Structure identification and content analysis of active components in Xiaoyao pills / 药学学报

Xiaoyao pills are a famous traditional Chinese medicine collected in Welfare Pharmacy, which is a classic prescription for treating liver depression and spleen deficiency. However, its composition is complex. In order to better control the quality of Xiaoyao pills, in this study, HPLC-ion-trap time-of-flight mass spectrometry (LC-IT-TOF/MS) was used to identify the main ingredients of Xiaoyao pills, paeoniflorin, albiflorin, glycyrrhizic acid, saikosaponin A and saikosaponin B2. Then a liquid chromatography tandem mass spectrometry (LC-MS/MS) was developed for simultaneous determination and quantification of the main compounds. Fragmentation pathways of five active components were obtained. The method was validated. Five active ingredients in Xiaoyao pills had a good linear relationship, and the values of RSD (%) of repeatability were all less than 5%, the recovery ranges were between 90% and 115%, and the values of RSD (%) of each substance were less than 10% after the sample solution is placed for 24 hours. Three batches of Xiaoyao pills (concentrated pellets) and two batches of Xiaoyao pills (water pellets) were determined, the contents of paeoniflorin in concentrated pills were more than 4.0 mg·g-1, and those in water pills were more than 2.5 mg·g-1, which was accordance with Chinese Pharmacopoeia. However, other compounds behave differently. This method has high sensitivity and reliable measurement results, which provides basis for quality control of Xiaoyao pills and material basis for pharmacology research.

Research on in vivo metabolites of Longxue Tongluo Capsules by liquid chromatography coupled with hybrid ion trap/time-of-flight mass spectrometry / 中国中药杂志

Longxue Tongluo Capsules(LTC) has good efficacy against blood stasis syndrome during the recovery period of ischemic stroke. Its main active ingredient is the phenolic extract of Chinese dragon's blood. In our previous study, the primary mass fragmentation pathways of phenolic derivatives from LTC were clarified. Herein, the metabolites in rat plasma were characterized following the oral administration of loureirin A and loureirin C using liquid chromatography coupled with hybrid ion trap/time-of-flight mass spectro-metry(LC-IT-TOF-MS), with 18 and 55 metabolites identified, respectively. On this basis, with the help of the obtained accurate molecular weight, characteristic fragment ions, reference comparison, combined with LTC database and natural products database self-created in our group, 18 prototypes and 106 metabolites were tentatively identified in rat plasma after oral gavage of LTC at a dose of 500 mg·kg~(-1). Glucuronidation, sulfonation, and methylation were major biotransformation pathways of LTC. This study preliminarily clarified the LTC constituents absorbed into blood and laid the foundation for clarifying the effective substances of LTC.

LCMS-IT-TOF Analysis of Retention Behavior of Sanguisorbae Radix Components on D101 Macroporous Resin and Polyamide Resin / 中国实验方剂学杂志

Objective:To study on the material basis of Sanguisorbae Radix by column chromatography and liquid chromatography-ion trap-time-of-flight mass spectrometry (LCMS-IT-TOF), and analyze the distribution of different components in Sanguisorbae Radix water extract on D101 macroporous resin and polyamide resin. Method:Sanguisorbae Radix water extract was separated by D101 macroporous resin and polyamide resin, and LCMS-IT-TOF was used for detection, chromatography separation was achieved on an ACQUITY UPLC HSS T3 column (2.1 mm×100 mm, 1.8 μm) with the mobile phase consisted of water (A) and acetonitrile (B) for gradient elution (0-10 min, 5%-20%B; 10-18 min, 20%-35%B; 18-23 min, 35%-50%B; 23-28 min, 50%-90%B; 28-30 min, 90%B; 30-33 min, 90%-5%B; 33-35 min, 5%B), the flow rate was 0.3 mL·min-1, the column temperature was 30 ℃. Data acquisition was carried out in electrospray ionization (ESI) under the positive and negative ion modes, the scanning range was m/z 100-1 200. According to mass spectrometry data such as accurate molecular mass and fragment information, combined with literature, different chemical components in loading effluents and ethanol eluents of Sanguisorbae Radix water extract were identified. A heat map of the distribution of components in each fraction was drawn by extracting mass spectrum peak intensity data of each sample. The elution rules of various components were compared visually. Result:The enrichment and separation of D101 macroporous resin and polyamide resin were obvious. Tannins in Sanguisorbae Radix water extract was mainly concentrated in loading effluent of macroporous resin and its water eluent, triterpenoids were mainly distributed in the 90% ethanol eluent of macroporous resin. In the above effluents and eluents, a total of 63 compounds (including isomers) were identified. Among them, 6 compounds, ellagic acid-4-pyranoarabinoside or its isomer, 6-O-galloylnorbergerin, 3-O-galloylnorbergerin, (6-acetyloxy-5,7-dihydroxy-8-methoxy-4-oxochromen-2-yl) acetate, ethyl 2-methyl-5,6-bis (sulfooxy) benzofuran-3- carboxylate were first discovered in Sanguisorbae Radix. Conclusion:The method can quickly and accurately identify the distribution of components in aqueous extract of Sanguisorbae Radix after column chromatography, providing experimental basis for exploring the pharmacodynamic components and mechanism of Sanguisorbae Radix.

Identification and Content Determination of Main Unknown Impurity in Oxiracetam Capsule / 中国药房

OBJECTIVE：To identify the main unknow impurity of Oxiracetam capsule and determine its content ，so as to improve the standard of quality control. METHODS ：Two-dimensional UPLC -IT-TOF-MS was adopted to qualitatively analyze the unknown impurity. One-dimensional liquid chromatogram analysis was performed on ST PAK C 18 ES column with mobile phase consisted of 0.02 mol/L sodium dihydrogen phosphate solution at the flow rate of 0.5 mL/min. The column temperature was set at 30 ℃，sample size was 20 μL. The detection wavelength was set at 210 nm. Two-dimensional liquid chromatogram analysis was performed on Techmate C 18-STⅡ column with mobile phase consisted of 0.02 mol/L ammonium acetate solution at the flow rate of 0.5 mL/min. The column temperature was 30 ℃. Mass spectrometry was adopted （electropray ionization source ，MS+ and MS － mode data acquisition ）. After the target impurity was located by one-dimensional liquid chromatography ，it was transferred to two-dimensional liquid chromatography-mass spectrometry system for qualitative analysis. The unknown impurity structure was inferred by means of molecular formula prediction module “Accurate Mass Calculator ”in LCMS Solution ，and the refined impurity products by preparation and purification were standardized and confirmed . The impurity content was determined by HPLC （with the same condition of one-dimensional liquid chromatography for qualitative analysis ）. RESULTS ：The main unknown impurity in Oxiracetam capsules is oxiracetam acid. The content of the refined product was 99.5％ after preparation and purification. The contents of oxiracetam acid in 9 batches of Oxiracetam capsules were 0.05％ -0.14％ . CONCLUSIONS ：The established two-dementional UPLC-IT-TOF-MS method can accurately locate the peak position of the impurity oxiracetam acid ，and analyze its structure，while the corresponding content determination method can better separate the impurity from the main drug and other components，with good sensitivity ，precision，repeatability，stability and accuracy. The quality of the finished product of Oxiracetam capsules can be well controlled by using above method .

Analysis of polarity components of Angelicae Sinensis Radix and its metabolites in rats by HPLC-MS~n / 中国中药杂志

This experiment aims to explore the metabolites of n-butanol and water soluble fraction of an ethanol extracts from Angelicae Sinensis Radix in rats. The chemical constituents of n-butanol and water extracts from Angelicae Sinensis Radix were identified by HPLC-DAD-ESI-IT-TOF-MS~n,and the in vivo metabolites of n-butanol and water extracts were analyzed. By analyzing n-butanol and water extracts from Angelicae Sinensis Radix,25 compounds were detected and identified,in which 11 phthalide glycosides were firstly reported. And 19 compounds were detected and identified in rat urine,including 2 prototype constituents and 17 metabolites,and the17 metabolites were new compounds. The method can identify the main constituents and metabolites of extracts from traditional Chinese medicine accurately and rapidly,and provide evidence for interpreting effective forms and pharmacodynamics substance( prototype,metabolites,or both) of Angelicae Sinensis Radix.

Reaction of Chlorogenic Acid with Sulfite and Its Application in Identification of Sulfur-fumigated Herbs / 中国实验方剂学杂志

Objective: To explore the structure and reaction sites of the reaction products of chlorogenic acid and sodium sulfite,and the chemical changes of chlorogenic acid in Lonicerae Japonicae Flos after sulfur fumigating. Method: Chlorogenic acid was reacted with sodium sulfite under mild conditions. Liquid chromatography-mass spectrometry-ion trap-time-of-flight (LC-MS-IT-TOF) and 1H nuclear magnetic resonance spectroscopy (1H-NMR) detection techniques were used to detect the reaction products,and the sulfur-fumigated and unsulfurized Lonicerae Japonicae Flos water extract was detected by LC-MS-IT-TOF. Result: After analyzing the mass spectrometry data of fragment ion,molecular cleavage and accurate molecular weight,according to the results of nuclear magnetic signals of chemical shift,peak intensity and peak splitting, the products of chlorogenic acid and sodium sulfite were preliminarily identified as chlorogenic acid α,β-unsaturated carbonyl addition product:3-((3-(3,4-dihydroxyphenyl)-2-sulfopropyl)oxy)-1,4,5-trihydroxycyclohexane-1-carboxylic acid or 3-((3-(3,4-dihy droxyphenyl)-3-sulfopropyl)oxy)-1,4,5-trihydroxycyclohexane-1-carboxylic acid,and the same characteristic fragments were detected as the addition product in the sulfur fumigated Lonicerae Japonicae Flos,but not found in the unsulfurized. Conclusion: It is the first time to demonstrate the structure and reaction sites of chlorogenic acid and sulfurous acid reaction products,and detect the chlorogenic acid sulfite addition product in sulfur-fumigated Lonicerae Japonicae Flos. Although it is still unclear how the sulfite addition compound produced by sulphur Lonicerae Japonicae Flos affects the efficacy and toxicological activity of Lonicerae Japonicae Flos,we shall still pay attention to the changes of active ingredients in sulphuric medicinal materials. Besides,this study can also provide reference for the studies of chemical composition changes after sulfuration of traditional Chinese medicine containing α,β-unsaturated carbonyl structure.

Analysis of flavonoids in Coreopsis tinctoria by integrating 2D-TLC and HPLC-IT-TOF-MS / 中国中药杂志

The qualitative analysis of flavonoids in Coreopsis tinctoria was carried out by a combination of 2 D-TLC and HPLC-IT-TOF-MS. The separation was conducted on 2 D-TLC and a Phenomenex Kinetex Evo C_(18) column(2.1 mm×100 mm, 2.6 μm) with methanol-0.05% aqueous formic acid by gradient elution. Electrospray ionization-(ESI) source was applied and operated in both positive and negative ionization modes. Eighteen flavonoids including three flavonoids, one flavonol, nine flavonones, one flavanonol and four chalcones, were putatively identified from the flavone-enriched fraction of C. tinctoria. 2 D-TLC could separate the flavonoids from C. tinctoria. HPLC-IT-TOF-MS was able to quickly and accurately analyze the flavonoids in C. tinctoria. The results would provide experimental information for the efficacy material basis clarification of C. tinctoria.

Chemical constituent cluster of decoction of Sanguisorbae Radix by HPLC-IT-TOF/MS / 中草药

Objective: To analyze the chemical constituent cluster of decoction of Sanguisorba Radix systemically by HPLC-IT-TOF/MS. Methods: The samples were scanned by broad spectrum of DAD detector and ionized in negative and positive environment of electron spray ionization. Results: A total of 82 chemical constituents (include isomers) were identified based on exact molecular mass, fragment, ultraviolet spectrum as well as the combination of the database. The chemical constituent cluster was composed of 69 phenolics, 8 triterpenes, 3 flavonoids, an organic acid, and a monoterpene glycoside, of which citric acid, brevifolin carboxylic acid, methoxybenzoic acid methyl ester-5-O-sulfate, isorhamnetin-sulfate, and methylellagic acid-sulfate, et al were firstly found in Sanguisorba Radix. Conclusion: The systemical analysis of the chemical constituent cluster of decoction of Sanguisorba Radix was able to supply a clear material basis for the further study of efficacy and pharmaceutical metabolism of Sanguisorba Radix.

Chemical profile characterization of Marsdenia tenacissima using HPLC-IT-TOF-MS / 中草药

Objective To in depth analyze the chemical profile of Marsdenia tenacissima using HPLC-IT-TOF-MS. Methods The pulverized materials were exacted with methanol in an ultrasonication manner and then separated on a Waters Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm, Milford, MA, USA) that was eluted in gradient with 0.1% formic acid-acetonitrile. The data was collected using automatically triggered tandem mass spectrometric mode in positive/negative ionization polarities. The mass fragmentation patterns of polyoxypregnane derivatives were proposed using some authentic compounds. Results Six chlorogenic acid derivatives and 15 polyoxypregnane derivatives were definitely assigned by referring to reference components, whereas the other signals, including 100 polyoxypregnane derivatives, four flavonoids, and two chlorogenic acid derivatives were tentatively annotated via matching the acquired information with those achieved information and the proposed mass fragmentation rules. Conclusion The research efficiently and accurately analyzed the chemical profile of M. tenacissima using HPLC-IT-TOF-MS, which will provide meaningful information for the quality evaluation and the therapeutic mechanism investigation of M. tenacissima as well as its preparation Xiao-Ai-Ping.

Identification of chemical constituents in Sinopodophylli Fructus by HPLC-DAD-ESI-IT-TOF-MSn / 中国中药杂志

This experiment was performed to analyze and identify the chemical constituents of Sinopodophylli Fructus by HPLC-DAD-ESI-IT-TOF-MSn. The analysis was performed on an Agilent Zorbax SB-C₁₈ (4.6 mm×250 mm, 5 μm) column.The mobile phase consisted of 0.1% formic acid was used for gradient at a flow rate of 1.0 mL·min⁻¹. Electrospray ionization ion trap time-of-flight multistage mass spectrometry was applied for qualitative analysis under positive and negative ion modes. The results indicated that 54 compounds consisted of 18 lignans and 36 flavonoids from Xiaoyelian had been detected by their HRMS data, the information of literature and reference substance. Among them, 27 compounds were reported in Sinopodophylli Fructus for the first time. In conclusion, an HPLC-DAD-ESI-IT-TOF-MSn method was established to qualitative analysis of Xiaoyelian in this study, which will provide the evidence for evaluating the quality of Xiaoyelian herbs, clarifying the mechanism, and guiding the development of pharmacological active ingredients.

Analysis of Impurity Profile of Cefotaxime Sodium by HPLC-MS / 中国药学杂志

OBJECTIVE: To establish an HPLC-MS method for the analysis of the impurity profile of cefotaxime sodium. METHODS: Shimadzu-LCMS-IT-TOF was used with Waters XBridge Shield (RP18, 4.6 mm×250 mm, 5 μm) column. Mobile phase A was 20 mmol·L-1 ammonium acetate (pH adjusted to 6.25)-methanol (92: 8), and mobile phase B was set at 20 mmol·L-1 ammonium acetate-methanol (60: 40) (pH adjusted to 6.25).Gradient elution was performed at a flow rate of 1.0 mL·min-1. ESI source was used.Positive and negative ion scanning was conducted in the range of m/z 150-900.The heating temperature was 200℃, CDL temperature was maintained at 200℃, atomization gas flow rate was 1.5 L·min-1, dry gas pressure was 94.0 kPa, and the post-column diversion ratio was 1: 4.Some related substances in cefotaxime sodium were identified by comparing the retention time in chromatography, [M+H]+ spectrum and MS2 spectrum with those of reference substances, the others which haven't reference substances were deduced or speculated by analyzing the MS2 or MSn fragmentation with the help of a rule summarized from the MS2 fragmentation of cefotaxime sodium and the reference substances of system suitability impurities. RESULTS: Twenty-six related substances were separated and detected in the sample, all of which were identified or deduced. They were cefotaxime sodium isomeric compounds and homologs generated during the production process or degradation products. CONCLUSION: The method can be applied in the identification and qualitative analysis of the related substances of cefotaxime sodium and the quality control and optimization of the synthesis of cefotaxime sodium.

Influence of ultrafiltration membrane on ophiopogonins and homoisoflavonoids in Ophiopogon japonicus as measured by ultra-fast liquid chromatography coupled with ion trap time-of-flight mass spectrometry / 中国天然药物

Ultrafiltration is one of the most fascinating technologies, which makes it possible to improve the quality of traditional medicines for application in the pharmaceutical industry. However, researchers have paid little attention to the effect of ultrafiltration membrane on traditional medicines chemical constituents. In this work, Ophiopogon japonicus (L.f) Ker-Gawl. was used as an example to illuminate the influence of ultrafiltration with different material and molecular weight cut-off (MWCO) membrane on natural chemical constituents as measured by ultra-fast liquid chromatography coupled with ion trap time-of-flight mass spectrometry (UFLC-IT-TOF/MS). Our results indicated that ultrafiltration membrane significantly impacted homoisoflavonoids, especially homoisoflavonoids that were almost completely retained on the polyethersulfone (PES) membrane. We also found that the larger number of aglycone hydroxy and sugar moiety in steroid saponins, the higher the transmittance. Furthermore, the passage rate (%) of ophiogenin type saponins was higher than that of others. The possible adsorptive mechanisms were hydrogen bonding, hydrophobic interactions, and benzene ring interaction by π-π stacking. In conclusion, it is crucial to choose appropriate ultrafiltration membrane based on the characteristics of produce products for application of ultrafiltration technique.

Characterization of metabolites in rats after gavage of two lignans from fruits of Vitex negundo var. cannabifolia by UFLC-IT-TOF-MS / 中草药

Objective: To investigate in vivo metabolic profiles of two lignans, 6-hydroxy-4-(4-hydroxy-3-methoxyphenyl)-3- hydroxymethyl-7-methoxy-3,4-dihydro-2-naphthaldehyde (VB-1) and vitedoin A (VB-2) in the rats. Methods: A UFLC-IT- TOF-MS method was applied to characterize the prototypes and metabolites of VB-1 and VB-2 in rat feces, urine, bile, and plasma after oral administration. Results: Eleven metabolites of the two parent compounds were detected and two prototypes were identified unambiguously by comparing with references. Analysis of metabolites revealed that glucuronidation, sulfation, and hydroxylation were major biotransformation pathways of two lignans. Conclusion: In this study, under the analysis of metabolites of two lignans, its in vivo metabolic process is basically clarified. The results could be helpful for the further pharmacokinetics and pharmacological evaluations of VB-1 and VB-2.

Identification of major constituents from propolis by LC-IT-TOF-MS / 中国药科大学学报

A qualitative analytical method of liquid chromatography coupled with ion trap time-of-flight mass spectrometry (LC-IT-TOF/MS) was developed for identification of major constituents in propolis from China (Shandong).The LC-IT-TOF-MS was performed on a Shim-pack VP-ODS column (2.0 mm× 150 mm,5 μm) with the mobile phase consisting of acetonitrile and water containing 0.2％ formic acid in gradient mode,and the flow rate was set at 0.3 mL/min.Negative ion mode was used for IT-TOF-MS.According to the accurate molecular weight,MS fragment pathway,comparison with the retention time of reference compounds,total 31 compounds,including twenty-five phenolic acids and six flavonoids were identified.The LC-IT-TOF-MS qualitative analysis method can be used for analyzing major components of propolis quickly and accurately.

Impurity profile study of raloxifene hydrochloride / 中国药学杂志

OBJECTIVE: To investigate the impurity profile of raloxifene thus to lay a foundation for the establishment of drug quality standard. METHODS: HPLC-IT-TOF-MS was adopted to analyze destroyed raloxifene and reference substances of impurities. According to the mass spectrometry fragmentation patterns of known impurities, the structures of unknown impurities were speculated. RESULTS: Destroyed raloxifene totally produced three impurities, one of which was unknown. Based on the regular mass spectrometry fragmentation patterns of raloxifene and known impurities, the unknown impurity was speculated to be a sulfone that was generated through further oxidation of raloxifene. CONCLUSION: The methods and results of this study could lay a foundation for the impurity control during production and in vivo analysis of raloxifene.

Identification of chemical constituents in Qiling Wenshen Xiaonang Formula by UPLC-ESI-IT-TOF/MS / 中草药

Objective: To establish a method for the content identification of chemical constituents in Qiling Wenshen Xiaonang Formula by using UPLC-ESI-IT-TOF/MS, which can infer the potential active ingredients in the formula, combining with literature. Methods: The chromatography separation was performed on a Waters ACQUITY UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm) with acetonitrile-0.1% formic acid in water by gradient elution. Electrospray ionization (ESI) source was applied and operated in both positive and negative ion mode. Results: A total of 36 chemical constituents were identified by ESI/MS through comparison on chemical standards and literature data, including 19 flavonoids, 11 phenolic acids, four triterpenoid saponins, one sesquiterpene glycoside, and one alkaloid. Conclusion: This study introduces a comprehensive analysis method for chemical ingredients in Qiling Wenshen Xiaonang Formula, and lays a foundation for the further analysis on the substance basis research. The results provide a significant guidance for the substance basis research and quality control of compound Chinese materia medica.

Identification and Analysis of Apigenin Metabolites in Rats / 中国药房

OBJECTIVE:To clarify the bio-transformation form of apigenin in rats,and to speculate its possible metabolic path-way. METHODS:Rats were divided into blank group and medication group(apigenin 200 mg/kg,i.g.)with 6 rats in each group. Urine and feces samples were collected from 2 groups within 24 h after medication. After corresponding treatment,urine and feces samples were analyzed and detected by HPLC-IT-TOF-MSn under cation mode and anion mode. RESULTS:9 metabolites were iden-tified in urine sample of rats from medication group,i.e. 2,3-double bond reduction of apigenin (U1,U7,U8,U9),bonded to glucuronic acid(U2,U3,U4),bonded to sulphate(U5,U6,U7,U8,U9)and bonded to glucose(U2). 4 metabolites were iden-tified in feces sample of rats from medication group,i.e. 2,3-double bond reduction of apigenin(F3),bonded to glucuronic acid (F2)and bonded to glucose(F1). CONCLUSIONS:Apigenin mainly exists in form of prototype drug in rats. The reduction hap-pens on 2,3-double bond by the intestinal bacteria,and the product of apigenin boned to glucuronic acid or glucose can be formed when excreting in intestinal tract and rats in vivo,while the product of apigenin boned to sulphate can be formed only when excret-ing in rats in vivo.

Identification of chemical constituents in Scrophulariae Radix by HPLC-IT-TOF-MS / 中国中药杂志

This experiment was performed to establish a qualitative analysis on chemical constituents of Scrophulariae Radix by HPLC-ESI-IT-TOF-MS．The analysis was conducted on a C₁₈ column (Kromasil 100-5, 4.6 mm×250 mm, 5 μm) with 0.1% formic acid-acetonitrile as the mobile phase for gradient elution; ESI ion source was used for mass spectra, and data were collected innegative and positive modes. The results showed that 64 compounds from Scrophulariae Radix had been identified by analyzing negative ion mass data including element composition and by comparing with data from literature. Two new compounds (4-hydroxy-6-O-methylcatalpol and acetylangoroside C) and seventeen known compounds were detected from Scrophulariae Radix for the first time. Seventeen known compounds included twelve iridoid glycosides, three phenylpropanoid glycosides and two other kind compounds. This study will provide chemical basis for elucidation of the effective substance in the Scrophulariae Radix.

A semimicroquality evaluation method on Panax notoginseng and its application in analysis of continuous cropping obstacles research samples / 中国中药杂志

Panax notoginseng is a commonly used traditional Chinese medicine with blood activating effect while has continuous cropping obstacle problem in planting process. In present study, a semimicroextraction method with water-saturated n-butanol on 0.1 g notoginseng sample was established with good repeatability (RSD<2.5%) and 9.6%-20.6% higher extraction efficiency of seven saponins than the conventional method. A total of 16 characteristic peaks were identified by LC-MS-IT-TOF, including eight 20(S)-protopanaxatriol (PPT) type saponins and eight 20(S)-protopanaxadiol (PPD) type saponins. The established method was utilized to evaluate the quality of notoginseng samples cultivated by manual intervened methods to overcome continuous cropping obstacles.As a result, HPLC fingerprint similarity, content of Fa and ratio of notoginsenoside K and notoginsenoside Fa (N-K/Fa) were found out to be as valuatable markers of the quality of samples in continuous cropping obstacle research, of which N-K/Fa could also be applied to the analysis of notoginseng samples with different growth years.Notoginseng samples with continuous cropping obstacle had HPLC fingerprint similarity lower than 0.87, in consistent with normal sample, and had significant lower content of notoginsenoside Fa and significant higher N-K/Fa (2.35-4.74) than normal group (0.45-1.33). All samples in the first group with manual intervention showed high similarity with normal group (>0.87), similar content of common peaks and N-K/Fa (0.42-2.06). The content of notoginsenoside K in the second group with manual intervention was higher than normal group. All samples except two displayed similarity higher than 0.87 and possessed content of 16 saponins close to normal group. The result showed that notoginseng samples with continuous cropping obstacle had lower quality than normal sample. And manual intervened methods could improve their quality in different levels.The method established in this study was simple, fast and accurate, and the markers may provide new guides for quality control in continuous cropping obstacle research of notoginseng.