Liver Protection of a Low-Polarity Fraction from Ficus pandurata Hance, Prepared by Supercritical CO2 Fluid Extraction, on CCl4-Induced Acute Liver Injury in Mice via Inhibiting Apoptosis and Ferroptosis Mediated by Strengthened Antioxidation.

Ficus pandurata Hance (FPH) is a Chinese herbal medicine widely used for health care. This study was designed to investigate the alleviation efficacy of the low-polarity ingredients of FPH (FPHLP), prepared by supercritical CO2 fluid extraction technology, against CCl4-induced acute liver injury (ALI) in mice and uncover its underlying mechanism. The results showed that FPHLP had a good antioxidative effect determined by the DPPH free radical scavenging activity test and T-AOC assay. The in vivo study showed that FPHLP dose-dependently protected against liver damage via detection of ALT, AST, and LDH levels and changes in liver histopathology. The antioxidative stress properties of FPHLP suppressed ALI by increasing levels of GSH, Nrf2, HO-1, and Trx-1 and reducing levels of ROS and MDA and the expression of Keap1. FPHLP significantly reduced the level of Fe2+ and expression of TfR1, xCT/SLC7A11, and Bcl2, while increasing the expression of GPX4, FTH1, cleaved PARP, Bax, and cleaved caspase 3. The results demonstrated that FPHLP protected mouse liver from injury induced by CCl4 via suppression of apoptosis and ferroptosis. This study suggests that FPHLP can be used for liver damage protection in humans, which strongly supports its traditional use as a herbal medicine.

Advances in Supercritical Carbon Dioxide Extraction of Bioactive Substances from Different Parts of Ginkgo biloba L.

Ginkgo biloba L. has always been a popular area of research due to its various active ingredients and pharmacological effects. Ginkgo biloba is rich in ginkgo flavonoids, ginkgolides, and ginkgolic acid, with anti-inflammation, antioxidation, neuroprotection, anti-platelet agglutination, hypolipidemic effect, anti-cancer, and anti-radiation properties. There are many methods to extract and separate the active components of ginkgo. Among them, supercritical carbon dioxide fluid extraction (SFE-CO2) is known for its green, clean, and environment-friendly properties. In this paper, the pharmacological activities, the active components, and structures of different parts of ginkgo, the extraction methods of its effective ingredients, and the application of the SFE-CO2 method for the extraction and separation of active ingredients in Ginkgo biloba from leaves, seeds, pollen, and roots were reviewed, in order to make best use of ginkgo resources, and provide support and references for the development of SFE-CO2 of active components from Ginkgo biloba.

Investigation of Commiphora myrrha (Nees) Engl. Oil and Its Main Components for Antiviral Activity.

The resinous exudate produced by Commiphora myrrha (Nees) Engl. is commonly known as true myrrh and has been used since antiquity for several medicinal applications. Hundreds of metabolites have been identified in the volatile component of myrrh so far, mainly sesquiterpenes. Although several efforts have been devoted to identifying these sesquiterpenes, the phytochemical analyses have been performed by gas-chromatography/mass spectrometry (GC-MS) where the high temperature employed can promote degradation of the components. In this work, we report the extraction of C. myrrha by supercritical CO2, an extraction method known for the mild extraction conditions that allow avoiding undesired chemical reactions during the process. In addition, the analyses of myrrh oil and of its metabolites were performed by HPLC and GC-MS. Moreover, we evaluated the antiviral activity against influenza A virus of the myrrh extracts, that was possible to appreciate after the addition of vitamin E acetate (α-tocopheryl acetate) to the extract. Further, the single main bioactive components of the oil of C. myrrha commercially available were tested. Interestingly, we found that both furanodienone and curzerene affect viral replication by acting on different steps of the virus life cycle.

Identification of lipophilic components in Citri Reticulatae Pericarpium cultivars by supercritical CO2 fluid extraction with ultra-high-performance liquid chromatography-Q Exactive Orbitrap tandem mass spectrometry.

To systematically identify the lipophilic constituents of Citri Reticulatae Pericarpium from different cultivars, supercritical CO2 fluid extraction and ultra-high-performance liquid chromatography-Q Exactive Orbitrap tandem mass spectrometry were integrated for the component analysis of 18 batches of Citri Reticulatae Pericarpium from 12 cultivars for the first time. A total of 57 components from the supercritical CO2 fluid extracts were demonstrably or tentatively identified by the obtained parent peaks, fragment peaks, and retention times. In total, two flavonoids, six organic acids, nine coumarins, three aldehydes, seven esters, three terpenes, one limonoid, and five other compounds were detected for the first time; notably, coumarin components have not yet been reported in Citri Reticulatae Pericarpium. Furthermore, the extract constituents differed between cultivars. In particular, organic acids were more abundant in Citrus reticulata "Chachi" than in other cultivars, and pterostilbene was exclusively found in Citrus reticulata "Yichangju". The results showed that a greater variety of compounds in Citri Reticulatae Pericarpium could be extracted by supercritical CO2 fluid extraction and detected by ultra-high-performance liquid chromatography-Q Exactive Orbitrap tandem mass spectrometry. This study provides a more scientific basis for further analysis of the pharmacological activity and quality of Citri Reticulatae Pericarpium components from different cultivars.

Eicosapentaenoic Acid Extraction from Nannochloropsis gaditana using Carbon Dioxide at Supercritical Conditions.

This research shows that carbon dioxide supercritical fluid (CO2-SF) is an emerging technology for the extraction of high interest compounds for applications in the manufacturing of pharmaceuticals, nutraceuticals, and cosmetics from microalgae. The purpose of this study is to recover fatty acids (FAs) and, more precisely, eicosapentaenoic acid (EPA) from Nannochloropsis gaditana biomass by CO2-SF extraction. In the paper, the effect of mechanical pre-treatment was evaluated with the aim of increasing FAs recovery. Extraction was performed at a pressure range of 250⁻550 bars and a CO2 flow rate of 7.24 and 14.48 g/min, while temperature was fixed at 50 or 65 °C. The effect of these parameters on the extraction yield was assessed at each extraction cycle, 20 min each, for a total extraction time of 100 min. Furthermore, the effect of biomass loading on EPA recovery was evaluated. The highest EPA extraction yield, i.e., 11.50 mg/g, corresponding to 27.4% EPA recovery, was obtained at 65 °C and 250 bars with a CO2 flow rate of 7.24 g/min and 1.0 g biomass loading. The increased CO2 flow rate from 7.24 to 14.48 g/min enhanced the cumulative EPA recovery at 250 bars. The purity of EPA could be improved by biomass loading of 2.01 g, even if recovery was reduced.

Extraction of Bioactive Compounds Using Supercritical Carbon Dioxide.

Microalgae Dunaliella salina contains useful molecules such as ß-carotene and fatty acids (FAs), which are considered high value-added compounds. To extract these molecules, supercritical carbon dioxide was used at different operative conditions. The effects of mechanical pre-treatment (grinding speed at 0⁻600 rpm; pre-treatment time of 2.5⁻7.5 min) and operating parameters for extraction, such as biomass loading (2.45 and 7.53 g), pressure (100⁻550 bars), temperature (50⁻75 °C) and CO2 flow rate (7.24 and 14.48 g/min) by varying the extraction times (30⁻110 min) were evaluated. Results showed that the maximum cumulative recovery (25.48%) of ß-carotene was achieved at 400 bars and 65 °C with a CO2 flow rate of 14.48 g/min, while the highest purity for stage (55.40%) was attained at 550 bars and 65 °C with a CO2 flow rate of 14.48 g/min. The maximum recovery of FAs, equal to 8.47 mg/g, was achieved at 550 bars and 75 °C with a CO2 flow rate of 14.48 g/min. Moreover, the lowest biomass loading (2.45 g) and the first extraction cycle (30 min) allowed the maximum extraction of ß-carotene and FAs.

Supercritical carbon dioxide extract of Cinnamomum cassia bark: toxicity and repellency against two stored-product beetle species.

The extract from Cinnamomum cassia Presl bark was obtained with supercritical CO2 fluid extraction (SFE). Chemical components of the SFE extract were characterized by GC-MS spectrometry. The repellency and contact toxicity of the SFE extract were evaluated against the adults of Tribolium castaneum and Lasioderma serricorne along with those of its two main compounds. The results of GC-MS analysis indicated that 33 volatile constituents were identified from the extract. The main components included trans-cinnamaldehyde (32.1%), 3,3-dimethylhexane (10.6%) and 2,4-di-tert-butylphenol (7.9%). Testing results showed that the SFE extract had potent contact toxicity against T. castaneum and L. serricorne with LD50 values of 3.96 and 23.89 µg/adult, respectively. LD50 values of trans-cinnamaldehyde against T. castaneum and L. serricorne were 5.78 and 3.24 µg/adult, respectively. Additionally, percentage repellency values of the SFE extract and trans-cinnamaldehyde against T. castaneum and L. serricorne were rather high (PR = 100% and PR > 90%, respectively) at 78.63 and 15.73 nL/cm2 at 2 h post-exposure. 2,4-Di-tert-butylphenol showed some repellency against both beetle species. Considering its insecticidal and repellent effects, the SFE extract from C. cassia bark might be used in integrated pest management programs for T. castaneum and L. serricorne.

Antioxidant and anti-inflammatory activities of Lonicera japonica Thunb. var. sempervillosa Hayata flower bud extracts prepared by water, ethanol and supercritical fluid extraction techniques.

Lonicera japonica Thunberg (LJ) has long been used as an antipyretic, anti-inflammatory and anti-infectious agent in East Asia. The subspecies L. japonica Thunb. var. sempervillosa Hayata (LJv) is a variant that mainly grows in Taiwan. This study examined the antioxidant and anti-inflammatory activities of the extracts from the flower buds of these two species. The extracts were obtained by three extraction methods: water extraction, ethanol extraction, and supercritical-CO2 fluid extraction (SFE). The antioxidant activities of dry LJ (dLJ) extracts were superior to those of LJv extracts. Water extracts possessed higher activities than that prepared by ethanol or SFE. The total polyphenols content, total flavonoids content, and the amount of chlorogenic acid and luteolin-7-O-glucoside were all higher in the water extracts compared to the other two. The SFE extracts of these two species all exhibited excellent anti-inflammatory activities. Although the water and ethanol extracts of dLJ extracts had higher anti-inflammatory activity than that of LJv extracts, the SFE extracts prepared from fresh LJv flower buds (fLJv) exhibited the highest activity among all extracts. The SFE effectively isolates the bioactive components of L. japonica and can obtain the L. japonica extracts with high anti-inflammatory activity.

Supercritical CO2 Fluid Extraction Technology Optimization of Volatile Oil from Hui Medicine Huihui Nao-gengtong and Its Component Analysis / 中国药房

OBJECTIVE:To optimize supercritical CO2 fluid extraction (SFE-CO2) technology of volatile oil from Hui medi-cine Huihui Naogengtong for stroke,and to analyze and identify its component. METHODS:With yield of volatile oil and the con-tent of cinnamaldehydeas indexes,and extraction pressure,temperature and time as factors,single factor test and orthogonal test were designed to optimize SFE-CO2 technology;optimized technology was also validated. Physical and chemical indexes of volatile oil were determined,and the constituents of volatile oil were analyzed by GC-MS and retention index. Relative percentage contents of each component were calculated by area normalization method. RESULTS:Optimal technology was as follows as extraction pres-sure 30 MPa,extraction temperature 50 ℃ and extraction time 100 min. In verification test,the yield of volatile oil was 4.8%(RSD=0.02%,n=3),and the content of cinnamylaldehyde was 2.0 mg/g (RSD=1.8%,n=3). Physicochemical properties of volatile oil were as follows as relative density 1.05,refractive index 1.52,acid value 28.128 mg(KOH)/g,iodine value 74.47 g(I)/100 g,saponification value 54.2 mg(KOH)/g,peroxide 19.42 meq/kg. GC-MS analysis showed the similarity with retention index in literatures of 31 compounds were more than 80%,21 of them were identified and accounted for 44% of total volatile oil, among which E-cinnamaldehyde (9.14%),β-caryophyllene (3.21%),α-bisabolol (3.11%) and β-bisabolene (2.56%) had high relative percentage content. CONCLUSIONS:Optimal extraction technology is stable and feasible. The results of extraction technol-ogy and component analysis can provide important trail base and reference data for the exploration and utilization of Huihui Nao-gengtong.

Analysis of the Volatile Constituents of the Compound Extract from 3 Medicinal Materials in Siji Ganmao Tablet by Supercritical CO2 Fluid Extraction / 中国药房

OBJECTIVE:To study the volatile constituents of the compound extract from Folium Perillae,Pericarpium Citri and Herb Schizonepetae in Siji Ganmao tablet by supercritical CO2 fluid extraction (SFE-CO2). METHODS:Supercritical CO2 ex-traction technology was used to extract the volatile constituents of 3 medical material in Siji Ganmao tablet. The chemical constitu-ents of the compound extract from 3 medicinal materials were analyzed by GC-MS. RESULTS:The extracting rate of SFE-CO2 was 2.21%,21 compounds in the compound extract were separated and 16 compounds were identified with the major components of si-nensetin(36.56%)and linolic acid(19.52%). The extracting rate of water steam distillation(SD)was 1.035%,51 compounds in the compound extract were separated and 32 compounds were identified with the major components of D-limonene(62.40%) and thujone(15.49%). CONCLUSIONS:The volatile constituents of F. perillae,P. cCitri and H. Schizonepetae can be compound ex-tracted by SFE-CO2,however,it is different from the constituents of the compound volatile oil by SD.

Comparative pharmacokinetics and bioavailability of four alkaloids in different formulations from Corydalis decumbens.

ETHNOPHARMACOLOGICAL RELEVANCE: Corydalis decumbens, a Traditional Chinese Medicine listed in Chinese Pharmacopoeia, is clinically used for the treatment of paralytic stroke, headache, rheumatic arthritis and sciatica in China. AIM OF THE STUDY: This study was aimed to compare the pharmacokinetics and bioavailability of protopine, tetrahydropalmatine, bicuculline, and egenine in three formulations prepared from the rhizomes of Corydalis decumbens. MATERIALS AND METHODS: Alkaloid extract (CDAs-SFE) was prepared from the rhizomes of Corydalis decumbens by supercritical CO2 fluid extraction; CDAs-SFE/HPßCD (hydroxypropyl-ß-cyclodextrin inclusion complex), and CDAs-SFE/HCl (hydrochloride freeze-dried powder) were resulted from CDAs-SFE through complexation with HPßCD and hydrochloride, respectively. An UFLC-MS/MS method was developed for quantitative analysis of protopine, tetrahydropalmatine, bicuculline and egenine simultaneously in rat plasma after oral administration. The differences of pharmacokinetics and bioavailability of the four alkaloids in three formulations were determined by pharmacokinetics analyses. RESULTS AND CONCLUSIONS: The Cmax, AUC and bioavailability of protopine and tetrahydropalamatine (bioactive components) in CDAs-SFE/HCl were significantly higher than in CDAs-SFE and in CDAs-SFE/HPßCD. In contrast, in CDAs-SFE/HPßCD, AUC and bioavailability of tetrahydropalamatine were significantly lower, while those of bicuculline (toxic compound) appeared to be higher than both in CDAs-SFE and in CDAs-SFE/HCl. The results indicated that CDAs-SFE/HCl was the best beneficial formulation among the three formulations for the alkaloid extract prepared from the rhizomes of Corydalis decumbens, in which protopine and tetrahydropalamatine displayed higher bioavailability, but lower for bicuculline.

The Effect of Cataplasma of Artemisia Argyi by Supercritical CO2 Fluid Extraction in the Treatment of Rheumatoid Arthritis Rats / 浙江中医药大学学报

[Objective] The aim of this study is to investigate the mechanism and curative effect of cataplasma of Artemisia Argyi by supercritical CO 2 fluid extraction to rat weight, foot swel ing, interleukin-1β, interleukin-17 and tumor necrosis factor-αin the adjuvant arthritis rat models;and afford theoreti-cal evidence for clinical work.[Methods]50 healthy male SD rats were randomly divided into 5 groups:Control group, Model group, Low-dose treatment group, Medium-dose treatment group, High-dose treatment group. Model and al treatment goups took CFA for model ing. After establishing animal models of AA rats, the first group and second group used cataplasm without active drug ingredients. Other groups were given treatment according to dif-ferent drug doses. Assess the joint swel ing degree at different times. Measure the arthritic index and rat weight in two weeks. After 21 days ,detect the cy-tokines level of IL-lβ,IL-17 and TNF-α from the rat's serum and examine the pathological changes of joint observed by HE . [Results]Cataplasma of Artemisia Argyi by supercritical CO2 fluid extraction can significantly reduce rat foot swel ing of the ankle joint pathology ,reduce arthritis score and slow down the weight loss,meanwhile,effectively improve the general daily behavior of rats. The standard of IL-1β,IL-17,TNF-α in serum can be decreased by cataplasma of Artemisia Argyi by supercritical CO2 fluid extraction. The pathological result showed: in the treatment group,the rats synovial hyperplasia the vesselwing reduced, the inflammatory cel infiltration was more decreased than the model group. [Conclusion]The result showed that the cataplasma of Artemisia Argyi by supercritical CO2 fluid extraction had an obvious effect in treating RA. The mechanism was probably that it regulated the immune dis-order of RA.

Extraction of the Volatile Oil from Huoxiang Zhengqi Prescription Process with Supercritical CO_2 Fluid:Optimization of the Technical Condition by Orthogonal Test / 中国药房

OBJECTIVE:To establish the optimal technical condition for the extraction of volatile oil from the volatile herbs of Huoxiang zhengqi prescription with supercritical CO2 fluid(SFE-CO2).METHODS:The technical condition for the extraction of volatile oil from the volatile herbs of Huoxiang zhengqi prescription were optimized using L9(34)orthogonal design table with extraction pressure,the extraction temperature,the pressure and temperature of separator Ⅰ as factors and with the yield of the volatile oil as index.RESULTS:A highest extraction yield of volatile oil from the volatile herbs of Huoxiang zhengqi prescription was achieved(2.057%)under an optimal technical condition as follows:with extraction pressure at 25 MPa,the extraction temperature at 30 ℃,the pressure for separator Ⅰ at 9 MPa and the temperature of separator Ⅰ at 45 ℃.CONCLUSION:Extraction of volatile oil from the volatile herbs of Huoxiang zhengqi prescription by SFE-CO2 is characterized by high extraction yield,stable,pure operation,safe,and by which the biological activity of the volatile oil can be maintained,thus it is a potential extraction and separation method.

GC-MS Analysis of the Liposoluble Constituents Extracted with Supercritical CO_2 Fluid Extraction from Aractylodes Mcrocephala / 中国药房

OBJECTIVE:To analysis and identify the liposoluble constituents extracted with supercritical CO 2 fluid ex-traction(SFE)from atractylodes macrocephala by gas chromatography-mass spectrometry(GC/MS).METHODS:Volatile oil was extracted from atractylodes macrocephala,to analsis and identify the chemical components in liposoluble constituents with GC/MS,DB-5MS capillary column was the chromatographic column and helium was carrier gas,the flow rate was 2.0ml/min,and the column temperature was raised from 70℃ to 250℃ at an increasing rate of 10℃/min.EI ion sources were adopted in mass spectra.RESULTS:A total of 19 chromatographic peaks were separated and identified,most of which were unsatu-rated fatty acids and esters,accounting for 96.2% of the total peak area.CONCLUSION:Using GC/MS,the chemical com-position of the liposoluble constituents in atractylodes macrocephala can be analyzed,which provides references for the further exploitation and utilization of atractylodes macrocephala.

Supercritical CO_2 extraction of atractylenolide Ⅰ from Atractylodes macrocephala / 中草药

Objective To investigate the extraction technique for atractylenolide Ⅰ in Atractylodes macrocephala by supercritical CO_2 fluid extraction and develop a method used for determining the content of atractylenolide Ⅰ in the extract by HPLC. Methods The effects of seven facters, such as the extracting pressure, resolving pressure etc, to the extraction rate of atractylenolide Ⅰ in A. macrocephala by supercritical CO_2 extraction were investigated. RP-HPLC was used to determine the content of atractylenolide Ⅰ in extraction of A. macrocephala. The separation was performed on Hypersil ODS2 column with methanol-water (70∶30) as mobile phase. The flow rate was 1.0 mL/min and the wavelength of UV detector was 220 nm. Results The optimal extracting conditions: taking 10% alcohol as entraiter, the particle size of medicinal substances was 60 screen meshes, extracting pressure 25 MPa, resolving pressure 5 MPa, extracting temperature 40 ℃, resolving temperature 30 ℃, and the extracting time 4 h. Conclusion Supercritical extraction is time-shorter and efficient in extracting atractylenolide Ⅰ from A. macrocephala. It is suitable to both trial and industrialized production. The method established to determine the content of atractylenolide Ⅰ of A. macrocephala by supercritical extraction is simple, sensitive, and reliable.

Effect of pressure in supercritical state on extraction and crystallization of andrographolide / 中草药

Object To separate and purify andrographolide by supercritical CO 2 extraction and crystallization. Methods The method of systematic observation was used to investigate the crystal appearance, purity and mass of andrographolide by supercritical CO 2 extraction and crystallization, analyzing by scanning electron microscope (SEM) and HPLC. Results The higher the pressure, the smaller and shorter the crystal shape, and the purer the purity, the more the mass of andrographolide. Conclusion This technique is a innovation of traditional supercritical fluid extraction (SCFE) technique, which realizes highly effectual separation of andrographolide by supercritical CO 2 extraction and synchronous crystallization, developes a new technology to separate andrographolide and other active ingredients from Chinese traditional herbs.