Simultaneous Quantification of Nine Target Compounds in Traditional Korean Medicine, Bopyeo-Tang, Using High-Performance Liquid Chromatography-Photodiode Array Detector and Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry.

Bopyeo-tang (BPT) is composed of six medicinal herbs (Morus alba L., Rehmannia glutinosa (Gaertn.) DC., Panax ginseng C.A.Mey., Aster tataricus L.f., Astragalus propinquus Schischkin, and Schisandra chinensis (Turcz.) Baill.) and has been used for the treatment of lung diseases. This study focused on establishing an analytical method that can simultaneously quantify nine target compounds (i.e., hydroxymethylfurfural, mulberroside A, chlorogenic acid, calycosin-7-O-glucoside, 3,5-dicaffeoylquinic acid, quercetin, kaempferol, schizandrin, and gomisin A) from a BPT sample using high-performance liquid chromatography with a photodiode array detector (HPLC-PDA) and ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). The separation of compounds in both analyses was performed on a C18 reversed-phase column using the gradient elution of water-acetonitrile as the mobile phase. In particular, the multiple reaction monitoring mode was applied for quick and accurate detection in UPLC-MS/MS analysis. As a result of analyzing the two methods, HPLC-PDA and UPLC-MS/MS, the coefficient of determination of the regression equation for each compound was ≥0.9952, and recovery was 85.99-106.40% (relative standard deviation (RSD) < 9.58%). Precision testing of the nine compounds was verified (RSD < 10.0%). The application of these analytical assays under optimized conditions for quantitative analysis of the BPT sample gave 0.01-4.70 mg/g. Therefore, these two assays could be used successfully to gather basic data for clinical research and the quality control of BPT.

Development of a HPLC-UV Method for the Separation and Quantification of Hesperidin, Neohesperidin, Neohesperidin Dihydrochalcone and Hesperetin.

An analysis method was developed for the separation and quantification of hesperidin, neohesperidin, neohesperidin dihydrochalcone and hesperetin by using HPLC-UV. Single factor experiments and Box-Behnken Designs were used to optimize separation of four flavonoids, in which a gradient elution method was adopted with 99% acetonitrile and 0.1% formic acid as mobile phases at a flow rate of 0.9 mL/min. A C18 column was used with a column temperature of 35 °C. LODs and LOQs were below 0.84 µg/mL and 2.84 µg/mL, respectively. Linearity with good correlation coefficients (r > 0.99, n = 5) was attained, recovery rate of four flavonoids ranged from 88% to 130%, the RSD indicating results precision for analyzing hesperidin, neohesperidin, neohesperidin dihydrochalcone and hesperetin ranged from 1.2% to 4.6%. Finally, the present method could be successfully applied to identify and quantify hesperidin, neohesperidin and hesperetin in Fructus Aurantii Immaturus and Pericarpium Citri Reticulatae.

Simultaneous quantification of cellulose and pectin in tobacco using a robust solid-state NMR method.

Cellulose and pectin are the important components of tobacco (Nicotiana tabacum L.) cell wall, which affect the formation of undesirable compounds. Their contents are closely related to the harmfulness of tobacco. But the simultaneous quantitative analysis of cellulose and pectin is hard to be achieved for traditional analytical methods. A solid-state 13C cross-polarization by multiple contact periods (multiCP) NMR method was developed for the simultaneous quantification of cellulose and pectin in tobacco. The multiCP spectrum at optimal parameters agreed well with the direct polarization (DP) spectrum within one-thirtieth of the measurement time and provided satisfactory signal to noise ratio (SNR). After three simple procedures of sample preparation and spectra deconvolution, simultaneous quantification of cellulose and pectin extracted from tobacco was effectively achieved. Compared with the chemical method, this interesting method was rapid, practicable, and very promising, which provided the technical support for the simultaneous quantification of cell wall substances in biological sample.

An LC-MS/MS method for simultaneous quantification of 11 components of Xian-Xiong-Gu-Kang in the plasma of osteoarthritic rats and pharmacokinetic analysis.

Xian-Xiong-Gu-Kang is composed of Epimedium brevicornu, Ligusticum chuanxiong, Radix clematidis, Cinnamomum cassia, and Fructus xanthii. It is used to treat numbness and pain of limbs. In this study, we developed a method to simultaneously quantify 11 components of Xian-Xiong-Gu-Kang (icarrin, epimedin A, epimedin B, epimedin C, icariside II, chlorogenic acid, ligustilide, senkyunolide A, senkyunolide I, ferulic acid, and cinnamic acid) in rat plasma using ultra-performance liquid chromatography coupled with quadrupole linear ion trap mass spectrometry. Chromatographic separation was performed on an ACQUITY UPLC BEH C18 column using gradient elution with a mobile phase comprising acetonitrile and 0.05% (v/v) formic acid aqueous solution. Mass spectrometry detection was performed using positive and negative electrospray ionization in the multiple reaction monitoring mode. The calibration curves of the 11 constituents were linear, with correlation coefficients > 0.99. The intra- and interday accuracy and precision values were within ±15.0%. The extraction recoveries of the 11 constituents and two internal standards were between 66.05 and 105.40%, and the matrix effects were between 86.74 and 112.86%. Using this method, the pharmacokinetic features of the 11 constituents were elucidated in the plasma of osteoarthritic rats after oral administration of the Xian-Xiong-Gu-Kang extract.

UHPLC-UV/PDA Method Validation for Simultaneous Quantification of Luteolin and Apigenin Derivatives from Elaeis guineensis Leaf Extracts: An Application for Antioxidant Herbal Preparation.

Luteolin and apigenin derivatives present in oil palm (Elaeis guineensis) leaves (OPL) are reported to possess excellent antioxidant properties relating to numerous health benefits. To meet the global demand for flavonoids, OPL, which is plentifully generated as an agricultural by-product from oil palm plantations, can be further exploited as a new source of natural antioxidant compounds. However, to produce a standardized herbal preparation, validation of the quantification method for these compounds is required. Therefore, in this investigation, we developed and validated an improved and rapid analytical method, ultra-high-performance liquid chromatography equipped with ultraviolet/photodiode array (UHPLC-UV/PDA) for the quantification of 12 luteolin and apigenin derivatives, particularly focusing on flavonoid isomeric pairs: orientin/isoorientin and vitexin/isovitexin, present in various OPL extracts. Several validation parameters were assessed, resulting in the UHPLC-UV/PDA technique offering good specificity, linearity, accuracy, precision, and robustness, where the values were within acceptable limits. Subsequently, the validated method was employed to quantify luteolin and apigenin derivatives from OPL subjected to different drying treatments and extraction with various solvent systems, giving total luteolin (TLC) and apigenin content (TAC) in the range of 2.04-56.30 and 1.84-160.38 µg/mg extract, respectively. Additionally, partial least square (PLS) analysis disclosed the combination of freeze dry-aqueous methanol yielded OPL extracts with high TLC and TAC, which are strongly correlated with antioxidant activity. Therefore, we provide the first validation report of the UHPLC-UV/PDA method for quantification of luteolin and apigenin derivatives present in various OPL extracts, suggesting that this approach could be employed in standardized herbal preparations by adopting orientin, isoorientin, vitexin, and isovitexin as chemical markers.

Simultaneous determination of 19 constituents in Cimicifugae Rhizoma by HPLC-DAD and screening for antioxidants through DPPH free radical scavenging assay.

Cimicifugae Rhizoma (sheng ma) is a well-known traditional Chinese medicine, which has been demonstrated to possess anti-inflammatory, antipyretic and analgesic activities. In the present study, a simple and efficient HPLC-DAD (high-performance liquid chromatography coupled with diode array detection) method was developed and validated for simultaneous quantification of 19 chemical components (including 16 phenolic acids, one coumarin and two alkaloids) in Cimicifugae Rhizoma. The result indicated that this method could effectively evaluate the quality of Cimicifugae Rhizoma and provide a valuable reference for further study. Additionally, the antioxidant activity of Cimicifugae Rhizoma was evaluated by DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging assay. The results showed that the content of phenolic acids and antioxidant activity exhibited significant correlation coefficients.

Validated LC-MS/MS method for simultaneous quantification of seven components of Naodesheng in rat serum after oral administration and its application to a pharmacokinetic study.

A simple, precise and reliable LC-MS/MS method was developed and validated for simultaneous quantification of vitexin, notoginsenoside R1, hydroxysafflor yellow A, ginsenoside Rd, puerarin, daidzein and senkyunolide I as components of Naodesheng (NDS) in rat serum. The Linearity ranges in rat serum were 0.045-4.5 µg/mL for vitexin, 0.0476-4.76 µg/mL for notoginsenoside R1, 0.0422-4.22 µg/mL for hydroxysafflor yellow A, 0.0426-4.26 µg/mL for ginsenoside Rd, 0.0436-4.36 µg/mL for puerarin, 0.026-2.6 µg/mL for daidzein, and 0.05-5 µg/mL for senkyunolide I, with the correlation coefficients greater than 0.99. The established method was validated in terms of intra- and inter-day precision and accuracy, recovery, matrix effect and stability. Furthermore, the method was successfully applied for pharmacokinetic study of these seven components in rat serum after oral administration of NDS.

Pharmacokinetic comparisons of major bioactive components after oral administration of raw and steamed rhubarb by UPLC-MS/MS.

A study was performed to compare the pharmacokinetics of major bioactive analytes in rhubarb plants to explore the pharmacological differences between raw pieces (RP) and steamed pieces (SP) of rhubarb. A rapid and efficient ultra-performance liquid chromatographic coupled with mass/mass spectrometry (UPLC-MS/MS) method was established to determine the concentrations of six analytes in rat plasma after oral administration of RP and SP. It was found that the AUC (area under the plasma concentration-time curve), Tmax (time to reach maximum plasma concentration) and Cmax (maximum plasma concentration) values were obviously increased in RP for rhein-8-O-ß-d-glucophyranoside, rhein and aloe-emodin. On the contrary, emodin, physcion and chrysophanol had higher bioavailability in SP. The significant differences indicated that steamed by wine may alter pharmacokinetic behaviors of analytes, providing theoretical basis of differences in pharmacological activity between SP and RP.

A new SE-HPLC method for simultaneous quantification of proteins and main phenolic compounds from sunflower meal aqueous extracts.

The aim of this research was to develop a method for simultaneous quantification of proteins and main polyphenolic compounds extracted from oleaginous meal by aqueous media. Size exclusion chromatography with a Biosep column (exclusion range from 1 to 300 kDa) and acetonitrile/water/formic acid (10:89.9:0.1 v/v) eluent at 0.6 mL min-1 yielded the most efficient separation of sunflower proteins and chlorogenic acid monoisomers (3-caffeoylquinic acid, 5-caffeoylquinic acid, and 4-caffeoylquinic acid). After a study of the stability of the extract components, the incorporation of a stabilization buffer (0.5 mol L-1 tris(hydroxymethyl)aminomethane-hydrochloric acid/1.0 mol L-1 sodium chloride at pH 7) was proposed to avoid polyphenol-protein interactions and/or isomeric transformation. The use of 214 nm as the wavelength for protein quantification was also included to minimize the effect of interference from polyphenol-protein interactions on the quantification. Under the used experimental conditions, the protein and chlorogenic acid monoisomer signals remained stable during 300 min at 20 °C (95-125% of the starting value). The developed method was validated and parameters such as specificity, sensitivity, precision, and accuracy were determined. The results from size exclusion chromatography correlated well with the results of protein determination by the reference Kjeldahl method. The proposed method was successfully applied for rapeseed extract analysis making simultaneous quantification of proteins and major rapeseed polyphenols (sinapine and sinapic acid) possible. Graphical abstract.

Quantification of the constituents of the traditional Korea medicine, Samryeongbaekchul-san, and assessment of its antiadipogenic effect.

Samryeongbaekchul-san (SBS) is a traditional herbal formula, which is used for the treatment of dyspepsia, chronic gastritis, and anorexia in Korea. To evaluate the quality of SBS decoction by quantifying its main constituents simultaneously using high-performance liquid chromatography coupled with photodiode array (HPLC-PDA) detection, and secondly to determine the antiadipogenic effect of SBS decoction. The main constituents in a 10-µL injection volume of the decoction were separated on Gemini C18 and Luna NH2 columns (both 250 mm × 4.6 mm, 5 µm) at 40 °C using a gradient of two mobile phases eluting at 1.0 mL/min. 3T3-L1 preadipocytes were differentiated into adipocytes for 8 days with or without SBS. After differentiation, accumulated triglyceride contents and leptin production were measured. The correlation coefficients of all constituents in a calibration curve were ≥0.9998 and showed good linearity in the tested concentration range after validation of the method established. The recovery of the four major compounds were 99.46-102.61% with intra- and interday precisions of 0.08-1.01% and 0.15-0.99%, respectively. The four compounds in the lyophilized SBS sample were detected up to 6.46 mg/g. SBS treatment of the differentiated adipocytes significantly inhibited lipid accumulation and leptin production without cytotoxicity. Optimized simultaneous determination of constituents by HPLC-PDA detection will help to improve quality assessment of SBS or related formulas. SBS has an antiadipogenic effect and further investigation to establish the mechanisms of action of its antiadipogenic effect is warranted.

Simultaneous Quantification of Nine New Furanocoumarins in Angelicae Dahuricae Radix Using Ultra-Fast Liquid Chromatography with Tandem Mass Spectrometry.

A series of new furanocoumarins with long-chain hydrophobic groups, namely andafocoumarins A-H and J, have been isolated from the dried roots of Angelica dahurica cv. Hangbaizhi (Angelicae Dahuricae radix) in our previous study, among which andafocoumarins A and B were demonstrated to have better anti-inflammatory activity than the positive controls. In this work, a sensitive, accurate, and efficient ultra-fast liquid chromatography coupled with triple quadrupole mass spectrometer (UFLC-MS/MS) method was developed and validated for simultaneous quantification of above-mentioned nine compounds in four cultivars of Angelicae Dahuricae Radix. Chromatographic separation was performed on a Kinetex 2.6u C18 100 Å column (100 × 2.1 mm, 2.6 µm). The mobile phases were comprised of acetonitrile and water with a flow rate of 0.5 mL/min. Using the established method, all components could be easily separated within 12 min. With the multiple reaction monitor mode, all components were detected in positive electrospray ionization. The method was validated with injection precision, linearity, lower limit of detection, lower limit of quantification, precision, recovery, and stability, respectively. The final results demonstrated that the method was accurate and efficient, which could be used to simultaneously quantify the nine andafocoumarins in Angelicae Dahuricae Radix. The results also indicated that in different batches of Angelicae Dahuricae Radix, some of the andafocoumarins were significantly different in terms of content.

Chemical constituents and quality control of two Dracocephalum species based on high-performance liquid chromatographic fingerprints coupled with tandem mass spectrometry and chemometrics.

Two similar Dracocephalum species, namely, Dracocephalum tanguticum Maxim and Dracocephalum moldavica L, are commonly used as ethnic medicines in China. Here we describe a strategy of combining high-performance liquid chromatography with quadrupole time-of-flight mass spectrometry, as well as fingerprints and chemometrics for characterization and discrimination of chemical constituents on the two herbs. A total of 49 compounds including 33 flavonoids, 5 phenylethanol glycosides, 1 coumarin glycoside, 8 organic acids, and 2 other types of compounds were unambiguously or tentatively identified from the two Dracocephalum species. Among the compounds identified, 26 were characterized for the first time and 4 compounds, rosmarinic acid (7), salvianolic acid B (10), luteoloside (22), diosmetin-7-O-glucoside (28), were inferred as common constituents for the two herbs. Flavonoids featured in these two Dracocephalum species while their types presented significant differences. Acacetin (45) and acacetin glycosides (acatetin-7-O-glucuronide (30), acacetin-7-O-(6"-O-malonyl) glucoside (33), buddleoside (34), tilianin (35), and agastachoside (42)) were detected only in D. moldavica, which can be used to discriminate two herbal medicines. In addition, six characteristic constitutes in D. tanguticum were simultaneously quantified. Moreover, the induced chemometrics methods including similarity analysis and hierarchical clustering analysis were successfully applied for origin discrimination and quality evaluation of D. tanguticum and D. moldavica.

Simultaneous Quantification of Seven Bioactive Flavonoids in Citri Reticulatae Pericarpium by Ultra-Fast Liquid Chromatography Coupled with Tandem Mass Spectrometry.

INTRODUCTION: Citri Reticulatae Pericarpium (CRP) is a commonly-used traditional Chinese medicine with flavonoids as the major bioactive components. Nevertheless, the contents of the flavonoids in CRP of different sources may significantly vary affecting their therapeutic effects. Thus, the setting up of a reliable and comprehensive quality assessment method for flavonoids in CRP is necessary. OBJECTIVE: To set up a rapid and sensitive ultra-fast liquid chromatography coupled with tandem mass spectrometry (UFLC-MS/MS) method for simultaneous quantification of seven bioactive flavonoids in CRP. METHODS: A UFLC-MS/MS method coupled to ultrasound-assisted extraction was developed for simultaneous separation and quantification of seven flavonoids including hesperidin, neohesperidin, naringin, narirutin, tangeretin, nobiletin and sinensetin in 16 batches of CRP samples from different sources in China. RESULTS: The established method showed good linearity for all analytes with correlation coefficient (R) over 0.9980, together with satisfactory accuracy, precision and reproducibility. Furthermore, the recoveries at the three spiked levels were higher than 89.71% with relative standard deviations (RSDs) lower than 5.19%. The results indicated that the contents of seven bioactive flavonoids in CRP varied significantly among different sources. Among the samples under study, hesperidin showed the highest contents in 16 samples ranged from 27.50 to 86.30 mg/g, the contents of hesperidin in CRP-15 and CRP-9 were 27.50 and 86.30 mg/g, respectively, while, the amount of narirutin was too low to be measured in some samples. CONCLUSION: This study revealed that the developed UFLC-MS/MS method was simple, sensitive and reliable for simultaneous quantification of multi-components in CRP with potential perspective for quality control of complex matrices. Copyright © 2016 John Wiley & Sons, Ltd.

Systematic characterization and simultaneous quantification of the multiple components of Rhododendron dauricum based on high-performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry.

Rhododendron dauricum L. has been used as a traditional Chinese medicine to treat cough and asthma and relieve phlegm and bronchitis. In this study, a reliable method based on high-performance liquid chromatography with diode array detection and quadrupole time-of-flight tandem mass spectrometry was established to systematically identify and quantify the components in this herb for the first time. A total of 33 compounds were identified, including 24 flavonoids, six phenolic acids, two coumarins and one terpene. Among them, poriolin (17), farrerol-7-O-ß-d-glucopyranoside (20), and syzalterin (30) were isolated from this plant for the first time, and quercetin-3-ß-d-(6-p-hydroxy benzoyl) galactoside (19), quercetin-3-ß-d-(6-p-coumaroyl) galactoside (21), and myrciacetin (23) were identified from this genus for the first time. Fragmentation pathways of flavonoids also have been investigated by electrospray ionization mass spectrometry. Moreover, seven bioactive constituents, namely, gallic acid (1), scopoletin (6), dihydroquercetin (7), quercetin (22), kaempferol (25), 8-desmethyl farrerol (27), and farrerol (28), were simultaneously quantified. The developed method has been validated and applied to analyze ten samples of R. dauricum from Hebei Province successfully. The contents of the seven compounds have been detected and compared.

Simultaneous quantitative determination of 20 active components in the traditional Chinese medicine formula Zhi-Zi-Da-Huang decoction by liquid chromatography coupled with mass spectrometry: application to study the chemical composition variations in different combinations.

Zhi-Zi-Da-Huang decoction (ZZDHD), a classical traditional Chinese medicine (TCM) prescription composed of four herbal medicines, has been widely used in treating various hepatobiliary disorders for a long time. The objective of this study was to develop a sensitive and efficient liquid chromatography coupled with mass spectrometry (LC-MS) method for quantitative determination of 20 active constituents, including three iridoid glycosides, 11 flavonoids, three anthraquinones and three tannins in ZZDHD. Separation was achieved on a phenomenex kinetex C18 column (150 × 4.6 mm, 2.6 µm) using gradient elution with a mobile phase consisting of acetonitrile and 0.1% formic acid in water. Detection was performed with electrospray ionization source in the negative ionization and selected ion monitoring mode. The established method was validated by determining the linearity (r(2) ≥ 0.9983), limit of quantification (0.16-300 ng/mL), precision (RSD ≤ 4.6%), average recovery (96.0-105.6%), repeatability (RSD ≤ 3.2%) and stability (RSD ≤ 4.5%). Then, the method was successfully applied to investigate the chemical composition variations owing to the interaction between the four component herbs of ZZDHD during the extraction process. It was found that different combinations of the herbs affect the extraction efficiency of chemical constituents in different ways. The validated LC-MS method provides a meaningful basis for quality control and further research on ZZDHD.

Chemical profile of the active fraction of Yi-Gan San by HPLC-DAD-Q-TOF-MS and its neuroprotective effect against glutamate-induced cytotoxicity.

Yi-Gan San (YGS), a traditional Chinese medicine for dementia-related symptoms, was previously fractionated. One active fraction, YGS40 exhibited a neuroprotective effect against glutamate-induced cytotoxicity. In the present study, high-performance liquid chromatography, coupled with diode-array detection and quadrupole time-of-flight mass spectrometry, was applied for the identification of its chemical constituents and for quantification studies. The chemical profile of YGS40 consisted of sixty-four identified or tentatively characterized compounds. The levels of the major marker compounds increased significantly in the mixed decoction compared with those in the single plant decoction. The results suggest the high precision of the analyses of most of the constituents in YGS40 and establish the quantitative variations of the major marker compounds between the single and mixed decoction processes.

Chemical constituents of Meconopsis horridula and their simultaneous quantification by high-performance liquid chromatography coupled with tandem mass spectrometry.

Meconopsis horridula Hook.f. Thoms has been used as a traditional Tibetan medicine to clear away heat, relieve pain, and mobilize static blood. In this study, a reliable method based on high-performance liquid chromatography with diode array detection and electrospray ionization quadrupole time-of-flight tandem mass spectrometry was established for the identification of components in this herb. A total of 40 compounds (including 17 flavonoids, 15 alkaloids, and eight phenylpropanoids) were identified or tentatively identified. Among them, 17 components were identified in the herb for the first time. Compound 39 appears to be a novel compound, which is confirmed as 3-(kaempferol-8-yl)-2,3-epoxyflavanone by NMR spectroscopy and mass spectrometry. Moreover, seven major constituents were simultaneously quantified by the developed high-performance liquid chromatography with tandem triple-quadrupole mass spectrometry method. The quantitative method was validated and quality parameters were established. The study provides a comprehensive approach for understanding this herbal medicine.

Chemical profile of the active fraction of Yi-Gan San by HPLC-DAD-Q-TOF-MS and its neuroprotective effect against glutamate-induced cytotoxicity / 中国天然药物

Yi-Gan San (YGS), a traditional Chinese medicine for dementia-related symptoms, was previously fractionated. One active fraction, YGS40 exhibited a neuroprotective effect against glutamate-induced cytotoxicity. In the present study, high-performance liquid chromatography, coupled with diode-array detection and quadrupole time-of-flight mass spectrometry, was applied for the identification of its chemical constituents and for quantification studies. The chemical profile of YGS40 consisted of sixty-four identified or tentatively characterized compounds. The levels of the major marker compounds increased significantly in the mixed decoction compared with those in the single plant decoction. The results suggest the high precision of the analyses of most of the constituents in YGS40 and establish the quantitative variations of the major marker compounds between the single and mixed decoction processes.

Analysis of the traditional medicine YiGan San by the fragmentation patterns of cadambine indole alkaloids using HPLC coupled with high-resolution MS.

YiGan San (YGS) has long been used in traditional Japanese and Chinese folk medicine and serves as a potent and novel therapeutic agent to treat Alzheimer's disease. In the present study, a rapid and sensitive method based on HPLC coupled with diode-array detection and quadrupole TOF MS (Q-TOF-MS) was designed to reveal the chemical constituents of YGS. Thirty-six compounds were identified and assigned in YGS, including 14 alkaloids, nine γ-lactones, six flavonoids, three triterpenoid saponinares, two small molecular organic acids, and two other types of compounds. In addition, the accurate fragment weight and MS/MS fragmentation reactions of a subtype indole alkaloid in Uncariae ramulus cum uncis were summarized for the first time to realize rapid identification without reference substances. For the first time, 11 major constituents were comprehensively quantified with a HPLC coupled with triple-quadrupole MS method. A three-section switch was used to realize such multicomponent identification. The contents of saikosaponin B2 and isoliquiritin, which produce anti-inflammatory and antidepressant-like effects, were extremely different, up to 700 times, in two sources of YGS. The developed qualitative and quantitative method was proved to be precise, accurate, and reproducible.

HPLC with quadrupole TOF-MS and chemometrics analysis for the characterization of Folium Turpiniae from different regions.

Folium Turpiniae has been used as a traditional Chinese medicine for the treatment of abscesses, fevers, gastric ulcers, and inflammations. This paper describes a strategy of combining HPLC with photodiode array detection and quadrupole TOF-MS, as well as phytochemical and chemometrics analysis for the characterization, isolation, and simultaneous quantification of the chemical constituents of Folium Turpiniae. 19 constituents were identified, namely, 11 flavonoids, seven gallic acid derivates, and quinic acid. Among them, 15 compounds were identified in this herbal medicine for the first time; compound 10 appears to be novel and was isolated and confirmed as ellagic acid-3-O-α-L-rhamnopyranoside by NMR spectroscopy and MS. In addition, nine marker compounds, namely gallic acid (2), ellagic acid-3-O-α-L-rhamnopyranoside (10), apigenin-7-O-(2''-rhamnosyl)gentiobioside (11), ellagic acid (12), luteolin-7-O-ß-D-neohesperidoside (13), ligustroflavone (14), 4'-O-methylellagic acid-3-O-α-L-rhamnopyranoside (16), rhoifolin (17), and neobudofficide (18), were quantified simultaneously in ten batches of Folium Turpiniae collected from different regions. Moreover, hierarchical clustering analysis and principal component analysis indicated that both samples from Hubei (S1) and Guangxi (S3) provinces showed apparent differences from the others. Samples from Jiangxi province (S2, S4, and S7-10) possessed similar properties and therefore belong to the same group.