CN1190394C - Pineapplie leaf extract, its preparation method and application - Google Patents
Pineapplie leaf extract, its preparation method and application Download PDFInfo
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Abstract
The present invention relates to a pineapple leaf extract, a preparation method thereof, and an application thereof, particularly a traditional Chinese medicine extract containing a plurality of active components and prepared from pineapple leaves used as raw material, and a usage thereof. The present invention has the technical scheme that the pineapple leaf extract containing 30 to 80% of total phenol components through separation and extraction; experimental researches show that the pineapple leaf extract has the functions of expanding cerebral vessels directly, relieving vascular spasm and diseases resulting from the vascular spasm better, regulating blood fat and resisting oxidation. More specifically, pineapple leaf extract can be used for preparing medicine for treating hyperlipemia, hyperlipemia of diabetes, cerebral ischemia, peroxide elevation of bodies and other diseases resulting from the peroxide raising.
Description
Technical field
The present invention relates to a kind of is the Chinese medical extract that contains the various active composition, preparation method of feedstock production and the new purposes aspect medical thereof with the pineapple leaves, belongs to technical field of traditional Chinese medicines.
Background technology
Pineapple is bromelia pineapple Ananas comosus (L.) Merr.[Bromelia comosa L.; A.sativusSchult.Et Schult.f.] fruit.Originate in America, existing extensively planting in the torrid areas, all there is plantation on ground such as China Fujian, Guangdong, Hainan, Guangxi, Yunnan.This product beginning is stated from clearly. and Wu Qirui " An Illustrated Book on Plants ", run after fame to reveal pocket.This secretary is carried: " reveal pocket and produce Guangdong, a POLO is given birth between hill, and is real in radish, the acropetal leaf cluster, and the long dark tooth of point, flavor look fragrant tool is good.The ripe golden yellow of fruit, Pi Jian such as fish scale shape, peeling food meat, fragrant and sweet no slag, June is ripe ".
Ancient times does not have clearly record for pineapple leaves, modern people arghan commonly used as the raw material of braiding and papermaking (" the Chinese Higher plant illustrated handbook. the 5th " Science Press 1976).Pineapple leaves on the books has and helps digestion and stomach when near, and the antidiarrheic effect cures mainly summer heat and rushes down, maldigestion, gastral cavilty distending pain etc. (" China's book on Chinese herbal medicine. the 8th volume " Shanghai science tech publishing house 1999).
Pineapple leaves is more rare as medicinal record, especially the pineapple leaves of not appearing in the newspapers or pineapple leaves extract are at preparation treatment disorders of lipid metabolism, cerebral vasospasm, the too much caused hyperlipidaemia of body endoperoxide, postprandial hyperlipemia of diabetes, functional disorders of brain, the application in the medicine of diseases such as cerebrovascular blood supply insufficiency.
Summary of the invention
The purpose of this invention is to provide pineapple leaves extract, pineapple leaves preparation method of extract and the pineapple leaves extract new purposes aspect medical.
Pineapple leaves extract provided by the invention is characterized in that: this extract contains 30～80% total phenol property composition, and described extract adopts following method to obtain:
(1) earlier 5～15 times of water of medicinal material weight or alcoholic solvent heating or room temperature are extracted, obtain extracting solution;
(2) with extracting solution behind normal pressure or concentrating under reduced pressure under 30～100 ℃ of temperature, with medicinal material weight 1～10 times of water dissolution, centrifugal or static filtration discards precipitation, gets supernatant liquor;
(3) supernatant liquor is passed through exchange column, first water wash-out, elutriant is discarded, uses concentration greater than 70% ethanol elution again, and with elutriant concentrating under reduced pressure under 30～90 ℃ of temperature, drying under reduced pressure or lyophilize obtain the pineapple leaves extract.
The invention provides a kind of pineapple leaves preparation method of extract, this method comprises the steps:
(1) earlier 5～15 times of water of medicinal material weight or alcoholic solvent are heated or room temperature is extracted at 30 ℃～100 ℃, obtain extracting solution;
(2) with extracting solution behind normal pressure or concentrating under reduced pressure under 30～100 ℃ of temperature, with medicinal material weight 1～10 times of water dissolution, centrifugal or static filtration discards precipitation, gets supernatant liquor;
(3) supernatant liquor is passed through exchange column, first water wash-out, elutriant is discarded, uses concentration greater than 70% ethanol elution again, and with elutriant concentrating under reduced pressure under 30～90 ℃ of temperature, drying under reduced pressure or lyophilize obtain the pineapple leaves extract.
The number of times that heating is extracted described in the above-mentioned steps (1) is 3 times, is 1～3 hour at every turn.Described room temperature was extracted as soaking at room temperature 2～10 days.
Exchange column of the present invention adopts macroporous adsorbent resin, ion exchange resin or polymeric amide.
Technical program of the present invention also lies in providing the new purposes of the medical aspect of pineapple leaves extract, this purposes is:
The application of pineapple leaves extract in the medicine of preparation treatment hyperlipidemia.
The pineapple leaves extract is in the preparation treatment diabetics application in the medicine of high fat after the meal.
The application of pineapple leaves extract in preparation treatment body endoperoxide raises the medicine of caused disease.
The application of pineapple leaves extract in the medicine of the caused disease of preparation treatment treatment cerebrovascular blood supply insufficiency.
When the disease for the treatment of disorders of lipid metabolism, superoxide rising, cerebral vasospasm and causing thus, with the pineapple leaves extract is activeconstituents, with the preparation process of routine, use separately or prepare the medicine of operable various different dosage forms clinically with other drug.
Experimental result of the present invention shows, the effect that the pineapple leaves extract is significantly increased mouse meninx volume of blood flow.Promptly there was cerebral blood flow to raise in 3 minutes behind the intravenously administrable, and shows certain dose-effect relationship.Show that the pineapple leaves extract has directly dilating effect to the cerebrovascular, mitigation is preferably arranged for the caused disease of vascular spasm.The pineapple leaves extract has and suppresses the diabetic mice effect that raises of blood triglyceride after the meal definitely, makes usually after the meal that blood fat remains on certain level, and unconspicuous persistence raises, and oral effective dose is 600mg/kg.The pineapple leaves extract gives rat the oral effect that can obviously suppress serum cholesterol and the obvious rising of triglyceride level simultaneously.Effective dose is 300mg/kg and 150mg/kg.Experiment also shows, the rising that pineapple leaves is extracted in the object, treated in vitro all can obviously suppress the brain superoxide, and the experiment in vitro minimal effective concentration is 1.2 * 10
-6G/ml.The vivo medicine-feeding subliminal dose is 300mg/kg.The above results prompting pineapple leaves extract has the blood fat of setting the tone and oxidation resistant effect.
Description of drawings
Fig. 1 is the typical curve of coffic acid reference substance concentration and absorbancy.
Embodiment
The preparation technology of pineapple leaves extract provided by the invention is as follows: at first with the 5-15 times of water of medicinal material weight or the alcohol of various concentration pineapple leaves is extracted 3 times 30 ℃～100 ℃ heating, be 1～3 hour at every turn; Or soaking at room temperature 2～10 days, obtain extracting solution.Used alcoholic solvent can adopt methyl alcohol, ethanol, Virahol, butanols etc. or aqueous alcoholic solvent.Then extracting solution is being reduced pressure (pressure 90-98Kpa), temperature concentrates down at 30～100 ℃, uses 1～10 times of water dissolution of medicinal material weight again, and centrifugal or static filtration discards precipitation, gets supernatant liquor; Make supernatant liquor pass through exchange column (exchange columns such as macroporous adsorbent resin, ion exchange resin, polymeric amide), first water wash-out, elutriant is discarded; Use concentration greater than 70% wash-out again, elutriant is 30～90 ℃ of (pressure 98Kpa) concentrating under reduced pressure in temperature, and enriched material decompression (pressure 98Kpa) drying or lyophilize promptly obtains the pineapple leaves extract.
The composition of pineapple leaves extract adopts ultraviolet spectrophotometry, colorimetry etc. to measure, and total phenol content is between 30～80%.Wherein main component is phenolic acid and flavones etc.
Be exactly individually or add that the preparation process with routine is prepared into medicament in the compound on medicament is used with the pineapple leaves extract; Its medicament can be the various different dosage forms that can use clinically, as powder, and pill, capsule, tablet, microcapsule, soft capsule, film, suppository, injection, paste, tincture, powder, electuary, aerosol, various external preparations etc.
Following embodiment and effect experiment data are confirmed preparation of the said pineapple leaves extract of the present invention and uses thereof, should not regard the following example as limitation of the present invention again simultaneously.
Embodiment 1:
Get 100 ℃ in the water heating of 5 times of amounts of 1 kilogram of usefulness of pineapple leaves and extract 3 times, each 3 hours, extracting solution decompression (pressure 98Kpa) temperature 100 ℃ concentrate after, static with 1 times of water dissolution of medicinal material weight, get supernatant, pass through polyamide column.Elder generation's water wash-out, elutriant is discarded, uses 70% ethanol elution again, and elutriant decompression (pressure 98Kpa) concentrates for 30 ℃, and lyophilize obtains extract.The total phenol property composition of pineapple leaves extract adopts colorimetry to measure, and the total phenol property component content is about 30% in the extract.
Embodiment 2:
Get 60 ℃ of the 70% ethanol heating of 15 times of amounts of 1 kilogram of usefulness of pineapple leaves and extract 3 times, each 2 hours, centrifugal after 60 ℃ of extracting solution decompression (pressure 98Kpa) heating concentrate with 5 times of water dissolution with medicinal material weight, get supernatant, pass through macroporous adsorbent resin.Elder generation's water wash-out, elutriant is discarded, uses 95% ethanol elution again, and elutriant decompression (pressure 98Kpa) concentrates for 90 ℃, and decompression (pressure 98Kpa) drying obtains extract.The total phenol property composition of pineapple leaves extract adopts colorimetry to measure, and the total phenol property component content is about 80% in the extract.
Embodiment 3:
Get 30 ℃ of the methyl alcohol heating of 5 times of amounts of 1 kilogram of usefulness of pineapple leaves and extract 3 times, each 1 hour, after 30 ℃ of extracting solution decompression (pressure 90-Kpa) heating concentrate, with 10 times of water dissolution with medicinal material weight, centrifugal, get supernatant, the accent pH value is a slightly acidic, passes through anionite-exchange resin.Elder generation's water wash-out, elutriant is discarded, uses 80% ethanol elution again, and elutriant decompression (pressure 98Kpa) concentrates for 30 ℃, and decompression (pressure 98Kpa) drying obtains extract.The total phenol property composition of pineapple leaves extract adopts colorimetry to measure, and the total phenol property component content is about 60% in the extract.
Embodiment 4:
Get the 95% ethanol soaking at room temperature 2 days of 15 times of amounts of 1 kilogram of usefulness of pineapple leaves, centrifugal after 50 ℃ of extracting solution decompression (pressure 90-Kpa) heating concentrate with 5 times of water dissolution of medicinal material weight, get supernatant, pass through macroporous adsorbent resin.Elder generation's water wash-out, elutriant is discarded, uses 80% ethanol elution again, and the elutriant decompression concentrates for 100 ℃, and lyophilize obtains the pineapple leaves extract, and the total phenol property composition in the extract adopts colorimetry to measure, and the total phenol property component content is about 50% in the extract.
The composition measurement of pineapple leaves extract total phenol property adopts colorimetry that its total phenol property composition is carried out assay.
1, reagent and instrument
Ultraviolet spectrophotometer (TU-1800PC); Analytical balance, coffic acid reference substance (available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute), the total phenol extraction thing of pineapple leaves; All the other reagent are analytical pure.
2, method and result
1. measure the selection of wavelength: dispose certain density coffic acid reference substance and the total phenol extraction thing of pineapple leaves trial-product respectively, before and after the colour developing, in 200-800nm, write down absorption spectrum respectively.The result shows that all there is maximum absorption the total phenol extraction thing of coffic acid reference substance and pineapple leaves trial-product colour developing back at the 720nm place.So select 720 ± nm as measuring wavelength.
2. the investigation in reaction times in the condition determination: draw certain density coffic acid reference substance, add methyl alcohol 3.5ml, 0.3% sodium dodecyl sulfate solution 2ml, 0.6% potassium ferricyanide solution 1ml, 0.9% liquor ferri trichloridi 1ml shakes up the dark place and leaves standstill 5min.Dilute hydrochloric acid with 0.1mol/L is diluted to 10ml.At the 720nm place, its optical density of different timings, result show that the 30min primitive reaction is complete, so selective reaction is 30min.
3. the configuration of reference substance solution: precision takes by weighing coffic acid reference substance 10mg, puts into the 10ml volumetric flask, uses the dissolve with methanol constant volume, shakes up.Precision is drawn 1ml, puts in the 50ml volumetric flask to add methanol constant volume, shakes up the standard substance that promptly get 20ug/ml.
4. the drafting of typical curve: precision takes by weighing coffic acid reference substance solution 0,1ml, 1.5ml, 2.0ml, 2.5ml, 3.0ml, 3.5ml puts respectively in the brown volumetric flask of 10ml, add methyl alcohol 3.5ml respectively, 2.5ml, 2.0ml, 1.5ml, 1ml, 0.5,0ml adds 0.3% sodium dodecyl sulfate solution 2ml again, 0.6% potassium ferricyanide solution 1ml, 0.9% liquor ferri trichloridi 1ml shakes up the dark place and leaves standstill 5min, and the dilute hydrochloric acid with 0.1mol/L is diluted to 10ml again.Shake up the dark place and leave standstill 30min.Compare with No. zero pipe, put in the cuvette of 1cm, measure its absorbancy, drawing standard curve at the 720nm place.Coffic acid reference substance concentration is respectively 2ug, 3ug, and 4ug, 5ug, 6ug, 7ug/ml, absorbancy is respectively 0.21,0.34, and 0.47,0.58,0.65,0.79.Because the absorbancy of coffic acid in the 2-7ug/ml scope is a straight line by initial point, so adopt the method for comparing to calculate total phenol content in the total phenol extraction thing of pineapple leaves.The results are shown in Figure 1.
5. the mensuration of samples contg: precision takes by weighing the total phenol extraction thing of pineapple leaves 10mg, puts into the brown volumetric flask of 10ml, uses the dissolve with methanol constant volume, shakes up.The accurate 0.05ml that draws adds methyl alcohol 3ml respectively, adds 0.3% sodium dodecyl sulfate solution 2ml again, 0.6% potassium ferricyanide solution 1ml, and 0.9% liquor ferri trichloridi 1ml shakes up the dark place and leaves standstill 5min, and the dilute hydrochloric acid with 0.1mol/L is diluted to 10ml again.Shake up the dark place and leave standstill 30min.Measure its absorbancy at the 720nm place, absorbancy is 0.35.Calculating content by the reference substance relative method is 62%.
Effect experiment example 1: to the effect of mouse meninx volume of blood flow
Observe the effect of pineapple leaves extract of the present invention to mouse meninx volume of blood flow.The pineapple leaves extract, lot number: 030311-2.Time spent is mixed with desired concn with physiological saline.Experimental animal is a male ICR mouse, non-inbred lines closed colony, and body weight 18-22g is provided by Beijing dimension tonneau China Experimental Animal Center.Laboratory room temperature 20-22 ℃, relative humidity 40%～60%, the ventilator ventilation, lamp 12h/ day, raise in cages, 8 in every cage, per three days cleaning cage houses are once.
After getting mouse urethane (1.0g/kg) anesthesia, fixing head cuts skin, in the motor area of cerebral cortex (AP
-1, L
2) settle laser probe (JI-200 laser microcirculation instrument, Tianjin Nankai University produces), treat stablize after 5-10 minute and begin to test cerebral blood flow, the variation of record administration front and back meninx volume of blood flow.
This experimental observation index and index are expressed: measured result is directly through the JI-200 software processes, and the meninx volume of blood flow is with KHz (kilohertz) expression, and the meninx volume of blood flow increases percentage value [volume of blood flow before=(the preceding volume of blood flow of volume of blood flow-administration after the administration)/administration].
This test dosage is respectively 25mg/kg, 12.5mg/kg and 6.25mg/kg; Route of administration is the tail intravenously administrable; The administration volume is the 0.05ml/10g mouse.
This test blank is an equal-volume physiological saline, and the positive drug contrast is ligustrazine injection, 40mg/2ml.Yongkang, Beijing pharmaceutical factory produces, lot number: 9811191, and the accurate word (1996) of medicine is defended No. 157008 in the capital, and the mouse consumption is 25mg/kg.
This testing data is all with the Excel software processes.
Table 1 pineapple leaves extract is to the effect of normal mouse brain volume of blood flow (x ± s)
Grouping agent cerebral blood flow (CBF) (KHz)
Amount
(mg/ 0135 10 15 (divides
Kg) clock)
Control group 31.32 31.84 31.42 31.58 31.58 31.45
±2.71 ±3.03 ±2.86 ±3.02 ±2.51 ±2.52
Ligustrazine 25 31.01 32.74 33.8 33.7 33.57 33.17
±2.37 ±1.99 ±1.94* ±2.12* ±1.9* ±2.02*
Extract 100 31.63 34.28 36.53 34.7 37.31 36.42
±2.6 ±3.46 ±5.23* ±3.76* ±3.18* ±3.18*
Extract 50 31.67 33.8 35.88 37.18 36.65 36.21
±3.18 ±2.89 ±2.72* ±2.43* ±3.7* ±1.9*
Extract 25 33.17 34.84 37.07 38.03 38.05 36.76
±6.61 ±7.35 ±8.24 ±7.85* ±8.39* ±8.43*
With compare P＜0.05 with the time control group.n＝8
Table 2 pineapple leaves extract increases percent value (x ± s) to normal mouse brain volume of blood flow
Grouping dosage cerebral blood flow (CBF) increases percent value (%)
(mg/ 0135 10 15 (divides
Kg) clock)
5.54 3.54 2.19 3.19 2.49
Ligustrazine 25 0 5.77 ± 9.17 ± 8.86 ± 8.52 ± 7.17 ±
2.97 3.77* 3.75* 4.38* 4.32*
Extract 100 0 8.31 ± 15.28 ± 18.18 ± 18.0 ± 15.2 ±
3.78* 9.83** 4.94** 3.36** 4.79**
Extract 50 0 6.91 ± 13.71 ± 17.99 ± 16.18 ± 14.96 ±
3.62* 7.16* 9.01* 7.86** 8.31*
Extract 25 0 4.96 ± 11.52 ± 14.69 ± 14.57 ± 10.59 ±
3.43 5.06* 4.3** 5.6* 6.28*
With compare * P＜0.05, * * P＜0.01 with the time control group.n＝8
Experimental result shows, the effect that the pineapple leaves extract is significantly increased mouse meninx volume of blood flow.Generally after administration, promptly there was cerebral blood flow to raise in 1 minute, and shows certain dose-effect relationship; Show that the pineapple leaves extract has directly dilating effect to the cerebrovascular, prompting has mitigation preferably for the caused disease of vascular spasm.
Effect experiment example 2: to the diabetic mice influence of blood fat after the meal
Observing pineapple leaves of the present invention extracts and gives birth to the diabetic mice restraining effect of high triglyceride after the meal.The used pineapple leaves extract of the present invention, lot number: 030311-2.Time spent is mixed with desired concn with physiological saline.Experimental animal is male ICR kind mouse, non-inbred lines closed colony, and body weight 18-22g is provided by Beijing dimension tonneau China Experimental Animal Center.Laboratory room temperature 20-22 ℃, relative humidity 40%～60%, the ventilator ventilation, lamp 12h/ day, raise in cages, 8 in every cage, per three days cleaning cage houses are once.
Get mouse and be divided into 4 groups at random, 12 every group, behind the fasting 24h, the quiet notes tetraoxypyrimidine of tail (lot number: 971201, the chemical plant produces in the west, Beijing.Chemical pure) (50mg/kg), blood sugar and blood triglyceride (TG) are surveyed in the blood sampling of fasting overnight after three days (behind about 8h) eye socket.Get blood sugar and be subjects the above person of 9mmoml/L.Irritate the stomach different pharmaceutical then, after the administration 0.5 hour, begin to irritate sweet oil (chemical pure, lot number: 980428, Beijing fragrant grass pharmaceutical ﹠ chemicall research development corporation) 0.15ml/10g to mouse, and pick up counting, take a blood sample through eye socket respectively at irritating behind the stomach sweet oil 1,2,4 and 8 hours, centrifugation serum detects blood sugar and triglyceride level.All quantitatively replenish physiological saline (0.1ml/10g body weight) after each blood sampling through gi tract.Measured result is through the Excel software processes.
This test dosage is 600mg/kg; The administration volume is the 0.2ml/10g mouse.Gastric infusion.
This test blank is an equal-volume physiological saline, and the positive drug contrast is gemfibrozil, lot number: 010401, and Yue Zhou pharmaceutical Co. Ltd in Zhejiang produces, and the mouse consumption is 600mg/kg.
Table 3 pineapple leaves extract to hyperglycemia mouse high-fat meal after the influence (x ± S) of blood sugar
Grouping drug dose blood sugar (mmol/L)
Mg/k 01248 is (little
During g)
Normal group physiology 5.21 7.24 6.602 6.081 6.615
Salt solution ± 2.91 ± 3.42 ± 3.041 ± 1.649 ± 1.735
Model group physiology 12.02 16.33 14.38 13.27 11.55
Salt solution ± 7.82 ± 7.94* ± 7.47 ± 9.73 ± 9.61
Model group lucky non-600 11.86 16.18 12.67 9.48 5.58
Luo Qi ± 6.79 ± 9.85* ± 10.03 ± 9.07 ± 2.45
Model group extracts 600 12.63 18.38 18.32 14.44 8.74
Thing ± 3.51 ± 7.52* ± 6.33 ± 5.68 ± 3.01
Compare * P＜0.05 with just organizing.n＝12
Table 4 pineapple leaves extract to hyperglycemia mouse high-fat meal after the influence (x ± S) of blood triglyceride
Grouping drug dose triglyceride level (mmol/L)
Mg/kg 01248 (hour)
Normal group physiology 1.038 1.972 ± 2.298 1.626 ± 1.605 ±
Salt solution ± 0.226 0.591 ± 0.471 0.675 0.668
Model group physiology 0.912 1.202 ± 1.881 2.682 ± 3.992 ±
Salt solution ± 0.347 0.352* ± 0.746 1.423* 2.886*
Model group lucky non-600 0.742 0.948 ± 1.545 1.525 ± 2.654 ±
Luo Qi ± 0.17 0.417 ± 1.263 1.533* 2.314
Model group extracts 600 0.746 0.972 ± 1.573 1.775 ± 1.736 ±
Thing ± 0.181 0.358 ± 0.725 0.499# 1.382#
Compare * P＜0.05 with normal group.Compare #P＜0.05 with model group.n＝10
Experimental result shows, feed raise the triglyceride level diet after 1-2 hour, triglyceride level raises in the normal mouse blood, descends later on, recovers normal after 4 hours.Model group is then fed and is raised behind the triglyceride level that triglyceride level raises gradually in the blood, reaches the peak, and maintains this high level later in 4 hours.Can suppress obviously after the administration of pineapple leaves extract after the meal that triglyceride level raises in the blood, raise the back 1-8 hour from feeding, triglyceride level all is in lower level in the blood.Compare with model group, notable difference (p＜0.05) is arranged.Show that the pineapple leaves extract has the obvious suppression effect of high triglyceride after the meal, and blood sugar is not had tangible influence.
Effect experiment example 3: to the regulating effect of disorders of lipid metabolism rat fat
The used male Wistar rat of the present invention, non-inbred lines closed colony, body weight 180-220g.Available from Chinese medical courses in general institute institute of lab animals.Conformity certification number: the capital is moving be betrothed to (036) number.Laboratory room temperature 20-22 ℃, relative humidity 40%～60%, the ventilator ventilation, lamp 12h/ day, raise in cages, 5 in every cage, per three days cleaning cage houses are once.
The used pineapple leaves extract of the present invention, lot number: 030311-2.Positive control drug be the fenofibrate capsule (sharp purport is flat, phenofibrate), the 100mg/ grain.Lot number: 0012162.The accurate word (1996) of medicine is defended No. 153001 in Guangdong.The used test instrument is the HITACHI-7060 automatic clinical chemistry analyzer.Rat high lipid food (prescription: 10% yolk powder, 5% big oil, 0.5% cholate, 0.2% cholesterol, 84.3% base-material) is available from Institute of Experimental Animals, Chinese Academy of Medical Sciences.
Get totally 60 of rats, be divided into 6 groups at random, 10 every group.Give high 2 weeks of fat material, eye socket venous plexus blood sampling in the 15th day, separation of serum is surveyed TC, TG.Do evenly adjustment between group according to TC and TG.Give the medicine of various dose simultaneously.Eye socket venous plexus blood sampling in the 14th day after administration, separation of serum is surveyed blood total cholesterol (TC) and triglyceride level (TG).The pineapple leaves extract is divided into heavy dose of 300mg/kg, and middle dosage group is 150mg/kg, and low dose is 75mg/kg, and fenofibrate is 70mg/kg, is equivalent to 2 times of clinical equivalent amount.Model control group is given equal-volume distilled water.The normal control group is given normal diet, and gives equal-volume distilled water.
The administration volume is 1mL/kg.The gastric infusion approach, every day 1 time.The gained data are by the Excel software processes, and t checks between group.
This experimental result is as table 5:
2 all rat TC after table 5 administration, and the TG variation (x ± s)
Grouping drug dose (mg/kg) TC (mg/dl) TG (mmol/L)
Normal group 104.12 ± 19.04 1.028 ± 0.219
Model group 172.97 ± 25.65# 1.967 ± 0.391#
Model group fenofibrate 70 153.15 ± 11.53* 1.448 ± 0.349*
The model group pineapple leaves is extracted 300 140.69 ± 26.82*, 1.458 ± 0.369*
Thing
The model group pineapple leaves is extracted 150 149.63 ± 26.83*, 1.58 ± 0.362*
Thing
The model group pineapple leaves is extracted 75 160.56 ± 33.74 1.609 ± 0.372*
Thing
Compare * P＜0.05 with model group.Compare * P＜0.05 with normal group.n＝10
Administration is during 2 weeks, and each organizes the equal blood TC of medicine and TG all shows tangible activity.The big or middle dosage group of pineapple leaves extract can obviously suppress the rising (P＜0.05) of TC and TG.The positive drug fenofibrate also has the obvious suppression effect to TC and TG.Prompting pineapple leaves extract has the effect of the blood fat of setting the tone.
Effect experiment example 4: to the influence of brain MDA generation
Observe the restraining effect that pineapple leaves extract of the present invention generates mouse brain MDA.The used pineapple leaves extract of the present invention, lot number: 030311-2.Time spent is mixed with desired concn with physiological saline.Testing used animal is male secondary ICR mouse, non-inbred lines closed colony, and Beijing dimension tonneau China Experimental Animal Center provides, and body weight is 18-22g.Laboratory room temperature 20-22 ℃, relative humidity 40%～60%, the ventilator ventilation, lamp 12h/ day, raise in cages, 10 in every cage, per three days cleaning cage houses are once.
Get normal mouse, get brain after the execution, add physiological saline do homogenate (with the brain ratio be 5: 1).4 ℃ of refrigerators left standstill 5 minutes, got the even muddy liquid in top.Every pipe adds cerebral tissue liquid 150uL, adds different concns soup 50ul, and 37 ℃ of water-baths were hatched 30 fens.Add 10%SDS 0.1ml successively, 50% Glacial acetic acid thiobarbituricacid liquid 0.5ml and 1% hydrochloric acid solution 2ml.Boiling water bath 15 minutes takes out back cold water and put 10 minutes, and 1500 left the heart 15 minutes, gets supernatant and surveys the absorbance A value in the 532nm place.Do three multiple pipes for every group.Calculate inhibiting rate=[(control group A-administration group A)/control group A * 100%].
The gained data are through the EXCEL software processes.T check between group.
This test pineapple leaves extract administration final concentration is respectively 1.2 * 10
-6G/ml, 1.2 * 10
-5G/ml, 1.2 * 10
-4G/ml, 1.2 * 10
-3G/ml, 1.2 * 10
-2G/ml.
This test is an experiment in vitro.This test blank is an equal-volume physiological saline. the positive drug contrast is for vitamin-E, and available from Sigma company, the experiment final concentration is 1.2 * 10
-2G/ml.
This testing data is all with the Excel software processes.
The external restraining effect that brain MDA is generated of table 6 pineapple leaves extract (x ± s)
Grouping medicine final concentration light absorption value (532nm)
(g/ml)
Physiological saline 0.598 ± 0.012
Vitamin-E 1.2 * 10
-20.273 ± 0.009**
Pineapple leaves extract 1.2 * 10
-20.32 ± 0.012**
Pineapple leaves extract 1.2 * 10
-30.391 ± 0.011**
Pineapple leaves extract 1.2 * 10
-40.422 ± 0.01**
Pineapple leaves extract 1.2 * 10
-50.481 ± 0.015*
Pineapple leaves extract 1.2 * 10
-60.554 ± 0.019*
Compare * P＜0.05, * * P＜0.01 with the physiological saline group.n＝3
The influence that table 7 pineapple leaves extract generates mouse brain MDA (x ± s)
Drug dose (mg/kg) light absorption value
532nm/ g albumen
Physiological saline 0.604 ± 0.177
Vitamin-E 50 0.422 ± 0.134*
Extract 600 0.396 ± 0.125*
Extract 300 0.437 ± 0.104*
Extract 150 0.525 ± 0.125
Compare * P＜0.05 with the physiological saline group.n＝10
This experimental result shows, the external effect that has inhibition strongly to lack sugared anoxic cerebral tissue MDA generation of pineapple leaves extract, and minimal effective concentration is 1.2 * 10
-5G/ml.Oral administration also can obviously suppress the generation of mouse brain MDA.Prompting pineapple leaves extract has certain oxidation resistant effect.
Claims (8)
1. pineapple leaves extract, it is characterized in that: this extract contains 30～80% total phenol property composition, and described extract adopts following method to obtain:
(1) earlier 5～15 times of water of medicinal material weight or alcoholic solvent are heated or room temperature is extracted at 30 ℃～100 ℃, obtain extracting solution;
(2) with extracting solution behind normal pressure or concentrating under reduced pressure under 30～100 ℃ of temperature, with medicinal material weight 1～10 times of water dissolution, centrifugal or static filtration discards precipitation, gets supernatant liquor;
(3) supernatant liquor is passed through exchange column, first water wash-out, elutriant is discarded, uses concentration greater than 70% ethanol elution again, and with elutriant concentrating under reduced pressure under 30～90 ℃ of temperature, drying under reduced pressure or lyophilize obtain the pineapple leaves extract.
2. pineapple leaves preparation method of extract as claimed in claim 1, this method comprises the steps:
(1) earlier 5～15 times of water of medicinal material weight or alcoholic solvent are heated or room temperature is extracted at 30 ℃～100 ℃, obtain extracting solution;
(2) with extracting solution in normal pressure or decompression, temperature 30～100 ℃ concentrate after, with 1～10 times of water dissolution of medicinal material weight, centrifugal or static filtration discards precipitation, gets supernatant liquor;
(3) supernatant liquor is passed through exchange column, first water wash-out, elutriant is discarded, uses concentration greater than 70% ethanol elution again, and at 30～90 ℃ of following concentrating under reduced pressure, drying under reduced pressure or lyophilize obtain the pineapple leaves extract with elutriant.
3. according to the described pineapple leaves preparation method of extract of claim 2, it is characterized in that: the number of times that the heating described in the step (1) is extracted is 3 times, is 1～3 hour at every turn; Described room temperature was extracted as soaking at room temperature 2～10 days.
4. according to the described pineapple leaves preparation method of extract of claim 2, it is characterized in that: described exchange column adopts macroporous adsorbent resin, ion exchange resin or polymeric amide.
5. the application of pineapple leaves extract as claimed in claim 1 in the medicine of preparation treatment hyperlipidaemia.
6. pineapple leaves extract as claimed in claim 1 is in the preparation treatment diabetics application in the medicine of hyperlipidemia after the meal.
7. the application of pineapple leaves extract as claimed in claim 1 in preparation treatment body endoperoxide raises the medicine of caused disease.
8. the application of pineapple leaves extract as claimed in claim 1 in the medicine of the caused disease of preparation treatment cerebrovascular blood supply insufficiency.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB031375774A CN1190394C (en) | 2003-06-18 | 2003-06-18 | Pineapplie leaf extract, its preparation method and application |
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| CNB031375774A CN1190394C (en) | 2003-06-18 | 2003-06-18 | Pineapplie leaf extract, its preparation method and application |
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| CN1190394C true CN1190394C (en) | 2005-02-23 |
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| CN101648980B (en) * | 2009-08-28 | 2013-01-30 | 广东轻工职业技术学院 | Method for combined preparation of pineapple polyphenol and juice by taking pineapple peel as main material and application thereof |
| DE102013208707A1 (en) * | 2013-05-13 | 2014-11-13 | Robert Bosch Gmbh | Overflow valve for a high pressure pump in a fuel injection system |
| CN106924141B (en) * | 2017-03-28 | 2020-06-23 | 海南师范大学 | Preparation method of pineapple leaf extract applied to whitening field |
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