CN112730496B - Method for measuring water content of gladiolus fresh cut flower - Google Patents
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Abstract
The invention discloses a method for measuring water content of gladiolus fresh cut flowers. The invention provides a water content determination method in a fresh cut flower senescence process by combining a low-field nuclear magnetic imaging method and a T2 relaxation method, and provides a standardized method for researching the physiological mechanism of fresh cut flower senescence, developing a preservation technology and delaying senescence. The method has the advantages of small sample amount and simple steps, and overcomes the defects that the detection by using the traditional drying method consumes long time and cannot meet the requirements of real-time feedback and real-time adjustment in the production process; and the method has the advantages of high determination speed, good signal-to-noise ratio and stable and accurate prediction result, and is suitable for identification, optimization, research and application of the fresh cut flower preservation technology.
Description
Technical Field
The invention belongs to the technical field of fresh cut flower detection, and particularly relates to a fresh cut flower moisture determination method.
Background
Fresh cut flowers are one of the important product types for flower production in the flower industry. Cut flowers can not be cut in the courtesy activities at home and abroad, and the fresh cut flower plays an important role in decorating the environment, beautifying the life and transmitting the emotion, so that the global fresh cut flower industry is rapidly developed along with the economic development. The fresh cut flower cells need to keep certain swelling pressure to maintain normal physiological metabolism and keep bright appearance. After the cut flowers are harvested, water and nutrition supply from the root systems of the parent bodies cannot be obtained, and the atmospheric relative humidity of the environment in which the cut flowers are located accelerates the water transpiration. When water is deficient, various soluble salt ions, nutrients, enzyme activities and the like are lost or reduced, and the cut flowers are low in openness rate, low in vividness and wilting and aging. Therefore, the water content is the most important physiological index for cut flower preservation and is also an indispensable measurement index in cut flower preservation technology. However, the freshness of cut flowers depends on the equilibrium relationship between water uptake and water loss and the normal metabolic processes affected thereby, rather than the amount of pure water content. Therefore, the moisture condition is an important factor for determining the aging process of the cut flowers, and the comparison of the moisture conditions of different components is more practical.
At present, the method for measuring the water content in the fresh cut flower industry is mainly a traditional drying method. The method usually needs the temperature of more than 70 ℃ for more than several hours, and needs cooling to room temperature, so that the detection is long in time consumption, and the requirements of real-time feedback and real-time adjustment in the production process cannot be met. Furthermore, the cut flowers contain volatile aromatic substances and lose the total amount of volatile substances which are not thermally stable at high temperature, not completely water, resulting in deviation of the obtained water content.
The nuclear magnetic resonance is a real-time, nondestructive and noninvasive quantitative measurement technology, and can reflect indexes such as water content and water occurrence state of materials from a microscopic angle. However, reports on the measurement of the water content, the water content state and the dynamic change of the fresh cut flowers are not seen yet. In addition, the flower organ of the fresh cut flower comprises different tissues such as bracts, petals, stamens and the like, the moisture content and the physicochemical property of each tissue are different, and the low-field nuclear magnetic resonance detection of specific protons is not influenced by the surface property in the measurement process, so that the low-field nuclear magnetic resonance technology has more advantages.
Disclosure of Invention
The invention aims to provide a fresh cut flower moisture determination method aiming at the defects of the fresh cut flower moisture determination method in the prior art.
The method is based on a nuclear magnetic resonance imaging technology, the water content is calculated by adopting NMR signal amplitude inversion, and the water state and the change in the fresh cut flowers are analyzed through a signal spectrogram; the method has the advantages of high signal-to-noise ratio, high detection speed and simple steps.
After the fresh cut flowers are picked, the fresh cut flowers are required to absorb water and nutrients through the conduction tissues of the stems so as to be kept undecided for a certain time. The main reason for the senescence of cut flowers is that the withering of cut flowers is aggravated by water loss inside petal tissues caused by microbial, physiological and physical blockage of flower stems. Therefore, the detection and analysis of the nuclear magnetic resonance imaging of the scape is beneficial to the research of the change rule of the water migration, which cannot be obtained only by the drying technology.
The method of the invention can not only measure the water content, but also can detect the water state and the dynamic change of the fresh cut flowers in the aging process; the preservative can also be applied to the technical field of preservation of fresh cut flowers, the effects of keeping the moisture content of different preservatives are compared, the ratio of the preservatives is improved, and the preservation technology is optimized.
The fresh cut flower moisture determination method comprises the following steps:
a. sample preparation: cutting fresh cut flower samples with proper sizes, weighing, and sealing in a nuclear magnetic sample tube;
b. detecting a sample by using low-field nuclear magnetic resonance, obtaining echo attenuation curve data of a CPMG sequence, placing the corresponding sample at 70 ℃, and drying to constant weight; in the drying process, collecting the CPMG signal of the corresponding sample every 1 hour;
the parameter setting range of the low-field nuclear magnetic resonance detection is as follows:
90-degree pulse width P1: 15 μ s, 180-degree pulse width P2: 29 μ s, oversampling waiting time Tw: 1000-10000ms, analog gain RG 1: 10 to 20, each being an integer; digital gain DRG 1: 2 to 5, each being an integer; receiver bandwidth SW: 100, 200, 300 KHz; control parameter RFD of start sampling time: 0.002-0.06 ms, echo number NECH: 1000-12000, repeated sampling waiting time Tw: 2000ms, echo time TE: 0.18-0.2 ms, and the repeated sampling times NS is 4/8/16;
c. b, immediately weighing the corresponding sample after the CPMG signal is collected at each time point in the step b, and calculating according to the mass of the corresponding sample to obtain the water content measured value of the corresponding sample;
d. processing the T2 echo attenuation curve signal of the CPMG sequence obtained in the step b, extracting a T2 initial point signal value of total water NMR, fitting the initial point signal value and the water content measured value of T2 corresponding to the water content measured value obtained in the step c, and establishing a fresh cut flower water content prediction model;
e. b, collecting T2 attenuation curve data of the fresh cut flower sample to be detected by adopting the same parameter setting as that in the step b, obtaining a first point signal value and T2 of different relaxation components, substituting the first point signal value into the fresh cut flower water content prediction model to obtain a water content prediction value corresponding to the fresh cut flower sample to be detected, and using T2 (inversion spectrum) of the different relaxation components for water state judgment;
f. low-field nuclear magnetic imaging: and (3) placing the fresh cut flower sample into a nuclear magnetic sample tube, performing imaging analysis by using a low-field nuclear magnetic resonance apparatus, and judging the water distribution condition according to the color depth in the image and the inversion spectrum of the transverse relaxation time T2.
Preferably, the step a of cutting a fresh cut flower sample with a suitable size is to cut not more than 1cm 2 Petals of a size or stem segments of no more than 2cm in length.
Preferably, the step b is repeated by using 3-6 samples collected each time by using low-field nuclear magnetic resonance detection.
Preferably, the NMR signals of T2 and T2 values of 1-10, 10-100 and 100-10000 ms of different relaxation components in the step e are respectively analyzed into bound water (T21), non-flowable water (T22) and free water (T23) in sequence.
In the method, according to the characteristic peaks of the obtained sample, NMR signals with T2 values of 1-10, 10-100 and 100-10000 ms are respectively analyzed into bound water (T21), non-flowing water (T22) and free water (T23), and the peak area of 75-90% of the inverted NRM signal corresponding to the water content of the fresh cut flower is located in 100-1000 ms. The adjusted parameters can be used for the measurement of the fresh cut flowers later. The fresh cut flower water content prediction model adopts a correlation coefficient R 2 Evaluation of the prediction model was performed.
Preferably, the low-field nuclear magnetic resonance apparatus in step f performs imaging analysis, specifically: adopting MRI imaging software and SE imaging sequence test to collect proton density image of sample; MRI imaging parameters: and accumulating and sampling 8 times, wherein GZ is-850, GY is-240, GX is-100, Tw is 2000.
Preferably, the fresh cut flower sample of the step f is a stem section with the length of not more than 2cm and taken at a position 20-22 cm away from the cut.
Preferably, the fresh cut flower is gladiolus.
Preferably, the instrument for detecting the low-field nuclear magnetic resonance is an NMI20 type low-field nuclear magnetic resonance detector, the diameter of the coil is 15mm, and the parameters are set as follows: the 90-degree pulse width P1 is 15us, the control parameter RFD of the start sampling time is 0.06ms, the analog gain RG1 is 20.0db, the digital gain DRG1 is 3, the receiver bandwidth SW is 200KHz, the repeated sampling waiting time Tw is 2000ms, the 180-degree pulse width P2 is 29us, the echo time TE is 0.2ms, the echo number NECH is 10000, and the repeated sampling number NS is 8.
The invention also provides application of the method in detecting the water content and the water migration change of the fresh cut flowers.
The invention has the advantages that:
1. by establishing the water content prediction model, the water content of the fresh cut flowers can be detected quickly and accurately in real time. Therefore, the method can meet the requirements of real-time feedback and real-time adjustment in the production process.
2. The moisture state and the dynamic change thereof can be detected through the transverse relaxation time, the comparison and improvement of different treated components can be realized, the comparison of the effects of keeping the moisture content of different preservatives can be realized, the ratio of the preservatives is improved, and the preservation technology is optimized.
3. The imaging observation is carried out on the flower stem, the main organ determining the water absorption of the fresh cut flower, and the water migration, so that the method is visual and clear, and cannot be obtained only by the traditional drying technology. Has very positive effect on researching the water migration rule of the fresh cut flowers and maintaining the intrinsic mechanism of the life.
Drawings
FIG. 1 is a water cut prediction model of example 1;
FIG. 2 shows the NMR signal amplitude during the opening of the flowers of gladiolus of example 1 and the water content during the opening of the flowers as measured by a conventional drying method;
FIG. 3 shows the change of different degrees of openness of gladiolus T2 (transverse relaxation time) in example 1;
FIG. 4 is the relative proportions of NMR signal amplitudes and different moisture states for gladiolus petals, stem segments and bracts of example 2;
FIG. 5 is MRI imaging of the gladiolus cut flower stem of example 2.
Detailed Description
The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
Gladiolus was selected as the sample material in the following examples. Gladiolus hvbridus Hort is a perennial herb of Gladiolus of Iridaceae, has various varieties, unique flower shapes and rich colors, is popular with consumers at home and abroad, and is one of four famous cut flowers in the world.
Example 1: water content of gladiolus florets in the process of blooming
Parameter debugging.
The first step is as follows: the height of the sample must be less than the set value of field Fov due to instrument field uniformity requirements. The diameter of a coil of the NMI20 type low-field nuclear magnetic resonance detector used in the test is 15mm, and the range of the sample loading height of the sample cannot exceed 2.5cm in order to ensure the accuracy of the detection result of the sample. The cut flowers were found to have a high fill value during actual testing and were difficult to compress to the detection height at sample levels above 0.6 g. Thus, the petal sample was cut into 1cm 2 And (3) taking 0.5g of the small pieces, putting the small pieces into a dry and clean nuclear magnetic sample tube with the diameter of 15mm, and sealing.
The second step is that: and exciting the sample by adopting a single 90-degree radio frequency pulse, and receiving a hard pulse FID signal for debugging. Accumulating the module values under 2 sampling records Tw, increasing Tw by 500ms, comparing the current module value with the previous module value, repeatedly increasing Tw until the module value variation range is less than 1%, and finally determining the proper Tw.
The third step: and acquiring data through a CPMG sequence, and debugging the number NECH of echoes to ensure that the transverse relaxation of the sample is basically finished within the sampling time. The accumulated samples are set to the power of 2 in the range of 2 to 4, i.e., the number of samples is 4 to 16. The number of samples with a signal-to-noise ratio greater than 50 is selected according to the signal attenuation curve. The 90-degree pulse width P1 is 15us, the control parameter RFD of the start sampling time is 0.06ms, the analog gain RG1 is 20.0db, the digital gain DRG1 is 3, the receiver bandwidth SW is 200KHz, the repeated sampling waiting time Tw is 2000ms, the 180-degree pulse width P2 is 29us, the echo time TE is 0.2ms, the echo number NECH is 10000, and the repeated sampling number NS is 8. The adjusted parameters can be used for the later determination of the fresh cut flowers of the same species.
And secondly, a low-field nuclear magnetic resonance detection sample is applied, and after echo attenuation curve data of the CPMG sequence are obtained, a first point signal value is extracted. And (3) putting the corresponding gladiolus floret sample at 115 ℃ for enzyme deactivation, drying at a constant temperature of 70 ℃ to constant weight, and collecting the CPMG signal of the corresponding sample every 1 hour in the drying process, wherein the CPMG signal is repeated at 6 time points.
Thirdly, measuring the moisture content of each sampling point in the drying process of the fresh cut flowers by adopting a drying constant weight method to obtain a moisture content measured value; the measurement of the water content was determined by the oven-dry constant weight method by taking the average value of the water content by repeating 6 times at each time point.
Fourthly, processing the obtained T2 echo attenuation curve signal of the CPMG sequence by adopting a chemometrics method, extracting a transverse relaxation time T2 initial point signal value of total moisture NMR, carrying out mass normalization on a corresponding dried water content measured value, fitting a T2 initial point signal value and the water content measured value, and establishing a fresh cut flower water content prediction model to obtain a standard curve y which is 0.0014 x-0.6118; wherein x is the initial signal value of the transverse relaxation time T2 attenuation curve, and y is the predicted water content value. Using a correlation coefficient R 2 Then, the prediction model is evaluated. Correlation coefficient R of the two 2 0.96428 (fig. 1). A sample to be tested is randomly taken and substituted into the formula to obtain a predicted value (prediction Y) of the water content, and the predicted value is compared with the measured value of the water content by a drying constant weight method (Table 1).
TABLE 1 comparison of measured and predicted moisture content of fresh cut flowers
And fifthly, taking materials and measuring the sample. Cutting small flowers which are opened for 1-6 days into about 1cm 2 After flaking, 0.5g of each of the above-mentioned small pieces was taken out, and 3 portions were repeated. Putting into a dry and clean nuclear magnetic sample tube with the diameter of 15mm and sealing. The first signal value and the transverse relaxation time T2 (fig. 2, fig. 3) were measured for each day. Different peaks of the T2 inversion spectrum represent different states of water, and NMR signals with T2 values of 1-10, 10-100 and 100-10000 ms are respectively analyzed into bound water (T21), non-flowing water (T22) and free water (T23).
Example 2: moisture state of different organs and tissues of gladiolus cut flower and distribution change after harvesting
The method comprises the steps of sampling and measuring samples. Cutting the bud and petal sample to about 1cm 2 Cutting about 0.5g of small pieces, cutting stem into pieces with a height of not more than 2cm, and sealing in a dry and clean nuclear magnetic sample tube with a diameter of 15 mm. And extracting a first point signal value through a CPMG sequence, inverting to obtain the NMR signal amplitude and transverse relaxation time T2 of each relaxation component, and collecting sample data.
And analyzing data. And obtaining characteristic peaks of the gladiolus cut flower, respectively analyzing NMR signals with T2 values of 1-10, 10-100 and 100-10000 ms into bound water (T21), non-flowing water (T22) and free water (T23), and analyzing the water content of the sample according to the signal amplitude and T2. The NMR signal amplitudes of different organs and tissues of the gladiolus cut flower are different, and the sizes and the occupation ratios of 3 peaks of combined water, difficult flowing water and free water are different (figure 4), which shows that the petals and the stem segments have high free water content, the free water content of the bracts is obviously lower than the former two, and the water content of different parts is different from the proportion of 3 types of state water. Therefore, the proportion of free water in the fresh cut flowers is far greater than that of the bound water and the water which is not easy to flow. According to the idea of regression analysis, the characteristic peaks of different transverse relaxation ranges and the water content are subjected to regression coefficient significance analysis, and the characteristic peaks of free water and the water content are found to be in a significant relation.
And imaging. And (3) taking a stem section (about 2cm) with the length of not more than 2cm at a position 20-22 cm away from the cut for each flower 0, 4 and 8 days after the flower is picked, and putting the stem section into a dry and clean nuclear magnetic sample tube with the diameter of 15mm for sealing. Based on the parameters, when the gradient is adjusted, the phase encoding gradient GA3 is equal to 0, so that other parameters can be optimized conveniently, and GS is sampled once; adjusting the frequency coding GA4 to maximize the echo amplitude; adjusting GA3, accumulating samples, and observing imaging effect. GA 0' 2 determines the layer selection, and the larger the value is, the thinner the layer is; and fourthly, regulating GA4 after regulating GA5 to see whether the signal value is not increased any more, and similarly, regulating GA0 and GA2 firstly and regulating GA1 secondly to see whether the signal value is not increased any more when regulating GA 1. MRI imaging parameters: GZ-850, GY-240, GX-100, Tw-2000, and 8 samples are accumulated. And acquiring data through the spin echo sequence by using the debugged parameters. It can be seen that the water content of the stem increases significantly from the time of bottle insertion to the fourth day (fig. 5), which is consistent with the appearance of the small flower opening process (fig. 2), and the water content of vascular bundle tissue in the stem is higher, and browning and marrow decay occur by the eighth day.
Claims (3)
1. The method for measuring the water content of the gladiolus fresh cut flower is characterized by comprising the following steps:
a. sample preparation: cutting and weighing a gladiolus fresh cut flower sample with proper size, and placing the cut flower sample into a nuclear magnetic sample tube for sealing; the sample is not more than 1cm 2 Petals of a size or stem sections of no more than 2cm in length;
b. detecting a sample by using low-field nuclear magnetic resonance, obtaining echo attenuation curve data of a CPMG sequence, placing the corresponding sample at 70 ℃, and drying to constant weight; in the drying process, collecting the CPMG signal of the corresponding sample every 1 hour;
the low-field nuclear magnetic resonance detector is an NMI20 type low-field nuclear magnetic resonance detector, the diameter of the coil is 15mm, and the parameters are set as follows:
the 90-degree pulse width P1 is 15 mus, the control parameter RFD of the start sampling time is 0.06ms, the analog gain RG1 is 20.0db, the digital gain DRG1 is 3, the receiver bandwidth SW is 200KHz, the repeated sampling waiting time Tw is 2000ms, the 180-degree pulse width P2 is 29 mus, the echo time TE is 0.2ms, the echo number NECH is 10000, and the repeated sampling time NS is 8;
c. b, immediately weighing the corresponding sample after the CPMG signal is collected at each time point in the step b, and calculating according to the mass of the corresponding sample to obtain the water content measured value of the corresponding sample;
d. processing the T2 echo attenuation curve signal of the CPMG sequence obtained in the step b, extracting a T2 initial point signal value of total water NMR, fitting a T2 initial point signal value and a water content measured value corresponding to the water content measured value obtained in the step c, and establishing a gladiolus fresh cut flower water content prediction model y of 0.0014x-0.6118, wherein x is the initial point signal value of a transverse relaxation time T2 attenuation curve, and y is a water content predicted value;
e. acquiring T2 attenuation curve data of the fresh cut flower sample to be detected by adopting the same parameter setting as that in the step b, obtaining a first point signal value and T2 of different relaxation components, substituting the first point signal value into the gladiolus fresh cut flower water content prediction model to obtain a predicted water content value of the gladiolus fresh cut flower sample to be detected in a corresponding state, using T2 inversion spectrums of the different relaxation components for water state judgment, and sequentially and respectively analyzing NMR signals with T2 values of 1-10, 10-100 and 100-10000 ms into combined water T21, non-flowing water T22 and free water T23;
f. low-field nuclear magnetic imaging: placing the gladiolus fresh cut flower stem section with the length not more than 2cm and taken at a position 20-22 cm away from the cut into a nuclear magnetic sample tube, and performing imaging analysis by using a low-field nuclear magnetic resonance apparatus: adopting MRI imaging software and SE imaging sequence test, collecting sample proton density image, MRI imaging parameters: accumulating and sampling 8 times, wherein GZ is-850, GY is-240, GX is-100, Tw is 2000; and (4) judging the moisture distribution condition according to the color depth in the image and the inversion spectrum of the transverse relaxation time T2.
2. The method of claim 1, wherein in step b, 3-6 samples are taken as biological replicates at a time using low field nuclear magnetic resonance detection.
3. Use of the method of any one of claims 1-2 for detecting changes in moisture content and moisture migration of fresh cut flowers of gladiolus hybridus.
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| Nano-silver treatments reduced bacterial colonization and biofilm formation at the stem-ends of cut gladiolus ‘Eerde’ spikes;Hongbo Li et al.;《Postharvest Biology and Technology》;20160927;第123卷;102-111 * |
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