Fifteen compounds were isolated from the 70% EtOH extract of leaves of Chinese hawthorn(Crataegus pinnatifida var. major) by various purification steps, and their structures were determined as 2α,3α,12β,19α,-tetrahydroxyursan-13β,28-olide(1),euscaphic acid(2), tormentic acid(3), ursolic acid(4), pomolic acid(5), corosolic acid(6), maslinic acid(7), linalyl rutinoside(8),(Z)-3-hexenyl β-D-glucoside(9),(3S, 6S)-cis-linalool-3,7-oxide-β-D-glucopyranoside(10), pisumionoside(11), icariside B6(12), byzantionoside B(13),(6R,7E,9R)-9-Hydroxy-4,7-megastigmadien-3-one 9-O-β-D-glucopyranoside(14) and(6S,7E,9R)-6,9-dihydroxy-4,7-megastigmadien-3-one 9-O-β-D-glucopyranoside(15) mainly based on the mass spectrum(MS) and nuclear magnetic resonance(NMR) spectroscopic techniques, of which compound 1 was a new pentacyclic triterpene, and compounds 2, 5, 6, 8, 10, 13 and 15 were isolated form this plant for the first time.
China ; Crataegus ; Molecular Structure ; Plant Leaves ; Terpenes ; Triterpenes

OBJECTIVETo develop an LC-MS method for identification of flavonoids in Chinese hawthorn fruits and an HPLC method for quantification of 5 flavonoids.

METHODAccurate molecular weights of the target flavonoids were acquired by time-of-flight mass spectrometer (TOF-MS) in both positive and negative ion modes. The flavoniods were then further confirmed by standards. An RP-HPLC method was established to quantitatively analysis five major flavonoids (chlorogenic acid, procyanidin B2, epicatechin, hyperoside and isoquercitrin) in hawthorn fruits from ten different regions.

RESULTSThe molecular formulae of 8 components were determined by TOF-MS. In the quantitative method validation, the method is linear over the studied range of 1.8-178.5, 2.2-284.8, 2.7-270. 6,1.5-150.0, 1.5-150.0 mg x L(-1) for chlorogenic acid, procyanidin B2, epicatechin, hyperoside and isoquercitrin, respectively. The correlation coefficient for each analyze was greater than 0.999. The inter-day precision of the analysis was lower than 4%, The RSDs of precision and stability were lower than 5%. The recovery rate was from 98.5% to 102.5%, and RSDs were less than 3.5%.

CONCLUSIONThe method is simple, specific and reliable, and is suitable for quality control of Chinese hawthorn.

Chromatography, High Pressure Liquid ; methods ; Crataegus ; chemistry ; Flavonoids ; analysis ; Fruit ; chemistry ; Mass Spectrometry ; Quality Control

OBJECTIVETo study the hypolipidemic active compounds from Crataegus pinnatifida and mechanism of action of those.

METHODGuided by the inhibitory activity to HMG-CoA reductase, the active compounds were separated and purified with macroporous resin and silica gel.

RESULTFour active compounds were obtained, which were quercetin, hyperoside, rutin and chlorogenic acid, the sum of their inhibitory rate was 50.01%, and the total inhibitory rate of the mixture of four active compounds matched was 79.48%.

CONCLUSIONQuercetin and hyperoside were the principle active components inhibiting HMG-CoA reductase in Hawthorn fruit, and there were synergistic action among them.

Crataegus ; chemistry ; Fruit ; chemistry ; Hydroxymethylglutaryl CoA Reductases ; analysis ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; pharmacology ; Plant Extracts ; pharmacology

This study was aimed at identifying the bioactive components of the crude and stir-baked hawthorn for invigorating spleen and promoting digestion, respectively, to clarify the processing mechanism of hawthorn by applying the partial least squares(PLS) algorithm to build the spectrum-effect relationship model. Firstly, different polar fractions of crude and stir-baked hawthorn aqueous extracts and combinations of different fractions were prepared, respectively. Then, the contents of 24 chemical components were determined by ultra-high performance liquid chromatography-mass spectrometry. The effects of different polar fractions of crude hawthorn and stir-baked hawthorn aqueous extracts and combinations of different fractions were evaluated by measuring the gastric emptying rate and small intestinal propulsion rate. Finally, the PLS algorithm was used to establish the spectrum-effect relationship model. The results showed that there were significant differences in the contents of 24 chemical components for different polar fractions of crude and stir-baked hawthorn aqueous extracts and combinations of different fractions, and the gastric emptying rate and small intestinal propulsion rate of model rats were improved by administration of different polar fractions of crude and stir-baked hawthorn aqueous extracts and combinations of different fractions. The bioactive components of crude hawthorn identified by PLS models were vitexin-4″-O-glucoside, vitexin-2″-O-rhamnoside, neochlorogenic acid, rutin, gallic acid, vanillic acid, citric acid, malic acid, quinic acid and fumaric acid, while neochlorogenic acid, cryptochlorogenic acid, rutin, gallic acid, vanillic acid, citric acid, quinic acid and fumaric acid were the bioactive components of stir-baked hawthorn. This study provided data support and scientific basis for identifying the bioactive components of crude and stir-baked hawthorn, and clarifying the processing mechanism of hawthorn.
Animals ; Rats ; Spleen ; Crataegus ; Quinic Acid ; Least-Squares Analysis ; Vanillic Acid ; Algorithms ; Digestion

In order to investigate the mechanism, the correlation between the odor change in Crataegi Fructus stir-fried process and 5-HMF were studied. Required samples were retrieved from Crataegi Fructus stir-fried process. Statistical quality control (SQC) was used to analyze the response values acquired by the electronic nose. At the same time, the content of 5-HMF was detected by high performance liquid chromatography (HPLC). Correlation analysis was used to analyze the relationship between the above two. Experimental results showed that SQC model established by response values of all samples could show the change law of odor in Crataegi Fructus stir-fried process and changes of 5-HMF content was dropped after the first increase. Correlation analysis showed that the odor change in Crataegi Fructus stir-fried process and 5-HMF were significantly correlated (P < 0.05). Sugar degradation reaction and the Maillard reaction may be one of the mechanisms of the odor change in Crataegi Fructus stir-fried process.
Chromatography, High Pressure Liquid ; Crataegus ; chemistry ; Furaldehyde ; analogs & derivatives ; analysis ; Hot Temperature ; Odorants ; analysis ; Plant Extracts ; analysis ; Technology, Pharmaceutical ; methods

OBJECTIVETo evaluate the stability of physical state with accelerate test and dropping in process before and after on compound hawthorn dropping pills.

METHODScanning electron microscope, TG-DTA, FT-IR and XRD were used.

RESULTThe active components presented amorphous, tiny crystal and molecular state in dropping pills, and it had no obvious reaction between PEG 4000 and active components. With time prolonging, a little of active components changed from amorphous state to tiny crystal or molecular state.

CONCLUSIONSolid dispersion improved the stability and dissolution of compound hawthorn dropping pills.

Crataegus ; chemistry ; Drug Compounding ; methods ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; Microscopy, Electron, Scanning ; Solubility ; Spectroscopy, Fourier Transform Infrared

OBJECTIVETo study the technical conditions of the extraction and purification of the active composition from Polygonum orientale and Crataegus pinnatifida Bge. in Hongye Xingtong soft capsules with the macroporous resin.

METHODThe orientm, isorientm and hyperoside were used as index to screen the five kinds of resins. And the technical conditions of the enrichment and purification of D101 resin selected out of above were all-round studied.

RESULTThe D101 was fit for adsorbing orientm, isorientm and hyperoside. Under the optimal conditions, the transfer rate of orientm, isorientm and hyperoside was above 91%, and the total solid was cut down by more than 60%.

CONCLUSIONThe D101 is greatly effective for the enrichment and purification of the active composition of P. orientale and C. pinnatifida Bge.

Crataegus ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Polygonum ; chemistry ; Quercetin ; analogs & derivatives ; chemistry ; isolation & purification ; Resins, Synthetic ; chemistry

In order to study the pesticide residues of the medicinal Crataegi Fructus,this study aims to establish an analysis method for pesticide residues( mainly containing insecticides and fungicides) suitable for the actual situation of medicinal Crataegi Fructus based on the survey of the pesticides of the Crataegi Fructus base,combined with the blind screening results of the LC-ESI-MS/MS pesticide screening platform established by the research team in the early stage. Then,the pesticide residues in medicinal Crataegi Fructus from Shandong,Hebei,Henan,Shanxi,and Liaoning( main cultivation areas) were analyzed. The samples were pretreated by the modified Qu ECh ERS method,i.e.,extracted with acetonitrile-water( 9 ∶1),purified by PSA,C_(18),GCB,silica gel. The detection of pesticides was performed by LC-MS/MS. The ion source was ESI with positive scanning mode,and the linearity of 11 kinds of pesticides in the range of 5-300 μg·kg~(-1) was acceptable( R~2>0. 996 9). All the recoveries of pesticides were within 70. 02%~(-1)12. 0% in the low,medium and high levels,with RSD≤17%. The results showed that the detection rate of carbendazim,chlorpyrifos and difenoconazole is 79%,82%,56%,respectively. Besides,the prohibition pesticide carbofuran were detected in some of the batches,indicating the security risk. This study provides methodological references and basic data for risk assessment of Crataegi Fructus and government regulation.
Chromatography, Liquid ; Crataegus/chemistry* ; Drug Contamination ; Drugs, Chinese Herbal/analysis* ; Pesticide Residues/analysis* ; Surveys and Questionnaires ; Tandem Mass Spectrometry

In order to understand the structural characteristics of squalene synthase genes in the triterpenoids biosynthesis pathway of Crataegus pinnatifida, the squalene synthase genes of C. pinnatifida was cloned and analyzed by bioinformatics and prokaryotic expression. Two squalene synthase genes CpSQS1 and CpSQS2 were cloned from C. pinnatifida fruit by RT-PCR. The ORF length of CpSQS1 and CpSQS2 were 1 239 bp and 1 233 bp respectively, encoding 412 aa and 410 aa respectively. CpSQS1 and CpSQS2 were predicted to be stable acidic proteins by online tools. The secondary structure was mainly composed of α-helix structure, and the tertiary structure was predicted by homology modeling. Structural functional domain analysis showed that 35-367 aa of CpSQS1 and CpSQS2 cDNA containing conserved trans-isoprenyl pyrophosphate synthase domains. Transmembrane domain analysis predicted that two transmembrane domains were founded in CpSQS1 and CpSQS2. The squalene synthase amino sequence of C. pinnatifida had higher homology with the known SQS of Salvia miltiorrhiza and Glycyrrhiza glabra. Phylogenetic tree analysis showed that CpSQS1 and CpSQS2 were clustered into one branch of MdSQS1 and MdSQS2, which were consistent with the phylogenetic rule. Prokaryotic expression vector pGEX-4 T-1-CpSQS1 and pGEX-4 T-1-CpSQS2 were transformed into Escherichia coli Transetta(DE3) for induction, and the target protein was successfully expressed at 65 kDa. The expression levels of CpSQS2 were significantly higher than that of CpSQS1 in three different developmental stages of C. pinnatifida. In this study, the full-length cDNA sequences of C. pinnatifida SQS1 and SQS2 were cloned and analyzed for the first time, which provided the foundation for further study on the metabolic pathway of C. pinnatifida triterpenoids.
Amino Acid Sequence ; Cloning, Molecular ; Crataegus/genetics* ; Farnesyl-Diphosphate Farnesyltransferase/genetics* ; Fruit/enzymology* ; Phylogeny ; Plant Proteins/genetics*

OBJECTIVETo study the relationship between HPLC fingerprint chromatogram and inhibitory effect on respiratory burst of rat PMN of leaves of crataegus L.

METHODHPLC fingerprint peaks of different species of hawthorn leaves were isolated and used for the effective experiment on the respiratory burst of rat PMN. The mathematic models of the relationship between the area and the effect of fingerprint peaks were established. According to the mathematic models, the HPLC fingerprint were change into bioactive fingerprint (include effective fingerprint and potency fingerprint) with the helps of mathematics, chemometrics, computer program simulation and etc.

RESULTThe chromatogram-effect relationship of leaves of crataegus. on respiratory burst of rat PMN was established. According to this relationship, the activities of fourteen samples of leaves of crataegus. were forecasted. It was positive correlation between the expected value and the practical value. And the correlation coefficients was 0.968 (P < 0.01).

CONCLUSIONAn all-around evaluative system, which includes not only chemical identification but also effective evaluation for traditional Chinese medicine was established. It will provide a new idea for study on fingerprint chromatogram of traditional Chinese medicine.

Animals ; Chromatography, High Pressure Liquid ; methods ; Crataegus ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Male ; Plant Leaves ; chemistry ; Rats ; Rats, Sprague-Dawley ; Respiratory Burst ; drug effects