CN107300554A - Rice manganese content simple and fast determination method - Google Patents
Rice manganese content simple and fast determination method Download PDFInfo
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- CN107300554A CN107300554A CN201710579508.5A CN201710579508A CN107300554A CN 107300554 A CN107300554 A CN 107300554A CN 201710579508 A CN201710579508 A CN 201710579508A CN 107300554 A CN107300554 A CN 107300554A
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Abstract
The invention discloses a kind of rice manganese content simple and fast determination method, paddy rice to be measured and standard items paddy rice are subjected to the steps 1 respectively)~5)；1), Brown Rice adds 4%NaOH solution, the gelatinization of 35% hydrogen peroxide；Then the extraction of 10%HCl solution is added；2), in step 1) obtained by solution in add 8M NaOH solutions, stand, then add absolute ethyl alcohol；3), in step 2) obtained by solution in add 1M EDTA solution；4), in step 3) obtained by solution in add formaldoxime solution, vibration shake up after stand；5), in step 4) obtained by solution in add ammonia hydrochloric acid hydroxylamine solution, vibration shake up after stand；6), by step 5) the brown rice reaction solution of gained paddy rice to be measured and the brown rice reaction solution of standard items paddy rice carry out color comparison；The color of paddy rice to be measured is deep, illustrates that the manganese content of Brown Rice to be measured is higher than standard items paddy rice.
Description
Technical field
The present invention relates to the simple and fast determination techniques and screening technique of Brown Rice manganese content, field is related generally to for agriculture
, breeding, paddy rice.
Background technology
Paddy rice is second-biggest-in-the-world cereal crops, is also the staple food crop of China, and its nutritional ingredient is directly connected to
Consumer's is healthy, but important mineral element necessary to some human bodies in many rice varieties promoted at present in production
Content is substantially not enough.Manganese is one of the essential trace elements of the human body, to promoting the normal growth and development of bone and maintaining brain function
Deng important role, appropriate manganese is beneficial to health.Human body manganese member can be supplemented by food additives or medicine etc.
Element, but there are problems that investing big, costly, coverage rate, it is impossible to the problem of manganese nutrition is bad is solved on the whole.By normal
The biological reinforced means such as breeding, mutation breeding are advised, the manganese nutrition that can be absorbed in rice for human body can be effectively improved
Constituent content.But cultivate and be rich in during the new rice variety (being) of manganese nutrition composition, it is necessary to determine a large amount of germ plasm resources, breeding
The rice manganese content of the individual plant such as colony or mutagenesis colony, high number.Therefore, the assay method and technology of manganese nutrition composition are sieves
Select the rich manganese individual plant of paddy rice, improve the most important restriction factor of rice manganese content, emphasis and difficult point are how simple, quick, warp
Ji and the content for effectively identifying rice manganese element.The assay method of current rice mineral content is typically necessary costliness
Instrument and cumbersome sample pre-treatments program；Not only determine costly, and can not fast and efficiently determine rice
Manganese content, thus it is not suitable for the high-volume individual plant screening during rice breeding.Due to the Single-plant selection mistake in rice breeding
Cheng Zhong, typically need not be accurately determined very much the rice manganese content level of paddy rice individual plant, need to only obtain between colony's individual plant or with it is right
The purpose of rich manganese individual plant screening is just can reach according to the relative different of rice manganese content between plant.It is therefore proposed that rice manganese content
Simple and fast determination techniques, for the rich manganese individual plant Rapid identification in rice breeding colony, can quickly filter out and meet the requirements
Rich manganese individual plant.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of rice manganese content simple and fast determination method, this method can be richness
Manganese rice breeding provides simple, quick, economically and efficiently individual plant screening technique.
In order to solve the above-mentioned technical problem, the present invention provides a kind of rice manganese content simple and fast determination method, by water to be measured
Rice and standard items paddy rice carry out the steps 1 respectively)~5)；
1) Brown Rice 4, is taken, 4%NaOH solution 1.0mL, 35% hydrogen peroxidase 10 .1mL is added, in 65 ± 5 DEG C of gelatinizations
3±0.1h；
Then 10%HCl solution 1.0mL extractions are added, 20 ± 2min of lower concussion in 50 ± 5 DEG C of thermostatic control oscillator vibrations,
Frequency of oscillation is 250 ± 10r/min；
Above-mentioned % is quality %；
2), in step 1) obtained by solution in add 8M (8mol/L) NaOH solution 0.4mL, stand 1h after mixing, then add
0.5mL absolute ethyl alcohol, vibration shakes up；
3), in step 2) obtained by solution in add 1M (1mol/L) EDTA solution 0.1mL, vibration shakes up；
4), in step 3) obtained by solution in add formaldoxime solution 0.1mL, vibration shakes up, and stands 20 ± 2min；
5), in step 4) obtained by solution in add ammonia-hydroxylamine hydrochloride solution 0.5mL, vibration shakes up, and stands 20min；
6), by step 5) the brown rice reaction solution (that is, the supernatant liquor of gained after standing) and standard items of gained paddy rice to be measured
The brown rice reaction solution of paddy rice carries out color comparison；
When the color of Brown Rice reaction solution to be measured is deeper than the color of standard items Brown Rice reaction solution, illustrate water to be measured
The manganese content of rice brown rice is higher than standard items paddy rice；
Conversely, when Brown Rice reaction solution to be measured color than standard items Brown Rice reaction solution it is of light color when, then say
The manganese content of bright Brown Rice to be measured is less than standard items paddy rice.
It is used as the improvement of the rice manganese content simple and fast determination method of the present invention：
The preparation method of the formaldoxime solution is：10g hydroxylamine hydrochlorides are dissolved in 50mL water, then add mass fraction be
35% formalin 5mL, is diluted with water to 100mL, is stored at least 30 days in -20 ± 2 DEG C, obtains formaldoxime solution.
Remarks explanation：Because formaldehyde and the speed that hydroxylamine hydrochloride reacts generation formaldoxime are slow, can not directly it make with postponing
With need to could be used after above-mentioned low tempertaure storage.
It is used as the further improvement of the rice manganese content simple and fast determination method of the present invention：The system of ammonia-hydroxylamine hydrochloride solution
Preparation Method is：By 4.7M ammonia solutions and 6M hydroxylamine hydrochloride solutions, mixing shakes up in equal volume.
It is used as the further improvement of the rice manganese content simple and fast determination method of the present invention：Standard items paddy rice educates 293 to be good.
In the present invention, rice material to be measured can be different types of rice varieties (being), breeding individual plant and standard pair
According to kind (being)；Brown rice is that paddy is sloughed after rice husk, the intact Rice Kernel of pericarp, kind skin, megarchidium layer；Brown Rice selection is big
Small basically identical ripe brown rice (paddy is through conventional heat treatment).In the present invention, color is deeper, illustrates paddy rice ratio to be measured
The manganese content of standard items paddy rice is higher.
Rich manganese individual plant in rice material to be measured can be filtered out using the method for the present invention.When the method for the present invention is amounted to
Between be only about 5h, operating process is simple, quick.
The present invention has following technical advantage：
(1) present invention can be detected to coloured and white brown rice, available for the extensive of rich manganese rice material or individual plant
Fast qualitative is screened.The brown rice method of double differences of most rice varieties not Ke Hulvebuji, size can be selected basically identical during screening
Ripe brown rice carry out Rapid identification.The paddy rice special material of control is significantly greater or less than for a small number of brown rice volumes, is carried out
It can increase during experiment or subtract 0.5~1 brown rice, make its brown rice total amount close, it may be necessary to be added using the method weighed
To correct.
Specifically：Weight difference with 4 standard items Brown Rices judges into total amount close ± 5%.
(2) Brown Rice reactant to be measured is contrasted with compareing the color depth of Brown Rice reactant, passes through simple mesh
Survey can be obtained, and color is deeper, and its manganese content is higher.Present invention is mainly used for the qualitative screening of rich manganese paddy rice, dyeing is mainly selected
The sample that colourity " obvious " is deeper than control is illustrated, and the colourity after the close paddy rice sample dyeing of manganese content is more or less the same.
(3) present invention is gelatinized 3h to Brown Rice using 4%NaOH in 65 DEG C, only needs brown rice just can qualitatively screen
Go out the higher rice varieties of manganese content (being) or individual plant.
That is, the present invention can directly utilize brown rice (that is, only needing 4 paddy can direct experiment after shelling), without milling,
Program, saving time can be simplified, the quick manganese content determined in rice.
And routine techniques such as ICP-MS at present about survey manganese is, it is necessary to which machine is milled after being shelled to paddy, and needs special
The instrument of door is cleared up to ground rice, and instrument is valuable and operating technology is strong.
(4) present invention is determined by contrast test：0.4mL 8M sodium hydroxide solutions, 0.1mL formaldoxime solution shows
Color is sensitive and stably.
(5) substantially, the cost that the present invention determines a sample only needs 0.2~0.3 yuan to cost advantage.And routine techniques is such as
The expenses such as ICP-MS are just very high, and detection unit, which surveys a sample, about needs 60~120 yuan.
In summary, rice manganese content simple and fast determination techniques proposed by the present invention only need brown rice can without milling
The higher rice varieties of manganese content (being) or individual plant are filtered out with directly qualitative, the screening of a large amount of samples can be completed in one day.
Compared with traditional ground rice assay method, the present invention can be provided for breeder one it is more simple, quick, economical, have
The rich manganese paddy rice individual plant screening technique of effect.
Brief description of the drawings
The embodiment to the present invention is described in further detail below in conjunction with the accompanying drawings.
Fig. 1 is that figure is compared in 5 kinds of different rice varieties dyeing：26715th, duck blood is glutinous, the early 94-10 of platform, good educate 293, Luo Kema
246。
Fig. 2 is influence figure of the sodium hydroxide concentration to absorbance.
Fig. 3 is influence figure of the formaldoxime developer consumption to absorbance.
Embodiment
With reference to specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in
This.
The rice varieties used in following examples：
26715：Early Xian glutinous rices, the seed selection of Jiaxing academy of agricultural sciences.
Duck blood is glutinous：Changshu, Jiangsu Province farm variety.
Platform morning 94-10 (platform early 9410)：Indica rice, the seed selection of City of Taizhou academy of agricultural sciences.
It is good to educate 293：Indica rice, the seed selection of Jiaxing City, Zhejiang Province academy of agricultural sciences.
Luo Kema 246：Yunnan long-grained nonglutinous rice farm variety.
Embodiment 1：So that 26715, duck blood is glutinous, the ripe Paddy rice of the early 94-10 and Luo Ke horses 246 of platform as detected materials, with
It is good to educate 293 standard varieties as control.After paddy rice to be measured and the shelling of standard items Paddy rice, the rough of different materials is obtained respectively
Rice；Take that the Brown Rice to be measured and standard items Brown Rice that size is basically identical proceed as follows respectively 1)~5)：
1), 4 paddy rice ripe brown rice are fitted into 5mL centrifuge tubes, addition mass fraction 4%NaOH solution 1mL, 35%
Hydrogen peroxidase 10 .1mL, 3h is gelatinized in 65 DEG C of baking ovens, 10%HCl solution 1mL extractions is added after being taken out from baking oven, in 50 DEG C
20min is shaken in thermostatic control oscillator vibration, frequency of oscillation is 250r/min；
2), in step 1) obtained by solution in add 8M NaOH solution 0.4mL, stand 1h after mixing, then add 0.5mL's
Absolute ethyl alcohol, vibration shakes up；
3), in step 2) obtained by solution in add 1M EDTA solution 0.1mL, vibration shakes up；
4), in step 3) obtained by solution in add formaldoxime solution 0.1mL, vibration shakes up, and stands 20min；
The preparation method of formaldoxime solution is：10g hydroxylamine hydrochlorides are dissolved in 50mL water, then add mass fraction to be 35%
Formalin 5mL, is diluted with water to 100mL, in being stored in -20 DEG C of refrigerator at least 30 days, obtains formaldoxime solution；
5), in step 4) obtained by solution in add ammonia-hydroxylamine hydrochloride solution 0.5mL, vibration shakes up, and stands 20min；
The preparation method of ammonia-hydroxylamine hydrochloride solution is：By 4.7M ammonia solutions and 6M hydroxylamine hydrochloride solutions, mixing is shaken in equal volume
It is even.
6), by step 5) the brown rice reaction solution and the brown rice reaction solution progress color ratio of standard items paddy rice of gained paddy rice to be measured
Compared with.
Remarks explanation：Layering occurs after standing, undissolved partly precipitated gets off, the present invention is carried out to the clear liquid on upper strata
Color compares.
As shown in figure 1, the brown rice coloration result of different manganese content rice varieties (being) is shown, 26715, duck blood is glutinous, platform is early
94-10, it is good educate 293, sieve can the color (rufous) of horse 246 successively decrease successively.In coloured picture, the reaction solution color of 26715 displays
For reddish brown, glutinous duck blood is rufous, and platform morning 94-10 is middle rufous, and good to educate 293 be pale red brown, sieve can horse 246 be micro-
Rufous.
With reference to HJ-T 344-2007《The measure of water quality manganese》, the manganese content of above-mentioned rice varieties (being) is measured.Knot
Fruit show glutinous 26715, duck blood, the early 94-10 of platform, the good manganese content for educating 293 and Luo Ke horses 246 be respectively 45.11,39.97,
29.05th, 23.64 and 19.49mg/kg.
Therefore, rice manganese content simple and fast determination techniques of the invention can be used for the breeding practice and kind of rich manganese paddy rice
(being) screens.
Contrast experiment 1：By the step 2 of embodiment 1) in 8M NaOH solutions be provided with altogether 5 gradients (0.30,0.35,
0.40th, 0.45,0.50mL), it have studied influence of the NaOH solution different amounts to absorbance.As a result show that it is inhaled in 0.40mL
Luminosity can reach maximum, therefore the present invention is from 0.40mL NaOH (Fig. 2).
Contrast experiment 2：The consumption of developer can influence the size of absorbance, as shown in Figure 3.When formaldoxime solution usage exists
During 0.05~0.20mL, absorbance is stable, and reaches maximum, and the present invention selects 0.10mL formaldoxime solution.
One of in addition, only list several specific embodiments of the present invention above.It is clear that the invention is not restricted to real above
Example is applied, there can also be many deformations.One of ordinary skill in the art directly can export or associate from present disclosure
All deformations arrived, are considered as protection scope of the present invention.
Claims (4)
1. rice manganese content simple and fast determination method, it is characterized in that：
Paddy rice to be measured and standard items paddy rice are subjected to the steps 1 respectively)~5)；
1), take Brown Rice 4, add 4%NaOH solution 1.0mL, 35% hydrogen peroxidase 10 .1mL, it is gelatinized 3 in 65 ± 5 DEG C ±
0.1h；
10%HCl solution 1.0mL extractions are added, 20 ± 2min of lower concussion, frequency of oscillation in 50 ± 5 DEG C of thermostatic control oscillator vibrations
For 250 ± 10r/min；
2), in step 1) obtained by solution in add 8M NaOH solution 0.4mL, stand 1h, then add 0.5mL absolute ethyl alcohol, shake
Swing and shake up；
3), in step 2) obtained by solution in add 1M EDTA solution 0.1mL, vibration shakes up；
4), in step 3) obtained by solution in add formaldoxime solution 0.1mL, vibration shakes up, and stands 20 ± 2min；
5), in step 4) obtained by solution in add ammonia-hydroxylamine hydrochloride solution 0.5mL, vibration shakes up, and stands 20min；
6), by step 5) the brown rice reaction solution of gained paddy rice to be measured and the brown rice reaction solution of standard items paddy rice carry out color comparison；
When the color of Brown Rice reaction solution to be measured is deeper than the color of standard items Brown Rice reaction solution, illustrate that paddy rice to be measured is rough
The manganese content of rice is higher than standard items paddy rice；
Conversely, when Brown Rice reaction solution to be measured color than standard items Brown Rice reaction solution it is of light color when, then explanation treat
The manganese content for surveying Brown Rice is less than standard items paddy rice.
2. rice manganese content simple and fast determination method according to claim 1, it is characterized in that：
The preparation method of the formaldoxime solution is：10g hydroxylamine hydrochlorides are dissolved in 50mL water, then add mass fraction to be 35%
Formalin 5mL, is diluted with water to 100mL, is stored at least 30 days in -20 ± 2 DEG C, obtains formaldoxime solution.
3. rice manganese content simple and fast determination method according to claim 2, it is characterized in that：
The preparation method of ammonia-hydroxylamine hydrochloride solution is：By 4.7M ammonia solutions and 6M hydroxylamine hydrochloride solutions, mixing shakes up in equal volume.
4. rice manganese content simple and fast determination method according to claim 3, it is characterized in that：
Standard items paddy rice educates 293 to be good.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108279232A (en) * | 2018-01-23 | 2018-07-13 | 浙江大学 | Rice phosphorus content rapid screening method |
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2017
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| CN101177459A (en) * | 2007-11-30 | 2008-05-14 | 四川大学 | High carbonyl content oxidation starch and method for making same |
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| CN105223142A (en) * | 2015-09-13 | 2016-01-06 | 浙江大学 | Rich magnesium paddy rice rapid screening method |
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| CN108279232A (en) * | 2018-01-23 | 2018-07-13 | 浙江大学 | Rice phosphorus content rapid screening method |
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