ± 7.2^**

517.8 ± 13.3^**

Palmatine

Capsule

3 006.3 ± 51.1

2 986.3 ± 91.6

2 964.0 ± 28.4

2 914.8 ± 19.8

2 936.8 ± 46.4

Without excipient

2 520.2 ± 52.2^**

2 607.5 ± 77.5^**

2 599.8 ± 57.2^**

2 564.7 ± 34.4^**

2 577.8 ± 51.7^**

Berberine

Capsule

21 251.7 ± 389.4

21 457.7 ± 337.8

21 304.2 ± 100.1

20 638.4 ± 186.3

21 129.0 ± 161.0

Without excipient

17 987.4 ± 259.4^**

17 958.2 ± 103.9^**

17 622.4 ± 695.9^**

17 912.3 ± 432.4^**

18 083.6 ± 310.2^**

Ononin

Capsule

60.0 ± 1.2

59.9 ± 0.7

60.1 ± 0.9

59.7 ± 1.4

Without excipient

58.8 ± 1.1^**

57.7 ± 1.2^**

58.4 ± 1.1^**

58.8 ± 1.2^**

58.6 ± 0.7^**

Calycosin

Capsule

57.7 ± 1.2

58.3 ± 0.6

58.4 ± 0.5

58.0 ± 0.4

58.4 ± 1.2

Without excipient

56.4 ± 1.1^**

56.5 ± 1.2^**

56.2 ± 0.8^**

56.4 ± 0.8^**

56.5 ± 0.6^**

Calycosin-7-glucoside

Capsule

152.1 ± 1.9

151.6 ± 2.9

150.5 ± 2.8

151.7 ± 1.1

150.6 ± 2.5

Without excipient

149.9 ± 2.8^**

148.4 ± 4.8^**

146.2 ± 2.4^**

146.6 ± 1.0^**

147.4 ± 1.2^**

Formononetin

Capsule

28.3 ± 1.0

27.8 ± 0.4

28.3 ± 0.3

28.0 ± 0.2

27.6 ± 0.4

Without excipient

26.8 ± 0.4^**

26.9 ± 0.5^**

27.2 ± 0.3^**

27.0 ± 0.2^**

27.1± 0.3^**

Astragaloside Ⅳ

Capsule

103.1 ± 3.0

100.3 ± 2.3

101.9 ± 3.7

98.6 ± 0.7

99.7 ± 2.0

Without excipient

95.2 ± 3.6^**

96.9 ± 3.7^**

97.4 ± 3.9^**

96.7 ± 2.9^**

97.6 ± 3.1^**

Chlorogenic acid

Capsule

14 671.6 ± 170.3

14 643.0 ± 330.0

14 972.4 ± 381.2

15 057.1 ± 325.9

15 095.4 ± 252.4

Without excipient

11 811.3 ± 577.7^**

11 630.7 ± 242.4^**

11 845.0 ± 255.4^**

11 840.7 ± 324.1^**

12 032.4 ± 416.1^**

Neochlorogenic acid

Capsule

1 164.8 ± 24.6

1 185.9 ± 22.9

1 162.0 ± 15.8

1 160.0 ± 15.6

1 183.3 ± 21.9

Without excipient

992.7 ± 10.3^**

992.0 ± 13.1^**

976.2 ± 8.3^**

978.0 ± 18.4^**

970.4 ± 30.9^**

Cryptochlorogenic acid

Capsule

2 088.6 ± 48.7

2 140.1 ± 64.6

2 108.2 ± 49.6

2 119.8 ± 62.1

2 115.7 ± 57.4

Without excipient

1 815.8 ± 38.8^**

1 841.3 ± 45.9^**

1 824.2 ± 41.9^**

1 807.9 ± 28.0^**

1 775.1 ± 46.0^**

金芪降糖胶囊多成分含量研究

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田甜 ¹ , 石荣 ² , 吴家胜 ¹ , 王天明 ¹ , 李建国 ³ , 夏国凤 ³ , 李安宁 ³ , 李园园 ¹^,^* , 马越鸣 ¹^,^*

药学学报 | 研究论文 2024,59(10): 2849-2856

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药学学报 | 研究论文 2024, 59(10): 2849-2856

金芪降糖胶囊多成分含量研究

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田甜¹, 石荣², 吴家胜¹, 王天明¹, 李建国³, 夏国凤³, 李安宁³, 李园园¹^,^*, 马越鸣¹^,^*

作者信息

1.上海中医药大学中药学院, 上海 201203

2.上海中医药大学科技实验中心, 上海 201203

3.吉林敖东延边药业股份有限公司, 吉林延边 133700

通讯作者:

^*李园园, Tel / Fax: 86-21-51322386, E-mail: yuanyuan60107@163.com;

马越鸣, E-mail: mayueming_117@126.com

Study on multi-component contents of Jinqi Jiangtang Capsule

Tian TIAN¹, Rong SHI², Jia-sheng WU¹, Tian-ming WANG¹, Jian-guo LI³, Guo-feng XIA³, An-ning LI³, Yuan-yuan LI¹^,^*, Yue-ming MA¹^,^*

Affiliations

1. School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

2. Science and Technology Experimental Center, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

3. Jilin Aodong Yanbian Pharmaceutical Company Limited, Yanbian 133700, China

出版时间: 2024-10-12 doi: 10.16438/j.0513-4870.2024-0361

文章导航

摘要

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金芪降糖胶囊(JQJTC) 临床上常用于2型糖尿病的预防与治疗, 但其中主要的化学成分含量尚不清楚。本研究首先建立JQJTC中新绿原酸、绿原酸、隐绿原酸、芒柄花素、芒柄花苷、毛蕊异黄酮、毛蕊异黄酮苷、黄芪甲苷、小檗碱、表小檗碱、小檗红碱、黄连碱、药根碱、巴马汀、木兰花碱的超高效液相-串联质谱联用的含量测定方法, 再采用所建立的方法测定JQJTC中15种成分的含量, 并考察辅料对JQJTC多成分含量的影响。使用ACQUITY UPLC BEH C₁₈ (100 mm × 2.1 mm, 1.7 μm) 色谱柱, 流动相为0.1%乙酸-5 mmol·L^-1乙酸铵水溶液(A)-乙腈(B), 流速0.3 mL·min^-1, 柱温40 ℃, 采用电喷雾离子源, 在正离子模式下进行检测。结果显示建立的方法符合中国药典含量测定方法学的要求; 所检测的15种成分在JQJTC中含量高低不等, 含量前5位依次是小檗碱、绿原酸、巴马汀、黄连碱、隐绿原酸, 共占87.31%; 未含辅料全方提取物中黄连生物碱类(小檗碱、表小檗碱、小檗红碱、黄连碱、药根碱、巴马汀、木兰花碱) 和金银花有机酸类(新绿原酸、绿原酸、隐绿原酸) 成分含量显著低于胶囊内容物含量, 这10种成分含量占测定成分总含量的99.20%。本研究建立了准确、灵敏和高效的JQJTC多成分超高效液相色谱-串联质谱测定方法, 稳定可靠地检测了胶囊中15种成分含量, 可为更全面的质量分析提供基础; 并发现辅料可使测定的生物碱类和有机酸类成分含量增加, 这可能有利于胶囊整体药效的发挥。

关键词

金芪降糖胶囊 / 多成分含量 / 辅料 / 超高效液相色谱-串联质谱

Abstract

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Jinqi Jiangtang Capsule (JQJTC) is clinically used for the prevention and treatment of type 2 diabetes, but the contents of its main chemical components are not yet clear. In this study, an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established for the determination of 15 components in JQJTC, including new chlorogenic acid, chlorogenic acid, cryptochlorogenic acid, formononetin, ononin, calycosin, calycosin-7-glucoside, astragaloside Ⅳ, berberine, epiberberine, berberrubine, coptisine, jatrorrhizine, palmatine and magnoflorine. The method was used to determine the contents of 15 components in the capsule and then to investigate the influence of excipients on the contents of the components in JQJTC. The separation was performed on a ACQUITY UPLC BEH C₁₈ column (100 mm × 2.1 mm, 1.7 μm) with a mobile phase consisting of 0.1% acetic acid and 5 mmol·L^-1 ammonium acetate (A) and acetonitrile (B) with gradient elution at a flow rate of 0.3 mL·min^-1 and a column temperature at 40 ℃. Electron spray ionization was used for mass spectrometry in positive ion mode. The established method meets the requirements of methodology of content determination in Chinese pharmacopoeia. The contents of 15 components in JQJTC varied from high to low. The top 5 contents were berberine, chlorogenic acid, magnoflorine, coptisine, and cryptochlorogenic acid, accounting for 87.31% of the total content. The contents of 10 components, including the alkaloids of coptidis rhizoma (berberine, epiberberine, berberrubine, coptisine, jatrorrhizine, palmatine and magnoflorine) and the organic acids of honeysuckle (new chlorogenic acid, chlorogenic acid, and cryptochlorogenic acid) in the whole formula extract without excipients was significantly lower than that in the capsule. These components accounted for 99.20% of the determined component contents. In this experiment, an accurate, sensitive and efficient UHPLC-MS/MS method for the determination of multi-components in JQJTC was established, which stably and reliably detected the contents of 15 components in the capsule and could provide the basis for more comprehensive quality analysis. It was also found that excipients had an increasing effect on the contents of detected alkaloid and organic acid components, which may be beneficial to the effectiveness of the capsules.

Key words

Jinqi Jiangtang Capsule / multi-component content / excipient / ultra performance liquid chromatography-tandem mass spectrometry

引用本文

田甜, 石荣, 吴家胜, 王天明, 李建国, 夏国凤, 李安宁, 李园园, 马越鸣. 金芪降糖胶囊多成分含量研究. 药学学报, 2024 , 59 (10) : 2849 -2856 . DOI: 10.16438/j.0513-4870.2024-0361

Tian TIAN, Rong SHI, Jia-sheng WU, Tian-ming WANG, Jian-guo LI, Guo-feng XIA, An-ning LI, Yuan-yuan LI, Yue-ming MA. Study on multi-component contents of Jinqi Jiangtang Capsule[J]. Acta Pharmaceutica Sinica, 2024 , 59 (10) : 2849 -2856 . DOI: 10.16438/j.0513-4870.2024-0361

正文

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糖尿病是一种常见的以高血糖为特征的代谢性疾病, 其发病率、并发症发生率和死亡率逐年上升^[1], 严重威胁人民健康。在传统中医中, 糖尿病属“消渴”范畴, 中医药在消渴的预防及治疗中具有悠久的历史。金芪降糖方源自唐代孙思邈所著《备急千金要方》中千金黄连丸, 由黄连(Rhizoma Coptidis)、黄芪(Astragalus)、金银花(Honeysuckle) 组成, 具有清热益气、生津止渴之功效, 主治消渴, 是现代临床广泛应用于2型糖尿病预防与治疗的中药复方^[2]。

金芪降糖方制剂有片剂和胶囊。研究表明金芪降糖片含有多种成分, 包括来源于黄连的生物碱类、黄芪的皂苷类及黄酮类成分、金银花的有机酸类成分等; 可通过增加葡萄糖摄取、促进糖原合成、保护胰岛和β细胞、减轻氧化应激、抗炎症、降低胰岛素抵抗和增强脂质代谢等多个环节改善糖尿病^[1]。这些结果都来源于金芪降糖片的研究, 关于金芪降糖胶囊(JQJTC) 研究报道较少。临床研究表明JQJTC对糖耐量异常患者有改善作用^[3], 可以降低2型糖尿病尿微量白蛋白排泄率^[4], 另外JQJTC可降低2型糖尿病大鼠血糖、血脂, 能改善糖脂代谢异常^[5], 值得临床推广使用。至今尚未有关于JQJTC成分分析研究, 关于JQJTC的化学成分含量尚不清楚。

本研究采用超高效液相色谱-三重四级杆串联质谱法(UHPLC-MS/MS) 建立JQJTC 15种成分(图 1) 的含量测定方法, 并对JQJTC内容物及未含辅料全方提取物中15种成分进行含量比较, 考察辅料对15种成分含量的影响, 为JQJTC后续的药代动力学及药效物质研究提供依据, 也为JQJTC更好的临床应用提供基础。

材料与方法

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试剂和药品 新绿原酸(neochlorogenic acid, 批号: N0805AS)、隐绿原酸(cryptochlorogenic acid, 批号: J0322AS)、小檗红碱(berberrubine, 批号: A1107AS)、盐酸巴马汀(palmatine, 批号: S0422AS)、盐酸黄连碱(coptisine, 批号: F1111AS) 均购自大连美仑生物科技有限公司; 卡马西平(carbamazepine, 批号: H21M7L11256)、盐酸小檗碱(berberine, 批号: S01A10K94340)、盐酸药根碱(jatrorrhizine, 批号: Z06S6S3041) 购于上海源叶生物科技有限公司; 表小檗碱(epiberberine, 批号: 121009)、木兰花碱(magnoflorine, 批号: 2141-09-5) 购于中国食品药品检定研究院; 芒柄花苷(ononin, 批号: PS000671)、芒柄花素(formononetin, 批号: PS000674)、绿原酸(chlorogenic acid, 批号: PS000629)、黄芪甲苷(astragaloside Ⅳ, 批号: PS013184)、毛蕊异黄酮(calycosin, 批号: PS010251)、毛蕊异黄酮苷(calycosin-7-glucoside, 批号: PS000687) 购自成都普思生物科技股份有限公司, 以上标准品纯度均大于98%。色谱级甲醇、乙腈购自Honeywell Burdick & Jackson公司; 乙酸、乙酸铵(色谱级) 均购于Tedia公司; 超纯水由Millipore Milli-Q纯水仪制备。JQJTC内容物(批号: 2403001、2403002、2403003、2404001、2404002)、未含辅料全方提取物(批号: 2403001、2403002、2403003、2404001、2404002)、辅料淀粉(批号: 2403001) 均由吉林敖东延边药业股份有限公司提供。

实验仪器 Agilent Infinity Ⅱ 6470-6495 QQQ UHPLC-MS/MS超高效液相色谱-串联质谱(美国Agilent公司); ScanSpeed1730MR冷冻离心机(丹麦LaboGene公司); USC-502超声波清洗器(上海波龙电子设备有限公司); QT-1涡旋混合器(上海琪特分析仪器有限公司)。

混合标准品储备液制备 分别精密称取木兰花碱、表小檗碱、盐酸黄连碱、盐酸药根碱、小檗红碱、盐酸巴马汀、盐酸小檗碱、芒柄花苷、毛蕊异黄酮、毛蕊异黄酮苷、芒柄花素、黄芪甲苷、新绿原酸、绿原酸、隐绿原酸对照品适量, 置于2 mL量瓶中定容得各标准品储备液。分别取各标准品储备液适量混合、定容至2 mL量瓶中, 得混合标准品储备液, 各成分浓度分别为1.6、1.6、1.6、0.8、1.6、1.6、3.2、1.0、1.0、1.0、1.0、20、80、10、10 μg·mL^-1, 于-20 ℃冰箱保存。

内标溶液制备 精密称取适量卡马西平标准品于2 mL量瓶中, 加入甲醇溶液配置成1 mg·mL^-1的内标储备液, 取适量内标储备液, 用甲醇溶液稀释为5 ng·mL^-1的内标溶液做沉淀试剂。将制备好的内标储备液和内标溶液转入-20 ℃冰箱储存备用。

混合标准品系列溶液与质控样本制备 混合标准品储备液分别用含辅料的50%的乙醇水溶液或纯水稀释20倍后为工作液, 再逐级稀释为标准品储备液的0.8、0.32、0.08、0.04、0.01倍, 共6个浓度, 得到50%乙醇水溶液或纯水配置的混合标准品系列溶液。分别取含辅料的50%的乙醇水溶液(针对生物碱类、黄酮类、皂苷类成分)/含辅料的纯水(针对有机酸类成分) 配置的50 μL不同浓度的混合标准品溶液, 按1∶3 (V/V) 加入内标溶液, 离心取上清100 µL按1∶1 (V/V) 加纯水, 混匀离心后取150 μL于进样瓶中待测。分别选取标准曲线从低浓度到高浓度中第2、3和5三个浓度点作为质控样品, 制备方式同前。

供试品溶液制备 分别取JQJTC内容物、未加辅料全方提取物, 精密称定200 mg于50 mL量瓶中, 加50%的乙醇水溶液(针对生物碱类、黄酮类、皂苷类成分)/纯水(针对有机酸类成分) 溶解定容到刻度配置为4 mg·mL^-1浓度, 摇匀; 超声溶解60 min (针对生物碱类、黄酮类、皂苷类成分)/水浴中热浸10 min (针对有机酸类成分); 冷却后用50%乙醇水溶液(针对生物碱类、黄酮类、皂苷类成分)/纯水(针对有机酸类成分) 补足失重, 14 000 r·min^-1 (4 ℃) 离心10 min, 取上层清液储存, 待后续处理。以上两种提取液分别以50%乙醇水或纯水稀释15倍和900倍, 得到JQJTC内容物和未加辅料全方提取物溶液稀释15倍或稀释900倍的样品; 按1∶3加内标溶液, 14 000 r·min^-1 (4 ℃) 离心10 min后取上清按1∶1加纯水, 取5 μL分别进样分析。

色谱条件 色谱柱: ACQUITY UPLC BEH C₁₈ (100 mm × 2.1 mm, 1.7 μm), 流动相: 0.1%乙酸-5 mmol·L^-1乙酸铵水溶液(A相)-乙腈(B相), 梯度洗脱0~2 min, 5%~10% B; 2~4 min, 10%~20% B; 4~8 min, 20%~50% B; 8~10 min, 50%~95% B; 10~10.1 min, 95%~5% B; 流速: 0.3 mL·min^-1, 柱温: 40 ℃, 进样量: 5 µL, 自动进样器温度: 6 ℃。

质谱条件 电喷雾离子源(ESI), 干燥气温度290 ℃, 干燥气流量14 L·min^-1, 毛细管电压4 000 V (+), 喷雾气压35 psi, 鞘气温度350 ℃, 鞘气流量11 L·min^-1, 加速电压5 V, 扫描方式为正离子多反应监测(MRM) 模式, 各成分及内标的质谱检测参数见表 1。

专属性考察 将含辅料的空白溶剂、含有15种成分及内标的混合标准品溶液和JQJTC溶液、未含辅料全方提取物溶液按“溶液配制”进样测定, 得各样品色谱图。

线性关系和定量下限考察 将混合标准品系列溶液(n = 5), 分别取5 μL进样分析。以15种待测物浓度为横坐标, 待测物与内标物的峰面积比值为纵坐标, 用加权(W = 1/x²) 最小二乘法进行回归运算, 求得的直线回归方程即为标准曲线。各成分的最低定量下限(LLOQ) 的信噪比不小于10, 并且空白样品中的干扰峰不大于LLOQ峰面积的20%。

精密度和准确度 取3个浓度质控样本(n = 3), 连续进样3天考察批内和批间精密度和准确度, 根据当日随行标准曲线计算质控样本中各成分浓度, 精密度和准确度以相对标准偏差(relative standard deviation, RSD) 和相对误差(relative error, RE) 表示。

加样回收率 将供试品样本稀释一半后, 分别加入三种不同浓度的混合标准品溶液, 为目标成分的60%、100%和140%, 按照“供试品溶液制备”项下的方法平行制备3份。计算各成分的加样回收率: 加样回收率/% = (实测量-样品所含被测成分量) / 加入对照品量× 100%。

重复性 取同一批JQJTC, 平行6份, 精密称定, 分别按“供试品溶液制备”项下的方法处理样品后进样分析, 以随行标准曲线测定各成分浓度, 考察测定结果的重复性。

稳定性 取“供试品溶液制备”下的样品(n = 3), 在进样器中放置24 h进样, 以随行标准曲线测定各成分浓度, 考察样品放置稳定性。

基质效应 根据胶囊中辅料配比称取适量淀粉, 加50%的乙醇水溶液或纯水定容至50 mL, 离心取上清, 加入适量标准品溶液, 制备低、中、高三个浓度的样品(n = 3), 分别进样分析, 记为面积A; 另取50%的乙醇水溶液或纯水代替作为对照, 同上操作, 进样测定获得面积记为B, 连续进样3批, 以A/B作为考察辅料对各成分检测是否有基质影响。

辅料对生物碱类与有机酸类成分单独及混合时的影响 用含辅料与不含辅料的溶剂分别配置接近样品浓度的生物碱类成分、有机酸类成分、生物碱类+ 有机酸类成分混合的标准品, 按“供试品溶液制备”项下方法处理, 各平行5份进样分析, 以含辅料溶剂配置的随行标准曲线计算各成分含量, 检测辅料对这两类成分单独及混合时的影响。

样品含量测定 分别取5批不同批次的JQJTC内容物及未含辅料全方提取物精密称取200 mg, 按“供试品溶液制备”项下方法处理, 平行5份进样分析, 以随行标准曲线计算各成分含量, 胶囊再扣除3%辅料以全方提取物所占实际比例进行化学成分含量计算。

数据处理 使用IBM SPSS Statistics 21.0统计软件进行数据分析, 结果以均值±标准差(x ± s) 表示。两组间比较采用T-TEST检验, 样品含量测定采用双因素(批次、是否有辅料) 方差分析, P < 0.05被认为存在显著性差异。

结果

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1 方法学

1.1 专属性

含辅料的空白样品、15种成分混合对照品溶液、JQJTC样品及未含辅料全方提取物的典型色谱图见图 2。结果表明, 各对照品溶液与样品溶液中相应组分对应的色谱峰具有相同保留时间, 空白样品不干扰待测成分及内标的测定。

1.2 线性关系和定量下限

15种成分的线性回归方程、线性范围和定量下限结果见表 2。

1.3 精密度和准确度

15种成分高、中、低QC的批内精密度(n = 3) RSD分别在0.1%~1.7%、0.1%~3.6%、0.3%~4.2%之间, 准确度RE分别在-1.7%~9.4%、-9.7%~3.7%、-9.8%~10.6%之间; 批间精密度(n = 9) 分别在1.1%~7.8%、1.8%~8.8%、1.8%~9.5%之间, 准确度分别在-4.4%~4.9%、-6.6%~4.1%、-7.4%~7.3%之间。

1.4 加样回收率

15种成分高、中、低浓度的加样回收率(n = 3) 分别在98.3%~106.1%、96.4%~106.9%、91.6%~106.0%之间, RSD在0.1%~1.7%、0.2%~5.3%、0.3%~2.7%之间。

1.5 重复性和稳定性

15种成分重复性(n = 6) 的RSD值在0.6%~4.4%之间。稳定性(n = 3) 结果表明, 样品制备后储存进样器条件下, 各成分至少在24 h内稳定, 其RE值范围在-2.3%~4.4%之间, RSD值在0.3%~2.6%之间。

1.6 基质效应

存在辅料时各成分标准品批内与批间的偏差均 < 10%, 15种成分高、中、低浓度(n = 3) 的批内RSD值分别在0.2%~1.7%、0.5%~4.7%、0.2%~4.1%之间, 批间(n = 9) RSD在0.8%~4.9%、0.9%~6.5%、0.5%~4.8%之间, 此结果表明辅料对成分测定无影响。

1.7 辅料对生物碱类与有机酸类成分单独及混合时的影响

单独生物碱类及单独有机酸类成分含辅料与不含辅料结果差异无显著意义。两类成分混合时, 不含辅料的溶剂配置的混合标准品浓度要显著低于含辅料溶剂配置的混合标准品浓度, 结果见表 3。

2 样品含量测定

5批次JQJTC内容物与未加辅料全方提取物中15种成分含量见表 4。结果表明, JQJTC中15种成分在样本中的含量高低不等, 其中小檗碱含量最高, 占42.69%, 含量前5位依次是小檗碱、绿原酸、巴马汀、黄连碱、隐绿原酸, 共占87.31%。双因素方差分析结果显示辅料对生物碱类和有机酸类成分含量有显著性影响(P < 0.01), 不同批次的含量差异无统计学意义。

5批JQJTC平均含量见图 3, 结果表明, 未加辅料全方提取物中10个成分包括木兰花碱、表小檗碱、黄连碱、药根碱、小檗红碱、巴马汀、小檗碱、绿原酸、新绿原酸、隐绿原酸的含显著低于胶囊内容物中的含量, 降低比例分别为21.37%、25.16%、18.33%、28.15%、28.03%、15.06%、18.11%、25.83%、19.28%、16.64%, 这10种成分的含量占测定成分总含量的99.20%。

讨论

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金芪降糖方是临床治疗和预防2型糖尿病常用的中药复方, 目前尚无关于JQJTC的成分含量测定研究。由于JQJTC中的中药提取方法、制剂工艺及辅料与片剂有区别, 而这些都会影响成药的有效成分含量, 因此, 有必要对于JQJTC多成分进行分析测定, 为其药代动力学与药效物质基础研究提供依据。

JQJTC中含有多种化学性质不同、含量差异显著的成分, 使得多成分的定量分析存在很大难度, 在建立方法过程中, 除了对色谱条件、质谱参数进行优化外, 还针对中药多成分的难点, 在成分选择、样品前处理方法和进样稀释倍数等方面, 进行了探索, 最终选择小鼠灌胃给予胶囊后体内可定量测定的15种成分作为含量测定的成分; 分别以50%乙醇水溶液(生物碱、黄酮和皂苷类成分) 和纯水(有机酸类成分) 作为提取溶剂可实现多种成分全部溶出; 选择15倍与900倍两种稀释倍数分别进样分析, 能使所有待测成分浓度在标曲范围内。方法学考察结果表明, 建立的含量测定方法符合中国药典的要求; 与前期的方法^[6]相比, 更灵敏、高效(检测时间减少一半), 可为更全面的质量分析提供基础。

JQJTC中多成分含量测定结果显示, 黄连生物碱类最高、其次是金银花有机酸类, 而黄芪皂苷及黄酮类成分较低。另外, JQJTC中的辅料即淀粉使得黄连生物碱类和绿原酸类成分的含量增加。为了分析辅料的影响, 在方法学考察时, 增加了辅料基质影响的观察, 结果表明辅料对所测成分在分析方法上无影响; 对接近样品浓度的单生物碱类或单有机酸类标准品没有影响, 但对两类成分混合后的标准品有显著性影响。有文献报道, 中药复方提取物中酸类与碱类成分的配伍可能产生酸碱沉淀^[7], JQJTC的辅料淀粉为天然淀粉, 其具有良好成膜能力与凝胶能力^[8], 作为大分子可能通过物理力与活性小分子结合, 所以推测辅料可能与生物碱类或有机酸类成分结合, 使酸碱沉淀减少, 从而使测定的生物碱类及有机酸类成分含量增高, 具体机制还需进一步研究。

有研究报道黄连中生物碱类成分如小檗碱能通过多个环节起主要降糖作用^[9-12]; 金银花中的绿原酸通过抑制α-葡萄糖苷酶降低餐后血糖水平^{[13, 14]}。JQJTC辅料即淀粉使测定的生物碱类及有机酸类成分含量增高, 可能有利于胶囊整体药效的发挥。本研究结果为JQJTC后续的药动学和药效物质研究提供了基础。

作者贡献: 田甜负责实验及撰写文章; 石荣对实验指导及数据处理方面提供了帮助; 吴家胜、王天明参与本文实验以及对文章修改提供了宝贵意见; 李建国、夏国凤、李安宁负责本文药物制备及质量控制; 李园园对本文实验思路、数据处理、方法建立及文章修改提供了宝贵意见; 马越鸣从选题的设计、实验思路、方法建立及文章修改提供了专业的指导。

利益冲突: 无任何利益冲突。

基金

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上海中医药大学基金会项目(202301003)

参考文献

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文献

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参考文献引证文献

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2024年第59卷第10期

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doi: 10.16438/j.0513-4870.2024-0361

接收时间：2024-04-12
首发时间：2025-11-24
出版时间：2024-10-12

补充材料

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出版历史

收稿日期：2024-04-12
修回日期：2024-07-29

基金

上海中医药大学基金会项目(202301003)

作者信息

1.上海中医药大学中药学院, 上海 201203

2.上海中医药大学科技实验中心, 上海 201203

3.吉林敖东延边药业股份有限公司, 吉林延边 133700

通讯作者:

^*李园园, Tel / Fax: 86-21-51322386, E-mail: yuanyuan60107@163.com;

马越鸣, E-mail: mayueming_117@126.com

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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Component	Precursor ion (m/z)	Product ion (m/z)	Dwell time/s	Fragmentor	Collision energy/eV	Polarity	Retention time/min
Magnoflorine	342.1	297.1	30	166	20	[M]⁺	4.62
Epiberberine	336.1	320.0	30	166	36	[M]⁺	6.48
Coptisine	320.1	292.1	30	166	32	[M]⁺	6.52
Jatrorrhizine	338.1	322.1	30	166	36	[M]⁺	6.52
Berberrubine	322.1	307.1	30	166	32	[M+H]⁺	6.76
Palmatine	352.1	336.1	30	166	36	[M]⁺	7.03
Berberine	336.1	320.1	30	166	36	[M]⁺	7.12
Ononin	431.1	269.1	30	166	17	[M+H]⁺	6.80
Calycosin	285.0	270.0	30	166	24	[M+H]⁺	7.21
Calycosin-7-glucoside	477.0	285.0	30	166	20	[M+H]⁺	5.45
Formononetin	269.0	197.0	100	166	48	[M+H]⁺	8.57
Astragaloside Ⅳ	807.5	807.5	100	166	0	[M+Na]⁺	8.43
Chlorogenic acid	355.1	162.9	100	166	10	[M+H]⁺	3.19
Neochlorogenic acid	355.1	162.9	100	166	10	[M+H]⁺	2.34
Cryptochlorogenic acid	355.1	162.9	100	166	10	[M+H]⁺	3.44
Carbamazepine	237.0	194.0	30	166	21	[M+H]⁺	7.88

Component

Precursor ion (m/z)

Product ion (m/z)

Dwell time/s

Fragmentor

Collision energy/eV

Polarity

Retention time/min

Magnoflorine

342.1

297.1

166

[M]⁺

4.62

Epiberberine

336.1

320.0

166

[M]⁺

6.48

Coptisine

320.1

292.1

166

[M]⁺

6.52

Jatrorrhizine

338.1

322.1

166

[M]⁺

6.52

Berberrubine

322.1

307.1

166

[M+H]⁺

6.76

Palmatine

352.1

336.1

166

[M]⁺

7.03

Berberine

336.1

320.1

166

[M]⁺

7.12

Ononin

431.1

269.1

166

[M+H]⁺

6.80

Calycosin

285.0

270.0

166

[M+H]⁺

7.21

Calycosin-7-glucoside

477.0

285.0

166

[M+H]⁺

5.45

Formononetin

269.0

197.0

100

166

[M+H]⁺

8.57

Astragaloside Ⅳ

807.5

100

166

[M+Na]⁺

8.43

Chlorogenic acid

355.1

162.9

100

166

[M+H]⁺

3.19

Neochlorogenic acid

355.1

162.9

100

166

[M+H]⁺

2.34

Cryptochlorogenic acid

355.1

162.9

100

166

[M+H]⁺

3.44

Carbamazepine

237.0

194.0

166

[M+H]⁺

7.88

Component	Regression equation	r	Linearity range/ng·mL^-1	LLOQ/ng·mL^-1
Magnoflorine	Y = 0.02X + 0.000 66	0.998 8	0.8-80	0.8
Epiberberine	Y = 0.16X + 0.012	0.997 5	0.8-80	0.8
Coptisine	Y = 0.05X + 0.003 4	0.997 6	0.8-80	0.8
Jatrorrhizine	Y = 0.05X + 0.003 5	0.996 5	0.4-40	0.4
Berberrubine	Y = 0.05X + 0.000 15	0.998 7	0.8-80	0.8
Palmatine	Y = 0.11X + 0.006 6	0.995 7	0.8-80	0.8
Berberine	Y = 0.11X - 0.002 8	0.998 0	1.6-160	1.6
Ononin	Y = 0.22X - 0.000 89	0.999 8	0.5-50	0.5
Calycosin	Y = 0.10X - 0.000 70	0.999 8	0.5-50	0.5
Calycosin-7-glucoside	Y = 0.15X - 0.001 6	0.999 8	0.5-50	0.5
Formononetin	Y = 0.24X + 0.005 4	0.998 7	0.5-50	0.5
Astragaloside Ⅳ	Y = 0.003 3X + 0.000 016	0.998 6	10-1 000	10.0
Chlorogenic acid	Y = 0.003 1X + 0.000 095	0.998 3	4-400	4.0
Neochlorogenic acid	Y = 0.000 87X - 0.000 029	0.997 1	5-500	5.0
Cryptochlorogenic acid	Y = 0.000 49X + 0.000 000 65	0.995 5	5-500	5.0

Component

Regression equation

Linearity range/ng·mL^-1

LLOQ/ng·mL^-1

Magnoflorine

Y = 0.02X + 0.000 66

0.998 8

0.8-80

0.8

Epiberberine

Y = 0.16X + 0.012

0.997 5

0.8-80

0.8

Coptisine

Y = 0.05X + 0.003 4

0.997 6

0.8-80

0.8

Jatrorrhizine

Y = 0.05X + 0.003 5

0.996 5

0.4-40

0.4

Berberrubine

Y = 0.05X + 0.000 15

0.998 7

0.8-80

0.8

Palmatine

Y = 0.11X + 0.006 6

0.995 7

0.8-80

0.8

Berberine

Y = 0.11X - 0.002 8

0.998 0

1.6-160

1.6

Ononin

Y = 0.22X - 0.000 89

0.999 8

0.5-50

0.5

Calycosin

Y = 0.10X - 0.000 70

0.999 8

0.5-50

0.5

Calycosin-7-glucoside

Y = 0.15X - 0.001 6

0.999 8

0.5-50

0.5

Formononetin

Y = 0.24X + 0.005 4

0.998 7

0.5-50

0.5

Astragaloside Ⅳ

Y = 0.003 3X + 0.000 016

0.998 6

10-1 000

10.0

Chlorogenic acid

Y = 0.003 1X + 0.000 095

0.998 3

4-400

4.0

Neochlorogenic acid

Y = 0.000 87X - 0.000 029

0.997 1

5-500

5.0

Cryptochlorogenic acid

Y = 0.000 49X + 0.000 000 65

0.995 5

5-500

5.0

Component	Content/μg·mL^-1
Magnoflorine	1 166.1 ± 35.1	983.6 ± 22.8^**
Epiberberine	1 643.7 ± 7.8	1 185.6 ± 32.1^**
Coptisine	2 100.6 ± 20.1	1 863.9 ± 11.7^**
Jatrorrhizine	1 187.0 ± 20	944.8 ± 16.0^**
Berberrubine	685.4 ± 26.4	501.6 ± 15.0^**
Palmatine	3 028.0 ± 15.5	2 550.8 ± 13.5^**
Berberine	20 912.4 ± 299.7	18 809.1 ± 524.6^**
Chlorogenic acid	13 795.4 ± 503.5	11 947.8 ± 688.3^**
Neochlorogenic acid	1 151.7 ± 24.8	973.1 ± 14.3^**
Cryptochlorogenic acid	2 121.8 ± 36.9	1 858.4 ± 28.8^**

Component

Content/μg·mL^-1

With excipient

Without excipient

Magnoflorine

1 166.1 ± 35.1

983.6 ± 22.8^**

Epiberberine

1 643.7 ± 7.8

1 185.6 ± 32.1^**

Coptisine

2 100.6 ± 20.1

1 863.9 ± 11.7^**

Jatrorrhizine

1 187.0 ± 20

944.8 ± 16.0^**

Berberrubine

685.4 ± 26.4

501.6 ± 15.0^**

Palmatine

3 028.0 ± 15.5

2 550.8 ± 13.5^**

Berberine

20 912.4 ± 299.7

18 809.1 ± 524.6^**

Chlorogenic acid

13 795.4 ± 503.5

11 947.8 ± 688.3^**

Neochlorogenic acid

1 151.7 ± 24.8

973.1 ± 14.3^**

Cryptochlorogenic acid

2 121.8 ± 36.9

1 858.4 ± 28.8^**

Component	Sample	Batch
Magnoflorine	Capsule	1 203.0 ± 16.8	1 178.0 ± 17.9	1 166.2 ± 17.6	1 155.4 ± 65.4	1 181.3 ± 21.9
Without excipient	968.6 ± 13.0^**	981.0 ± 24.0^**	971.1 ± 22.8^**	951.1 ± 33.4^**	976.0 ± 18.8^**
Epiberberine	Capsule	1 539.7 ± 41.8	1 562.0 ± 50.0	1 550.2 ± 39.0	1 545.1 ± 43.0	1 570.5 ± 57.7
Without excipient	1 250.0 ± 146.1^**	1 258.0 ± 103.2^**	1 253.1 ± 51.7^**	1 219.9 ± 73.1^**	1 225.3 ± 81.1^**
Coptisine	Capsule	2 097.7 ± 31.8	2 071.3 ± 60.8	2 096.5 ± 19.6	2 102.2 ± 13.7	2 094.3 ± 35.6
Without excipient	1 744.1 ± 87.0^**	1 790.5 ± 24.7^**	1 789.7 ± 40.0^**	1 745.6 ± 18.6^**	1 771.2 ± 22.1^**
Jatrorrhizine	Capsule	1 166.3 ± 25.1	1 163.9 ± 16.7	1 176.4 ± 11.1	1 160.9 ± 17.6	1 190.3 ± 15.4
Without excipient	914.6 ± 6.7^**	924.1 ± 25.1^**	914.6 ± 19.6^**	903.7 ± 16.4^**	913.8 ± 19.6^**
Berberrubine	Capsule	653.2 ± 17.8	666.9 ± 14.4	673.2 ± 25.3	661.6 ± 12.4	650.1 ± 15.7
Without excipient	513.2 ± 15.8^**	515.7 ± 15.5^**	518.0 ± 20.8^**	516.7 ± 7.2^**	517.8 ± 13.3^**
Palmatine	Capsule	3 006.3 ± 51.1	2 986.3 ± 91.6	2 964.0 ± 28.4	2 914.8 ± 19.8	2 936.8 ± 46.4
Without excipient	2 520.2 ± 52.2^**	2 607.5 ± 77.5^**	2 599.8 ± 57.2^**	2 564.7 ± 34.4^**	2 577.8 ± 51.7^**
Berberine	Capsule	21 251.7 ± 389.4	21 457.7 ± 337.8	21 304.2 ± 100.1	20 638.4 ± 186.3	21 129.0 ± 161.0
Without excipient	17 987.4 ± 259.4^**	17 958.2 ± 103.9^**	17 622.4 ± 695.9^**	17 912.3 ± 432.4^**	18 083.6 ± 310.2^**
Ononin	Capsule	60.0 ± 1.2	59.9 ± 0.7	60.1 ± 0.9	60.1 ± 0.9	59.7 ± 1.4
Without excipient	58.8 ± 1.1^**	57.7 ± 1.2^**	58.4 ± 1.1^**	58.8 ± 1.2^**	58.6 ± 0.7^**
Calycosin	Capsule	57.7 ± 1.2	58.3 ± 0.6	58.4 ± 0.5	58.0 ± 0.4	58.4 ± 1.2
Without excipient	56.4 ± 1.1^**	56.5 ± 1.2^**	56.2 ± 0.8^**	56.4 ± 0.8^**	56.5 ± 0.6^**
Calycosin-7-glucoside	Capsule	152.1 ± 1.9	151.6 ± 2.9	150.5 ± 2.8	151.7 ± 1.1	150.6 ± 2.5
Without excipient	149.9 ± 2.8^**	148.4 ± 4.8^**	146.2 ± 2.4^**	146.6 ± 1.0^**	147.4 ± 1.2^**
Formononetin	Capsule	28.3 ± 1.0	27.8 ± 0.4	28.3 ± 0.3	28.0 ± 0.2	27.6 ± 0.4
Without excipient	26.8 ± 0.4^**	26.9 ± 0.5^**	27.2 ± 0.3^**	27.0 ± 0.2^**	27.1± 0.3^**
Astragaloside Ⅳ	Capsule	103.1 ± 3.0	100.3 ± 2.3	101.9 ± 3.7	98.6 ± 0.7	99.7 ± 2.0
Without excipient	95.2 ± 3.6^**	96.9 ± 3.7^**	97.4 ± 3.9^**	96.7 ± 2.9^**	97.6 ± 3.1^**
Chlorogenic acid	Capsule	14 671.6 ± 170.3	14 643.0 ± 330.0	14 972.4 ± 381.2	15 057.1 ± 325.9	15 095.4 ± 252.4
Without excipient	11 811.3 ± 577.7^**	11 630.7 ± 242.4^**	11 845.0 ± 255.4^**	11 840.7 ± 324.1^**	12 032.4 ± 416.1^**
Neochlorogenic acid	Capsule	1 164.8 ± 24.6	1 185.9 ± 22.9	1 162.0 ± 15.8	1 160.0 ± 15.6	1 183.3 ± 21.9
Without excipient	992.7 ± 10.3^**	992.0 ± 13.1^**	976.2 ± 8.3^**	978.0 ± 18.4^**	970.4 ± 30.9^**
Cryptochlorogenic acid	Capsule	2 088.6 ± 48.7	2 140.1 ± 64.6	2 108.2 ± 49.6	2 119.8 ± 62.1	2 115.7 ± 57.4
Without excipient	1 815.8 ± 38.8^**	1 841.3 ± 45.9^**	1 824.2 ± 41.9^**	1 807.9 ± 28.0^**	1 775.1 ± 46.0^**

Component

Sample

Batch

2403001

2403002

2403003

2404001

2404002

Magnoflorine

Capsule

1 203.0 ± 16.8

1 178.0 ± 17.9

1 166.2 ± 17.6

1 155.4 ± 65.4

1 181.3 ± 21.9

Without excipient

968.6 ± 13.0^**

981.0 ± 24.0^**

971.1 ± 22.8^**

951.1 ± 33.4^**

976.0 ± 18.8^**

Epiberberine

Capsule

1 539.7 ± 41.8

1 562.0 ± 50.0

1 550.2 ± 39.0

1 545.1 ± 43.0

1 570.5 ± 57.7

Without excipient

1 250.0 ± 146.1^**

1 258.0 ± 103.2^**

1 253.1 ± 51.7^**

1 219.9 ± 73.1^**

1 225.3 ± 81.1^**

Coptisine

Capsule

2 097.7 ± 31.8

2 071.3 ± 60.8

2 096.5 ± 19.6

2 102.2 ± 13.7

2 094.3 ± 35.6

Without excipient

1 744.1 ± 87.0^**

1 790.5 ± 24.7^**

1 789.7 ± 40.0^**

1 745.6 ± 18.6^**

1 771.2 ± 22.1^**

Jatrorrhizine

Capsule

1 166.3 ± 25.1

1 163.9 ± 16.7

1 176.4 ± 11.1

1 160.9 ± 17.6

1 190.3 ± 15.4

Without excipient

914.6 ± 6.7^**

924.1 ± 25.1^**

914.6 ± 19.6^**

903.7 ± 16.4^**

913.8 ± 19.6^**

Berberrubine

Capsule

653.2 ± 17.8

666.9 ± 14.4

673.2 ± 25.3

661.6 ± 12.4

650.1 ± 15.7

Without excipient

513.2 ± 15.8^**

515.7 ± 15.5^**

518.0 ± 20.8^**

516.7 ± 7.2^**

517.8 ± 13.3^**

Palmatine

Capsule

3 006.3 ± 51.1

2 986.3 ± 91.6

2 964.0 ± 28.4

2 914.8 ± 19.8

2 936.8 ± 46.4

Without excipient

2 520.2 ± 52.2^**

2 607.5 ± 77.5^**

2 599.8 ± 57.2^**

2 564.7 ± 34.4^**

2 577.8 ± 51.7^**

Berberine

Capsule

21 251.7 ± 389.4

21 457.7 ± 337.8

21 304.2 ± 100.1

20 638.4 ± 186.3

21 129.0 ± 161.0

Without excipient

17 987.4 ± 259.4^**

17 958.2 ± 103.9^**

17 622.4 ± 695.9^**

17 912.3 ± 432.4^**

18 083.6 ± 310.2^**

Ononin

Capsule

60.0 ± 1.2

59.9 ± 0.7

60.1 ± 0.9

59.7 ± 1.4

Without excipient

58.8 ± 1.1^**

57.7 ± 1.2^**

58.4 ± 1.1^**

58.8 ± 1.2^**

58.6 ± 0.7^**

Calycosin

Capsule

57.7 ± 1.2

58.3 ± 0.6

58.4 ± 0.5

58.0 ± 0.4

58.4 ± 1.2

Without excipient

56.4 ± 1.1^**

56.5 ± 1.2^**

56.2 ± 0.8^**

56.4 ± 0.8^**

56.5 ± 0.6^**

Calycosin-7-glucoside

Capsule

152.1 ± 1.9

151.6 ± 2.9

150.5 ± 2.8

151.7 ± 1.1

150.6 ± 2.5

Without excipient

149.9 ± 2.8^**

148.4 ± 4.8^**

146.2 ± 2.4^**

146.6 ± 1.0^**

147.4 ± 1.2^**

Formononetin

Capsule

28.3 ± 1.0

27.8 ± 0.4

28.3 ± 0.3

28.0 ± 0.2

27.6 ± 0.4

Without excipient

26.8 ± 0.4^**

26.9 ± 0.5^**

27.2 ± 0.3^**

27.0 ± 0.2^**

27.1± 0.3^**

Astragaloside Ⅳ

Capsule

103.1 ± 3.0

100.3 ± 2.3

101.9 ± 3.7

98.6 ± 0.7

99.7 ± 2.0

Without excipient

95.2 ± 3.6^**

96.9 ± 3.7^**

97.4 ± 3.9^**

96.7 ± 2.9^**

97.6 ± 3.1^**

Chlorogenic acid

Capsule

14 671.6 ± 170.3

14 643.0 ± 330.0

14 972.4 ± 381.2

15 057.1 ± 325.9

15 095.4 ± 252.4

Without excipient

11 811.3 ± 577.7^**

11 630.7 ± 242.4^**

11 845.0 ± 255.4^**

11 840.7 ± 324.1^**

12 032.4 ± 416.1^**

Neochlorogenic acid

Capsule

1 164.8 ± 24.6

1 185.9 ± 22.9

1 162.0 ± 15.8

1 160.0 ± 15.6

1 183.3 ± 21.9

Without excipient

992.7 ± 10.3^**

992.0 ± 13.1^**

976.2 ± 8.3^**

978.0 ± 18.4^**

970.4 ± 30.9^**

Cryptochlorogenic acid

Capsule

2 088.6 ± 48.7

2 140.1 ± 64.6

2 108.2 ± 49.6

2 119.8 ± 62.1

2 115.7 ± 57.4

Without excipient

1 815.8 ± 38.8^**

1 841.3 ± 45.9^**

1 824.2 ± 41.9^**

1 807.9 ± 28.0^**

1 775.1 ± 46.0^**