rapid screening and identification of sesquiterpene ...€¦ · quiterpene lactones were...

– 150 –

Chinese Journal of Natural Medicines 2018, 16(2): 01500160

doi: 10.3724/SP.J.1009.2018.00150

Chinese Journal of Natural Medicines

Rapid screening and identification of sesquiterpene lactones in Kudiezi injection based on high-performance liquid chromatog-raphy coupled with linear ion trap-orbitrap mass spectrometry

LIU Rong-Rong1∆, ZHANG Xiu-Ping1∆, WANG Fang1, SHANG Zhan-Peng1, WANG Fei1, LIU Ying2, LU Jian-Qiu3*, ZHANG Jia-Yu2*

1 School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 100029, China; 2 Beijing Research Institution of Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China;

3 Library of Beijing University of Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China

Available online 20 Feb., 2018

[ABSTRACT] Sesquiterpene lactones are considered as the major active compounds in Kudiezi injection in virtue of their special structures and activities. Herein, an analytical method was developed for rapid screening and identification of sesquiterpene lac-tones in Kudiezi injection using high-performance liquid chromatography coupled with linear ion trap-orbitrap mass spectrometry (HPLC-LTQ-Orbitrap) in negative ion mode. First, two sesquiterpene lactone reference standards were analyzed to obtain their characteristic ESI-MS/MS fragmentation patterns. Second, based on extracted ion chromatography (EIC) data-mining method and characteristic fragmentation pathways analysis, sesquiterpene lactones in Kudiezi injection were rapidly screened and identified. Finally, an important parameter Clog P was adopted to discriminate the isomers of sesquiterpene lactones. As a result, 50 ses-quiterpene lactones were characterized, including 9 sesquiterpene lactone aglycones, 39 sesquiterpene lactone glycosides, and 2 amino acid-sesquiterpene lactone conjugates. Among them, 13 compounds were tentatively identified as new compounds. The results demonstrated that the established method would be a rapid, effective analytical tool for screening and identification of ses-quiterpene lactones in the complex system of natural medicines.

[KEY WORDS] HPLC-LTQ-Orbitrap; Sesquiterpene lactones; Characteristic fragmentation pathways; Kudiezi injection

[CLC Number] R917 [Document code] A [Article ID] 2095-6975(2018)02-0150-11

Introduction

Ixeris sonchifolia (Bge.) Hance, belonging to the compo-sitae family, is a kind of bitter perennial herb that is distrib-uted and cultivated widely in northeastern China. It has been traditionally used as folk medicine for invigorating blood circulation, dissipating blood stasis to relieve pain, and nor-malizing menstruation [1]. As a preparation extracted and re-fined from the whole herb of I. sonchifolia, Kudiezi injection [Received on]16-June-2017 [Research funding] This work was supported by Beijing Nova Pro-

gram (No. Z171100001117029) and the National Natural Science

Foundation of China (No. 81503244).

[*Corresponding author] Tel: 86-10-64287540, E-mail: [email protected] (ZHANG Jia-Yu); Tel: 86-10-64287502, E-mail: [email protected] (LU Jian-Qiu). ∆These two authors contributed equally to this work.

These authors have no conflict of interest to declare.

has been widely used to treat coronary heart disease, effort angina, and acute cerebral infarction [2-3]. The chemical con-stituents of Kudiezi injection are quite complicated, including sesquiterpene lactones, flavonoids, triterpenes and amino acids, which have been reported to possess anti-inflammatory, anti-atherosclerotic, antioxidant and anticancer activities [4–7]. Until now, few reports are available on systematic analysis of sesquiterpene lactones in Kudiezi injection.

Among many different LC-MS platforms, high-resolution

mass spectrometry (HRMS) has exhibited excellent perfor-

mances for components identification because of its high

efficiency, sensitivity, and selectivity [8-9]. The hybrid linear

ion trap-orbitrap mass spectrometer (LTQ-Orbitrap MS) com-

bined high trapping capacity and MSn scanning function of

linear ion trap along with accurate mass measurements within

5 ppm and a resolving power of up to 100 000 over a wider

dynamic range than many other mass spectrometers [10–12].

Meanwhile, the combined application of tandem mass spec-

LIU Rong-Rong, et al. / Chin J Nat Med, 2018, 16(2): 150160

– 151 –

trometry for identifying the complicated constituents in tradi-

tional Chinese medicines (TCMs) would generate a large

quantity of information data, such as elemental compositions

and fragmentation patterns of multiple-stage. In addition,

off-line data processing also plays an important role in the

identification of components, such as extracted ion chroma-

tography (EIC) data-mining method with the remarkable mer-

its of purifying total ion chromatography (TIC) spectra and

facilitating the identification of unknown chromatographic

peaks [13-14]. These advantages have made LTQ-Orbitrap MS

one of the most powerful approaches for the rapid identifica-

tion of multiple constituents in TCMs.

In the present study, an HPLC-LTQ-Orbitrap data-acqui-sition approach combined with EIC data-mining technique was established for comprehensive identification of ses-quiterpene lactones and their derivatives in Kudiezi injection. In addition, an important parameter Clog P was used to dis-criminate the isomers of sesquiterpene lactones.

Materials and Methods

Chemicals The reference standards of Ixerin Z and 11, 13α-dihydroi-

xerin Z (purity greater than 98%) were previously extracted, isolated and identified from I. sonchifolia in our laboratory [15]. Their structures (Fig. 1) were fully elucidated by 1H, 13C NMR and ESI-MS data [16-17]. The commercial products of Kudiezi injection produced by Tonghua Huaxia Pharmaceuti-cal Co., Ltd. (Jilin, China) were purchased by prescription from Dongfang Hospital in Beijing, China. HPLC grade for-mic acid, acetonitrile, and methanol were purchased from Merck (Darmstadt, Germany). Ultra-pure water was produced by a Milli-Q purification system (Millipore, Bedford, MA, USA) in the experiment. Sample preparation

The standard solutions of Ixerin Z and 11, 13α-dihydroi-xerin Z were prepared in methanol at concentrations of 100 µg·mL−1. All the standard solutions were stored at 4 ºC until use. Kudiezi injection was filtered through a 0.22 µm membrane, and then an aliquot of 10 µL of the subsequent filtrate was injected into LC-MS system for analysis. Instrumentation and conditions

HPLC analysis was carried out on an Accela HPLC system equipped with a binary pump and an autosampler (Thermo Scientific, Bremen, Germany). The compounds were sepa-rated on a Thermo Hypersil BDS C18 column (250 mm × 4.6 mm i.d., 5 μm) at room temperature. The mobile phase consisted of 0.1% (V/V) formic acid in water (A) and acetonitrile (B) with the elution gradient set as follows: 0−18 min, 2%−8% B; 18−36 min, 8%−12% B; 36−55 min, 12%−18% B; 55−70 min, 18%−25% B; 70−80 min, 25%−30% B; 80−85 min, 30%−40% B. The flow rate was set at 1.0 mL·min–1.

LTQ-Orbitrap XL mass spectrometer (Thermo Scientific, Bremen, Germany) was connected to the LC system via an electrospray ionization (ESI) interface. HPLC effluent was

introduced into the ESI source in a post-column splitting ratio of 1 : 4. Full scan data acquisition was performed from m/z 100 to 600 in negative ion mode. The important ESI parame-ters were set as follows: sheath gas (nitrogen) flow rate of 30 arb.; auxiliary gas (nitrogen) flow rate of 10 arb.; capillary temperature of 350 ºC; electrospray voltage of 3.0 kV; capil-lary voltage of −35 V; and tube lens voltage of −110 V. The resolution of Orbitrap analyzer was set at 30 000 with data-dependent ESI-MS2 analysis triggered by the three most abundant ions from one-stage mass spectrometry scanning, followed by ESI-MS3 analysis of the most abundant product ions. Collision-induced dissociation (CID) was performed in LTQ with an activation q of 0.25 and activation time of 30 ms. The isolation width was 2 amu, and the normalized collision energy was set to 35%. Data were acquired and analyzed using Xcalibur data 2.1 software (Thermo Scientific, Bremen, Ger-many).

Results and Discussion

Overview of analysis approaches of experimental results Owing to the low content of sesquiterpene lactones in

Kudiezi injection, a sensitive and reliable HPLC-LTQ-Orbi-trap approach was established for achieving their accurate mass and molecular formula. Meanwhile, considering the lack of reference standards, the characteristic fragmentation path-ways of sesquiterpene lactones were concluded and then used for rapid identification of the other sesquiterpene lactones in Kudiezi injection. Furthermore, sesquiterpene lactone isomers were differentiated by Clog P parameter. As a result, a total of 50 sesquiterpene lactones in Kudiezi injection were screened and divided into sesquiterpene lactone aglycones, sesquiter-pene lactone glycosides, and conjugates of amino acid-sesqui-terpene lactones. The fragmentation patterns of reference standards

Ixerin Z exhibited its [M – H]– ion at m/z 421.149 3 (C21H25O9) with mass errors within 5 ppm. It produced the base peak ion at m/z 259 by neutral loss of a dehydrated glu-cose. The ion at m/z 259 generated prominent ions at m/z 215 and m/z 241 through losing CO2 and H2O, respectively. Moreover, the product ion at m/z 241 further generated the predominant ion at m/z 197 by loss of one molecular of CO2. The ion at m/z 215 was also followed by loss of H2O or CO, yielding two minor ions at m/z 197 ([M – H – 162 – CO2 – H2O]–) and m/z 187 ([M – H – 16 – CO2 – CO]–) (Fig. 2).

11, 13α-Dihydroixerin Z firstly produced its base peak ion at m/z 261 via a glycosidic bond cleavage, and then gen-erated two minor ions at m/z 217 ([M – H – 162 – CO2]

–) and m/z 199 ([M – H – 162 – H2O]–). Meanwhile, the ion at m/z 187 was also observed in its MS3 spectrum, corresponding to [M – H – 162 – C3H6O2]

– due to the fragmentation of the α-methyl-γ-lactone in 11, 13α-dihydroixerin Z with saturated C11-13 bond (Fig. 3).

Therefore, the characteristic fragment ions of reference standards were deduced, such as [M – H – 44]– generated by


– 152 –

loss of CO2 from α-methyl-γ-lactone moiety, as well as [M – H – 162]–, [M – H – 162 – 44]–, [M – H – 162 – 44 – 28]– and [M – H – 162 – 44 – 18]–, etc (Figs. 2 and 3). According to

the above characteristic fragmentation patterns, sesquiterpene lactones in Kudiezi injection could be rapidly screened and identified.

Fig. 1 The identified chemical structures of sesquiterpene lactones in Kudiezi injection

Identification of sesquiterpene lactone aglycones The molecular formula of sesquiterpene lactones were

predicted by high resolution mass spectrometry data built in-house. Combined with multi-stage mass spectrometry fragmentation information and bibliography data, three cate-gories of sesquiterpene lactone and its derivatives were screened and determined, including 9 sesquiterpene lactone aglycones, 39 sesquiterpene lactone glycosides, and 2 amino acid-sesquiterpene lactone conjugates. The EIC chroma-tograms are shown in Fig. 4.

Compounds 1 and 2 yielded identical [M – H]– ions at m/z 247.096 2 (C14H15O4) with mass errors within 5 ppm in negative ion mode. Both of their deprotonated molecular ions generated a serial of fragment ions at m/z 203, 219, 229, 185 and 187, corresponding to [M – H – CO2]

–, [M – H – CO]–, [M – H – H2O]–, [M – H – CO2 – H2O]– and [M – H – C2H4O2]

–. Compound 1 generated the fragment ion at m/z 187 due to the overall fracture of α-methyl-γ-lactone ring in ses-

quiterpene lactones. The parent nucleus of sesquiterpene lac-tones (C12H11O3) was usually substituted in C-3, C-4, C-9, C-11 positions and the substituents were -OH, -CH3, -CH2OH, -Glc, etc (Fig. 1). Therefore, it could be deduced that one -OH and two -CH3 or one -CH2OH and one -CH3 could be added to their structures. To the best of our knowledge, these two sesquiterpene lactones were identified as new compounds and their structures are shown in Fig. 1.

Compound 3 produced its [M – H]– ion at m/z 249.148 2 (C15H21O3) with mass errors within 5 ppm. Upon CID mode, it further generated [M – H – CO2]

– ion at m/z 205, consistent with the characteristic fragmentation pathway of α-methyl-γ- lactone ring. Besides, its [M – H]– ion also yielded fragment ions at m/z 187 [M – H – CO2 – H2O]–, m/z 221 [M – H – CO]–, m/z 175 [M – H – C3H6O2]

– in its MS3 spectrum. The presence of m/z 175 showed that there was one -CH3 linked with C-11. According to the literature [18], compound 3 was tentatively characterized as Sonchifoliasolide M.


– 153 –

Fig. 2 The proposed fragmentation pathways of Ixerin Z

Compounds 4, 5, and 6 were all observed with the same [M – H]– ions at m/z 259.096 5 (C15H15O4) with mass errors within 5 ppm. They all produced the base peak [M – H – CO2]

– ions at m/z 215 (C15H15O4) in the ESI-MS2 spectra, corresponding to the characteristic fragmentation pathway of α-methyl-γ-lactone ring. Combined with bibliography data [18-20] and the values of Clog P, these three compounds were tenta-tively deduced as 8β-Hydroxydehydrozaluzanin C, 15-hydroxy- 1(10), 11(13), 3-guaiatriene-12, 6-olide-2-one, and 3-Hydro-xydehydroleucodin, respectively.

Compounds 7, 8, and 9 generated the identical [M – H]– ions at m/z 277.107 0 (C15H17O5) with mass errors within 5 ppm. After the CID cleavage, further fragmentation resulted

in [M – H – CO2 – H2O]– ion at m/z 215, [M – H – H2O]– ion at m/z 259 and [M – H – CO2]

– ion at m/z 233, consistent with the characteristic fragmentation pathway of sesquiterpene lac-tones. According to the literature [21] and the values of Clog P, compounds 7, 8, and 9 were plausibly characterized as 11β, 13- dihydrolacurin, sonchifoliactone E, and sonchifoliactone E isomer. Identification of sesquiterpene lactone glycosides

In this experiment, five kinds of elemental composition of sesquiterpene lactone glycosides were screened and char-acterized, including C21H25O9, C21H27O9, C21H29O9, C21H27O10 and C21H27O11 by LTQ-Orbitrap MS spectra. After losing of 162 Da or 180 Da, they produced aglycone ions at m/z 259, m/z 261, m/z 259, m/z 263, m/z 277, and m/z 293, respectively,


– 154 –

Fig. 3 The proposed fragmentation pathways of 11, 13α-dihydroixerin Z

indicating that there was a glucose group in their respective molecular structures. Furthermore, their fragment ions in the ESI-MSn spectra were in line with the fragmentation path-ways of sesquiterpene lactones.

Compounds 10−16 showed the same [M – H]– ions at m/z 421.149 3 (C21H25O9) with mass errors within 5 ppm. First, they were divided into two categories based on the character-istic fragmentation pathways deduced above. Compounds 11, 12, and 13 yielded [M – H – 44 – 74 – 18 – 28]– at m/z 257, [M – H – 74 – 18 – 28]– at m/z 301 and [M – H – 44 – 74 – 18 – 28]– at m/z 257 in their MS2 spectra, respectively, suggest-ing the presence of one -CH3 group in their molecular struc-tures. Combined the basic nuclear parent of sesquiterpene lactones (C12H11O3) with their accurate molecules, it could be

deduced that there were one molecule of C12H12O6, two groups of CH3, and one double bond in their structures. Fur-thermore, the fragment ions validated the above deduction. Finally, according to their Clog P values, they were tenta-tively identified and differentiated. Taking compound 11 as an example, it generated the base peak ion [M – H – 44]– at m/z 377 and yielded a serial of fragment ions, such as [M – H – 162]– at m/z 259, [M – H – 162 – 44]– at m/z 215, [M – H – 44 – 74 – 18 – 28]– at m/z 257 in its MS2 spectrum. As far as we know, there were no related literatures reported. And thus, compound 11 was deduced as a new compound (Fig. 1). For compounds 10 and 14−16, [M – H – C3H6O2]

– ion was not observed in their ESI-MS/MS spectra, suggesting there was only one double bond linked to C-11 position. Compound 15


– 155 –


– 156 –

Fig. 4 The EIC spectra of 50 detected sesquiterpene lactones, including 9 sesquiterpene lactone aglycones (a, b, c and d), 39 ses-quiterpene lactone glycosides (e, f, g, h and i), and 2 amino acid-sesquiterpene lactone conjugates (j)

was identified as Ixerin Z by comparing with the reference standard. Compared to the nucleus structure, it could be speculated that there were one C12H12O6 and two double bonds in the molecular structures of compounds 10, 14, and 16. Therefore, compounds 10 and 14 were tentatively as-signed as 15-desoxylactucin-α-D-glucopyranoside and 11, 13-dehydrolactuside C, respectively [22-23]. Meanwhile, com-pound 16 was finally identified to be a new compound.

Compounds 17−27 all gave the similar [M – H]– ions at m/z 423.165 0 (C21H27O9) with mass errors within 5 ppm. First, they were discriminated into two categories based on whether they produced the [M – H – C3H6O2]

– fragment ion. For compounds 21, 23 and 24, all of them produced the char-acteristic fragmentation ions, such as the [M – H – 74]– ion at m/z 347 in their MS2 spectra and the [M – H – 162 – 74]– ion at m/z 187 in their MS3 spectra, indicating the presence of one -CH3 group. Taking compound 21 for an example, in addition to [M – H – 74]– ion at m/z 349, it also generated a series of characteristic fragment ions, such as [M – H – 162]– ion at m/z 261, [M – H – 44]– ion at m/z 199, [M – H – 162 – 44 – 74]– ion at m/z 143. Therefore, compound 21 and 24 were plausibly characterized as 11, 13β-dihydroixerin Z and 11β, 13-dihydroixerin Z, respectively [24]. By comparing with the reference standard, compound 23 was unambiguously as-signed as Ixerin Z. In the same way, compounds 17−20, 22, and 24−27 were inferred with a double bond in C-11 position without the characteristic fragment ion [M – H – C3H6O2]

–. For example, compound 17 yielded the base peak ion at m/z

243 ([M – H – 162]–) by loss of a glucose residue. In its ESI-MS/MS spectra, it generated [M – H – 162 – 18 – 44]– at m/z 199 corresponding to the characteristic fragmentation pathway of sesquiterpene lactones. Compound 25 generated the base peak ion at m/z 261 ([M – H – 180]–) by loss of a dehydrated glucose. Further fragmentation of the ion at m/z 261 yielded two minor ions [M – H – 162 – 44]– at m/z 217 and [M – H – 162 – 44 – 18]– at m/z 199. Among them, m/z 217 was a characteristic fragment ion by the loss of CO2 from the α-methyl-γ-lactone. Besides, the [M – H – 44]– at m/z 379 ion was also observed. According to the above method, the other compounds were all identified (Table 1) [25-27].

Compounds 28−35 produced the same [M – H]– ions at m/z 425.180 6 (C21H29O9) with mass errors within 5 ppm. Based on the fragmentation patterns, they were divided into two categories. None but compound 31 exhibited the charac-teristic fragment ion [M – H – 162 – 74]– at m/z 189, sug-gesting the presence of one -CH3 group. It was plausibly de-scribed as 11, 13-dihydroixerinoside or Sonchifoliasolide J isomer [18]. For compounds 28−30 and 32−35, it could be concluded that there was a double bond in their C-11 position. For instance, compounds 29, 34, and 35 generated character-istic fragment ions [M – H – 162]– at m/z 263, [M – H – 44]– at m/z 381, [M – H – 28]– at m/z 397, [M – H – 162 – 28]– at m/z 235, [M – H – 162 – 18 – 44]– at m/z 201, etc. With reference to literature [18, 27] and the Clog P values, they were tentatively identified as 3-O-β-D-glucopyranosyl-8β- hydroxyguauan-10(14)-ene-6, 12-olide, Sonchifoliasolide


– 157 –


– 158 –


– 159 –

N or their isomer Sonchifoliasolide L and Ixerisoside C, respectively. According to the parent nucleus and fragment ions, compounds 30, 32, and 33 were deduced and as-signed as new compounds (Fig. 1).

Compounds 36−47 generated the same [M – H]– ions at m/z 439.159 9 (C21H27O10) with mass errors within 5 ppm. In the same way, they were divided into two groups. Compounds 36, 37, 39, 42, and 45 generated the same characteristic frag-ment ions [M – H – 74]– at m/z 365, [M – H – 74 – 44]– at m/z 321, [M – H – 162 – 74]– at m/z 203 in their ESI-MS/MS spectra, indicating the presence of one -CH3 group in their molecular structures. For others, it could be deduced that there was a double bond in C-11 position. For compound 45, besides the characteristic fragment ion ([M – H – 162 – 74]–), the base peak ion [M – H – 162 – 44]– at m/z 233, the minor ions [M – H – 44]– at m/z 395, [M – H – 162]– at m/z 277, [M – H – 180]– at m/z 259 were also detected. It was plausibly as-signed as Sonchifolactone D [28]. For compound 43, the char-acteristic ion [M – H – 74]– was not detected, suggesting there was a double bond linked in its C-11 position. Based on its fragment ions, such as [M – H – 162]– at m/z 277, [M – H – 162 – 44]– at m/z 233, [M – H – 18]– at m/z 421, [M – H – 44]– at m/z 395, it was tentatively deduced as 3, 10β-dihydroxy-2-oxo-guaia-3, 11(13)-dien-1α, 5α, 6α, 7αH-12, 6-olide-10-O-β-D-glucopyranoside [17]. Similarly, other compounds were successively identified (Table 1).

Compound 48 yielded [M – H]– ion at m/z 455.154 79 (C21H27O11) with mass errors within 5 ppm. The deprotonated molecular ion yielded the base peak ion [M – H – 162]– at m/z 293, suggesting the presence of one molecular of glucose group. In its ESI-MS/MS spectra, it generated the characteris-tic ion [M – H – 162 – 74]– at m/z 219. It was tentatively as-signed as Sonchifolactone C [28]. Identification of amino acid-sesquiterpene conjugates

Compounds 49 and 50 exhibited their respective [M – H]– ion at m/z 536.212 5 and m/z 536.211 0, which were con-sistent with the elemental composition of C26H34NO11 within 5 ppm mass errors. In their ESI-MS/MS spectra, they both generated the base peak at m/z 421, which was identical to the fragment ion at m/z 421 of sesquiterpene lactone glycosides, suggesting that they were sesquiterpene lactone glycoside derivatives and the elemental composition of the derivative group was C5H9NO2. Several amino acids have been reported from I. sonchifolia, and the content of proline (molecular formula C5H9NO2) was highest. Finally, compounds 49 and 50 were tentatively identified as amino acid-sesquiterpene lactone conjugates [29-30].

Conclusion

In the present study, a sensitive HPLC-LTQ-Orbitrap coupled with EIC data-mining method was established for the rapid characterization of sesquiterpene lactones in Kudiezi injection. According to the characteristic fragmentation path-ways of the reference standards, the sesquiterpene lactones

were divided into 3 categories, i.e., sesquiterpene lactone aglycones, sesquiterpene lactone glycosides and amino acid-sesquiterpene lactone conjugates. Furthermore, an im-portant parameter Clog P was used to differentiate the iso-mers of sesquiterpene lactones. As a result, 50 sesquiterpene lactones were preliminarily identified, including 37 known compounds and 13 new compounds. These compounds in-cluded 9 sesquiterpene lactone aglycones, 39 sesquiterpene lactone glycosides and 2 amino acid-sesquiterpene lactone conjugates. It was the first systematic report of sesquiterpene lactones in Kudiezi injection. The results indicated that the established method could be employed as a rapid, effective technique to screen and identify sesquiterpene lactones in Kudiezi injection. The study also provided a significant clue for analysis of the other herbal medicines or preparations.

References

[1] Jiangsu Medical College Encyclopedia of Chinese Materia Medica [M]. Shanghai Science and Technology Press, 1977: 1300-1301.

[2] Chang YP, Li L, Xie YM, et al. Clinical outcomes research on parenterally administered kudiezi in treatment of cerebral in-farction [J]. Chin J Chin Mater Med, 2013, 38(18): 3155-3160.

[3] Liao X, Zeng XB, Xie YM, et al. Use propensity score method to analyze data about effectiveness of coronary heart disease treated by kudiezi injection [J]. Chin J Chin Mater Med, 2013, 38(18): 3172-3179.

[4] Zhang YC, Zhou L, Ng KY. Sesquiterpene lactones from Ixeris sonchifolia Hance and their cytotoxicities on A549 human non-small cell lung cancer cells [J]. J Asian Nat Prod Res, 2009, 11(4): 294-298.

[5] Lu JC, Feng XZ, Sun QS, et al. Effect of six flavonoid com-pounds from Ixeris sonchifolia on stimulus-induced superoxide generation and tyrosyl phosphorylation in human neutro-phils [J]. Clin Chim Acta, 2002, 316(1-2): 95-99.

[6] Zhang YC, He CN, Chew EH. Studies on the chemical con-stituents and biological activities of Ixeris [J]. Chem Biodiverse, 2013, 10(8): 1373-1391.

[7] Yee SB, Lee JH, Chung HY, et al. Inhibitory effects of luteolin isolated from Ixeris sonchifolia Hance on the proliferation of hepg2 human hepatocellular carcinoma cells [J]. Arch Phar-macal Res, 2003, 26(2): 151-156.

[8] Zhang JY, Wang F, Zhang H, et al. Rapid identification of polymethoxylated flavonoids in traditional Chinese medicines with practical strategy of stepwise mass defect filtering cou-pled to diagnostic product ions analysis based on a hybrid LTQ-Orbitrap mass spectrometer [J]. Phytochem Anal, 2014, 25(5): 405-414.

[9] Zhang QQ, Dong X, Liu XG, et al. Rapid separation and iden-tification of multiple constituents in Danhong Injection by ul-tra-high performance liquid chromatography coupled to elec-trospray ionization quadrupole time-of-flight tandem mass spectrometry [J]. Chin J Nat Med, 2016, 14(2): 147-160.

[10] Li Y, Liu Y, Liu RR, et al. HPLC-LTQ-Orbitrap MSn profiling method to comprehensively characterize multiple chemical constituents in xiao-er-qing-jie granules [J]. Anal Methods, 2015, 7(18): 7511-7526.

[11] Chen LW, Wang Q, Qin KM, et al. Chemical profiling of


– 160 –

Qixue Shuangbu Tincture by ultra-performance liquid chro-matography with electrospray ionization quadrupole-time-of- flight high-definition mass spectrometry(UPLC-QTOF/MS) [J]. Chin J Natural Med, 2016, 14(2): 141-146.

[12] Wang F, Zhang JY, Yin PH, et al. Rapid identification of poly-phenols in Kudiezi injection with a practical technique of mass defect filter based on high-performance liquid chromatography coupled with linear ion trap/orbitrap mass spectrometry [J]. Anal Methods, 2014, 6(10): 3515-3523.

[13] Zhang JY, Li N, Che YY, et al. Characterization of seventy polymethoxylated flavonoids (PMFs) in the leaves of Murraya paniculata by on-line high-performance liquid chromatography coupled to photodiode array detection and electrospray tandem mass spectrometry [J]. J Pharm Biomed Anal, 2011, 56(5): 950-961.

[14] Zhang JY, Lu JQ, Wang F, et al. A strategy of EIC-MS coupled with diagnostic product ions analysis for efficient discovery of new hydroxylated polymethoxyflavonoid glycosides from the leaves of Murraya paniculata L. using HPLC-DAD-MS/MS [J]. Anal Methods, 2013, 5(5): 2880-2891.

[15] Cai W, Zhang JY, Li GL, et al. Isolation and purification of sesquiterpene lactones from Ixeris sonchifolia (Bunge) Hance by high-speed counter current chromatography and semi-pre-parative high performance liquid chromatography [J]. Trop J Pharm Res, 2014, 13(12): 2065-2069.

[16] Ma JY, Wang ZT, Xu LS, et al. Sesquiterpene lactones from Ixeris sonchifolia [J]. Phytochemistry, 1998, 48(1): 201- 203.

[17] Suh JY, Jo YM, Kim ND, et al. Cytotoxic constituents of the leaves of Ixeris sonchifolia [J]. Arch Pharmacal Res, 2002, 25(3): 289-292.

[18] Zhang WZ. Studies on Chemical constituents and bioactivity of Ixeris Sonchifolia (Bge.) Hance and Xanthium Mongolicum Kitag [D]. Dalian University of Technology, 2010.

[19] Khalil AT, Shen YC, Guh JH, et al. Two new sesquiterpene lactones from Ixeris chinensis [J]. Chem Pharm Bull, 2005,

53(1): 15-17. [20] Zhang YC, Zhou L, Ng KY. Sesquiterpene lactones from Ixeris

sonchifolia Hance and their cytotoxicities on A549 human non-small cell lung cancer cells [J]. J Asian Nat Prod Res, 2009, 11(4): 294-298.

[21] Zhang N, Lv AL, Zheng Z, et al. Two new compounds from Ixeris sonchifolia [J]. J Asian Nat Prod Res, 2008, 10(3-4): 211-213.

[22] Abdel-Mogib M, Awad SN, Abou-Elzahab MM, et al. A ses-quiterpene glucoside from Reichardia tingitana [J]. Phytochem, 1993, 34(5): 1434-1435.

[23] Michalska K, Szneler E, Kisiel W. Sesquiterpene lactones from Lactuca canadensis and their chemotaxonomic significance [J]. Phytochem, 2013, 90(6): 90-94.

[24] Song SJ, Zhou LY, Li LZ, et al. Two new sesquiterpene lac-tones from Ixeris sonchifolia [J]. Nat Prod Commun, 2011, 6(8): 1055-1057.

[25] Kisiel W, Michalska K. Sesquiterpenoids and phenolics from Taraxacum hondoense [J]. Fitoterapia, 2005, 76(6): 520-524.

[26] Zhang WZ, Li XL, Wang MJ, et al. Sesquiterpene lactones from Ixeris sonchifolia (Bge.) Hance [J]. J Asian Nat Prod Res, 2008, 10(8): 753-758.

[27] Ma JY, Wang ZT, Xu LS, et al. A sesquiterpene lactone gluco-side from Ixeris denticulata f. pinnatipartita [J]. Phytochem, 1999, 50(1): 113-115.

[28] Feng XZ. Studies on the constitutions and biological activities of Ixeris sonchifolia [D]. Shenyang Pharmaceutical University, 2001.

[29] Young HS, Im KS, Choi JS. The pharmaco-chemical study on the plant of Ixeris spp. 2. flavonoids and free amino acid com-position of Ixeris sonchifolia [J]. J Korean Soc Food Nutr, 1992, 21(3): 296-301.

[30] Shi PY, Zhang YF, Qu HB, et al. Systematic characterisation of secondary metabolites from Ixeris sonchifolia by the combined use of HPLC-TOF-MS and HPLC-IT-MS [J]. Phytochem Anal, 2011, 22(1): 66-73.

Cite this article as: LIU Rong-Rong, ZHANG Xiu-Ping, WANG Fang, SHANG Zhan-Peng, WANG Fei, LIU Ying, LU Jian-Qiu, ZHANG Jia-Yu. Rapid screening and identification of sesquiterpene lactones in Kudiezi injection based on high-per-formance liquid chromatography coupled with linear ion trap-orbitrap mass spectrometry [J]. Chin J Nat Med, 2018, 16(2): 150-160.

rapid screening and identification of sesquiterpene ...€¦ · quiterpene lactones were...

Documents