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

TOOLS
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

Copyright © 2016 The Korean Society of Pharmacognosy

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 40oC 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.

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

REFERENCES

(1). Qiu J.Nat. Rev. Drug Discov. 2007; 6:506–507.
(2). Wang L., Zhou G. B., Liu P., Song J. H., Liang Y., Yan X. J., Xu F., Wang B. S., Mao J. H., Shen Z. X., Chen S. J., Chen Z.Proc. Natl. Acad. Sci. U. S. A. 2008; 105:4826–4831.
(3). Chen H., Wu M., Kubo K. Y. J.Ethnopharmacol. 2012; 142:80–85.
(4). Lee S. R.Korean J. Orient. Physiol. Pathol. 2009; 23:374–380.
(5). Heo J.Dongeuibogam; Namsandang: Seoul. 2004; 440.
(6). Yang Q. H., Xu Y. J., Liu Y. Z., Liang Y. J., Feng G. F., Zhang Y. P., Xing H. J., Yan H. Z, Li Y. Y.Evid. Based Complement. Alternat. Med. 2014; 2014:671013–671021.
(7). Yang Q. H., Xu Y. J., Feng G. F., Hu C. F., Zhang Y. P., Cheng S. B., Wagn Y. P., Gong X. W. Afr. J.Tradit. Complement. Altern. Med. 2013; 11:213–221.
(8). Liu X., Jia Y., Iiu Q., Wu J.Lishizhen Med. Mater. Med. Res. 2011; 22:2671–2673.
(9). You Y., Liu U., Gao S.Chinese J. Experiment. Trad. Med. Formulae. 2012; 18:136–140.
(10). Turner R., Stamp L. K., Kettle A. J. J.Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2012; 891-892:85–89.
(11). Ren Y., Wang P., Wu C., Zhang J., Niu C.Biomed. Chromatogr. 2013; 27:17–26.
crossref
(12). Wang Y., Yang L., He Y. Q., Wang C. H., Welbeck E. W., Bligh S. W., Wang Z. T.Rapid Commun. Mass Spectrom. 2008; 22:1767–1778.
(13). Zhao J., Su C., Yang C., Liu M., Tang L., Su W., Liu Z. J.Pharm. Biomed. Anal. 2012; 64-65:94–97.
(14). Tan G., Zhu Z., Jing J., Lv L., Lou Z., Zhang G., Chai Y.Biomed. Chromatogr. 2011; 25:913–924.
(15). Na Y. C., Ha Y. W., Kim Y. S., Kim K. J. J.Chromatogr. A. 2008; 1189:467–475.
(16). Chen L., Qi J., Chang Y. X., Zhu D., Yu B. J.Pharm. Biomed. Anal. 2009; 50:127–137.

Fig. 1.
Chemical structures of the compounds 1–13 in Samryeongbaekchul-san.
nps-22-93f1.tif
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).
nps-22-93f2a.tif
Fig. 2.
contineud
nps-22-93f2b.tif
Fig. 2.
contineud
nps-22-93f2c.tif
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).
nps-22-93f3.tif
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 equationa Correlation coefficient LODb (ng/mL) LOQc (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
Mean SD RSD (%)
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.

TOOLS
Similar articles