CN110618159A - LF-NMR determination method for water content of nanmu seeds - Google Patents
LF-NMR determination method for water content of nanmu seeds Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
- G01N24/08—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
- G01N24/082—Measurement of solid, liquid or gas content
Abstract
The invention belongs to the technical field of seed determination, and discloses an LF-NMR determination method for the water content of a nanmu seed, which comprises the following steps of: picking seeds of the superior nanmu trees by using high-branch scissors in a nanmu seed maturation season, washing the seeds with deionized water for 3-5 times, removing pulp, removing surface moisture of the seeds on dust-free absorbent paper, selecting 500g of nanmu seeds with complete and undamaged surfaces, and placing the seeds in a refrigerator at 4 ℃ to be tested; detecting a nanmu seed sample by using a low-field nuclear magnetic resonance technology; based on the obtained seed T2And spectrum data, namely adding the 3 nuclear magnetic resonance relaxation peaks by utilizing a linear relation between the water content of the seeds and the total signal value to obtain the water content of the seeds, and calculating the water content of the nanmu seeds. The method is scientific, simple and easy to operate, is not influenced by human factors and environmental factors, has accurate and objective results, saves more time than the traditional drying method, and has more accurate measurement results.
Description
Technical Field
The invention belongs to the technical field of seed determination, and particularly relates to an LF-NMR determination method for water content of nanmu seeds.
Background
Currently, the current state of the art commonly used in the industry is such that:
the nanmu (Phoebe zhennan) (Phoebe) is a big tree of Lauraceae (Lauraceae) nanmu (Phoebe) and belongs to subtropical evergreen broad-leaf tall trees, is a unique rare endangered protective tree species in China, has excellent wood material, is hard and corrosion-resistant, and is a famous and excellent tree species with high economic value. The Phoebe is mainly distributed in western Hubei province, northwest Guizhou province, Sichuan province and Chongqing city in China, has tall and straight tree forms, luxuriant branches and leaves and evergreen seasons, and is a main tree species for forming evergreen broad-leaved forests. The nanmu is scattered in the mixed forest with the elevation of less than 1000 m, has the biological characteristics of neutral and partial negative, developed root system, long service life and slow growth speed, and can enter the vigorous growth stage only in 50 to 60 years. The nanmu seed fructification has an obvious big-year-old phenomenon, and the nanmu seed has a dormancy characteristic and is not easy to germinate under natural conditions, so that the nanmu seed is required to be picked and stored when the nanmu seed fructification is old for later use when the seeds are young. When storing seeds, the moisture content in the seeds is the most critical index related to the storage quality of the seeds.
Moisture is a medium for metabolism in seed cells, and the physical properties and chemical processes of seeds and the yield and quality of seeds during development, maturation, harvesting and storage of seeds are closely related to the state and content of moisture. The water in the seeds is a complex system and is generally divided into three states of bound water, non-flowing water and free water, the existing state of the water in the seeds is closely related to the internal activity of the seeds, when the seeds only contain the bound water, the enzymes in the seeds, particularly the hydrolytic enzymes, are in an inactive state, the metabolism of the seeds is extremely weak, and the vitality maintenance and the service life of the seeds are favorably prolonged; the non-flowing water belongs to a transition state between the bound water and the free water; when free water is present, the enzymes in the seeds are converted from a deactivated state to an activated state, and the respiration of the seeds is rapidly increased. With the increase of the free water content in the seeds, not only the metabolism of the seeds becomes vigorous, but also the activity of microorganisms is caused, the seeds can generate heat when the water content is serious, and the seeds can germinate when the water content exceeds a certain limit. Thus ensuring that the seeds are in a safe moisture content range is of great importance in maintaining the life of the seeds.
The Low Field Nuclear Magnetic Resonance (LF-NMR) technique can study the distribution, migration, moisture content and other properties related thereto of the moisture in the material by measuring the transverse and longitudinal relaxation times of hydrogen nuclei in the material under the action of a Magnetic Field and a radio frequency signal, and has the advantages of non-loss, greenness, rapidness and the like. The technology uses a specific pulse sequence Carr-Purcell-Meiboom-Gill (CPMG), the process of enabling hydrogen protons in a sample to restore from an unbalanced state to an balanced state after resonance is called a relaxation process, and a time constant required by the transverse vector direction to restore the balance in the process is called transverse relaxation time (T2). The hydrogen proton has extremely high content in plant seeds and can generate strong nuclear magnetic resonance signals. The presence state of hydrogen protons tends to have a better correlation with relaxation time,
the relaxation time of the plant seed sample is changed due to different physicochemical environments, so that a great deal of internal information of the plant seed sample can be acquired. At present, the method becomes one of important technologies for detecting the internal moisture of the materials, and the LF-NMR is widely applied to the determination of the moisture content in materials such as fruits, vegetables, meat, grain and oil and the like.
According to the water content determination regulation of forest seeds in International seed inspection Association (ISTA) International seed inspection regulations and China national Standard GB 2772 and 1999 forest seed inspection regulations, large-grain seeds with the water content higher than 17% are firstly sliced and then determined by a low constant temperature drying method, namely, the seed samples are placed in a drying oven with a method of 103 +/-2 ℃ for drying for 17 +/-1 hours, and the factors of more samples, uncertain sample delivery time and the like often appear when the seeds are harvested, tedded and warehoused. The method needs one working day to complete the moisture determination of one nanmu seed, the determination time is too long, if the sample sending time is improper, the detection result can not be obtained on the same day, and the time for putting the seeds in storage is delayed.
In summary, the problems of the prior art are as follows:
the conventional forest seed measuring method has long measuring time and is easily influenced by human and environmental factors.
The method for measuring the water content of the seeds in the prior art is a drying method, and has the main problems that the time consumption is too much, the time of about one day is needed for completing the measurement, and when the number of the seeds is extremely large, the warehousing time of the seeds is delayed, so that the efficiency of the seed purchasing time is low.
The difficulty of solving the technical problems is as follows:
in the traditional drying method, the determination result is time-consuming; in the traditional rapid determination method, the measurement result is not accurate enough; the above problems are difficult to solve by the prior art.
The significance of solving the technical problems is as follows:
the method can overcome the defects in the existing seed measurement process, and has important significance for quickly and accurately measuring the water content of the seeds.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides an LF-NMR determination method for the water content of nanmu seeds. The invention can save a large amount of time for purchasing seeds by using a low-field nuclear magnetic resonance technology, thereby improving the efficiency. The method is used for measuring by using a low-field nuclear magnetic resonance technology, and the method can be used for quickly and accurately detecting the moisture content of the nanmu seeds.
The invention is realized in such a way that the LF-NMR determination method of the water content of the nanmu seeds comprises the following steps:
step one, sampling a nanmu seed sample: picking seeds of the superior nanmu trees by using high-branch scissors in a nanmu seed maturation season, washing the seeds with deionized water for 3-5 times, removing pulp, removing surface moisture of the seeds on dust-free absorbent paper, selecting 500g of nanmu seeds with complete and undamaged surfaces, and placing the seeds in a refrigerator at 4 ℃ to be tested;
detecting a nanmu seed sample by using a low-field nuclear magnetic resonance technology;
step three, based on the obtained seeds T2Spectrum data, the linear relation between the water content of the seeds and the total signal value is utilized, the water content of the seeds is obtained by adding 3 nuclear magnetic resonance relaxation peaks, and the nanmu is calculatedSeed moisture content.
Further, in the first step, picking the seeds of the nanmu superior tree by using the high branch scissors in the nanmu seed maturation season specifically comprises:
the collection time of the nanmu seeds is 9-10 months of the mature season of the nanmu seeds every year;
the nanmu superior tree is a nanmu with the age of more than 30 years.
Further, in the second step, the low-field nuclear magnetic resonance detection method specifically includes:
(1) collecting a signal of a seed by using a Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence; setting pulse sequence parameters: the sampling frequency is 1200kHz, the width of a 180-degree hard pulse is 10.2 mus, the width of a 90-degree hard pulse is 5.85 mus, the number of echoes is 520, the half-echo time is 120 mus, and the accumulation frequency is 12 times;
(2) measuring the moisture content of the nanmu seeds by using a low-field nuclear magnetic resonance instrument LF-NMR to obtain echo curve data; the resonance frequency of the low-field nuclear magnetic resonance instrument is set to be SF (sulfur factor) 22MHz (hydrogen proton), the magnet temperature is 32 ℃, the receiver bandwidth is 200kHz, the sampling time control parameter is 0.15ms, and the signal is repeatedly measured for 5 times;
(3) obtaining transverse relaxation time inversion spectrum (T) by WinDXP software carried by low-field nuclear magnetic resonance apparatus by using inversion algorithm2A spectrum;
(4) repeatedly measuring each nanmu seed sample for 5 times, inverting each echo data, recording the 5 times of inverted data, carrying out arithmetic mean, and taking the mean value as the final T of the sample2Spectral data.
Further, T of the seed2The spectral data includes:
the nuclear magnetic relaxation inversion spectra of the nanmu seed sample have 3 peaks, which respectively represent three water states in the seeds: bound water, non-flowing water and free water;
the seed T2The spectral data includes free water relaxation signal values, stagnant water relaxation signal values, and bound water relaxation signal values.
Further, in the third step, the method for calculating the water content of the nanmu seeds specifically comprises the following steps:
processing the measured data by SPSS18.0 statistical analysis software; seed moisture content was determined using the following technique:
the water content of the nanmu seeds is the free water relaxation signal value + the non-flowing water relaxation signal value + the bound water relaxation signal value.
The invention also aims to provide a nanmu seed moisture content LF-NMR measuring device for implementing the nanmu seed moisture content LF-NMR measuring method.
In summary, the advantages and positive effects of the invention are:
the invention provides a low-field nuclear magnetic resonance (LF-NMR) measuring method for nondestructively and rapidly measuring the water content of nanmu seeds, which is strong in scientificity, simple and easy to operate, free from the influence of human factors and environmental factors, accurate and objective in result, more time-saving than a traditional drying method and more accurate in measuring result.
According to the invention, the nanmu seeds are washed by deionized water for 3-5 times, the pulp is removed, the surface moisture of the seeds is removed on the dust-free absorbent paper, and the nanmu seeds are placed in a refrigerator at 4 ℃ to be measured, so that the moisture of the sample is not lost, and the accuracy of the sample measurement is ensured.
The method comprises the steps of collecting seed signals by using a Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence, measuring the moisture content of a nanmu seed by using a low-field nuclear magnetic resonance instrument produced by Shanghai Nymei electronic technology company Limited, and obtaining a transverse relaxation time inversion spectrum (T2 spectrum) by using a WinDXP software running inversion algorithm carried by the low-field nuclear magnetic resonance instrument; obtaining echo curve data; the method can make the measuring process of the sample more controllable, and ensure the accuracy of the measuring result of the sample.
Selecting a nanmu superior tree with the age of 30 years, wherein the collection time of the nanmu seeds is about 10 months of the mature season of the nanmu seeds.
The water content of the seeds was measured using a low-field nuclear magnetic resonance apparatus and the results were as follows:
drawings
Fig. 1 is a flow chart of an LF-NMR measurement method for determining the moisture content of nanmu seeds provided by an embodiment of the present invention.
Fig. 2 is a schematic diagram of the relationship between total signal quantity and dry-based moisture content of seeds provided by the embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The conventional forest seed measuring method has long measuring time and is easily influenced by human and environmental factors.
To solve the above problems, the present invention will be described in detail with reference to the accompanying drawings.
As shown in fig. 1, the LF-NMR measurement method for determining the moisture content of the nanmu seeds provided by the embodiment of the present invention includes:
s101, sampling a nanmu seed sample: picking seeds of the superior nanmu trees by using high-branch scissors in a nanmu seed maturation season, washing the seeds with deionized water for 3-5 times, removing pulp, removing surface moisture of the seeds on dust-free absorbent paper, selecting 500g of nanmu seeds with complete and undamaged surfaces, and placing the seeds in a refrigerator at 4 ℃ to be tested.
And S102, detecting a nanmu seed sample by using a low-field nuclear magnetic resonance technology.
S103, based on the obtained seeds T2And spectrum data, namely adding the 3 nuclear magnetic resonance relaxation peaks by utilizing a linear relation between the water content of the seeds and the total signal value to obtain the water content of the seeds, and calculating the water content of the nanmu seeds.
In step S101, the picking of the seeds of the nanmu superior tree by using the high branch shears in the nanmu seed maturing season provided by the embodiment of the present invention specifically includes:
the collection time of the nanmu seeds is 9-10 months of the mature season of the nanmu seeds every year.
The nanmu superior tree is a nanmu with the age of more than 30 years.
In step S102, the low-field nuclear magnetic resonance detection method provided in the embodiment of the present invention specifically includes:
(1) collecting a signal of a seed by using a Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence; setting pulse sequence parameters: the sampling frequency is 1200kHz, the width of a 180-degree hard pulse is 10.2 mus, the width of a 90-degree hard pulse is 5.85 mus, the number of echoes is 520, the half-echo time is 120 mus, and the accumulation frequency is 12 times.
(2) Measuring the moisture content of the nanmu seeds by using a low-field nuclear magnetic resonance instrument LF-NMR to obtain echo curve data; the resonance frequency of the low-field nuclear magnetic resonance apparatus is set to be SF 22MHz (hydrogen proton), the magnet temperature is 32 ℃, the receiver bandwidth is 200kHz, the sampling time control parameter is 0.15ms, and the signal is repeatedly measured for 5 times.
(3) Obtaining transverse relaxation time inversion spectrum (T) by WinDXP software carried by low-field nuclear magnetic resonance apparatus by using inversion algorithm2Spectra.
(4) Repeatedly measuring each nanmu seed sample for 5 times, inverting each echo data, recording the 5 times of inverted data, carrying out arithmetic mean, and taking the mean value as the final T of the sample2Spectral data.
The T of the seed provided by the embodiment of the invention2The spectral data includes:
the nuclear magnetic relaxation inversion spectra of the nanmu seed sample have 3 peaks, which respectively represent three water states in the seeds: bound water, non-flowing water and free water.
The seed T2The spectral data includes free water relaxation signal values, stagnant water relaxation signal values, and bound water relaxation signal values.
In step S103, the method for calculating the moisture content of the nanmu seeds provided by the embodiment of the present invention specifically includes:
processing the measured data by SPSS18.0 statistical analysis software; seed moisture content was determined using the following technique:
the water content of the nanmu seeds is the free water relaxation signal value + the non-flowing water relaxation signal value + the bound water relaxation signal value.
The technical solution and technical effects of the present invention are further described below with reference to specific embodiments.
Example 1:
the embodiment provides an LF-NMR determination method for water content of nanmu seeds, which includes a nanmu seed sample sampling step, a low-field nuclear magnetic resonance technology (LF-NMR) detection step, and a seed water content calculation step:
firstly, sampling a nanmu seed sample: the method comprises the steps of collecting machilus nanmu seeds for 9-10 months every year, picking the seeds of machilus nanmu superior trees (machilus nanmu-1) by using high-branch scissors in machilus nanmu seed maturation season, washing the seeds for 3-5 times by using deionized water, removing pulp, removing surface moisture of the seeds on dust-free absorbent paper, selecting 500g of machilus nanmu seeds with intact surfaces, and placing the machilus nanmu seeds in a refrigerator at 4 ℃ to be measured.
(II) a low-field nuclear magnetic resonance technology (LF-NMR) detection step: the process comprises the following steps:
firstly, collecting a signal of a seed by using a Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence; setting pulse sequence parameters: SW (sampling frequency) 1200kHZP180(180 DEG hard pulse width) 10.2. mu.s, P90(90 DEG hard pulse width) 5.85. mu.s, NECH (echo count) 520, TAU (half echo time) 120. mu.s, NS (cumulative count) 12 times.
In a second step, the seeds are measured by low-field nuclear magnetic resonance (LF-NMR): measuring the water content of the nanmu seeds by using a low-field nuclear magnetic resonance instrument of Shanghai Newman electronic technology Co., Ltd to obtain echo curve data; the resonance frequency of a low-field nuclear magnetic resonance (LF-NMR) is set to SF 22MHZ(Hydrogen protons), magnet temperature 32 ℃, receiver bandwidth 200kHZThe time control parameter was sampled for 0.15ms and the signal was measured 5 times repeatedly.
Thirdly, obtaining a transverse relaxation time inversion spectrum (T) by running an inversion algorithm through WinDXP software carried by the low-field nuclear magnetic resonance instrument2Spectra).
Fourthly, repeating the operations: repeatedly measuring each nanmu seed sample for 5 times, inverting each echo data, recording the 5 times of inverted data, carrying out arithmetic mean, and taking the mean value as the final T of the sample2Spectral data.
(III) calculating the water content of the nanmu seeds: according to the relationship between the transverse relaxation signal quantity of the nanmu seeds and the moisture content of different dry bases, the dry base moisture content and the nuclear magnetic resonance total signal amplitude in the seed drying process are in an obvious linear relationship, the linear equation is y which is 154.09x +1547.7, the correlation coefficient R2 which is obtained by fitting is 0.9912, and the fact that the total signal value has good correlation with the moisture content of the dry bases is explained. The total signal value according to the nmr transverse relaxation time can therefore represent the moisture content of the seed, as shown in figure 2.
The water content of the seeds is the free water relaxation signal value + the non-flowing water relaxation signal value + the bound water relaxation signal value.
The moisture content of the nanmu seeds is tested by the LF-NMR measurement method for the moisture content of the nanmu seeds of this embodiment, and the test results are shown in the following table:
the above experimental results are the results of the LF-NMR method for determining the moisture content of the nanmu superior tree (nanyou-1) seed sample.
Meanwhile, the water content of the seeds of the nanmu superior tree (nanyou-1) is measured by a conventional drying method as a control, and the result is as follows:
as can be seen from the above table, the moisture content of the seeds of the naughty tree (naughty-1) measured by the drying method is 33.66% to 35.01%, while the moisture content of the seeds of the naughty tree (naughty-1) measured by the LF-NMR method is 36.34% to 39.58%. The paired t test shows that the difference between the two is very significant (P < 0.001). Therefore, the determination result of the low-field nuclear magnetic resonance technology on the water content of the nanmu seeds is obviously higher than that of a direct drying method.
In the LF-NMR measurement method for the moisture content of the nanmu seeds of this embodiment, the step of sampling the nanmu seed sample(s) may slightly differ the result of the measured moisture content due to the different mass of the seeds at each measurement.
The moisture content of the nanmu seeds measured by the LF-NMR measuring method for the moisture content of the nanmu seeds in the embodiment is directly expressed in a quantitative manner, so that the time is saved and the measuring result is more accurate and objective compared with the traditional drying method. In addition, the determination method also has the advantages of simple method, easy operation and no influence of human factors and environmental factors.
Example 2:
this example is substantially the same as example 1, except that the nanmu seeds are seeds collected from nanyou-2-best trees, and the tree age is 50 years.
Example 3:
this example is substantially the same as example 1, except that the nanmu seeds are seeds collected from nanyou-3-quality trees, and the tree age is 75 years.
Example 4:
this example is substantially the same as example 1, except that the nanmu seeds are seeds collected from nanyou-4-quality trees, and the tree age is 75 years.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (6)
1. An LF-NMR determination method for the water content of nanmu seeds is characterized by comprising the following steps:
step one, sampling a nanmu seed sample: picking seeds of the superior nanmu trees in a nanmu seed maturing season, washing with deionized water, removing pulp, removing surface water on the seeds, selecting the nanmu seeds with intact surfaces, and testing to be tested;
detecting a nanmu seed sample by using a low-field nuclear magnetic resonance technology;
step three, based on the obtained seeds T2Spectral data, using a linear relationship between seed water content and total signal valueAnd adding the multiple nuclear magnetic resonance relaxation peaks to obtain the water content of the seeds, and calculating the water content of the nanmu seeds.
2. The LF-NMR measurement method for determining moisture content of nanmu seeds of claim 1, wherein in the first step, the picking of seeds of a nanmu superior tree by using averruncators in a nanmu seed maturation season specifically comprises:
the collection time of the nanmu seeds is 9-10 months of the mature season of the nanmu seeds every year;
the nanmu superior tree is a nanmu with the age of more than 30 years.
3. The LF-NMR determination method for the water content of the nanmu seeds of claim 1, wherein in the second step, the low-field nuclear magnetic resonance detection method specifically comprises the following steps:
(1) collecting a signal of a seed by using a Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence; setting pulse sequence parameters: the sampling frequency is 1200kHz, the width of a 180-degree hard pulse is 10.2 mus, the width of a 90-degree hard pulse is 5.85 mus, the number of echoes is 520, the half-echo time is 120 mus, and the accumulation frequency is 12 times;
(2) measuring the moisture content of the nanmu seeds by using a low-field nuclear magnetic resonance instrument LF-NMR to obtain echo curve data; the resonance frequency of the low-field nuclear magnetic resonance instrument is set to be SF (sulfur factor) 22MHz (hydrogen proton), the magnet temperature is 32 ℃, the receiver bandwidth is 200kHz, the sampling time control parameter is 0.15ms, and the signal is repeatedly measured for 5 times;
(3) obtaining transverse relaxation time inversion spectrum (T) by WinDXP software carried by low-field nuclear magnetic resonance apparatus by using inversion algorithm2A spectrum;
(4) repeatedly measuring each nanmu seed sample for 5 times, inverting each echo data, recording the 5 times of inverted data, carrying out arithmetic mean, and taking the mean value as the final T of the sample2Spectral data.
4. The LF-NMR method for determining water content of nanmu seeds of claim 3, wherein T of the seeds is2The spectral data includes:
the nuclear magnetic relaxation inversion spectra of the nanmu seed sample have 3 peaks, which respectively represent three water states in the seeds: bound water, non-flowing water and free water;
the seed T2The spectral data includes free water relaxation signal values, stagnant water relaxation signal values, and bound water relaxation signal values.
5. The LF-NMR measurement method for the water content of the nanmu seeds of claim 1, wherein in step three, the calculation method for the water content of the nanmu seeds specifically comprises:
processing the determined data by statistical analysis software; seed moisture content was determined using the following technique:
the water content of the nanmu seeds is the free water relaxation signal value + the non-flowing water relaxation signal value + the bound water relaxation signal value.
6. An LF-NMR measurement apparatus for determining the moisture content of a nanmu seed, which implements the LF-NMR measurement method for determining the moisture content of a nanmu seed according to claim 1.
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Application publication date: 20191227 |