Homogenate-assisted high-pressure disruption extraction for determination of phenolic acids in Lonicerae Japonicae Flos.

An effective method based on the combined homogenate-assisted high-pressure disruption extraction (HHPDE) was applied to the extraction and determination of the main phenolic acid compounds from Lonicerae Japonicae Flos. The optimized HHPDE showed competitive advantage in yield (The extraction yields of NCA, CA, 3,5DCA and 4,5DCA in HHPDE were 1.21, 1.08, 1.06 and 1.17 fold higher than those in UAE), time-saving (<5 min) and relative low temperature requirement (4-16 °C) compared to HRE and UAE. Furthermore, the HHPDE method behaved a good repeatability and reproducibility according to the HPLC. The mentioned HHPDE method is firstly applied in the extraction and quantification of neochlorogenic acid chlorogenic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in Lonicerae Japonicae Flos. This work provided an excellent alternative for the extraction and quantification of thermosensitive from plants.

Qualitative and Quantitative Characterization of Monosaccharide Components of Salvia miltiorrhiza, Liguspyragine Hydrochloride, and Glucose Injection.

Salvia miltiorrhiza, liguspyragine hydrochloride, and glucose injection (SLGI) was made of Salvia miltiorrhiza Bge., liguspyragine hydrochloride, glucose, and glycerin. There were many kinds of monosaccharide components in SLGI, which might be from the raw material and Salvia miltiorrhiza Bge. Separation was performed on a Phenomenex Luna C18 analytical column (250 mm × 4.6 mm i.d., 5 µm, AccuStandard Inc., USA) at 30°C. The mobile phase consisted of two solvents: 0.1 mol/L phosphate-buffered saline (pH 6.7) (solvent A) and acetonitrile (solvent B) with gradient elution. The flow rate was maintained at 1.0 mL/min. Five kinds of monosaccharide components, glucose, D-mannose, L-rhamnose monohydrate, galactose, and xylose, were detected by precolumn derivatization HPLC, and their contents were compared with each other. And finally, concentrations of glucose in SLGI were determined and they were higher than the values of marked amount, which showed that one source of glucose might be from Salvia miltiorrhiza Bge. in SLGI. The average concentration of glucose was 5.18 g/100 mL, which was near the average value at 5.25 g/100 mL detected by ultraviolet spectrophotometry and also close to the marked amount (5.00 g/100 mL) on the label.

Ganoderin A, a novel 9,11-secosterol from Ganoderma lucidum spores oil.

In this study, four sterols were isolated from the Ganoderma lucidum spores oil obtained via supercritical CO2 extraction. Four chemical constituents were ganoderin A (1), chaxine B (2), ergosterol, (3) and stellasterol (4). All the separated ingredients were characterized using spectral data interpretation and by comparing with reported data. Noticeably, stellasterol and chaxine B were both firstly isolated from Ganoderma lucidum spores oil and ganoderin A was shown to bear an unprecedented skeleton.

[Extraction of 10-Deacetyl Baccatin by Supercritical CO2 from Taxus yunnanensis Branches and Leaves].

OBJECTIVE: To study the supercritical CO2 fluids extraction (SFE) method to extract the components from Taxus yunnanensis. METHODS: Medicinal meterials were extracted by supercritical CO2, and then purified by industrial chromatography. Using the extraction yield of 10-DAB as the index,single factor test was carried out to investigate the effect of co-solvent, extraction time, extraction pressure, extraction temperature, pressure and temperature of separation kettle I. Then orthogonal experiment was used to optimize the best extraction condition. RESULTS: The suitable extraction condition was as follows: the ratio of co-solvent (80% ethanol) amount and the madicinal materials was 3: 1, Separation kettle I pressure was 14 MPa, separation kettle I temperature was 40 °C, extraction pressure was 25 MPa, extraction temperature was 60 T and extraction time was 90 min. The extract was separated by industrial chromatographic and then crystallized. CONCLUSION: The supercritical CO2 extraction and purification process of 10-DAB were simple and feasible.

[Fingerprint and Simultaneous Determination of Multi-components in Water-soluble Components of Salvia miltiorrhiza in Miao Autonomous County of Songtao, Guizhou].

OBJECTIVE: To establish the HPLC fingerprint of water-soluble components of Salvia miltiorrhiza in Songtao, Guizhou, and to perform simultaneous determination of six components in it, so as to provide analytical method for its quality control. METHODS: The analyses were performed on a Phenomenex Luna C18 (2) (250 mm x 4. 6 mm, 5µm) column eluted with 0. 4% formic acid(A) - acetonitrile(B) in a gradient mode. The flow rate was 1. 0 mL/min, column temperature was set at 30 °C. RESULTS: Eleven common peaks were identified form the HPLC fingerprint of Salvia miltiorrhiza from 10 batches, the HPLC fingerprint similarities of 10 batches were not less than 0. 999. The linear ranges of danshensu, protocatechuic aldehyde, caffeic acid, rosmarinic acid, lithospermic acid and salvianolic acid B were 0. 0680 ~ 1. 3583 mg/mL, 0. 0008 ~ 0. 3967 mg/mL, 0. 0005 ~ 0. 2660 mg/mL, 0. 0020 ~ 0. 3992 mg/mL, 0. 0063 ~ 0. 6311 mg/mL and 0. 0097 ~ 1. 9306 mg/mL with r ≥ 0. 9999, respectively. The recovery rates were 100. 84%,102. 44%, 100. 53% ,100. 63%, 100. 83% and 100. 35% with RSD <2. 3%, respectively. CONCLUSION: The established method is simple, accurate and can provide reference for quality control of Salvia miltiorrhiza.

[Study on Chemical Constituents of Fat-soluble Extraction from Lepidium meyenii].

OBJECTIVE: To study the chemical constituents of the fat-soluble extraction from Lepidium meyenii root. METHODS: Different extraction methods were studied, including supercritical carbon dioxide extraction, circumfluence extraction and steam distillation. Chemical constituents of the fat-soluble extraction from Lepidium meyenii were analyzed by GC/MS. RESULT: The number of compounds isolated by the above four methods were 38, 31, 14, 21 (specific gravity less than 1 in steam distillation) , and 25 (specific gravity greater than 1 in steam distillation), accounting for 85.79%, 81.18%, 62.08%, 98.36% (specific gravity less than 1 in steam distillation) and 81.54% (specific gravity greater than 1 in steam distillation) of each total peak area, respectively. CONCLUSION: This study lays a certain foundation for further study and development of functional factors in Lepidium meyenii root.

[Chemical components from essential oil of Pandanus amaryllifolius leaves].

OBJECTIVE: To analyze the chemical compositions of Pandanus amaryllifolius leaves essential oil extracted by steam distillation. METHODS: The essential oil of Pandanus amaryllifolius leaves was analyzed by gas chromatography-mass spectrum, and the relative content of each component was determined by area normalization method. RESULTS: 128 peaks were separated and 95 compounds were identified, which weighed 97.75%. The main chemical components of the essential oil were phytol (42.15%), squalene (16.81%), what's more pentadecanal (6.17%), pentadecanoic acid (4.49%), 3, 7, 11, 15-tetramethyl-2-hexadecen-1-ol (3.83%), phytone (2.05%) and the other 74 chemical compositions were firstly identified from the essential oil of Pandanus amaryllifolius leaves. CONCLUSION: The chemical compositions of Pandanu samaryllifolius leaves essential oil was systematically, deeply isolated and identified for the first time. This experiment has provided scientific foundation for further utilization of Pandanus amaryllifolius leaves.

[Supercritical CO2 extraction and component analysis of Aesculus wilsonii seed oil].

OBJECTIVE: To research the optimal extraction process of supercritical CO2 extraction and analyze the component of the oil extracted from Aesculus wilsonii seed. METHODS: Using the yield of Aesculus wilsonii seed oil as the index, optimized supercritical CO2 extraction parameter by orthogonal experiment methodology and analysed the compounds of Aesculus wilsonii seed oil by GC-MS. RESULTS: The optimal parameters of the supercritical CO2 extraction of the oil extracted from Aesculus wilsoniit seed were determined: the extraction pressure was 28 MPa and the temperature was 38 degrees C, the separation I pressure was 12 MPa and the temperature was 40 degrees C, the separation II pressure was 5 MPa and the temperature was 40 degrees C, the extraction time was 110 min. The average extraction rate of Aesculus wilsonii seed oil was 1.264%. 26 kinds of compounds were identified by GC-MS in Aesculus wilsonii seed oil extracted by supercritical CO2. The main components were fatty acids. CONCLUSION: Comparing with the petroleum ether extraction, the supercritical CO2 extraction has higher extraction rate, shorter extraction time, more clarity oil. The kinds of fatty acids with high amounts in Aesculus wilsonii seed oil is identical in general, the kinds of fatty acids with low amounts in Aesculus wilsonii seed oil have differences.

[Study on the immunocompetence of polysaccharide extracted from root of Salvia miltiorrhiza].

OBJECTIVE: To study the immune activity of polysaccharide extracted from root of Salvia miltiorrhiza. METHODS: Investigated the effects of polysaccharide extracted from root of Salvia miltiorrhiza on lymphocyte proliferation response of mouse induced by LPS (the lipopolysaccharide LPS), phagocytosis of the peritoneal macrophage of mice to chick erythrocytes and the mouse models of delayed type hypersensitivity (DTH) response induced by DNFB. RESULTS: Lymphocyte proliferation and phagocytosis of the peritoneal macrophage of mice could be promoted by the polysaccharide, which could inhibit ear edema and capillary permeability increase induced by DNFB and enlarged the thymus and splenic index in mice. The expression of iNOS, IFN-alpha and IL-1beta was inhibited significantly in the treatment group. CONCLUSION: The polysaccharide extracted from Salvia miltiorrhiza can improve immune function.

[Optimization for supercritical CO2 extraction with response surface methodology of Prunus armeniaca oil].

OBJECTIVE: To optimize the extraction conditions of Prunus armeniaca oil by Supercritical CO2 extraction and identify its components by GC-MS. METHODS: Optimized of SFE-CO extraction by response surface methodology and used GC-MS to analysis Prunus armeniaca oil compounds. RESULTS: Established the model of an equation for the extraction rate of Prunus armeniaca oil by supercritical CO2 extraction, and the optimal parameters for the supercritical CO2 extraction determined by the equation were: the extraction pressure was 27 MPa, temperature was 39 degrees C, the extraction rate of Prunus armeniaca oil was 44.5%. 16 main compounds of Prunus armeniaca oil extracted by supercritical CO2 were identified by GC-MS, unsaturated fatty acids were 92.6%. CONCLUSION: This process is simple, and can be used for the extraction of Prunus armeniaca oil.

[Optimization for supercritical CO2 extraction with response surface methodology and component analysis of Sapindus mukorossi oil].

OBJECTIVE: To study the extraction conditions of Sapindus mukorossi oil by Supercritical CO2 Extraction and identify its components. METHODS: Optimized SFE-CO2 Extraction by response surface methodology and used GC-MS to analysie Sapindus mukorossi oil compounds. RESULTS: Established the model of an equation for the extraction rate of Sapindus mukorossi oil by Supercritical CO2 Extraction, and the optimal parameters for the Supercritical CO2 Extraction determined by the equation were: the extraction pressure was 30 MPa, temperature was 40 degrees C; The separation I pressure was 14 MPa, temperature was 45 degrees C; The separation II pressure was 6 MPa, temperature was 40 degrees C; The extraction time was 60 min and the extraction rate of Sapindus mukorossi oil of 17.58%. 22 main compounds of Sapindus mukorossi oil extracted by supercritical CO2 were identified by GC-MS, unsaturated fatty acids were 86.59%. CONCLUSION: This process is reliable, safe and with simple operation, and can be used for the extraction of Sapindus mukorossi oil.

[Research of the essential oil of Plumeria rubra var. actifolia from Laos by supercritical carbon dioxide extraction].

The orthogonal test and the supercritical carbon dioxide fluid extraction were used for optimizing the extraction of the essential oil from Plumeria rubra var. actifolia for the first time. Compared with the steam distillation, the optimal operation parameter of extraction was as follows: extraction pressure 25 MPa, extraction temperature 45 degrees C; separator I pressure 12 MPa, separator I temperature 55 degrees C; separator II pressure 6 MPa, separator II temperature 30 degrees C. Under this condition the yield of the essential oil was 5.8927%. The components were separated and identified by GC-MS. 53 components of Plumeria rubra var. actifolia measured by SFE method were identified and determined by normalization method. The main components were 1, 6, 10-dodecatrien-3-ol, 3, 7, 11-trimethyl, benzoic acid, 2-hydroxy-, phenylmethyl ester, 1, 2-benzenedicarboxylic acid, bis(2-methylpropyl) ester,etc.. 1, 2-Benzenedicarboxylic acid, bis (2-methylpropyl) este. took up 66.11% of the total amount, and there was much difference of the results from SD method.

[Purification of Ganoderma lucidum polysaccharide by ultrafiltration technology].

OBJECTIVE: To investigate the feasibility and technology for purifying Ganodermna lucidum polysaccharide (GLP) by ultrafiltration. METHODS: A spire roll ultrafiltration unit was selected to purify GLP. RESULTS: Purity of 73.5% and recovery ratio of 69.4% for GLP were obtained by using a membrane with cut-off molecular mass of 6000, while operating temperature, pressure and time were 45-50 degrees C, 0.6 MPa and 45 min, respectively. CONCLUSION: Ultrafiltration is one of the new feasible methods for the purification of GLP.

[Study on isolation of Chlorogenic acid from Eucommia ulmoides leaves by ultrafiltration technique].

OBJECTIVE: To isolate and purify Chlorogenic acid from Eucommia ulmoides leaves by ultrafiltration technique. METHODS: Chlorogenic acid transferring ratio was investigated by temperature, pressure, time, cut-off molecular mass. RESULTS: The optimal conditions for the ultrafiltration technique were finally determined. It was found that, for the membrane with a cut-off molecular mass of 20000, when the temperature, pressure, time were respectively 40 degrees C, 0.36 MPa, 20 minutes and adding 24L water, yield of Chlorogenic acid was 99.38%. CONCLUSION: High Chlorogenic acid transfering ratio can be gotten by ultrafiltration technique, which is helpful to the further isolation and purification of Chlorogenic acid in the subsequent process.

[Study on the technique of ultrasonic wave extraction of triterpeneclucoside].

The extraction technique of ultrasonic wave on increasing the triterpeneclucoside was studied. Based on single factor experiments and the orthogonal design introduced,the optimized extracting conditions of triterpeneclucoside were investigated. The results showed that the best extraction technique conditons are 65% alcohol-water, the ratio of solid to liquid 1:30,dousing over 30 minutes,ultrasonic wave lasting for 40 minutes, ultrasonic frequency 26.8 KHZ, ultrasonic (output) power 1080 kw and extraction temperature 40 degrees C, respectively.

[Study on supercritical CO2 extraction of fatty oils from the seed of Akebia trifoliata (Thunb) Koidz].

The extraction of fatty oils from the seed of Akebia trifoliata (Thunb) Koidz with supercritical CO2 was studied. The effects of extraction pressure, extraction temperature on the yields were discussed. The optimal condition of this method was as follow: extraction pressure 30 MPa, extraction temerature 45 degrees C, separator I pressure 11 MPa, separator I temperature 50 degrees C, separator II pressure 6MPa, separator II temperature 45 degrees C, extraction period 2 hours. Compared with the traditional solvent extraction, with a GC-MS analysis, it revealed that the component extracted with supercritical CO2 was basically consistent with that extracted with petroleum ether, and it was rich unsaturated fatty acid.

[Analysis of the essential oils from Amomun kravak Pierre ex Gagnep by GC-MS].

OBJECTIVE: To establish the fingerprint chromatogram of Amomun kravak Pierre ex Gagnep. METHODS: The composition of the essential oil was isolated by PR solvent and determined by GC-MS. Forty-two species of compounds were identified, of which the relative contents were calculated using square peaks to normalization, among which the character components were 1,8-cineole, beta-pinene, alpha-pinene, p-cymene, etc. EMIC software was used and EMIC fingerprint chromatogram was established by selecting the character ion of those 10 compounds. Different extraction methods were also compared. RESULTS: The GC-MS fingerprint chromatogram can provide qualitative information confirming compounds of the corresponding chromatographic peaks. EMIC was first used to reason the quality control of the medicine quality and it can establish the foundation for safe and effective quality control of the medicine and its product. PR solvent can easily extract the essential oil from the medicine. CONCLUSION: This method has the advantage of high efficiency and low cost, and the GC-MS fingerprint chromatogram provided the ascription and quality of the medicine and will play an important role in promoting the modernization of Chinese traditional medicine.

[Study on ingredients of essential oils of Curcuma wenyujin extracted by supercritical-CO2 fluid extraction and steam distillation].

OBJECTIVE: To compare the ingredients of essential oils of Curcuma wenyujin extracted by supercritical-CO2 fluid extraction and by steam distillation. METHOD: GC-MS was applied in this experiment. RESULT: The ingredients and physical and chemical properties of essential oils of C. wenyujin extracted by supercritical-CO2 fluid extraction and by steam distillation are similar. CONCLUSION: Supercritical-CO2 fluid extraction is better than steam distillation in extraction time, power consumption, recovery and purity.

[Study on the supercritical carbon dioxide extraction of Brucea Javanica oil].

The technics of supercritical carbon dioxide extraction of Brucea Javanica for injection oil was studied. The effects of the conditions of extraction and separation on extraction oil yield and quality were investigated and the effects of extraction pressure and separation pressure on acid number of the oil were discussed. So the optimal condition of this method was determined. With a quality evaluation, the Brucea Javanica oil extracted with supercritical CO2 agreed with the quality standard of refined Brucea Javanica oil for injection. This method shows sufficiently the advantages of supercritical CO2 extraction applied in the extraction of traditional Chinese medicine.

[Determination of artemisinin in Artemisia annuna by HPLC-ELSD].

OBJECTIVE: A HPLC method was first established to determine artemisinin in Artemisia annuna by HPLC-ELSD. Artemisinin in Herba of Artemisia annuna from different places was determined by this new method. METHODS: The method utilized MeOH-H2O (75: 25) as mobile phase with flow rate of 1 ml/min, C18 (250 x 4.6 mm, 5 microm) column. The temperature of detector was 40 degrees C, the pressure of N2 was 3.5 bar, and gain value was 9. RESULTS: The calibration curves were linear within the range of 1 - 5 microg, and the average recovery was 99.33% (RSD = 1.97%). CONCLUSION: The method was rapid, convenient, and accurate. It can be used for the quality control of this herbal medicine.