The effect of soil environmental factors on the yield and quality of Pueraria lobata.

Pueraria lobata is a typical medicinal and edible plant with great market value and demand, thus exploring the relationship between soil environmental factors and the yield and quality of Pueraria lobata is of great significance for its high-value cultivation. In this study, using the Guige 1 variety (Pueraria montana var. Thomsonii) selected by our research group as the material to compare the effects of five soil types, endophytes in three parts of Pueraria lobata and two fertilizers on its yield and quality. The results showed that the comprehensive evaluation effect of five soil types on the yield and quality of Guige 1 was as follow: red-yellow mixed soil (RYMS) > black loam soil (BLS) > sandy loam soil (SLS) > sandy loam soil waterlogging (SLSW) > yellow soil compaction soil (YSCS); the descending order of endophyte types and quantities is in BLS > RYMS > SLS > YSC > SLSW; applying General Compound Fertilizers (GCF) in RYMS is more suitable for the rapid expansion of Guige 1 than Organic-Slow-Release-Fertilizers (OSRF). The high potassium content in RYMS and high effective phosphorus content in BLS are positively correlated with the content of starch and isoflavone in Pueraria lobata. The conclusion is that the high potassium and available phosphorus content in RYMS and BLS, as well as the rich types and quantities of endophytic bacteria, are positively correlated with the yield and quality of Pueraria lobata. The research results have important guiding significance for the high-value cultivation of Pueraria lobata.

An integrated strategy for characterization of chemical constituents in Stephania tetrandra using LC-QTOF-MS/MS and the target isolation of two new biflavonoids.

LC-MS has been a widely used analytical technique for identification of natural compounds. However, sophisticated and laborious data analysis is required to identify chemical components, especially new compounds, from a large LC-MS dataset. The aim of this study is to develop an integrated data-mining strategy that combines molecular networking (MN), in-house polygonal mass defect filtering (MDF), and diagnostic fragment ion filtering (DFIF) to identify phytochemicals in Stephania tetrandra based on LC-MS data. S. tetrandra samples were prepared by matrix solid-phase dispersion extraction methods and then raw MS spectra were acquired using LC-QTOF-MS/MS. MN and in-house polygonal MDF classified the compounds roughly. Modified DFIF were then used in succession to place each spectrum into a specific class. Finally, the exact structures were deduced by fragmentation pathways and related botanical biogenesis, with the help of the narrowed classification from MN and MDF. The total workflow was a combination of data filtering and identification methods for rapid characterization of known compounds (dereplication) and discovery of new compounds. Consequently, 144 compounds were identified or tentatively identified in the aerial parts and roots of S. tetrandra, including 11 potentially new compounds and 63 compounds first identified in this species. Among 144 compounds, 61 were from the aerial parts exclusively, 8 were from the roots exclusively, and 75 were found in both parts. Furthermore, two new biflavonoids were isolated with the guide of LC-MS analysis and structurally elucidated by spectroscopic methods. In conclusion, the proposed data-mining strategy based on LC-MS can be used to profile chemical constituents with high efficiency and guide the isolation of new compounds from medicinal plants. The comparison of the components of the aerial parts and roots of S. tetrandra would be helpful for the rational utilization of the medicinal plant.

Cardiac safety analysis of first-line chemotherapy drug pegylated liposomal doxorubicin in ovarian cancer.

Pegylated liposomal doxorubicin (PLD) is a nano-doxorubicin anticancer agent. It was used as early as 2014 to treat ovarian and breast cancer, multiple myeloma and Kaposi's sarcoma. The 2018 National Comprehensive Cancer Network guidelines listed PLD as first-line chemotherapy for ovarian cancer. PLD has significant anticancer efficacy and good tolerance. Although PLD significantly reduces the cardiotoxicity of conventional doxorubicin, its cumulative-dose cardiotoxicity remains a clinical concern. This study summarizes the high-risk factors for PLD-induced cardiotoxicity, clinical dose thresholds, and cardiac function testing modalities. For patients with advanced, refractory, and recurrent malignant tumors, the use of PLD is still one of the most effective strategies in the absence of evidence of high risk such as cardiac dysfunction, and the lifetime treatment dose should be unlimited. Of course, they should also be comprehensively evaluated in combination with the high-risk factors of the patients themselves and indicators of cardiac function. This review can help guide better clinical use of PLD.

Identification of alkaloidal compounds from leaves and roots of Stephania succifera by HPLC-QTOF-MS and prediction of potential bioactivity with PharmMapper.

INTRODUCTION: The roots of Stephania succifera are used in traditional medicine for the treatment of several diseases. Research on this plant has mainly focused on bioactive alkaloids from the roots, and no previous work on compounds from the abundant leaves has yet been reported. OBJECTIVE: To identify and compare alkaloidal compounds in S. succifera roots and leaves and to predict the potential bioactivity of some alkaloids. METHODS: High-performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry (HPLC-QTOF-MS/MS) was employed to identify alkaloidal compounds from S. succifera. The potential targets and bioactivities of most alkaloids were predicted using the PharmMapper server. RESULTS: Fifty-six alkaloidal compounds, including protoberberine-, aporphine-, proaporphine-, benzylisoquinoline-, and lactam-type alkaloids, were identified or tentatively identified in S. succifera roots and leaves based on the HPLC-MS data. Forty-one compounds have not been previously reported in S. succifera and eight of them have not been previously reported in the literature. Twenty-four alkaloidal compounds were found in both roots and leaves. Twelve potential targets with different indications were predicted for some alkaloids. CONCLUSION: Comparison of chemical constituents and their potential bioactivities for S. succifera roots and leaves indicated that diverse bioactive alkaloids were present in the leaves as well as the roots. PharmMapper provided new directions for bioactivity screening. This study will be helpful for further understanding the medicinal components of S. succifera and the rational utilisation of plant resources.

Comparative Transcriptome Analysis Reveals Candidate Genes Involved in Isoquinoline Alkaloid Biosynthesis in Stephania tetrandra.

The roots of Stephania tetrandra are used as a traditional Chinese medicine. Isoquinoline alkaloids are considered to be the most important and effective components in this herb, but little is known about the molecular mechanism underlying their biosynthesis. In this context, this study aimed to reveal candidate genes related to isoquinoline alkaloid biosynthesis in S. tetrandra. Determination of tetrandrine and fangchinoline in the roots and leaves of S. tetrandra by HPLC showed that the roots had much higher contents of the two isoquinoline alkaloids than the leaves. Thus, a comparative transcriptome analysis of the two tissues was performed to uncover candidate genes involved in isoquinoline alkaloid biosynthesis. A total of 71 674 unigenes was obtained and 31 994 of these were assigned putative functions based on BLAST searches against 6 annotation databases. Among the 79 isoquinoline alkaloid-related unigenes, 51 were differentially expressed, with 42 and 9 genes upregulated and downregulated, respectively, when the roots were compared with the leaves. The upregulated differentially expressed genes were consistent with isoquinoline alkaloid accumulation in roots and thus were deemed key candidate genes for isoquinoline alkaloid biosynthesis in the roots. Moreover, the expression profiles of 10 isoquinoline alkaloid-related differentially expressed genes between roots and leaves were validated by quantitative real-time polymerase chain reaction, which indicated that our transcriptome and gene expression profiles were reliable. This study not only provides a valuable genomic resource for S. tetrandra but also proposes candidate genes involved in isoquinoline alkaloid biosynthesis and transcription factors related to the regulation of isoquinoline alkaloid biosynthesis. The results lay a foundation for further studies on isoquinoline alkaloid biosynthesis in this medicinal plant.

DNA barcodes for the identification of Stephania (Menispermaceae) species.

Stephania is a medicinal plants-rich genus of Menispermaceae. However, the identification of morphologically-similar species in Stephania is difficult using the currently reported methods. The indiscriminate overexploitation of Stephania plants has resulted in clinical misuse and endangerment of many species, which necessitates the development of an efficient and reliable method for species authentication. Therefore, six candidate DNA barcode sequences (ITS, ITS2, psbA-trnH, matK, rbcL, and trnL-F) were tested for their capacity to identify Stephania species. The barcodes were analyzed either as a single region or in combination by tree-based [neighbor-joining (NJ) and Bayesian inference (BI)], distance-based (PWG-distance), and sequence similarity-based (TaxonDNA) methods. Amplification and sequencing success rates were 100% for all six candidate barcodes. A comparison of six barcode regions showed that ITS exhibited the highest number of variable and informative sites (182/179), followed by psbA-trnH (173/162). DNA barcoding gap assessment showed that interspecific distances of the six barcodes were greater than intraspecific distances. The identification results showed that species discrimination rates of combination barcodes were higher than those of single-region barcodes. Based on best match and best close match methods, the ITS+psbA-trnH combination exhibited the highest discrimination power (93.93%). Further, all Stephania species could be resolved in the phylogenetic trees based on ITS+psbA-trnH (NJ, BI). This study demonstrates that DNA barcoding is an efficient method to identify Stephania species and recommends that the ITS+psbA-trnH combination is the best DNA barcode for the identification of Stephania species.

Comprehensive identification of minor components and bioassay-guided isolation of an unusual antioxidant from Azolla imbricata using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry combined with multicomponent knockout and bioactivity evaluation.

Azolla imbricata (Roxb.) Nakai is used as a traditional Chinese medicine. However, very limited information is available on its effective components. Traditional procedures for discovering natural bioactive compounds, especially for minor ones, are usually time-consuming and labor-intensive. Therefore, an efficient approach using ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UHPLC-QTOF-MS/MS) combined with multicomponent knockout and bioactivity evaluation was developed to obtain more information about the bioactive constituents in A. imbricata. A total of 93 compounds were identified or tentatively identified, including five major components knocked out by semi-preparative HPLC and 88 minor components in the extract with the major components' knockout. These compounds involved chlorogenic acid derivatives, flavonoids, cinnamoyltyrosine derivatives, cinnamic acid derivatives, fatty acids and their derivatives, coumarins, lignans, and chromones. Eighty-two compounds have not been previously reported in the literature for this species, including a new flavanol derivative named brainin D. Among them, 64 compounds, including brainin D, exhibited antioxidant activities using 1,1-diphenyl-2-picrylhydrazyl radical (DPPH)-UHPLC-MS as guidance. The new antioxidant, brainin D, was concomitantly isolated and unequivocally identified by spectroscopic methods, and it showed good DPPH radical activity with an IC50 value of 9.3 ±â€¯0.6 µg/mL. In conclusion, the proposed combination approach can be used for systematic identification of minor constituents and guided isolation of new compounds from natural sources with high efficiency. The comprehensive understanding of the minor constituents and antioxidants in A. imbricata lays the foundation for further rational utilization of this medicinal herb.

[Simultaneous Nitrification and Denitrifying Phosphorus Removal in Continuous Flow Reactor with Intermittent Aeration].

A continuous flow reactor (TCFR) with 10 compartments was used to treat domestic sewage. The anaerobic compartments of TCFR were kept at 3. The anoxic compartments of TCFR were reduced from 2 to 0. Therefore, the aerobic compartments of TCFR were increased gradually from 5 to 7. The aerobic compartments were set to continual aeration in Run1 and intermittent aeration from Run2 to Run4. The aeration/non-aeration ratios were 40 min/20 min,40 min/30 min, and 40 min/40 min, respectively. The nitrification liquid reflux ratios were reduced gradually from 150% to 0%. When the average influent concentrations of COD, NH4+-N, TN, and PO43--P were 259.34, 60.26, 64.42, and 6.10 mg·L-1, respectively, the corresponding effluent concentrations were 26.40, 1.03, 5.84, and 0.3 mg·L-1, respectively in Run4. The nitrogen removal amounts increased gradually from 192.30 mg·h-1 in Run1 to 244.00 mg·h-1 in Run4, and the corresponding removal rates increased from 65.40% to 95.30%. The activity of denitrifying phosphorus accumulating organisms (DPAOs) and phosphorus accumulating organisms (PAOs) increased from 36.05% and 38.20% in Run1 to 140.50% and 133.40% in Run4, respectively. Simultaneous nitrification and denitrifying phosphorus removal was achieved in TCFR by adopting intermittent aeration, which provided a reference for the reformation of sewage treatment plants.

Determination of caffeoylquinic acid derivatives in Azolla imbricata by chitosan-based matrix solid-phase dispersion coupled with HPLC-PDA.

Five main components of Azolla imbricata were isolated and identified as caffeoylquinic acid derivatives, four of which were isolated from this species for the first time. The five compounds exhibited strong antioxidant activities and were used as chemical markers for quantitative analysis. A chitosan-based matrix solid-phase dispersion (MSPD) extraction coupled with high-performance liquid chromatography (HPLC) was developed for the determination of the five analytes in A. imbricata. The optimal conditions for the chitosan-based MSPD extraction were as follows: low viscosity chitosan HL-1 as the dispersant, sample-to-dispersant mass ratio of 1:1, 10 mL of a methanol-sulfuric acid aqueous solution (0.2 M) (7:3, v/v) as the elution solvent, and a grinding time of 1 min. Compared with ultrasonic assisted extraction (UAE), the chitosan-based MSPD extraction exhibited higher extraction efficiency, consumed less solvent and time, and provided a cleaner extract. Compared with C18-based MSPD, the developed method was less expensive and more environmentally-friendly. The validated MSPD-HPLC method was efficient, reliable, and applicable to the quantification of caffeoylquinic acids in A. imbricata.

[Characteristics of Denitrifying Phosphorus Removal by A2/O-BAF at Low Temperatures].

Low C/N domestic sewage was treated by an A2/O-biological aerated filter (BAF) system at low temperatures (11-14℃). The characteristics of pollutant removal, the ratio of denitrifying phosphorus to nitrogen (ΔPO43-/ΔNO3-N) and effects of aeration flow and effective packing height on nitrification in BAF were studied. The results showed that when the average influent concentrations of COD, NH4+-N, TN and PO43- were 193.1, 58.6, 60.3 and 5.1 mg·L-1 respectively, their effluent concentrations were 46.3, 2.5, 13.4 and 0.3 mg·L-1 respectively, which met the first level A criteria specified in the discharge standard of pollutants for municipal wastewater treatment plant (GB 18918-2002). The linear fitting of ΔPO43-/ΔNO3--N was between 0.47 and 1.75. The normal distribution of mathematical statistics was applied-and the average standard deviation for ΔPO43-/ΔNO3--N were 1.20 and 0.29 respectively. When the aeration flows were 60 L·h-1 and 100 L·h-1, the effluent concentration of NH4+-N was less than 5.0 mg·L-1, corresponding to the effective packing heights in the BAF of 1.8 m and 1.0 m respectively. However, when the aeration flow was increased to 120 L·h-1, the air-water flow led to biofilm detachment, which caused the effluent concentration of NH4+-N to increase beyond 5.0 mg·L-1.

Structural Characterisation of Alkaloids in Leaves and Roots of Stephania kwangsiensis by LC-QTOF-MS.

INTRODUCTION: The tuberous roots of Stephania kwangsiensis, which contain bioactive alkaloids, are used as a traditional Chinese medicine. Overexploitation of the roots has made the plant increasingly rare, and the abundant leaves of the same plant may offer a potential alternative. However, there is insufficient phytochemical information for a comparison of alkaloid compositions in the two parts. OBJECTIVE: To characterise and compare the alkaloids in the leaves and roots of S. kwangsiensis. METHODS: The alkaloids in S. kwangsiensis were characterised using high pressure liquid chromatography coupled with positive electrospray ionisation quadrupole time-of-flight tandem mass spectrometry (HPLC-(+)ESI-QTOF-MS/MS). The alkaloid compositions in the leaves and roots were compared by visual inspection combined with principal component analysis (PCA) of the HPLC-MS data. RESULTS: Seventy-five alkaloids comprising aporphine-, proaporphine-, protoberberine-, benzylisoquinoline-, bisbenzylisoquinoline- and morphine-type alkaloids were identified or tentatively identified in the roots and leaves of S. kwangsiensis. Sixty-three of these alkaloids have not been previously reported in this species, and three have not been previously reported in the literature. The roots and leaves had similarities in alkaloid composition but differences in the peak intensities of most alkaloids. The PCA revealed that the samples were clustered into two distinct groups, which corresponded to leaves and roots. CONCLUSION: This study further clarified the chemical constituents in the roots of S. kwangsiensis, and revealed that diverse alkaloids were also present in the leaves. The comparative chemical profiling of the two parts provides useful information on their potential medicinal use. Copyright © 2017 John Wiley & Sons, Ltd.

Identification of Alkaloids in Stephania hainanensis by Liquid Chromatography Coupled with Quadrupole Time-of-flight Mass Spectrometry.

INTRODUCTION: Plants in the genus Stephania can produce diverse bioactive alkaloids. Stephania hainanensis is a medicinal plant that contains effective alkaloids. However, only 10 alkaloids have been reported in this species. OBJECTIVE: To characterise the alkaloids in Stephania hainanensis using liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (LC-QTOF-MS/MS). METHODS: An LC-QTOF-MS/MS method was developed for structural characterisation of the alkaloids in Stephania hainanensis. The chromatographic separation was performed on a phenyl column with gradient elution, and the tandem mass spectra were obtained by using an electrospray ionisation (ESI) interface in positive ionisation mode. Compound identification was based on the exact masses, fragmentation pathways, retention behaviours and related botanical biogenesis. RESULTS: A total of 37 tetrahydroprotoberberine-, quaternary protoberberine-, aporphine-, proaporphine-, benzylisoquinoline- or bisbenzylisoquinoline-type alkaloids were identified or tentatively identified in a single LC run. Twenty-seven of these alkaloids, including the benzylisoquinoline-type of alkaloids, have not been previously reported in Stephania hainanensis. The possible fragmentation pathways of different types of alkaloids were proposed. Besides the general fragmentations, the characteristic losses of CH3 N = CH2 were observed for the benzylisoquinoline and aporphine alkaloids with two methyl groups on the nitrogen. CONCLUSION: The LC-QTOF-MS/MS method enabled profiling and rational, but tentative, identification of diverse alkaloids in Stephania hainanensis. The results obtained may be helpful for understanding the bioactivity of S. hainanensis and evaluating the quality of this plant. Copyright © 2016 John Wiley & Sons, Ltd.

Cytotoxic oxepinochromenone and flavonoids from the flower buds of Rosa rugosa.

A new oxepinochromenone, rugosachromenone A (1), seven new flavonoids, rugosaflavonoids A-G (2-8), and 11 known compounds (9-19) were isolated from the flower buds of Rosa rugosa. Compound 1 is found from Nature for the first time. Compound 2 displayed cytotoxicity against NB4, SHSY5Y, and MCF7 cells with IC50 values of 2.2, 2.5, and 2.3 µM, respectively, and 3 was toxic to A549 and MCF7 cells with IC50 values of 1.2 and 2.8 µM, respectively.

Antiviral phenolic compounds from Arundina gramnifolia.

Five new phenolic compounds, gramniphenols C-G (1-5), and eight known compounds (6-13) were isolated from the whole plant of Arundina gramnifolia. Compounds 1, 4, and 5 showed anti-tobacco mosaic virus activity, with IC(50) values of 20.8, 40.8, and 57.7 µM, respectively. Compounds 1-10 were also tested for their anti-HIV-1 activity; compounds 2, 3, and 6 displayed anti-HIV-1 activity with therapeutic index values above 100:1.

Intestinal permeability of forskolin by in situ single pass perfusion in rats.

The intestinal permeability of forskolin was investigated using a single pass intestinal perfusion (SPIP) technique in rats. SPIP was performed in different intestinal segments (duodenum, jejunum, ileum, and colon) with three concentrations of forskolin (11.90, 29.75, and 59.90 µg/mL). The investigations of adsorption and stability were performed to ensure that the disappearance of forskolin from the perfusate was due to intestinal absorption. The results of the SPIP study indicated that forskolin could be absorbed in all segments of the intestine. The effective permeability (P (eff)) of forskolin was in the range of drugs with high intestinal permeability. The P (eff) was highest in the duodenum as compared to other intestinal segments. The decreases of P (eff) in the duodenum and ileum at the highest forskolin concentration suggested a saturable transport process. The addition of verapamil, a P-glycoprotein inhibitor, significantly enhanced the permeability of forskolin across the rat jejunum. The absorbed fraction of dissolved forskolin after oral administration in humans was estimated to be 100 % calculated from rat P (eff). In conclusion, dissolved forskolin can be absorbed readily in the intestine. The low aqueous solubility of forskolin might be a crucial factor for its poor oral bioavailability.

Isoprenylated flavonoids and adipogenesis-promoting constituents from Morus nigra.

Ten new isoprenylated flavonoids, nigrasins A-J (1-10), and three known compounds were isolated from the twigs of Morus nigra. Compounds 8 and 9 promoted adipogenesis, characterized by increased lipid droplet and triglyceride content in 3T3L1 cells, and induced up-regulation of the expression of adipocyte-specific genes, aP2 and GLUT4.

A fast and sensitive HPLC-MS/MS analysis and preliminary pharmacokinetic characterization of cudratricusxanthone B in rats.

A method for the quantitative analysis of cudratricusxanthone B (CXB) in rat plasma by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS) has been developed and validated. The method involved liquid-liquid extraction from plasma, simple chromatographic conditions on a Venusil XBP-PH C(18) column with the mobile phase of 0.5% formic acid in methanol, and mass spectrometric detection using an API-3000 instrument. Multiple reaction monitoring (MRM) mode was used to monitor precursor/product ion transitions of m/z 397.1/285.0 for CXB and m/z 381.6/269.2 for the internal standard (I.S.) cudraxanthone H. The standard curves were linear over the concentration range of 1-500 ng/mL for CXB in rat plasma. The intra- and inter-batch accuracy for CXB at four concentrations was 89.4-99.5% and 89.4-100.8%, respectively. The RSDs were less than 7.92%. The lower limit of quantification for CXB was 1.0 ng/mL using 100 microL of plasma. The average extraction recoveries of CXB ranged from 80.1 to 95.4% at the concentrations of 2, 50 and 500 ng/mL, respectively. This method was successfully applied to the pharmacokinetic study after an intravenous administration of CXB in male Sprague-Dawley (SD) rats.

[Studies on the chemical constituents of Coleus forskohlii].

OBJECTIVE: To study the chemical constituents in the aerial parts of Coleus forskohlii. METHODS: The compounds were isolated by various column chromatographic methods, and their structures were identified by spectroscopic methods. RESULTS: Twelve compounds were isolated and identified as chamaecydin (1), 6 alpha-hydroxydemethylcryptojaponol (2), alpha-cedrene (3), oleanolic acid (4), forskolin G (5), forskolin J (6), 1,6-diacetyl-9-deoxyforskolin (7), forskolin A (8), forskolin H (9), 6-acetyl-1-deoxyforskolin (10), betulinic acid (11), beta-sitosterol (12). CONCLUSION: Compounds 1 - 3 are isolated from Coleus genus for the first time, and compound 4 is isolated from C. forskohlii for the first time.

Determination of aglycone conjugated metabolites of scutellarin in rat plasma by HPLC.

A specific, precise and accurate high-performance liquid chromatography (HPLC) method for the determination of aglycone conjugated metabolites of scutellarin in plasma after enzymolysis to scutellarein (the aglycone of scutellarin) was developed and validated. The chromatographic separation was performed on a Lunar C18(2) reversed-phase column at a column temperature of 40 degrees C. The mobile phase, delivered at 1.0 ml/min, consisted of acetonitrile-KH2PO4 buffer (40 mM, pH 2.5) (33:67, v/v). The detection wavelength was set at 335 nm. Scutellarein and I.S. (quercetin) were isolated by a liquid-liquid extraction after incubating the plasma samples with beta-glucuronidase/sulfatase. The method was validated using scutellarin spiked plasma as standards. Linearity was confirmed in the concentration range of 0.2165-4.329 nmol/ml, R.S.D.s were within 8.32%, and the recoveries of scutellarein ranged from 101.2 to 108.6%. The method is applicable to the pharmacokinetic study of aglycone conjugated metabolites of scutellarin in rats after oral administration of scutellarin.

[The method for determination of aconitum alkaloids in Radix Aconiti Kusnezoffii].

The methods for the determination of Aconitum alkaloids in Radix Aconiti Kusnezoffii were established. Volumetric analysis with the indicator of methyl red-bromcresol green and colorimetric analysis with acid dye of bromecresol green under the wavelength of 416 nm were used for the assay of total alkaloids. Colorimetric analysis of hydroxamic acid-Fe was for the assay of total ester-diterpene type Aconitum alkaloids. Three kinds of diester-diterpene type Aconitum alkaloids (aconitine, hypaconitine and mesaconitine) were determined by HPLC method. The HPLC system consisted of C18 as the stationary phase and acetonitrile: buffer (acetic acid-triethylamine, pH 6.25) = 71:29 as the mobile phase. The wavelength was 235 nm.