CN107132141B - Method for rapidly determining content of waxiness on pear fruit epidermis - Google Patents
Method for rapidly determining content of waxiness on pear fruit epidermis Download PDFInfo
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- CN107132141B CN107132141B CN201710289003.5A CN201710289003A CN107132141B CN 107132141 B CN107132141 B CN 107132141B CN 201710289003 A CN201710289003 A CN 201710289003A CN 107132141 B CN107132141 B CN 107132141B
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Abstract
The invention discloses a method for rapidly measuring the content of wax on the surface of pear fruit. In the method, the wax content of the pear is measured by changing 'whole fruit soaking' into 'quantitative peel soaking', and the wax content is quickly converted according to a relation coefficient between the whole fruit soaking and the quantitative peel soaking, so that the surface area of the pear does not need to be estimated. Liquid transfer is not needed in the operation process, reagents are saved to the maximum extent, time is saved, the operation is simple and convenient, the harm of toxic reagents to experimenters is reduced, and the rapid determination of the content of the wax on the surface of the pear fruit can be realized.
Description
Technical Field
The invention belongs to the technical field of plant epidermis wax content determination, and particularly relates to a method for rapidly determining pear fruit epidermis wax content.
Background
The wax layer is a layer of hydrophobic protective substance covering the surface of the plant, is used as a first barrier for the contact of the plant and the outside, has the functions of keeping the surface of the plant clean, resisting adverse environment, preventing plant diseases and insect pests, preventing non-porous water loss of the plant and the like, and also has important influence on the shape of leaves and fruits of the plant and the development of pollen. In addition, the wax can be used as an important raw material of paint, lubricant, medicine, cosmetics and biofuel, and some contained terpenoids contain components beneficial to human health and have the functions of diminishing inflammation, resisting tumors and the like. Therefore, the research on the plant wax has potential health-care benefits and economic values, and the wax is becoming one of important contents of plant research. The pear tree cultivation history in China is long, resources are rich, the cultivation area and the yield are always the first in the world, and the pear tree cultivation method is of necessary significance for researching pear fruit waxiness at the third position of the total area and the total yield of fruit cultivation in China.
At present, most researchers extract wax by soaking plant tissues and organs in organic solvents. The organic solvent includes chloroform, dichloromethane, methanol, n-hexane, etc. In practice, to prevent the extraction of fat-soluble substances from the cells inside the plant, it is usually necessary to immerse the whole plant organ in the extraction liquid. Wax content is expressed by wax content per unit area, and in the actual operation process, the difficulty of accurately and rapidly measuring the wax content of the epidermis of some plant organs is higher due to different tissue forms of various plants. The plant leaves have small volume, the surface area is easy to calculate, the wax is easy to extract, and the content calculation is more accurate. The shape of fruits such as oranges and grapes is correct, and the wax content can be estimated according to spheres. But the wax content of the surface of the pear fruit with irregular shape and large size is accurately and rapidly determined, so that the difficulty is high. At present, the following problems mainly exist in the rapid determination of the waxy content of the surface skin of the fruit with irregular fruit shape:
(1) at present, the calculation of the pear surface area is mostly estimated by measuring data such as longitudinal and transverse diameters by referring to a method reported by researchers of Yuan Kejun et al (2000). Different varieties of pears have larger fruit shape difference, such as oval, round, gourd-shaped and the like, and the surface area of all pears is estimated by using the same formula, so that the experimental error is larger;
(2) too much chloroform is difficult to evaporate, and too little chloroform affects the completeness of wax extraction. Generally, the pear is relatively large, and about 500mL of organic solvent is required for extracting wax from 500g of pear. So survey the pear epidermis wax content and need consume a large amount of organic solvents, and a large amount of organic solvents distill and weather and need longer time to the solvent is too much to need to borrow instruments such as rotary evaporator and carry out the rapid distillation, this just leads to need shift many times to the wax extract in the extraction process, causes the medicament to waste, the time increase, and the testing cost increases, and experimental error increases. The trichloromethane is used for extracting the plant epidermis wax, although the effect is better, the trichloromethane belongs to a toxic reagent, and the trichloromethane is easily in contact with the plant epidermis wax for a long time in the test process, so that the health of test personnel and the environment are greatly damaged.
These factors all restrict researchers from rapidly determining the content of the wax in the pear fruit.
Disclosure of Invention
In order to solve the problems, the applicant combines the existing wax determination method, and after exploration and practice, particularly provides a method for rapidly determining the wax content of pear fruits, and can rapidly determine the wax content of the pear fruits by using a small amount of chloroform.
The purpose of the invention can be realized by the following technical scheme:
a method for rapidly measuring the content of waxy substances on the surface of a pear fruit comprises the following steps:
(1) cutting a plurality of peels with the thickness of 0.5-1 mm from the surfaces of the pears; (ii) a
(2) Immersing the peel in the step (1) into chloroform with the mass of W1Soaking the substrate in the container for 30-90 seconds;
(3) after the immersion, the chloroform in the vessel was evaporated to dryness, and the vessel at that time was weighed to a mass of W2；
(4) Calculating the wax content of the pear fruit according to the following formula;
M(mg/cm2)＝(W2-W1)/(S×K)×103
wherein the total surface area of the peel is S; m: wax content (mg/cm) of the extract2) (ii) a The K value is 3.8-4.2.
The technical scheme of the invention is as follows: in the step (1), the total surface area S of the pear fruit surface is 5-15 cm2The fruit peel of (1). Preferably, the method comprises the following steps: step (1) is to obtain pomeThe total surface area S taken on a solid surface is 9cm2The fruit peel of (1).
The technical scheme of the invention is as follows: the dosage of the trichloromethane in the step (2) is 5-15 mL. Preferably, the method comprises the following steps: the dosage of the trichloromethane in the step (2) is 8 mL.
The technical scheme of the invention is as follows: and (3) evaporating the trichloromethane at 40 ℃.
The technical scheme of the invention is as follows: the K value in the step (4) is 4.
In the wax extraction method of the technical scheme of the invention, the whole fruit soaking is changed into the fruit peel soaking: the existing method for extracting the waxiness of the pear fruits is to completely soak the pear fruits in trichloromethane, and the changed method is to use a scalpel to take a certain area of peel on the surface of the pear fruits for measurement.
In the pear surface area calculation method of the technical scheme of the invention, the integral estimation is changed into accurate calculation: the existing method for measuring the waxiness on the surface of the pear fruit is to estimate the surface area of the whole pear fruit, and the method after the change is to directly obtain the surface area of the pear fruit.
According to the technical scheme of the invention, the peel is used for extracting the wax, and fat-soluble substances in the intradermal side cells of the pome are extracted, but the substance is related to the wax content extracted by soaking the whole fruit. Correlation analysis is carried out on two groups of data of the epidermis waxy content measured after the whole pear fruit is soaked and the waxy content extracted from the peel with a certain area, and the correlation between the two groups of data is judged.
Advantageous effects
According to the method, the wax content of the pear is measured without soaking the whole pear, the peel is directly taken from the surface of the pear to be measured, the complexity is simplified, and the surface area calculation is accurate. 500mL of trichloromethane is needed for soaking one fruit in the whole fruit, and only 8mL of trichloromethane is needed by adopting the method, the excessive trichloromethane is completely evaporated for a long time, and liquid transfer is not needed in the whole process, so that medicines are saved to the maximum extent, the time is saved, and the workload and the injury of toxic reagents to experimenters are reduced.
Drawings
FIG. 1 is a linear correlation diagram of wax content of pear fruit extracted by whole fruit soaking and pericarp soaking;
FIG. 2 is a box diagram of the ratio of wax content in pear fruit extracted by whole fruit soaking and pericarp soaking;
FIG. 3 is a flow chart of the whole fruit soaking and pericarp soaking processes for extracting waxiness from pear fruit.
Wherein, A: a flow chart of extracting the waxiness of the pear fruits by soaking the whole fruits; b: a flow chart for extracting the waxiness of the pear fruits by soaking the peels and the whole fruits; a: a rotary evaporator; b: a nitrogen blowing instrument; c: a chloroform reagent; d: extracted wax; a-1: soaking the whole pear fruit in an organic solvent; a-2: evaporating by a rotary evaporator; 3: evaporating and drying by a nitrogen blowing instrument; 4: obtaining wax; b-1: cutting a plurality of pear peels with specifications by using a scalpel; b-2: soaking the cut pericarp in chloroform
Detailed Description
The following description will take the application of the method of the present invention in the determination of the waxy content in the epidermis of pear fruit as an example.
(1) Cleaning fructus Pyri with clear water, and cutting into three pieces (1 × 1 cm) at upper, middle and lower parts of fructus Pyri with surgical knife2The fruit peel of (1). In order to ensure the result to be accurate as much as possible and the peel to contain less pulp as much as possible, 1 × 1cm can be prepared in advance2The hard paper sheets are stuck on the surfaces of the pears for cutting;
(2) the taken peel is quickly immersed in 8mL of trichloromethane with the mass of W1The fruit peel is taken out after the fruit peel is lightly shaken for 1 minute in a screw reagent bottle (the volume is 10 mL);
(3) placing the screw reagent bottle on a JHD-001S nitrogen blowing instrument, evaporating chloroform to dryness at constant temperature of 40 ℃ under stable nitrogen flow, and weighing the container at the moment2；
(4) Calculating the wax content of the pear fruit according to the following formula:
M2(mg/cm2)＝M1/4＝(W2-W1)/9×103
wherein M is2: waxy content (mg/cm) extracted from whole fruit by soaking2)；M1: waxy content (mg/cm) extracted from 9 pieces of pericarp by soaking2)；W2: total weight (g) of screw reagent bottle and wax; w1: screw reagent bottle weight (g).
The applicant respectively carries out whole fruit soaking and pericarp taking soaking on 35 different pear fruit samples to measure the content of the epidermal waxy substances, then carries out correlation analysis on two groups of data (figure 1), and the result shows that the two groups of data are obviously in positive correlation (R)20.9853). And the content of the waxiness extracted by soaking the peel is found to be 4 times of the content of the waxiness extracted by soaking the whole fruit through box diagram analysis, so the method is suitable for rapidly determining the content of the waxiness on the pear peel.
In the experimental process, conversion can be carried out according to the linear relation between the wax contents extracted by the peel and whole fruit soaking methods, the conversion is flexible, and the wax contents of a large number of pears can be rapidly measured.
Fruit wax content extraction result of pear by whole fruit soaking and pericarp soaking
Note: m1Representative is 9 blocks of 1X 1cm2Waxy content, M, from peel soaking2Representative is the waxy content of the whole fruit extract. In table M1And M2Values are averaged over three replicates. Wherein K is M2/M1。
(5) The method can also be applied to the rapid determination of the wax content of other fruits.
Claims (3)
1. A method for rapidly measuring the content of waxy substances on the surface of a pear fruit is characterized by comprising the following steps: the method comprises the following steps:
(1) cutting a plurality of peels with the thickness of 0.5-1 mm from the surfaces of the pears;
(2) immersing the peel in the step (1) into chloroform with the mass of W1Soaking the substrate in the container for 30-90 seconds;
(3) after the immersion, the chloroform in the vessel was evaporated to dryness, and the vessel at that time was weighed to a mass of W2；
(4) Calculating the wax content of the pear fruit according to the following formula;
M(mg/cm2) = (W2-W1)/(S×K)×103
wherein the total surface area of the peel is S; m: wax content (mg/cm) of the extract2) (ii) a The K value is 4;
in the step (1), the total surface area S of the pear fruit surface is 5-15 cm2The peel of (a); the dosage of the trichloromethane in the step (2) is 5-15 mL;
and (3) evaporating the trichloromethane at 40 ℃.
2. The method for rapidly determining the content of waxy substances on the pear fruit skin according to claim 1, is characterized in that: step (1) intercepting the total surface area S of 9cm on the surface of the pear fruit2The fruit peel of (1).
3. The method for rapidly determining the content of waxy substances on the pear fruit skin according to claim 1, is characterized in that: the dosage of the trichloromethane in the step (2) is 8 mL.
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| CN109470655A (en) * | 2018-12-14 | 2019-03-15 | 南京航空航天大学 | The measuring system and method for high reflectance fruit surface wax content based on light scattering |
| CN114814056A (en) * | 2022-04-24 | 2022-07-29 | 南京农业大学 | Application of dimethyl carbonate in extracting plant epidermis wax |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101162191A (en) * | 2007-11-08 | 2008-04-16 | 武汉科技学院 | Method for determining pectins content in china grass |
| CN105132185A (en) * | 2015-08-19 | 2015-12-09 | 山东师范大学 | Method for rapidly extracting wax from sweet sorghum stem |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101162191A (en) * | 2007-11-08 | 2008-04-16 | 武汉科技学院 | Method for determining pectins content in china grass |
| CN105132185A (en) * | 2015-08-19 | 2015-12-09 | 山东师范大学 | Method for rapidly extracting wax from sweet sorghum stem |
Non-Patent Citations (4)
| Title |
|---|
| 库尔勒香梨表皮蜡质成分分析及其贮藏中的影响研究;李珍慈;《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》;20170215(第02期);第7-8页 * |
| 枸杞表皮蜡质层成分及显微结构的研究;杨爱梅等;《食品工业科技》;20111231;第32卷(第12期);第112-114页 * |
| 红枣不同部位的脂溶性成分分析;游凤 等;《食品与发酵工业》;20131231;第39卷(第11期);第241-244页 * |
| 苹果果实表皮蜡质结构观察与组分分析;杨艳青;《中国优秀硕士学位论文全文数据库 农业科技辑》;20140215(第02期);第14-15页 * |
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