An accurate, cost-effective and simple colorimetric method for the quantification of total triterpenoid and steroidal saponins from plant materials.

Saponins are heterosides widely distributed in the plant kingdom. Their properties are used in many industrial sectors, such as food, cosmetics, agriculture, and pharmaceuticals, and their use is increasing due to the market trend to use natural ingredients. Although many techniques exist to quantify saponins (e.g., gravimetric, foaming, spectrophotometric or chromatographic), none of these allow simultaneous accurate, rapid and inexpensive analysis of both triterpenoid and steroidal saponins. A new colorimetric method constituted of p-anisaldehyde and sulfuric acid was developed and avoids all of the above disadvantages. Parameters used in this method allow a similar molar absorptivity for steroidal and triterpenoid saponins with high specificity in complex matrices reducing the sample preparation step and allowing quantification of saponins blends.

Four new cycloartane-type triterpenoids from the leaves of Combretum mellifluum Eichler: assessment of their antioxidant and antileishmanial activities.

The beneficial pharmacological actions including antioxidant effects as an antileishmanial, antibacterial, antifungal, antidiabetic, anti-inflammatory, antitumor, antiviral, and analgesic of compounds isolated from Combretum mellifluum Eichler (Combretaceae) are well established. The aim of the present study was to determine the phytochemistry as well as assess the antioxidant and antileishmanial activities of the leaves from Combretum mellifluum Eichler (Combretaceae). Analysis of ethanolic extract resulted in isolation and identification of two epimeric mixtures of four previously unknown cycloartane-type triterpenoids, methyl quadrangularate M and methyl 24-epiquadrangularate M, and 2α,3ß,24ß-trihydroxy-cycloart-25-ene and 2α, 3ß, 24α-trihydroxy-cycloart-25-ene, and eight known compounds. Their structures were using one-dimensional nuclear magnetic resonance (1D NMR), 2D NMR and high-resolution electrospray ionization mass spectroscopy (HRESIMS) analysis. Further, the extract and fractions were tested for antioxidant potential. The ethyl acetate and aqueous fractions demonstrated the highest antioxidant activity against 2,2-dipheny-1-picrylhydrazl (DPPH) free radicals, which correlated directly with total flavonoid content. All extracts and fractions from C. mellifluum Eichler were assessed for antileishmanial activity. The supernatant fraction exhibited highest potential, inhibiting the growth of Leishmania amazonensis with IC50 value 31.29 µg/ml. Our findings provide information on the chemical composition of C. mellifluum and the potential beneficial therapeutic usefulness as an antioxidant agent in various diseases.

Safety assessment and antioxidant evaluation of betulin by LC-MS combined with free radical assays.

Betulin, as a new type of natural food preservative, is widely used in various kinds of meat products. However, its detailed mechanism of action and metabolism have not been clarified. In this study, for further gain insight of the mechanism of betulin as a preservative, an efficient method has been applied for measuring the antioxidant capacity of betulin, based on the absorbance of the DPPH• and ABTS• radical cation. When the concentration of betulin was more than 2.0 mg/mL, the scavenging rate of ABTS and DPPH radical reached over 90%, which was equivalent to the antioxidant capacity of Trolox. It is indicated that betulin has significant DPPH and ABTS free radical scavenging ability. This should be one of the important mechanisms for betulin as a preservative. A sensitive method using UHPLC-Q-TOF-MS/MS was established to determine the metabolite profile in vivo and in vitro of betulin. 32 phase I and 2 phase II metabolites were structurally characterized. This study will provide theoretical support for the safety and effectiveness of betulin in the field of preservatives and provide theoretical basis for the further study of betulin and the other natural preservatives. This research also contributes to the development of the food industry.

Triterpenoids from Quercus petraea: Identification in Wines and Spirits and Sensory Assessment.

Eight new triterpenoids (1-8), the known genin (9), and two known functionalized triterpenoids (10 and 11) were isolated from a Quercus petraea heartwood extract. The structures of the new compounds were unequivocally elucidated using HRESIMS and 1D/2D NMR experiments. Sensory analyses were performed in a non-oaked wine on the pure compounds 1-11. Except compounds 1 and 11, all molecules exhibited a sweet taste at 5 mg/L that was particularly intense for compounds 3 and 9. Using LC-HRMS, compounds 1-11 were observed in an oak wood extract and in oaked red wine and cognac. They were also semiquantified in several samples of sessile ( Q. petraea) and pedunculate ( Q. robur) oak wood extract. All compounds were found in quantities significantly higher in sessile than in pedunculate oak wood. These results support the hypothesis of their contribution to the increase in sweetness during oak aging and show that they can be used as chemical markers to identify the species of oak used for cooperage.

Pharmacokinetics Comparison, Intestinal Absorption and Acute Toxicity Assessment of a Novel Water-Soluble Astragaloside IV Derivative (Astragalosidic Acid, LS-102).

BACKGROUND AND OBJECTIVES: Astragaloside IV (AGS IV) is the most important bioactive constituent of Radix Astragali. However, its disappointing clinical application is mainly caused by its very low solubility in biologic fluids, resulting in poor bioavailability after oral administration. We recently obtained a novel water-soluble derivative of AGS IV (astragalosidic acid, LS-102) that displayed significant cardioprotective potential against hypoxia-induced injury. The objective of this study was to investigate the intestinal absorption, main pharmacokinetic parameters and acute toxicity of LS-102 in rodents compared with AGS IV. METHODS: An oral dose of LS-102 and AGS IV (20 mg/kg) was administered to Sprague-Dawley (SD) rats, and blood samples were collected at predetermined time points. The plasma concentrations were detected by a validated UHPLC-MS/MS method, and pharmacokinetic parameters were calculated using a compartmental model. In the intestinal permeability study, the transport of LS-102 across Caco-2 cell monolayers was investigated at six concentrations from 6.25 to 250 µM. Moreover, the acute toxicity of LS-102 (40-5000 mg/kg) via a single oral administration was investigated in BALB/c mice. RESULTS: LS-102 was rapidly absorbed, attaining a maximum concentration of 248.7 ± 22.0 ng/ml at 1.0 ± 0.5 h after oral administration. The relative bioavailability of LS-102 was twice that of AGS IV. LS-102 had a Papp (mean) of 15.72-25.50 × 10-6 cm/s, which was almost 500-fold higher than that of AGS IV, showing that LS-102 had better transepithelial permeability and could be better absorbed in the intestinal tract. The acute toxicity study showed no abnormal changes or mortality in mice treated with LS-102 even at the single high dose of 5000 mg/kg body weight. CONCLUSIONS: Oral LS-102 produced a pharmacokinetic profile different from AGS IV with higher bioavailability, while the toxic tolerance was similar to previous estimates. Thus, we speculated that LS-102 might provide better clinical efficacy and be a potential candidate for the new drug development of Radix Astragali.

Rapid differentiation of Ganoderma species by direct ionization mass spectrometry.

In this study, direct ionization mass spectrometry (DI-MS) has been developed for rapid differentiation of Ganoderma (known as Lingzhi in Chinese), a very popular and valuable herbal medicine. Characteristic mass spectra can be generated by DI-MS directly from the raw herbal medicines with the application of a high voltage and solvents. Rapid differentiation of the Ganoderma species that are officially stated in the Chinese pharmacopoeia from easily confused Ganoderma species could be achieved based on this method, as the acquired DI-MS spectra showed that ganoderic acids, the major active components of Ganoderma, could be found only in the official Ganoderma species but not in the confused Ganoderma species. In addition, classification of wild and cultivated Ganoderma and potential differentiation of Ganoderma from different geographical locations could be accomplished based on principal component analysis (PCA) or hierarchical clustering analysis (HCA). The method is rapid, simple and reproducible, and can be further extended to analysis of other herbal medicines.

NIR Spectroscopy of Actaea racemosa L. rhizome - En Route to Fast and Low-Cost Quality Assessment.

Rhizomes of Actaea racemosa L. (formerly Cimicifuga racemosa) gained increasing interest as a plant-derived drug due to its hormone-like activity and the absence of estrogenic activity. According to the Current Good Manufacturing Practices guidelines and pharmacopeial standards, quality assessment of herbal starting materials includes tests on identity and substitution, as well as quantification of secondary metabolites, usually by HPTLC and LC methods. To reduce the laboratory effort, we investigated near-infrared spectroscopy for rapid species authentication and quantification of metabolites of interest.Near-infrared spectroscopy analysis is carried out directly on the milled raw plant material. Spectra were correlated with reference data of polyphenols and triterpene glycosides determined by LC/diode array detection and LC/evaporative light scattering detection, respectively. Quantification models were built and validated by cross-validation procedures. Clone plants, derived by vegetative propagation, and plants of a collection from different geographical origins cultivated in Berlin were analysed together with mixed batches from wild harvests purchased at wholesalers.Generally, good to excellent correlations were found for the overall content of polyphenols with coefficients of determination of R2 > 0.93. For individual polyphenols such as fukinolic acid, only models containing clone plants succeeded (R2 > 0.92). For the total content of triterpene glycosides, results were generally worse in comparison to polyphenols and were observed only for the mixed batches (R2 = 0.93).Next to quantitative analysis, near-infrared spectroscopy was proven as a rapid alternative to other, more laborious methods for species authentication. Near-infrared spectroscopy was able to distinguish different Actaea spp. such as the North American Actaea cordifolia and the Asian Actaea cimicifuga, Actaea dahurica, Actaea heracleifolia, and Actaea simplex.

Ultraviolet spectroscopy combined with ultra-fast liquid chromatography and multivariate statistical analysis for quality assessment of wild Wolfiporia extensa from different geographical origins.

A quality assessment system comprised of a tandem technique of ultraviolet (UV) spectroscopy and ultra-fast liquid chromatography (UFLC) aided by multivariate analysis was presented for the determination of geographic origin of Wolfiporia extensa collected from five regions in Yunnan Province of China. Characteristic UV spectroscopic fingerprints of samples were determined based on its methanol extract. UFLC was applied for the determination of pachymic acid (a biomarker) presented in individual test samples. The spectrum data matrix and the content of pachymic acid were integrated and analyzed by partial least squares discriminant analysis (PLS-DA) and hierarchical cluster analysis (HCA). The results showed that chemical properties of samples were clearly dominated by the epidermis and inner part as well as geographical origins. The relationships among samples obtained from these five regions have been also presented. Moreover, an interesting finding implied that geographical origins had much greater influence on the chemical properties of epidermis compared with that of the inner part. This study demonstrated that a rapid tool for accurate discrimination of W. extensa by UV spectroscopy and UFLC could be available for quality control of complicated medicinal mushrooms.

New dimension of slow food movement using supercritical fluid technology and methods to influence society by effective marketing strategies.

Although slow food movement is a well-known movement nowadays, in order to make it more widespread to the society, necessity to develop and to adapt new techniques has become inevitable for healthier consumption age. For this purpose, possibility of increased usage of healthy foods with addition of natural extracts using new techniques came out from relevant questionaries applied to people of different age groups. In this study, specific properties of supercritical carbon dioxide at distinct temperatures and water in subcritical conditions were used to obtain extracts rich in water-soluble organic compounds. Experiments were carried out at pressures of 10, 20, 30, and 40 MPa and temperatures ranging from 40 to 200 ℃ with and without modifier for 2 h of extraction time. The flow rate was kept at 4 and 1 ml/min for CO2 and water, respectively. The highest water-soluble organic compound recovery yield was 78.10%. Results were supported by marketing strategies to announce this new application and products to the society. Group of sample questions was prepared to investigate (a) frequency of staple food usage, (b) the brand names and relevant reasons that bring up consumers to buy specifically same branded products, (c) knowledge about the ingredients and how advertising effects purchasing decision, etc. Finally, efficiency increase in slow food consumption was proved with supercritical fluid technology to draw attention to the health of consumers with newer and functional healthy foods.

Quantitative assessment of bioactive compounds and the antioxidant activity of 15 jujube cultivars.

Fifteen jujube cultivars late in their maturation were analysed in the red stage for bioactive compounds; including total phenolics (bound/free), total flavonoids, total polysaccharides, ascorbic acid, total triterpenes, proanthocyanidins and cyclic adenosine monophosphate (cAMP). The antioxidant activity was evaluated using the 2,2-diphenyl-1-picrylhydracyl (DPPH) and 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonicacid) (ABTS(+)) scavenging methods and the ferric reducing antioxidant power (FRAP) assay. The Order Performance by Similarity to Ideal Solution method (TOPSIS) was employed to evaluate the nutrition of different jujube cultivars based on their bioactive compounds. The results indicated that the contents of bioactive compounds and antioxidant capacities vary between different jujube cultivars. Correlation analysis revealed that ascorbic acid, polyphenols and proanthocyanidins were the 3 main components responsible for the antioxidant activity of jujubes. TOPSIS analysis indicated that Zyzyphus jujube cv. Nanjingyazao ranks the highest of the 15 jujube fruits with regards to nutritional value.

Assessment of in-depth degradation of artificially aged triterpenoid paint varnishes using nonlinear microscopy techniques.

The present work investigates the applicability of nonlinear imaging microscopy for the precise assessment of degradation of the outer protective layers of painted artworks as a function of depth due to aging. Two fresh and artificially aged triterpenoid varnishes, dammar and mastic, were tested. Nonlinear imaging techniques have been employed as a new diagnostic tool for determination of the exact thickness of the affected region due to artificial aging of the natural varnishes. The measured thicknesses differ from the calculated mean penetration depths of the samples. These nondestructive, high resolution modalities are valuable analytical tools for aging studies and they have the potential to provide unique in-depth information. Single photon laser induced fluorescence measurements and Raman spectroscopy were used for the integrated investigation and analysis of aging effects in varnishes.

Fast analysis of triterpenoids in Ganoderma lucidum spores by ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry.

A rapid and reliable method was established for simultaneous determination of main triterpenoids in Ganoderma lucidum spores using ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-TQ-MS). The established method was validated in terms of linearity, sensitivity, precision, accuracy and stability, and was successfully applied to determine the contents of 10 main triterpenoids in different batches of G. lucidum spores. The analysis results showed that moderate levels of triterpenoids were found in G. lucidum spores. In addition, a MS full scan with a daughter ion scan experiment was performed to identify the potential derivatives of triterpenoids present in G. lucidum spores. As a result, a total of 22 triterpenoids from different G. lucidum spores were unequivocally or tentatively identified via comparisons with authentic standards and literatures. This method provides both qualitative and quantitative results without the need for repetitive UPLC-MS analyses, thereby increasing efficiency and productivity, making it suitable for high-throughput applications.

A simple GC-MS method for the screening of betulinic, corosolic, maslinic, oleanolic and ursolic acid contents in commercial botanicals used as food supplement ingredients.

The occurrence of triterpene pentacyclic acids in plants is extensive, but little is known about their availability in commercial extracts. A simple GC-MS method for the simultaneous determination of betulinic, corosolic, maslinic, oleanolic and ursolic acids was developed and applied to 38 different commercial plant extracts sold as ingredients for dietary supplements. A suitable protocol was set up to perform routine control of a diverse array of samples with different botanical, chemical and physical characteristics. Remarkable quantities of corosolic acid were found in dried extracts from aerial parts of Lagerstroemia speciosa and Ortosiphon stamineus (14233 and 1132 mg/kg, respectively), while oleanolic acid was abundant in O. stamineus and Crataegus monogyna flowers (2774 and 2339 mg/kg); ursolic was identified in O. stamineus, C. monogyna, L. speciosa and Arctostaphylos uva-ursi leaves (7773, 4165, 2108 and 1034 mg/kg). Only L. speciosa was rich in maslinic acid (4958 mg/kg), while minor amounts of betulinic acid (257 and 80 mg/kg) were detected in L. speciosa and C. monogyna extracts. Lower quantities of triterpenic acids were identified in dried extracts of Harpagophyton procumbens root, propolis, Punica granatum root, Styrax benzoin, Vaccinium myrtillus fruits and Vitis vinifera seeds. Decoctions and fluid extracts lacked or contained very low amounts of triterpenic acids. Results are discussed in terms of quality and safety of these ingredients.

Chemical and biological assessment of Angelica herbal decoction: comparison of different preparations during historical applications.

The commonly used Angelica herbal decoction today is Danggui Buxue Tang (DBT), which is a dietary supplement in treating menopausal irregularity in women, i.e. to nourish "Qi" and to enrich "Blood". According to historical record, many herbal decoctions were also named DBT, but the most popular formulation of DBT was written in Jin dynasty (1247 AD) of China, which contained Astragali Radix (AR) and Angelicae Sinensis Radix (ASR) with a weight ratio of 5:1. However, at least two other Angelica herbal decoctions recorded as DBT were prescribed in Song (1155 AD) and Qing dynasties (1687 AD). Although AR and ASR are still the major components in the DBT herbal decoctions, they are slightly varied in the herb composition. In order to reveal the efficiency of different Angelica herbal decoctions, the chemical and biological properties of three DBT herbal extracts were compared. Significantly, the highest amounts of AR-derived astragaloside III, astragaloside IV, calycosin and formononetin and ASR-derived ferulic acid were found in DBT described in 1247 AD: this preparation showed stronger activities in osteogenic, estrogenic and erythropoetic effects than the other two DBT. The current results supported the difference of three DBT in chemical and biological properties, which could be a result of different herbal combinations. For the first time, this study supports the popularity of DBT described in 1247 AD.

Oxidative stability of virgin olive oil as affected by the bene unsaponifiable matters and tertiary-butylhydroquinone.

During 16 h heating at 180 °C, the oxidative stability (OS) of virgin olive oil (VOO) as affected by the same concentrations (200 ppm) of tertiary-butylhydroquinone (TBHQ) and unsaponifiable matters of bene kernel (UKO) and hull (UHO) oils in terms of the inhibitory effect on the formation of conjugated diene hydroperoxides (OS(CDV)) and off-flavor carbonyl compounds (OS(CV)) was investigated. TBHQ was not able to considerably increase the OS(CDV) (7.51) of the VOO (7.2) and showed no synergistic effect with indigenous antioxidative compounds of the VOO (IOV) in this respect. However, it could significantly improve the OS(CV) (from 2.49 to 4.52), which was mainly due to its synergism with the IOV. The UKO increased considerably the OS(CDV) (to 11.8), and its OS(CV) (4.22) was nearly the same as that of TBHQ. The IOV still had marked contributions to the prevention of VOO oxidation but the majority of stabilizing effect was related to the UKO and its synergism with the IOV. The OS(CDV) in presence of the UHO was less than that of the VOO (5.96), although it significantly increased the OS(CV) (to 5.2), mainly due to the stabilizing effect of UHO and its synergism with the IOV.

Chemical composition of South American Burseraceae non-volatile oleoresins and preliminary solubility assessment of their commercial blend.

INTRODUCTION: Non-volatile oleoresins from neotropical Burseraceae are traditionally used for craft, technological and medicinal purposes. The crude resin is usually sold in popular markets of the forest communities. Adding value to this rainforest raw material requires establishing its composition. OBJECTIVE: To analyse the resin composition from different Burseraceae species and establish a minimally reproducible profile by gas chromatography, in order to parameterise its quality control. METHODOLOGY: Crude oleoresin samples of 10 Protium and Trattinnickia species and a commercial blend were subjected to hydrodistillation to remove volatile compounds. The chloroform-soluble residues were methylated, analysed by GC-FID (flame ionisation detection), and individual components were identified by analysing their mass fragmentation pattern in GC-MS and comparison with data from the literature. The blend solubility was assayed in 30 non-chlorinated solvents at three different proportions. RESULTS: The resins consisted exclusively of triterpenes, showing a common predominance of four major compounds in all the samples, corresponding to α-amyrin, ß-amyrin, α-amyrenone and ß-amyrenone. This profile was complemented with minor amounts of the tetracyclic ß-elemolic and α-elemolic acids, maniladiol, brein and other oxidised trace compounds. The better solvents for the resin were those chemically bearing more than four carbon atoms, as n-butyl acetate, 2-ethoxyethanol and isopropanol. The crude resin blend sold contained around 10% of insoluble material that was constituted by up to 70% inorganic residues mixed with humic acid derivatives, as attested by ash analysis and IR spectroscopy, respectively. CONCLUSION: The experimental results, complemented by a general inspection of the literature, demonstrated a systematically reproducible triterpene profile in Protium and Trattinnickia species.

[Species differentiation and quality assessment of Ziziphus jujuba var. spinosa based on the HPLC fingerprint and quantitative analysis].

OBJECTIVE: To establish an HPLC method of a characteristical chemical fingerprint analysis in combination with simultaneous determination of four bioactive components for species differentiation and quality assessment of Ziziphus jujuba var. spinosa. METHODS: The chromatographic separation was performed on an Agilent TC-C18 BDS (250 mm x 4.6 mm, 5 microm) column. The mobile phase consisted of acetonitrile and water in a linear gradient elution procedure. The evaporator tube temperature of ELSD was set at 110.5 degrees C with the nebulizing gas flow rate of 3.1 mL/min and the flow rate of mobile phase was 1.0 mL/min. The column was maintained at 30 degrees C. The injection volume was 20 microL. RESULTS: HPLC methodology for both chemical fingerprint analysis and quantitative determination of four active ingredients were validated, respectively. According to the contents of the four ingredients and the chemical fingerprints of Ziziphus jujuba var. spinosa using principal component analysis, Ziziphus jujuba var. spinosa was different from the fake derived from the seeds of Ziziphus mauritiana. CONCLUSION: The developed HPLC method is exclusive and repetitive for the species identification and quality evaluation of Ziziphus jujuba var. spinosa.

[Study on industrialization production technology of decoction pieces shuangfei of Platycodon grandiflorum].

OBJECTIVE: To establish industrialization production technology parameter for decoction pieces shuangfei of Platycodon grandiflorum. METHODS: Taking platycodin D, polysaccharide and the appearance of the decoction pieces shuangfei of Platycodon grandiflorum as the quality evaluation indexes, we investigated the size of Platycodon grandiflorum, the best slicing extent, the best slicing thickness, drying temperature and drying time. RESULTS: The optimal processing was as follows: the diameter of Platycodon grandiflorum was about 1.5 cm, moistend the selected Platycodon grandiflorum until the water content was 55% - 60%. Then rolled them into 1.5 cm thickness pieces. Dryed them at 60 degrees C for 8 - 9 h and packed them after they were cold. CONCLUSION: The industrialization production technology for decoction pieces shuangfei of Platycodon grandiflorum is stable and the efficiency is improved.

A rapid LC/MS/MS method for the analysis of nonvolatile antiinflammatory agents from Mentha spp.

UNLABELLED: Mints (Mentha spp.), aromatic crops grown largely for their essential oils, also are rich sources of nonvolatile antiinflammatory agents. Identification and quantitation of the constituents responsible for their antiinflammatory activity is challenging owing to the lack of suitable chromatographic methodology. In the present research, the simultaneous quantitation of antiinflammatory constituents rosmarinic acid, oleanolic acid, and ursolic acid in mints was attained by using a unique tandem HPLC column system coupled with an electrospray ionization mass detection (MRM mode). The ion mode optimization for rosmarinic acid under negative and triterpenoid acids under positive was achieved by setting 2 time segments in a single run where the polarity mode was switched from negative (0 to 10 min) to positive (10 to 40 min). For the investigated concentration ranges of antiinflammatory agents in mints, good linearities (r² ≥ 0.998) were obtained for each calibration curve. Validation of precision and accuracy for this method showed that intra- and inter-day repeatabilities for all analytes were less than 5.51%, and the recoveries varied from 97.8% to 99.3%. The developed LC/MS/MS assay provides a suitable quality control method for the determination of antiinflammatory constituents in Mentha spp. There is a wide range of diversity in the natural product composition for these acids across the Mentha germplasm collection evaluated. The presence of these antiinflammatory acids in post-distilled mints shows that value-added nutraceutical enriched products can be developed with proper processing and recovery systems in addition to the distillation and capture of the valuable volatile essential oils. PRACTICAL APPLICATION: Results from this research would benefit both commercial farmers growing mint for essential oil and those in the food industry where value-added phytopharmaceutical enriched products can be developed with proper processing, quality control, and recovery systems during mint essential oil distillation.

Quality assessment of Cortex Phellodendri by high-performance liquid chromatography coupled with electrospray ionization mass spectrometry.

A simple method based on liquid chromatography coupled with diode array detection and electrospray ionization mass spectrometry (LC-DAD-ESI-MS) was developed for the quality assessment of Cortex Phellodendri (CP), which was mainly derived from two species of Phellodendron chinense Schneid and Phellodendron amurense Rupr. Total 41 compounds, including 14 phenols, 24 alkaloids and three liminoidal triterpenes were identified or tentatively characterized from the 75% methanol extract of CP samples by online ESI-MS(n) fragmentation and UV spectra analysis. Among them, two phenols and six alkaloids were simultaneously quantified using HPLC-DAD method. The validated HPLC-DAD method showed a good linearity, precision, repeatability and accuracy for the quantification of eight marker compounds. Furthermore, the plausible fragmentation pathway of the representative compounds were proposed in the present study. The differences of the chemical constituents content and the comprehensive HPLC profiles between the two CP species using LC-DAD-ESI-MS method are reported for the first time, indicating that the CP drugs from different resources should be used separately in the clinic.