CN107361130A - A kind of method for suppressing Apple greasy and occurring - Google Patents

A kind of method for suppressing Apple greasy and occurring Download PDF

Info

Publication number
CN107361130A
CN107361130A CN201710474332.7A CN201710474332A CN107361130A CN 107361130 A CN107361130 A CN 107361130A CN 201710474332 A CN201710474332 A CN 201710474332A CN 107361130 A CN107361130 A CN 107361130A
Authority
CN
China
Prior art keywords
fruit
greasy
egcg
apple
analysis
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201710474332.7A
Other languages
Chinese (zh)
Inventor
刘翠华
王聪
杨艳青
周彬
张玉洁
任小林
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Northwest A&F University
Original Assignee
Northwest A&F University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Northwest A&F University filed Critical Northwest A&F University
Priority to CN201710474332.7A priority Critical patent/CN107361130A/en
Publication of CN107361130A publication Critical patent/CN107361130A/en
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
    • A23B7/00—Preservation or chemical ripening of fruit or vegetables
    • A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
    • A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
    • A23B7/154—Organic compounds; Microorganisms; Enzymes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/02—Food
    • G01N33/025—Fruits or vegetables

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Polymers & Plastics (AREA)
  • Medicinal Chemistry (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The invention discloses a kind of method for suppressing Apple greasy and occurring, adopting the accumulation of rear apple epicutile wax causes the generation of greasy, fruit quality is influenceed, fatty acid synthase is the key enzyme of wax synthesis, and Epigallo-catechin gallate (EGCG) is the inhibitor of fatty acid synthase.To probe into effects of the EGCG in apple greasy, respectively with 0,1.5,3.0gL‑1EGCG aqueous solution soakings ' Pink Lady ' Apple, is stored at 25 DEG C, results of regular determination greasy degree, waxy component content and physical signs.As a result the fruit surface main fatty acid and esters content for showing to handle through EGCG are below control group, and reduce the occurrence degree of pericarp greasy.This explanation EGCG reduces the accumulation of waxy component, so as to slow down the generation of greasy phenomenon by suppressing the activity of fatty acid synthase.EGCG does not have a significant impact only for fatty acid synthase to the quality trait of fruit.

Description

A kind of method for suppressing Apple greasy and occurring
Technical field
The invention belongs to agricultural technology field, is related to a kind of method for suppressing Apple greasy and occurring.
Background technology
The same with most of higher plants, Apple epidermis outermost covers one layer of lipid for being soluble in organic solvent and mixed Compound, we are called wax.Wax is divided into the inside wax that is filled in cuticula and positioned at cuticula outermost layer and direct The epicutile wax contacted with external environment, its key component include over-long chain fatty acid and its derivative, such as:Hydrocarbon, aldehyde, alcohol, (the Millar etc. 1999 such as ketone, ester；Jenks etc. 2002), and some triterpene compounds and other small molecule secondary metabolisms production Thing (Bianchi etc. 1990).But at present to detailed pathway, Control factors, the mechanism that influences each other and the transporter of wax synthesis System etc. still imperfectly understands (Wang Dongyang etc. 2016；Chen Wei etc. is 2016).Epicutile wax in the growing of plant, adapt to the external world Environment etc. plays an important role (superfine 2011) of Song.For after fruit is adopted, wax coat can effectively prevent non-stomata Property moisture loss (Premachandra etc. 1992), regulation gas exchanges (Baur etc. 1996), resist microorganism infringement (Fan etc. 1999；Yin etc. is 2011) and other adverse environmental factors have vital effect (Li Ling grade 2011).But in storage The change of waxy component also results in the disorderly generation of Fruit physiology, and the greasy as caused by apple epicutile wax change of component drops Low fruit appearance quality (Richardson-Harman etc. 1998) and commodity value (Dadzie etc. 1995；Veraverbeke Deng 2001；High China etc. 2005；Curry 2008), so as to cause larger economic loss.
At present, the research of the generating process and pathogenesis of pericarp greasy, which has obtained adopting rear field, more widely closes Note, work (Christeller and the Roughan 2016 of correlation are all carried out on the fruits such as apple, pears；Jia Xiaohui etc. 2016), existing research shows that the accumulation of aliphatic acid and greasy have important relation.By long-term storage, apple Content of fatty acid understands persistent accumulation and causes exacerbation (Dadzie etc. 1995 of greasy in fruit；Christeller and Roughan 2016), and as the synthesis precursor of esters etc., the increase of free fatty acids also results in the product of Ester It is tired, a large amount of unsaturated oleic acid and linoleic esters are such as identified in ' loud, high-pitched sound ' apple epicutile wax that greasy occurs (Christeller and Roughan2016).In order to reduce the loss brought after greasy phenomenon is adopted to apple, forefathers pass through 1- MCP is handled Apple, it is found that 1-MCP can keep the hardness of fruit and titratable acid content etc. (Fan etc. 1999), and Greasy degree is reduced, its effect is mainly (Wang Xiaofei etc. 2014) to realize by the signal transduction path of ethene suppressing. In addition, 1-MCP use has a negative impact to part fruit, such as colours uneven, part fruit resistance ability and declines, disease increase, Aromatic substance is produced and is suppressed, so as to influence flavor of product etc. (Wei Haocheng etc. 2003).Therefore, find more single-minded suppression Agent suppresses greasy, while seems without influenceing other qualities and its necessary.
The synthesis of Apple wax originates in the de novo formation of C16 and C18 aliphatic acid, and the reaction is in epidermal cell Completed in plastid, wherein fatty acid synthase (Fatty Acid Synthase, FAS) is the key in fatty acid synthesis pathway Enzyme.Researcher has found a kind of natural active matter --- Epigallo-catechin gallate (EGCG) (epigallocatechin- 3-gallate, EGCG) it can effectively suppress activity (Wang and the Tian 2001 of fatty acid synthase in zooblast；Wang Deng 2003).EGCG is a kind of catechin compounds, is extracted from tealeaves, has and removes interior free yl, anti-inflammatory, resists The bioactivity such as mutation, anti-aging and improvement liver function, have been listed in potential cancer therapy drug in states such as Sino-U.S. and have been studied (Wang Xia etc. 2005).But the relevant report that EGCG is not applied in terms of apple greasy at present.
The content of the invention
It is an object of the invention to provide a kind of method for suppressing Apple greasy and occurring, this method is directed to Apples Key component fitter acids and its ester analog derivative in real wax, this research and utilization various concentrations EGCG processing ' Pink Lady ' apple Fruit, it is intended to influences of the fatty acid sythetase inhibitor EGCG to greasy phenomenon is inquired into, so as to further disclose pericarp greasy With the relation of fatty acid metabolism, theoretical foundation on the one hand is provided to study the regulation and control of greasy and its genesis mechanism, on the other hand Also laid the foundation to find a kind of commercial, nontoxic antistaling agent for effectively suppressing greasy generation on apple.
Its concrete technical scheme is：
A kind of method for suppressing Apple greasy and occurring, comprises the following steps：
Step 1, fruit processing
Experiment sets 3 processing:(1) (distilled water) is compareed；(2)1.5g·L-1EGCG；(3)3.0g·L-1EGCG.At three Fruit using vacuum infiltration method processing fruit, i.e., is placed in vacuum desiccator, sealed, then using SHZ-3 recirculated waters by reason It is 0.06Mpa that multiplex vavuum pump, which is evacuated to vacuum, maintains 60s, is slowly gone up to normal pressure.Fruit taking-up is rearmounted to dry in the air at room temperature Dry, the fruit after drying is placed at 25 DEG C and stored.Each processing sets 3 repetitions, each repeats 30 fruits.
Step 2, physical signs and fruit quality measure
With TRACE GC ULTRA type gas chromatograph for determination Ethylene Production Rates.Method is as follows:Taken out after the closed 1h of fruit Take gas, GDX-502 chromatographic columns, carrier gas N2, 70 DEG C of column temperature, 70 DEG C of injector temperature, hydrogen flowing quantity 0.7kgcm-2, air stream Measure 0.7kgcm-2, nitrogen flow 1.0kgcm-2, sample size 1mL, flame ionization ditector detection, detect room temperature 150℃。
Respiratory intensity:With the type CO of TELAIRE 70012Analysis-e/or determining；
The hardness of fruit is determined using GS-15 type fruit texture analyzer；
Step 3, greasy degree evaluation
The degree of fruit greasy is judged using the method for subjective appreciation, greasy degree is divided into 4 grades:0 is Do not occur；1 is slight；2 be moderate；3 be severe.Subjective appreciation group is formed by 3 by training and experienced student, is Reduce error, it is selected the apple (0 grade) of greasy not to occur and the fruit (3 grades) of greasy most serious compares, it is slight to touch Fruit surface evaluation greasy degree is touched, and every time before touch fruit surface by hand wiped clean.Greasy degree is 3 people The average value of results of sensory evaluation, subjective appreciation fruit are 15 fixed fruits.
Step 4, the processing of fruit waxy component；
Step 5, data processing
Test data analyzer uses Excel 2007, SAS 9.4 to carry out statistical analysis, carries out variance analysis and uses ANOVA Method, Multiple range test use DUNCAN methods, significant difference P<0.05.
Further, in step 2, the hardness of fruit is determined using GS-15 type fruit texture analyzer.Design parameter is set as: Probe diameter 10mm；Activation threshold value 0.10kg；Pop one's head in fall off rate 10mms-1；Probe returns to speed 10mms-1；Measurement speed Rate 5mms-1；Measurement distance 10.0mm.Titratable acid content is determined by GMK-835F fruit acidity analyzer.Solubility is solid Shape thing content holds saccharometer using PAL-1 types and measured.
Further, in step 3, the measure work of an index of correlation is carried out every 3d, fruit greasy changes after 9d Relatively slowly, the measure of an index of correlation is carried out every 4 or 5d.To sum up, physical signs and fruit quality measure, greasy degree Evaluation is carried out in fruit storage 0,3,6,9,13,17,21 and 26d respectively.
Further, step 4 is specially：
The extraction of the total wax of 4.1 fruits
Respectively in fruit storage 0,6,13,21 and 26d extraction waxs.Whole fruit is immersed under normal temperature and fills 400mL's 45s is soaked in the beaker of chloroform, and each fruit extracts 3 times, the leachate of 3 times is mixed after extraction, filtering, is transferred to Concentrated in 3000mL round bottom distilling flasks using RE-2000 Rotary Evaporators under the conditions of 40 DEG C, concentrate is transferred to In 50mL serum bottle.Wax extract solution is dried up with MD200-1 nitrogen evaporators under stable nitrogen stream, then with chloroform and Methanol is with 10:1 ratio is settled to 20mL, then is heated with 50 DEG C of water-baths, wax is completely dissolved, mixed, is eventually adding 1mL internal standard (heptadecane) is standby.
Esters and the processing method and analysis condition of aliphatic acid in 4.2 waxy components
Silylation reaction is first carried out before esters analysis:Drawing 500 μ L, the standby wax extract solution of constant volume has been extremely In 1.5mL QSP centrifuge tubes, 4h is to being completely dried for vacuum drying concentration.Add double (trimethyl silicon substrate) trifluoroacetamides of 400 μ L (BSTFA) the Silylation reaction that 70 DEG C of baking ovens carry out 45min is put into after, sample uses n-hexane constant volume after nitrogen evaporator dries up To 500 μ L, (made through in 0.45 μm of organic system membrane filtration to 1.5mL sample injection bottles, finally carrying out gas chromatographic analysis with heptadecane For internal standard).
Ester analysis uses gas chromatography.Condition is as follows:The DB-35MS low pole capillary chromatographic columns (U.S. Agilent J＆W scientific companies, 30m × 0.32mm, 0.25 μm), 70 DEG C of column temperature, 230 DEG C of injector temperature, detection 300 DEG C of device temperature, carrier gas is high purity N2.Heating schedule:70 DEG C of initial temperature, 1min is kept, with 10 DEG C of min-1Heating Speed rises to 200 DEG C, then with 5 DEG C of min-1Heating rate rise to 300 DEG C, keep 30min.
Esterification is first carried out before total fatty acids analysis, method is as follows:Draw the 500 μ L standby wax extractions of constant volume Liquid, add 50 μ L Heptadecanoic acides and dried up as internal standard with nitrogen evaporator, add 200 μ L potassium hydroxide-methanol solution (0.4mol mL-1), shaken up after hydrolyzing 15min at 70 DEG C, add 300 μ L boron trifluorides (BF3) 30min is esterified under the conditions of 70 DEG C, finally 500 μ L are settled to n-hexane.About 500 μ L of supernatant liquid are drawn to 1.5mL sample injection bottles with liquid-transfering gun.
Total fatty acids analysis uses gas chromatography.Condition:KB-FFAP capillary chromatographic columns (U.S. Agilent J＆W Scientific companies, 30m × 0.32mm, 0.25 μm), 70 DEG C of column temperature, 230 DEG C of injector temperature, 300 DEG C of detector temperature； Carrier gas is the N of high-purity2.Heating schedule:70 DEG C of initial temperature, 1min is kept, with 10 DEG C of min-1Heating rate rises to 200 DEG C, then with 5 DEG C of min-1Heating rate rises to 230 DEG C, keeps 20min.
Compared with prior art, beneficial effects of the present invention：
The study result show that：As a result fruit surface main fatty acid and the esters content handled through EGCG is shown Control group is below, and reduces the occurrence degree of pericarp greasy.This explanation EGCG by suppressing the activity of fatty acid synthase, The accumulation of waxy component is reduced, so as to slow down the generation of greasy phenomenon.EGCG is only for fatty acid synthase, to the product of fruit Matter character does not have a significant impact.
Brief description of the drawings
Fig. 1 is influence of the EGCG processing to ' Pink Lady ' fruit greasy grade；
Fig. 2 is influence of the EGCG processing to ' Pink Lady ' fruit aliphatic acid, and Fig. 2-A are EGCG on linoleic influence； Fig. 2-B are influences of the EGCG to oleic acid；Fig. 2-C are influences of the EGCG to palmitoleic acid；Fig. 2-D are shadows of the EGCG to tetradecylic acid Ring；Fig. 2-E are influences of the EGCG to eicosenoic acid；Fig. 2-F are influences of the EGCG to lauric acid/dodecanoic acid.
Fig. 3 is influence of the EGCG processing to ' Pink Lady ' fruit esters, and Fig. 3-A are EGCG to linoleic acid farnesol ester Influence；Fig. 3-B are influences of the EGCG to oleic acid farnesol ester；Fig. 3-C are influences of the EGCG to linoleic acid butyl ester；Fig. 3-D are Influences of the EGCG to linoleic acid propyl ester.
Fig. 4 is that EGCG processing refers to ' Pink Lady ' fruit Ethylene Production Rate (A), respiration of fruits speed (B) and quality Mark the influence of (C, D, E).Fig. 4-A are influences of the EGCG to fruit Ethylene Production Rate；Fig. 4-B are EGCG to respiration of fruits speed Influence；Fig. 4-C are influences of the EGCG to content of fruit soluble solids；Fig. 4-D are influences of the EGCG to fruit titratable acid；Figure 4-E is influences of the EGCG to the hardness of fruit.
In Fig. 1-Fig. 4, different lowercase letters on the contemporaneity data post significant difference in P=0.05 levels.
Embodiment
Technical scheme is described in more detail with specific embodiment below in conjunction with the accompanying drawings.
1 material and reagent
' Pink Lady ' apple (Malus domestica Borkh. ' Pink Lady ') is purchased from Shaanxi in December, 2015 The commercial freezers of province Fuping County one.Greasy phenomenon does not occur for selection, and uniform coloring is in the same size, and the fruit having no mechanical damage is Try material.
Reagent:EGCG is purchased from Dalian U.S. logical sequence Technology Co., Ltd., purity 95%；Chloroform, n-hexane, methanol are (pure Degree ＞ 99.9% Tianjin Kermel Chemical Reagent Co., Ltd.) distill after use.
Instrument:U.S. GC company's T RACE GC ULTRA type gas chromatographs；Hangzhou is difficult to understand to contain MD-200 nitrogen evaporators；Good fortune is stood GC9790II gas chromatographs；Jiangsu hundred million leads to 7001 serial capnographs of Electronics Co., Ltd. TELAIRE；South Africa Guss Manufacturer company GS-15 type fruit texture analyzers；South Korea's G-WON Hitech companies GMK-835F type apples Acidometer；Japanese ATAGO companies PAL-1 types hold saccharometer；Henan Lan Fan trade Co., Ltds RE-2000 Rotary Evaporators.
2 methods
The processing of 2.1 fruits
Experiment sets 3 processing:(1) (distilled water) is compareed；(2)1.5g·L-1EGCG；(3)3.0g·L-1EGCG.At three Fruit using vacuum infiltration method processing fruit, i.e., is placed in vacuum desiccator, sealed, then using SHZ-3 recirculated waters by reason It is 0.06Mpa that multiplex vavuum pump, which is evacuated to vacuum, maintains 60s, is slowly gone up to normal pressure.Fruit taking-up is rearmounted to dry in the air at room temperature Dry, the fruit after drying is placed at 25 DEG C and stored.Each processing sets 3 repetitions, each repeats 30 fruits.
2.2 physical signs and fruit quality measure
With TRACE GC ULTRA type gas chromatograph for determination Ethylene Production Rates.Method is as follows:Taken out after the closed 1h of fruit Take gas, GDX-502 chromatographic columns, carrier gas N2, 70 DEG C of column temperature, 70 DEG C of injector temperature, hydrogen flowing quantity 0.7kgcm-2, air stream Measure 0.7kgcm-2, nitrogen flow 1.0kgcm-2, sample size 1mL, flame ionization ditector detection, detect room temperature 150℃。
Respiratory intensity:With the type CO of TELAIRE 70012Analysis-e/or determining (Dong Xiaoqing etc. 2009).
The calculating of fruit surface area is implemented according to Yuan Kejun etc. (1995) method.
The hardness of fruit is determined using GS-15 type fruit texture analyzer.Design parameter is set as:Probe diameter 10mm；Touch Send out threshold value 0.10kg；Pop one's head in fall off rate 10mms-1；Probe returns to speed 10mms-1；Measure speed 5mms-1；Measurement Distance 10.0mm.Titratable acid content is determined by GMK-835F fruit acidity analyzer.Soluble solid content uses PAL-1 types hold saccharometer measurement.
3 greasy degree evaluations
The degree of fruit greasy is judged using the method for subjective appreciation, method is with reference to (2005) and Dadzie such as Liu Chunxiang Deng (1995).Greasy degree is divided into 4 grades:0 is not occur；1 is slight；2 be moderate；3 be severe.Subjective appreciation Group is formed by 3 by training and experienced student, to reduce error, the selected apple (0 grade) that greasy does not occur with And the fruit (3 grades) of greasy most serious compares, slight touch fruit surface evaluation greasy degree, and fruit is touched every time By hand wiped clean before real surface.Greasy degree is the average value of 3 people's results of sensory evaluation, and subjective appreciation fruit is fixed 15 fruits.
Trial test early stage result shows to store fruit greasy degree change early stage comparatively fast, therefore carries out once phase every 3d The measure work of index is closed, the change of fruit greasy is slower after 9d, and the measure of an index of correlation is carried out every 4 or 5d. To sum up, physical signs and fruit quality measure, greasy degree evaluation are respectively in fruit storage 0,3,6,9,13,17,21 and 26d Shi Jinhang.
4 fruit waxy components
The extraction of the total wax of 4.1 fruits
The extraction of pericarp wax with reference to (2012), Wang Meifang etc. (2008) such as Dong method, respectively fruit storage 0, 6th, 13,21 and 26d extracts wax.Whole fruit is immersed under normal temperature in the beaker for the chloroform for filling 400mL and soaks 45s, Each fruit extracts 3 times, mixes the leachate of 3 times after extraction, filtering, is transferred in 3000mL round bottom distilling flasks and uses RE-2000 Rotary Evaporators are concentrated under the conditions of 40 DEG C, and concentrate is transferred in 50mL serum bottle.Wax extract solution Dried up with MD200-1 nitrogen evaporators under stable nitrogen stream, then with chloroform and methanol with 10:1 ratio is settled to 20mL, then heated with 50 DEG C of water-baths, wax is completely dissolved, mixed, the internal standard (heptadecane) for being eventually adding 1mL is standby.
Esters and the processing method and analysis condition of aliphatic acid in 4.2 waxy components
Silylation reaction is first carried out before esters analysis:Drawing 500 μ L, the standby wax extract solution of constant volume has been extremely In 1.5mL QSP centrifuge tubes, 4h is to being completely dried for vacuum drying concentration.Add double (trimethyl silicon substrate) trifluoroacetamides of 400 μ L (BSTFA) the Silylation reaction that 70 DEG C of baking ovens carry out 45min is put into after, sample uses n-hexane constant volume after nitrogen evaporator dries up To 500 μ L, (made through in 0.45 μm of organic system membrane filtration to 1.5mL sample injection bottles, finally carrying out gas chromatographic analysis with heptadecane For internal standard).
Ester analysis uses gas chromatography.Condition is as follows:The DB-35MS low pole capillary chromatographic columns (U.S. Agilent J＆W scientific companies, 30m × 0.32mm, 0.25 μm), 70 DEG C of column temperature, 230 DEG C of injector temperature, detection 300 DEG C of device temperature, carrier gas is high purity N2.Heating schedule:70 DEG C of initial temperature, 1min is kept, with 10 DEG C of min-1Heating Speed rises to 200 DEG C, then with 5 DEG C of min-1Heating rate rise to 300 DEG C, keep 30min.
Esterification is first carried out before total fatty acids analysis, method is as follows:Draw the 500 μ L standby wax extractions of constant volume Liquid, add 50 μ L Heptadecanoic acides and dried up as internal standard with nitrogen evaporator, add 200 μ L potassium hydroxide-methanol solution (0.4mol mL-1), shaken up after hydrolyzing 15min at 70 DEG C, add 300 μ L boron trifluorides (BF3) 30min is esterified under the conditions of 70 DEG C, finally 500 μ L are settled to n-hexane.About 500 μ L of supernatant liquid are drawn to 1.5mL sample injection bottles with liquid-transfering gun.
Total fatty acids analysis uses gas chromatography.Condition:KB-FFAP capillary chromatographic columns (U.S. Agilent J＆W Scientific companies, 30m × 0.32mm, 0.25 μm), 70 DEG C of column temperature, 230 DEG C of injector temperature, 300 DEG C of detector temperature； Carrier gas is the N of high-purity2.Heating schedule:70 DEG C of initial temperature, 1min is kept, with 10 DEG C of min-1Heating rate rises to 200 DEG C, then with 5 DEG C of min-1Heating rate rises to 230 DEG C, keeps 20min.
5 data processings
Test data analyzer uses Excel 2007, SAS 9.4 to carry out statistical analysis, carries out variance analysis and uses ANOVA Method, Multiple range test use DUNCAN methods (significant difference P<0.05).
Experimental result
Influence of the 1 EGCG processing to ' Pink Lady ' fruit greasy degree
With the extension of storage period, there is greasy phenomenon in control group and EGCG treatment groups fruit, and constantly aggravate. Although three groups of processing greasy degree do not have significant difference statistically, the time that greasy occurs for treatment group is obvious It is later than control group, greasy growth trend takes place in 6d for control group fruit, and treatment group fruit just starts to occur until 9d Greasy increases.During 13d, greasy phenomenon, 13~21d control groups and 1.5gL have all occurred for whole fruits-1EGCG treatment groups Greasy degree be significantly higher than 3.0gL always-1EGCG treatment groups；During 26d, the greasy degree of three groups of processing is without conspicuousness Difference.Control group and 1.5gL during wherein 17d-1EGCG treatment group greasy degree reaches highest level (Fig. 1).In summary, EGCG processing can postpone the time of origin of greasy, 3.0gL-1The inhibition that greasy occurs for EGCG processing is optimal, but It is to completely inhibit.
Influence of the 2 EGCG processing to ' Pink Lady ' fruit wax content and component
6 kinds of higher aliphatic acid of content, wherein control group fruit Linoleic acid content highest are detected in this research altogether, can Up to 75.12 μ gcm-2, oleic acid takes second place up to 12.61 μ gcm-2, 3 kinds of lauric acid/dodecanoic acid, tetradecylic acid, eicosenoic acid aliphatic acid Content is in 0.20~0.45 μ gcm-2Between, palmitoleic acid content is minimum.Fruit Linoleic acid and oleic acid after EGCG is handled Content is below control group, wherein during 6 and 13d, 3.0gL-1The content of EGCG treatment groups Linoleic acid and oleic acid is substantially less than Control group and 1.5gL-1EGCG groups (Fig. 2-A and B).Palmitoleic acid 1.5 and 3.0gL when whole storage period removes 6d- 1Content in EGCG groups is substantially less than control group, and there was no significant difference (Fig. 2-C) between three groups of other times.Lauric acid/dodecanoic acid, 14 Acid, eicosenoic acid are not significantly different (Fig. 2-D~F) all the time between whole three groups of storage period.In summary, EGCG is not to The inhibition of class aliphatic acid of the same race is different, and the EGCG of various concentrations is also different to the inhibition of same aliphatic acid.But Generally EGCG can suppress the most linoleic plus oleic acid of the accumulation of aliphatic acid, especially content, and 3.0gL-1EGCG's Best results, there is the time point of significant difference in 6d and/or 13d.
The main esters that this research detects have 4 kinds, including linoleic acid farnesol ester, oleic acid farnesol ester, linoleic acid fourth Ester, linoleic acid propyl ester；Control group fruit Linoleic acid farnesol ester content highest, up to 6.71 μ gcm-2, oleic acid farnesol ester Take second place (3~4 μ gcm-2), the content of linoleic acid butyl ester reaches as high as 0.92 μ gcm-2, oleic acid propyl ester content is minimum (to be less than 0.4μg·cm-2).Content sustainable growth between 0~13d of linoleic acid farnesol ester in three groups of processing fruits, control group and place There was no significant difference between reason group.The content of linoleic acid farnesol ester reduces between 13~26d, and in 21d, the content in control group shows Work is higher than EGCG treatment groups, and 1.5gL-1The content of EGCG treatment groups is significantly higher than 3.0gL-1EGCG treatment groups (Fig. 3- A).The trend fallen after rising also is presented in the content of oleic acid farnesol ester, the content highest in 13d, and the content in control group It is significantly higher than EGCG treatment groups, 1.5 and 3.0gL-1There was no significant difference (Fig. 3-B) between EGCG processing.Linoleic acid butyl ester Content reaches maximum in 21d, but there was no significant difference between three groups of processing.In 6d, 1.5gL-1EGCG treatment groups contain Amount is significantly higher than control group and 3.0gL-1EGCG treatment groups, in 26d, the content conspicuousness of control group is less than EGCG treatment groups (Fig. 3-C).There was no significant difference between three groups of processing for content between 0~21d for linoleic acid propyl ester content, and control group contains during 26d Amount is substantially less than EGCG treatment groups (P ＜ 0.05) (Fig. 3-D).In summary, with the extension of storage time, content highest two Kind material, the i.e. content of linoleic acid farnesol ester and oleic acid farnesol ester, which are presented, first increases the trend reduced again, and linoleic acid fourth Trend stable after lasting rise increases or increased is presented in ester and linoleic acid propyl ester.Although linoleic acid farnesol ester and oleic acid method Buddhist nun The variation tendency of alcohol ester is different from linoleic acid butyl ester and linoleic acid propyl ester, but linoleic acid farnesol ester and oleic acid farnesol ester Content proportion in 4 kinds of esters is larger.It thus is seen that EGCG processing is different to the action effect of different Esters, but It is mainly to make the reduction of its content.
Influence of the 3 EGCG processing to ' Pink Lady ' fruit Ethylene Production Rate, respiratory rate and fruit quality
Control group, 1.5gL-1EGCG groups, 3.0gL-1There is ethene peak, peak value point in 6d in EGCG groups fruit Wei not 57.9,61.1,41.2 μ Lkg-1·h-1, now control group compare 3.0gL-1The acetate releasing quantity of EGCG groups is high 28.8%, there is significant difference, but control group and 1.5gL-1Without significant difference (Fig. 4-A) between EGCG treatment groups.Control When the respiratory climacteric of group comes across 3d, and the no obvious respiratory climacteric of EGCG treatment groups fruit, and at control group and two There is no significant difference (Fig. 4-B) all the time in whole storage period between reason group.Soluble solid and titratable acid content exist Downward trend is presented in whole storage period, but there was no significant difference all the time between three groups of processing, and the former is in control group, 1.5gL- 1EGCG、3.0g·L-1EGCG groups fruit have dropped 3.37%, 6.61%, 5.38% (Fig. 4-C) respectively；The latter have dropped respectively 42.87%th, 47.94%, 50.98% (Fig. 4-D).The variation tendency unobvious of the hardness of fruit, it is overall in 7.8~9.2kgcm-2 In fluctuation status, and the difference between three kinds of processing is not notable (Fig. 4-E).In summary, only 3.0gL-1EGCG treatment groups exist The rate of release of ethene is reduced during 6d, EGCG handles the Ethylene Production Rate to the other times point of fruit, whole storage period Respiratory rate, soluble solid, titratable acid and the hardness of fruit have no significant effect.
The foregoing is only a preferred embodiment of the present invention, protection scope of the present invention not limited to this, any ripe Those skilled in the art are known in the technical scope of present disclosure, the letter for the technical scheme that can be become apparent to Altered or equivalence replacement are each fallen within protection scope of the present invention.

Claims (4)

  1. A kind of 1. method for suppressing Apple greasy and occurring, it is characterised in that comprise the following steps：
    Step 1, fruit processing
    Experiment sets 3 processing:(1) distilled water is compareed；(2)1.5g·L-1EGCG；(3)3.0g·L-1EGCG；Three processing make Fruit is handled with vacuum infiltration method, i.e., fruit is placed in vacuum desiccator, is sealed, it is then multi-purpose true using SHZ-3 recirculated waters Empty pumping to vacuum is 0.06Mpa, maintains 60s, is slowly gone up to normal pressure；Fruit taking-up is rearmounted to dry at room temperature, dries Fruit afterwards is placed at 25 DEG C and stored；Each processing sets 3 repetitions, each repeats 30 fruits；
    Step 2, physical signs and fruit quality measure
    With TRACE GC ULTRA type gas chromatograph for determination Ethylene Production Rates；Method is as follows:Gas is extracted after the closed 1h of fruit Body, GDX-502 chromatographic columns, carrier gas N2, 70 DEG C of column temperature, 70 DEG C of injector temperature, hydrogen flowing quantity 0.7kgcm-2, air mass flow 0.7kg·cm-2, nitrogen flow 1.0kgcm-2, sample size 1mL, flame ionization ditector detection, detect room temperature 150 ℃；
    Respiratory intensity:With the type CO of TELAIRE 70012Analysis-e/or determining；
    The hardness of fruit is determined using GS-15 type fruit texture analyzer；
    Step 3, greasy degree evaluation
    The degree of fruit greasy is judged using the method for subjective appreciation, greasy degree is divided into 4 grades:0 is not send out It is raw；1 is slight；2 be moderate；3 be severe；Subjective appreciation group is formed by 3 by training and experienced student, to reduce Error, it is selected the apple of greasy not to occur and the fruit of greasy most serious compares, the evaluation of slight touch fruit surface Greasy degree, and every time before touch fruit surface by hand wiped clean；Greasy degree is 3 people's results of sensory evaluation Average value, subjective appreciation fruit are 15 fixed fruits；
    Step 4, the processing of fruit waxy component；
    Step 5, data processing
    Test data analyzer uses Excel 2007, SAS 9.4 to carry out statistical analysis, carries out variance analysis and uses ANOVA methods, more Compare again using DUNCAN methods, significant difference P<0.05.
  2. 2. the method according to claim 1 for suppressing Apple greasy and occurring, it is characterised in that in step 2, use GS-15 type fruit texture analyzer determines the hardness of fruit；Design parameter is set as:Probe diameter 10mm；Activation threshold value 0.10kg； Pop one's head in fall off rate 10mms-1；Probe returns to speed 10mms-1；Measure speed 5mms-1；Measurement distance 10.0mm；Can Acid content is titrated to determine by GMK-835F fruit acidity analyzer；Soluble solid content holds pol using PAL-1 types Meter measurement.
  3. 3. the method according to claim 1 for suppressing Apple greasy and occurring, it is characterised in that in step 3, every 3d carries out the measure work of an index of correlation, and the change of fruit greasy is slower after 9d, is carried out once every 4 or 5d related The measure of index；To sum up, physical signs and fruit quality measure, greasy degree evaluation respectively fruit storage 0,3,6,9, 13rd, carried out during 17,21 and 26d.
  4. 4. the method according to claim 1 for suppressing Apple greasy and occurring, it is characterised in that step 4 is specially：
    The extraction of the total wax of 4.1 fruits
    Respectively in fruit storage 0,6,13,21 and 26d extraction waxs；Whole fruit is immersed under normal temperature the trichlorine for filling 400mL 45s is soaked in the beaker of methane, and each fruit extracts 3 times, the leachate of 3 times is mixed after extraction, filtering, is transferred to 3000mL Concentrated in round bottom distilling flask using RE-2000 Rotary Evaporators under the conditions of 40 DEG C, concentrate is transferred to 50mL's In serum bottle；Wax extract solution is dried up with MD200-1 nitrogen evaporators under stable nitrogen stream, then with chloroform and methanol with 10:1 ratio is settled to 20mL, then is heated with 50 DEG C of water-baths, wax is completely dissolved, mixed, is eventually adding the interior of 1mL It is standby to mark heptadecane；
    Esters and the processing method and analysis condition of aliphatic acid in 4.2 waxy components
    Silylation reaction is first carried out before esters analysis:Draw 500 μ L the standby wax extract solution of constant volume to 1.5mL In QSP centrifuge tubes, 4h is to being completely dried for vacuum drying concentration；70 are put into after adding double (trimethyl silicon substrate) trifluoroacetamides of 400 μ L DEG C baking oven carries out 45min Silylation reaction, and sample is settled to 500 μ L after nitrogen evaporator dries up with n-hexane, through 0.45 μ In m organic systems membrane filtration to 1.5mL sample injection bottles, gas chromatographic analysis is finally carried out, internal standard is used as using heptadecane；
    Ester analysis uses gas chromatography；Condition is as follows:70 DEG C of DB-35MS low pole capillary chromatographic columns column temperature, enters 230 DEG C of sample mouth temperature, 300 DEG C of detector temperature, carrier gas is high purity N2；Heating schedule:70 DEG C of initial temperature, 1min is kept, With 10 DEG C of min-1Heating rate rise to 200 DEG C, then with 5 DEG C of min-1Heating rate rise to 300 DEG C, keep 30min；
    Esterification is first carried out before total fatty acids analysis, method is as follows:The 500 μ L standby wax extract solutions of constant volume are drawn, 50 μ L Heptadecanoic acides are added as internal standard to be dried up with nitrogen evaporator, add 200 μ L 0.4molmL-1Potassium hydroxide-methanol is molten Liquid, shaken up after hydrolyzing 15min at 70 DEG C, add 300 μ L boron trifluorides and 30min is esterified under the conditions of 70 DEG C, finally with just oneself Alkane is settled to 500 μ L；500 μ L of supernatant liquid are drawn to 1.5mL sample injection bottles with liquid-transfering gun；
    Total fatty acids analysis uses gas chromatography；Condition:KB-FFAP capillary chromatographic columns, 70 DEG C of column temperature, injector temperature 230 DEG C, 300 DEG C of detector temperature；Carrier gas is the N of high-purity2；Heating schedule:70 DEG C of initial temperature, 1min is kept, with 10 ℃·min-1Heating rate rises to 200 DEG C, then with 5 DEG C of min-1Heating rate rises to 230 DEG C, keeps 20min.
CN201710474332.7A 2017-06-21 2017-06-21 A kind of method for suppressing Apple greasy and occurring Pending CN107361130A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710474332.7A CN107361130A (en) 2017-06-21 2017-06-21 A kind of method for suppressing Apple greasy and occurring

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710474332.7A CN107361130A (en) 2017-06-21 2017-06-21 A kind of method for suppressing Apple greasy and occurring

Publications (1)

Publication Number Publication Date
CN107361130A true CN107361130A (en) 2017-11-21

Family

ID=60305551

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710474332.7A Pending CN107361130A (en) 2017-06-21 2017-06-21 A kind of method for suppressing Apple greasy and occurring

Country Status (1)

Country Link
CN (1) CN107361130A (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108548871A (en) * 2018-02-01 2018-09-18 大连大学 Korea's blueberry branch cuticula waxy component and its detection method
CN111296561A (en) * 2020-03-03 2020-06-19 中国农业科学院农产品加工研究所 Method and composition for preventing apple fruits from becoming greasy
CN115943990A (en) * 2023-02-10 2023-04-11 浙江大学 Method for delaying fruit ripening of tomatoes

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
王聪，刘翠华，杨艳青，周彬，闫丹: "《表没食子儿茶素没食子酸酯(EGCG)对‘粉红女士’苹果果皮油腻化的影响》", 《植物生理学报》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108548871A (en) * 2018-02-01 2018-09-18 大连大学 Korea's blueberry branch cuticula waxy component and its detection method
CN111296561A (en) * 2020-03-03 2020-06-19 中国农业科学院农产品加工研究所 Method and composition for preventing apple fruits from becoming greasy
CN111296561B (en) * 2020-03-03 2022-04-15 中国农业科学院农产品加工研究所 Method and composition for preventing apple fruits from becoming greasy
CN115943990A (en) * 2023-02-10 2023-04-11 浙江大学 Method for delaying fruit ripening of tomatoes

Similar Documents

Publication Publication Date Title
CN107361130A (en) A kind of method for suppressing Apple greasy and occurring
Du et al. Determination of phthalate esters in teas and tea infusions by gas chromatography–mass spectrometry
Maes et al. Dissection of the phytohormonal regulation of trichome formation and biosynthesis of the antimalarial compound artemisinin in Artemisia annua plants
Pereira et al. Towards chemical and nutritional inventory of Portuguese wild edible mushrooms in different habitats
Xu et al. Optimization of supercritical carbon dioxide extraction of sea buckthorn (Hippophae thamnoides L.) oil using response surface methodology
CN103543233B (en) The assay method of short chain or medium chain chlorinated paraffin content in a kind of coating
Brock et al. Biosynthesis of Sesqui‐and Diterpenes by the Gibberellin Producer Fusarium fujikuroi
Li et al. Chemical composition and antioxidant activity of Tuber indicum from different geographical regions of China
Berke et al. Characterization of the last deglacial transition in tropical East Africa: Insights from Lake Albert
CN105738504A (en) Method for measuring hickory fatty acid content through gas chromatograph mass spectrometry
Sarria-Villa et al. Characterization and determination of the quality of rosins and turpentines extracted from Pinus oocarpa and Pinus patula resin
Wang et al. Identification of volatile compounds and metabolic pathway during ultrasound-assisted kombucha fermentation by HS-SPME-GC/MS combined with metabolomic analysis
Chung et al. Inhibition of diacylglycerol acyltransferase by betulinic acid from Alnus hirsuta
Khan et al. Regulation of vascular endothelial function by red wine procyanidins: implications for cardiovascular health
Wu et al. Comprehensive metabolic analyses provide new insights into primary and secondary metabolites in different tissues of Jianghua Kucha tea (Camellia sinensis var. assamica cv. Jianghua)
Yansong et al. Response of soil nutrients and their stoichiometric ratios to stand density in Pinus tabuliformis and Robinia pseudoacacia plantations in the loess region of western Shanxi Province, northern China
CN102590382A (en) Determination method of phthalic acid ester in printing ink for cigarette
Xin-Hua et al. Essential oils composition from roots of Santalum album L.
Li et al. Analysis of sugars and fatty acids during haustorium development and seedling growth of coconut
CN104569266B (en) A kind of method of 2-dodecyl cyclobutanone content in quick detection irradiation fat-containing food
Qiu et al. Study on migration of two acrylate monomers in plastic food contact materials
CN104807916A (en) Method for detecting rongalit in food by gas chromatograph mass spectrometer
Tociu et al. Preliminary investigations for the valorization of grape seeds after the oil extraction
Kilic Essential Oil and Fatty Acid Composition of Leaves of Some Lamiaceae Taxa FromTurkey
Pańczak et al. Biomarkers in fossil resins and their palaeoecological significance

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20171121