Qualitative and Quantitative Analysis of Thirteen Marker Components in Traditional Korean Formula, Samryeongbaekchul-san using an Ultra-Performance Liquid Chromatography Equipped with Electrospray Ionization Tandem Mass Spectrometry

Chang-Seob Seo; Hyeun-Kyoo Shin

doi:10.20307/nps.2016.22.2.93

Journal List > Nat Prod Sci > v.22(2) > 1060648

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Seo and Shin: Qualitative and Quantitative Analysis of Thirteen Marker Components in Traditional Korean Formula, Samryeongbaekchul-san using an Ultra-Performance Liquid Chromatography Equipped with Electrospray Ionization Tandem Mass Spectrometry

Original Article

Natural Product Sciences 2016;22(2):93-101.

Published online: 17 December 2016

DOI: https://doi.org/10.20307/nps.2016.22.2.93

Qualitative and Quantitative Analysis of Thirteen Marker Components in Traditional Korean Formula, Samryeongbaekchul-san using an Ultra-Performance Liquid Chromatography Equipped with Electrospray Ionization Tandem Mass Spectrometry

Chang-Seob Seo, Hyeun-Kyoo Shin^∗

K-her Research Center, Korea Institute of Oriental Medicine, 1672 Yuseongdae-ro, Yuseong-gu, Daejeon 34054, Korea

^∗Author for correspondence Hyeun-Kyoo Shin, K-herb Research Center, Korea Institute of Oriental Medicine, 1672 Yuseong-daero, Yuseong-gu, Daejeon 34054, Korea Tel: +82-42-868-9464; E-mail: hkshin@kiom.re.kr

Received 2 November 2015 Revised 26 November 2015 Accepted 2 December 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

For efficient quality control of the Samryeongbaekchul-san decoction, a powerful and accurate an ultra-performance liquid chromatography (UPLC) coupled with electrospray ionization (ESI) tandem mass spectrometry (MS) method was developed for quantitative analysis of the thirteen constituents: allantoin (1), spinosin (2), liquiritin (3), ginsenoside Rg1 (4), liquiritigenin (5), platycodin D2 (6), platycodin D (7), ginsenoside Rb1 (8), glycyrrhizin (9), 6-gingerol (10), atractylenolide III (11), atractylenolide II (12), and atractylenolide I (13). Separation of the compounds 1–13 was performed on a UPLC BEH C₁₈ column (2.1 × 100 mm, 1.7 μm) at a column temperature of 40^oC with a gradient solvent system of 0.1% (v/v) formic acid aqueous-acetonitrile. The flow rate and injection volume were 0.3 mL/min and 2.0 μL. Calibration curves of all compounds were showed good linearity with values of the correlation coefficient ≥ 0.9920 within the test ranges. The values of limits of detection and quantification for all analytes were 0.04–4.53 ng/mL and 0.13–13.60 ng/mL. The result of an experiment, compounds 2, 6, 12, and 13 were not detected while compounds 1, 3–5, and 7–11 were detected with 1,570.42, 5,239.85, 299.35, 318.88, 562.27, 340.87, 12,253.69, 73.80, and 115.01 μg/g, respectively.

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Keywords: Qualitative analysis, Quantitative analysis, Samryeongbaekchul-san, UPLC, LC-MS/MS

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Fig. 1.

Chemical structures of the compounds 1–13 in Samryeongbaekchul-san.

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Fig. 2.

M ass spectra of the precursor ion (Q1, A) and product ion (Q3, B) for LC-MS/MS MRM mode of the compounds 1–13. Allantoin (1), spinosin (2), liquiritin (3), ginsenoside Rg1 (4), liquiritigenin (5), platycodin D2 (6), platycodin D (7), ginsenoside Rb1 (8), glycyrrhizin (9), 6-gingerol (10), atractylenolide III (11), atractylenolide II (12), and atractylenolide I (13).

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Fig. 2.

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Fig. 2.

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Fig. 3.

Total ion chromatograms of the standard solution (A) and Samryeongbaekchul-san sample (B) by LC-MS/MS MRM mode. Allantoin (1), spinosin (2), liquiritin (3), ginsenoside Rg1 (4), liquiritigenin (5), platycodin D2 (6), platycodin D (7), ginsenoside Rb1 (8), glycyrrhizin (9), 6-gingerol (10), atractylenolide III (11), atractylenolide II (12), and atractylenolide I (13).

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Table 1.

Composition of Samryeongbaekchul-san

Herbal medicine	Scientific name	Family	Origin	Amount (g)
Ginseng Radix	Panax ginseng C. A. Meyer	Araliaceae	Yeongju, Korea	11.250
Atrctylodes Rhizoma Alba	Atractylodes macrocephala Koidzumi	Compositae	China	11.250
Poria Sclerotium	Poria cocos Wolf	Polyporaceae	Pyeongchang, Korea	11.250
Dioscoreae Rhizoma	Dioscorea batatas Decaisne	Dioscoreaceae	Andong, Korea	11.250
Glycyrrhizae Radix et Rhizoma	Glycyrrhiza uralensis Fischer	Leguminosae	China	11.250
Coicis Semen	Coix lacryma-jobi L. var. Ma-yuen Stapf	Gramineae	Muju, Korea	5.625
Nelumbinis Semen	Nelumbo nucifera Gaertner	Nymphaeaceae	Vietnam	5.625
Platycodonis Radix	Platycodon grandiflorum A. De Candolle	Campanulaceae	Muju, Korea	5.625
Dolichoris Semen	Dolichos lablab L.	Leguminosae	China	5.625
Amomi Fructus	Amomum villosum Loureiro	Zingiberaceae	Laos	5.625
Zingiferis Rhizoma Recnes	Zingiber officinale Roscoe	Zingiberaceae	Ulsan, Korea	3.750
Zizyphi Fructus	Zizyphus jujube Miller var. inermis Rehder	Rhamnaceae	Yeongcheon, Korea	3.750
Total amount				91.875

Table 2.

Conditions for the LC-MS/MS analysis of Samryeongbaekchul-san

HPLC condition
Column	ACQUITY UPLC BEH C₁₈ (100 × 2.1 mm, 1.7 µ m)
Flow rate	0.3 mL/min
Injection volume	2.0 µ L
Column temperature	45°C
Sample temperature	5°C
Mobile phase	Time (min)	A (%)^a	B (%)^b
	0	80	20
	0.1	80	20
	14.0	5	95
	15.0	0	100
	15.1	80	20
	18.1	80	20
MS condition
Capillary voltage (kV)	3.3
Extract voltage (V)	3.0
Source temp. (°C)	120
RF lens (V)	0.3
Desolvation temp. (°C)	300
Desolvation gas (L/h)	600
Cone gas (L/h)	50
Collision gas (mL/min)	0.14

^a 0.1% (v/v) formic acid in water,

^b Acetonitrile

Table 3.

Linearities, regression equation, correlation coefficients, LOD, and LOQ for the compounds 1–13

Analyte	Linear range (ng/mL)	Regression equation^a	Correlation coefficient	LOD^b (ng/mL)	LOQ^c (ng/mL)
1	0–500	y = 0.50 x – 9.16	0.9922	4.53	13.60
2	0–500	y = 4.02 x – 9.16	0.9987	0.18	0.53
3	0–500	y = 5.62x – 11.57	0.9997	0.18	0.53
4	0–500	y = 1.03 x – 12.52	0.9974	1.11	3.33
5	0–500	y = 15.87 x + 28.64	0.9996	0.06	0.18
6	0–500	y = 0.29 x – 3.61	0.9923	2.79	8.37
7	0–500	y = 1.44 x – 2.69	0.9978	0.96	2.89
8	0–500	y = 0.70 x – 4.92	0.9920	0.94	2.81
9	0–500	y = 1.56 x – 18.10	0.9964	0.46	1.38
10	0–500	y = 5.72 x – 43.06	0.9982	0.04	0.13
11	0–500	y = 21.20 x + 41.06	0.9997	0.28	0.85
12	0–500	y = 28.32 x + 172.36	0.9993	0.12	0.37
13	0–500	y = 35.25 x + 133.65	0.9996	0.06	0.17

^a y: peak area of compounds; x: concentration (ng/mL) of compounds,

^b LOD = 3 × signal-to-noise ratio,

^c LOQ = 10 × signal-to-noise ratio.

Table 4.

Mass detection condition of the compounds 1–13

Analyte	Molecular weight (Da)	Ionization mode	Retention time (min)	Precursor ion (m/z)	Product ion (m/z)	Cone voltage (V)	Collision energy (eV)
1	158.12	[M− H]⁻	0.80	157.0	96.9	20	15
2	608.54	[M+H]⁺	1.36	609.6	327.1	40	25
3	418.39	[M− H]⁻	1.65	417.4	255.3	30	15
4	801.01	[M− H]⁻	2.69	800.4	637.0	50	20
5	256.25	[M+H]⁺	2.98	257.1	137.0	35	25
6	1387.48	[M− H]⁻	3.29	1386.4	843.6	45	48
7	1225.32	[M− H]⁻	3.38	1224.1	469.4	45	48
8	1109.29	[M− H]⁻	4.34	1107.4	178.8	50	45
9	822.93	[M− H]⁻	5.20	821.8	351.0	45	40
10	294.39	[M+H]⁺	6.16	295.2	177.2	13	10
11	248.32	[M+H]⁺	6.71	249.3	231.2	25	10
12	232.32	[M+H]⁺	8.24	233.2	187.1	35	15
13	230.13	[M+H]⁺	9.33	231.2	185.1	35	20

Table 5.

Contents of the compounds 1–13 in Samryeongbaekchul-san (n = 3)

Compound	Amount (µ g/g)			Source
Compound	Mean	SD	RSD (%)	Source
1	1,570.42	94.55	6.02	D. batatas
2	N.D.	–	–	Z. jujube
3	5,239.85	87.79	1.68	G. uralensis
4	299.35	9.78	3.27	P. ginseng
5	318.88	25.30	7.94	G. uralensis
6	ND	–	–	P. grandiflorum
7	562.27	22.83	4.06	P. grandiflorum
8	340.87	115.46	33.87	P. ginseng
9	12,253.69	437.80	3.57	G. uralensis
10	73.80	10.44	14.14	Z. officinale
11	115.01	4.16	3.62	A. macrocephala
12	N.D.	–	–	A. macrocephala
13	N.D.	–	–	A. macrocephala

^a N.D. means not detected.

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