CN103416165A - Method for collecting rosemary and method for preparing rosemary antioxidant extractives - Google Patents
Method for collecting rosemary and method for preparing rosemary antioxidant extractives Download PDFInfo
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- CN103416165A CN103416165A CN2012101588045A CN201210158804A CN103416165A CN 103416165 A CN103416165 A CN 103416165A CN 2012101588045 A CN2012101588045 A CN 2012101588045A CN 201210158804 A CN201210158804 A CN 201210158804A CN 103416165 A CN103416165 A CN 103416165A
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Abstract
The invention discloses a method for collecting rosemary and method for preparing rosemary antioxidant extractives. The method includes the steps of collecting rosemary in summer in a place with the east longitude from 103 degrees 36 minutes to 109 degrees 35 minutes and the northern latitude from 24 degrees 37 minutes to 29 degrees 13 minutes or carrying out secondary rosemary collection in winter after collecting rosemary in summer. Experiments show that the collection method can be applied to collecting rosemary in mountainous areas in Guizhou Province in summer, and secondary collection of leaves of rosemary in winter also can be achieved when the rosemary grows and recovers in autumn after the summer collection, the antioxidant activity of obtained ethanol extracts is higher than others, and the lipid oxidation inhibition performance is inconsistent with free radical scavenging capacity. The restriction capacity of extracts in stems is higher than that of extracts in leaves, and the activity of rosemary with 10-year age of plant and collected in winter is high.
Description
Technical field
The present invention relates to biological technical field, relate in particular to a kind of rosemary, Xue MingRosma rinus officinalis of gathering the preparation method of method and rosemary, Xue MingRosma rinus officinalis antioxidant extract.
Background technology
Rosemary, Xue MingRosma rinus officinalis (Rosmarinusofficinalis L.) is Labiatae Rosmarinus undershrub, originates in Mediterranean Region, is distributed widely in the ground such as Central Europe, West Europe, north African, is a kind of famous and precious natural perfume plant.The essential oil and the antioxidant that in rosemary, Xue MingRosma rinus officinalis flower and leaf, extract, have significantly biologically active and the value medical health cares such as antibiotic, anti-oxidant, anti-inflammatory, as spices and essence, anticorrosive additive, agrochemical, is widely used in daily use chemicals, food and medicine industry.This makes rosemary, Xue MingRosma rinus officinalis become one of resource plant of Mediterranean Region most worthy.
Rosmarinus officinalis extract is as antioxidant, and energy Scavenger of ROS and free radical, effectively suppress lipid oxidation, thereby it is rotten to postpone food spoilage, keeps good colour and taste, is the natural additive for foodstuff of safety non-toxic.The active ingredient of rosemary antioxidant is mainly carnosic acid, carnosol and Rosmarinic acid.The antioxidation activity of the content of these three kinds of phenols components and ratio and extract is closely related.Research shows, many factors can affect the antioxidation activity of Rosmarinus officinalis extract, cultivation condition (temperature, light, water, fertilizer) for example, plantation area, breed difference and the position etc. of drawing materials.Therefore, during the new exotic plant of introduction of use, the impact of the factor such as the environmental condition, vegetation growth state, cultivation step on ground on plant product introduced in research, for successfully introducing a fine variety, adaptive training and obtain high income, significant.
Summary of the invention
An object of the present invention is to provide a kind of rosemary, Xue MingRosma rinus officinalis of gathering method.
Method provided by the invention, comprise the steps: 103 ° 36 '-109 ° 35 of east longitude ', 24 ° 37 '-29 ° 13 of north latitude ' locate, the annual rosemary, Xue MingRosma rinus officinalis of gathering in summer and/or winter.
In said method, be June or July described summer; The monthly mean temperature 20-22 ℃ in described June or July, monthly mean rainfall 150-200mm;
Be December described winter; The monthly mean temperature 5-7 ℃ in described December, monthly mean rainfall 40-50mm.
In said method, the monthly mean temperature 20-22 ℃ in described June or July, the monthly total precipitation 150-200mm in described June or July;
The monthly mean temperature 5-7 ℃ in described December, the monthly total precipitation 40-50mm in described December;
During described gathering rosemary, Xue MingRosma rinus officinalis strain age for not higher than 10 years, during described gathering rosemary, Xue MingRosma rinus officinalis strain be specially 10 years age, 4 years or 2 years;
103 ° 36 '-109 ° 35 of described east longitude ', 24 ° 37 '-29 ° 13 of north latitude ' locate to economize into Guizhou In China;
Described picking methods is for retaining described rosemary, Xue MingRosma rinus officinalis acrial part 25-35cm, other acrial parts of gathering;
Described other acrial parts are leaf and/or stem.
The rosemary, Xue MingRosma rinus officinalis of being gathered by above-mentioned method is also the scope of protection of the invention.
The application that above-mentioned rosemary, Xue MingRosma rinus officinalis has in antioxidation activity rosemary, Xue MingRosma rinus officinalis ethanol extract preparation is also the scope of protection of the invention.
Second purpose of the present invention is to provide a kind of method for preparing the rosemary, Xue MingRosma rinus officinalis ethanol extract.
Method provided by the invention, comprise the steps:
By above-mentioned rosemary, Xue MingRosma rinus officinalis tissue 95%(volumn concentration) ethanol multi-reflow extraction, collect extract, obtain the rosemary, Xue MingRosma rinus officinalis ethanol extract.
In said method, above-mentioned rosemary, Xue MingRosma rinus officinalis tissue and above-mentioned 95%(volumn concentration) proportioning of ethanol is 1g: 8ml;
The number of times of described reflux extraction is 3 times;
The temperature of each described reflux extraction is 85 ℃, and the time of described reflux extraction is 1h.
In said method, before described reflux extraction, also comprise by above-mentioned rosemary, Xue MingRosma rinus officinalis dry, the step pulverized of tissue;
After described reflux extraction, also comprise described extract is filtered, concentrates, obtain the step of rosemary, Xue MingRosma rinus officinalis ethanol extract;
Described rosemary, Xue MingRosma rinus officinalis be organized as stem or leaf.
The rosemary, Xue MingRosma rinus officinalis ethanol extract prepared by said method is also the scope of protection of the invention; Above-mentioned Rosmarinus officinalis extract specifically has antioxidation activity.
It is also the scope of protection of the invention that above-mentioned rosemary, Xue MingRosma rinus officinalis ethanol extract has application in the antioxidation activity product in preparation.In above-mentioned application, described antioxidation activity is specially to be removed the DPPH ability or removes the ABTS ability.
Of the present invention experimental results show that, collecting method of the present invention can be gathered in Guizhou mountainous area summer or winter, the more important thing is can be through the growth recovery in autumn after gathering summer, winter the secondary recovery rosemary, Xue MingRosma rinus officinalis, the rosemary, Xue MingRosma rinus officinalis ethanol extract of gathering for twice in 1 year inherent June and December is low unlike the antioxidation activity of the rosemary, Xue MingRosma rinus officinalis ethanol extract once of gathering 1 year inherent September, high on the contrary, the rosemary, Xue MingRosma rinus officinalis of gathering for twice in 1 year inherent June and December is described, and not only biomass is many, and the antioxidation activity of rosemary, Xue MingRosma rinus officinalis ethanol extract is higher; Further selecting not homophyletic to be tested age, can find out 2,4,10 and there is every year antioxidation activity, therefore illustrate this collecting method can make rosemary, Xue MingRosma rinus officinalis plant to degenerate year after year not obvious, can sustainable utilization at least ten years, and all there is antioxidation activity.Therefore, this be a kind of take obtain the high activity antioxidant and gather and extracting method as target, efficient, continuable rosemary, Xue MingRosma rinus officinalis.
The accompanying drawing explanation
Fig. 1 removes the impact of ability season on DPPH
Fig. 2 removes the impact of ability season on ABTS
Fig. 3 is the impact of season on the beta carotene bleaching
Embodiment
The experimental technique used in following embodiment if no special instructions, is conventional method.
In following embodiment, material used, reagent etc., if no special instructions, all can obtain from commercial channels.
Experiment in following embodiment is triplicate, results averaged.
Gather groping of opportunity and the extraction of embodiment 1, rosemary, Xue MingRosma rinus officinalis has antioxidative active extractive
One, the rosemary, Xue MingRosma rinus officinalis collecting time gropes
1, the different time rosemary, Xue MingRosma rinus officinalis of gathering
Guizhou province is located in southern plateau, winter gentleness, summer is damp and hot, rosemary, Xue MingRosma rinus officinalis can be grown under wild state, is applicable to commerial growing for the production of antibiotic and oxidation resistant product, obtains the ecological benefits of green barren hill, soil and slope protection simultaneously.
2009, in Guizhou (103 ° 36 '-109 ° 35 of east longitude ', 24 ° 37 '-29 ° 13 of north latitude '), respectively in summer (June 25,21.3 ℃ of monthly mean temperatures, monthly total precipitation 195.4mm), (September 10 in autumn, 20.2 ℃ of monthly mean temperatures, monthly total precipitation 156.7mm) and pluck 2 years strain rosemary, Xue MingRosma rinus officinaliss in age winter (December 15,6.7 ℃ of monthly mean temperatures, monthly total precipitation 47.1mm), retain rosemary, Xue MingRosma rinus officinalis acrial part 30cm, other acrial parts of gathering.And the plant of gathering winter is the plant after the growth in autumn after gathering summer, the plant for not gathering in summer of gathering autumn.
2, extract
By the above-mentioned 3 groups rosemary, Xue MingRosma rinus officinaliss of plucking stem, leaf process with the following method respectively: by rosemary, Xue MingRosma rinus officinalis stem, leaf dried in the shade, pulverize respectively, get again 30g and pulverize rear rosemary, Xue MingRosma rinus officinalis cured leaf and the rear rosemary, Xue MingRosma rinus officinalis scapus of 50g pulverizing, be respectively charged in the round-bottomed flask of 2000ml, by solid-liquid ratio 1: ethanol 8(quality (g): volume (ml)) add the 95%(volumn concentration), 85 ℃ of reflux extraction 1h, filter, add 95% ethanol, repeat twice of reflux extraction.Merge extract, add the 5g activated carbon decolorizing, filter, obtain deep orange look clarified solution.Vacuum-concentrcted, the product obtained finally by drying is rosemary, Xue MingRosma rinus officinalis ethanol extract (leaf) and rosemary, Xue MingRosma rinus officinalis ethanol extract (stem), the said extracted thing is yellow powder.
3, the active detection
Above-mentioned rosemary, Xue MingRosma rinus officinalis ethanol extract (leaf) and rosemary, Xue MingRosma rinus officinalis ethanol extract (stem) are carried out respectively to following antioxidation activity detection:
1), DPPH removes experimental technique (removing free radical)
Accurately take DPPH(1,1-diphenyl-2-trinitrophenyl-hydrazine) (sigma, article No.: D9132) 39.4mg, dissolve also constant volume with 95% ethanol and, in the 1000mL volumetric flask, obtain 1 * 10
-4The DPPH reagent of mol/L, keep in Dark Place.Add 3.9mlDPPH reagent in test tube, 0.1ml rosemary, Xue MingRosma rinus officinalis ethanol extract (leaf or stem), fully mix.The lower lucifuge of room temperature (25 ℃) is placed 30min, measures light absorption value under the 515nm wavelength, is designated as Ax.95% ethanol of 0.1ml of usining replaces sample as blank, and the light absorption value of mensuration is A
0, 95% ethanol of 3.9ml of usining replaces DPPH ﹒ test solution as the contrast of sample liquid background, records light absorption value Ax
0.
Calculation sample is removed the clearance rate E of DPPH free radical: clearance rate
A wherein
0For blank absorbency; Ax is the sample absorbance; Ax
0For the absorbance of sample solution background, using clearance rate and sample concentration as calibration curve, ask IC50.
2) ABTS removes experimental technique (removing free radical)
Take 0.0384g ABTS(sigma, article No.: A1888) volumetric flask is settled to 10ml, and 0.0134g potassium peroxydisulfate water is settled to 10ml, and the two fully mixes, standing over night under the condition of room temperature lucifuge, Xing Cheng ﹒ ABTS
+The free radical storing solution.Be diluted to working solution with absolute ethyl alcohol before using, making the absorbance of this solution under the 734nm wavelength is 0.70 ± 0.02, and 30 ℃ of balances.Test need be carried out under 30 ℃ of conditions, and the Rosmarinus officinalis extract of 40 μ L is joined to 4.0mL De ﹒ ABTS
+In working solution, with 40 μ L ethanol, replace the rosemary, Xue MingRosma rinus officinalis extract to join 4.0mL De ﹒ ABTS
+In working solution, as blank, the 4.0mL absolute ethyl alcohol of usining replaces 4.0mL De ﹒ ABTS+ working solution as sample liquid background test solution, reacts 30min under 30 ℃, under the 734nm wavelength, measures absorbance.Calculation sample Qing Chu ﹒ ABTS
+The clearance rate of free radical: clearance rate
Wherein: A
0For the reacted absorbance of blank, Ax is the reacted absorbance of sample liquid, A
X0Absorbance for sample liquid background.Using clearance rate and sample concentration as calibration curve, ask IC50.
3), carotin bleaching experiment method (inhibition lipid oxidation)
0.5mg beta carotene (Fluka-22040) is dissolved in the 1ml chloroform, adds afterwards 25 μ l linoleic acid and 200mg polysorbate40, is mixed to get beta carotene-linoleic acid solution.Use Rotary Evaporators to remove the chloroform in mixed liquor, add afterwards 120ml distilled water, acutely shake and mix.Get test tube, add the above-mentioned mixed liquor of 2.5ml and 350 μ l rosemary, Xue MingRosma rinus officinalis ethanol extracts (3mg/ml), 50 ℃ of water-bath 3h.Measure absorbance every half an hour under the 470nm wavelength.Vitamin C and TBHQ are as positive control, and 350 μ l distilled water replace antioxidant solution as blank.Inhibiting rate: AA%=[1-(A
0-A
t)/(A
0 s-A
t s)] * 100.Wherein, A
0 sAnd A
0Be respectively sample and the blank absorbance at 0min, A
t sAnd A
tBe respectively sample and the blank absorbance after water-bath t min.
Shown in the testing result of above-mentioned 3 experiments following Fig. 1-3 and table 1-4:
Table 1 is season DPPH to be removed the impact of ability
Table 2 is season ABTS to be removed the impact of ability
Table 3 is the impact of season on the beta carotene bleaching
The activity of the extract that the rosemary, Xue MingRosma rinus officinalis that table 4 obtains for different collecting seasons prepares
From Fig. 1 and Fig. 2, can find out, from leaf or stem, can find out, the curve in June and December is all on September, the IC50 value of associative list 1, explanation is high at the antioxidation activity of the rosemary, Xue MingRosma rinus officinalis ethanol extract of gathering in June and December, and can find out, the rosemary, Xue MingRosma rinus officinalis ethanol extract of gathering for twice in 1 year inherent June and December is low unlike the antioxidation activity of the rosemary, Xue MingRosma rinus officinalis ethanol extract once of gathering 1 year inherent September, high on the contrary, the rosemary, Xue MingRosma rinus officinalis of gathering for twice in 1 year inherent June and December is described, and not only biomass is many, and the antioxidation activity of rosemary, Xue MingRosma rinus officinalis ethanol extract is higher.
Two, rosemary, Xue MingRosma rinus officinalis the groping of strain age of gathering
1, the homophyletic rosemary, Xue MingRosma rinus officinalis of gathering age not
2010, in Guizhou (103 ° 36 '-109 ° 35 of east longitude ', 24 ° 37 '-29 ° 13 of north latitude '), in summer (June 25,21.3 ℃ of monthly mean temperatures, monthly total precipitation 195.4mm), pluck 2 years, 4 years and 10 years strain rosemary, Xue MingRosma rinus officinaliss in age.
2, extract
Method with above-mentioned one 2;
3, the active detection
Method with above-mentioned one 3;
As a result shown in following table 5:
The activity of the extract that the rosemary, Xue MingRosma rinus officinalis that table 5 is gathered for homophyletic not age prepares
Can find out, the rosemary, Xue MingRosma rinus officinalis in 2,4,10 years ages of strain, the antioxidation activity no significant difference of ethanol extract, illustrate rosemary, Xue MingRosma rinus officinalis in 10 years antioxidation activity all less than significantly changing.
Therefore, as can be seen from the above, in Guizhou (103 ° 36 '-109 ° 35 of east longitude ', 24 ° 37 '-29 ° 13 of north latitude '), June and December gather rosemary, Xue MingRosma rinus officinalis the antioxidation activity of ethanol extract all high.
Claims (10)
- The rosemary, Xue MingRosma rinus officinalis of gathering method, comprise the steps: 103 ° 36 '-109 ° 35 of east longitude ', 24 ° 37 '-29 ° 13 of north latitude ' locate, the annual rosemary, Xue MingRosma rinus officinalis of gathering in summer and/or winter.
- 2. method according to claim 1, it is characterized in that: be June or July described summer;Be December described winter.
- 3. method according to claim 1 and 2 is characterized in that:The monthly mean temperature 20-22 ℃ in described June or July, the monthly total precipitation 150-200mm in described June or July;The monthly mean temperature 5-7 ℃ in described December, the monthly total precipitation 40-50mm in described December;During described gathering rosemary, Xue MingRosma rinus officinalis strain age for not higher than 10 years, during described gathering rosemary, Xue MingRosma rinus officinalis strain be specially 10 years age, 4 years or 2 years;103 ° 36 '-109 ° 35 of described east longitude ', 24 ° 37 '-29 ° 13 of north latitude ' locate to economize into Guizhou In China;Described picking methods is for retaining described rosemary, Xue MingRosma rinus officinalis acrial part 25-35cm, other acrial parts of gathering;Described other acrial parts are leaf and/or stem.
- 4. the rosemary, Xue MingRosma rinus officinalis that arbitrary described method is gathered in claim 1-3.
- 5. rosemary, Xue MingRosma rinus officinalis claimed in claim 4 has the application in antioxidation activity rosemary, Xue MingRosma rinus officinalis ethanol extract preparation.
- 6. a method for preparing the rosemary, Xue MingRosma rinus officinalis ethanol extract, comprise the steps:By rosemary, Xue MingRosma rinus officinalis claimed in claim 4 tissue 95%(volumn concentration) ethanol multi-reflow extraction, collect extract, obtain the rosemary, Xue MingRosma rinus officinalis ethanol extract.
- 7. method according to claim 6 is characterized in that:Described rosemary, Xue MingRosma rinus officinalis claimed in claim 4 tissue and described 95%(volumn concentration) proportioning of ethanol is 1g: 8ml;The number of times of described reflux extraction is 3 times;The temperature of each described reflux extraction is 85 ℃, and the time of described reflux extraction is 1h.
- 8. according to the described method of claim 6 or 7, it is characterized in that:Before described reflux extraction, also comprise by rosemary, Xue MingRosma rinus officinalis claimed in claim 4 dry, the step pulverized of tissue;After described reflux extraction, also comprise described extract is filtered, concentrates, obtain the step of rosemary, Xue MingRosma rinus officinalis ethanol extract;Described rosemary, Xue MingRosma rinus officinalis be organized as stem or leaf.
- 9. the rosemary, Xue MingRosma rinus officinalis ethanol extract that in claim 6-8 prepared by arbitrary described method; Described Rosmarinus officinalis extract specifically has antioxidation activity.
- 10. rosemary, Xue MingRosma rinus officinalis ethanol extract claimed in claim 9 has in the antioxidation activity product and applies in preparation; Described antioxidation activity is specially to be removed the DPPH ability or removes the ABTS ability.
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Cited By (3)
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| CN106360609A (en) * | 2016-08-19 | 2017-02-01 | 天津春发生物科技集团有限公司 | Vanilla flavor liquid seasoning and production method thereof |
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| CN115337334A (en) * | 2022-09-22 | 2022-11-15 | 杭州师范大学 | Application of rosemary alcohol extract in inhibiting AGEs (advanced glycation end products) induced by Methylglyoxal (MGO) |
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| CN106490156A (en) * | 2016-09-30 | 2017-03-15 | 张文 | Herba Rosmarini Officinalis breast |
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| CN106360609A (en) * | 2016-08-19 | 2017-02-01 | 天津春发生物科技集团有限公司 | Vanilla flavor liquid seasoning and production method thereof |
| CN114887350A (en) * | 2022-04-18 | 2022-08-12 | 中国科学院植物研究所 | Method for extracting rosemary extract serving as strong antioxidant |
| CN115337334A (en) * | 2022-09-22 | 2022-11-15 | 杭州师范大学 | Application of rosemary alcohol extract in inhibiting AGEs (advanced glycation end products) induced by Methylglyoxal (MGO) |
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