Method for rapidly detecting protein content in tobacco leaves

A protein content and protein precipitation technology, applied in the direction of color/spectral characteristic measurement, etc., can solve the problems of large amount of reagents, time-consuming and complicated operation, and achieve the effect of shortening digestion time, less overall time-consuming and accurate results

Pending Publication Date: 2022-04-29
GANSU TOBACCO IND
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  • Application Information

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Problems solved by technology

The above methods all have their own advantages, but to a certain extent, there are problems such as cumbersome operation, long detection cycle, and unfriendly to the environment.
[0003] However, the existing technology has problems such as long detection period, unfriendly environment, and high requirements for detection ins

Method used

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  • Method for rapidly detecting protein content in tobacco leaves
  • Method for rapidly detecting protein content in tobacco leaves
  • Method for rapidly detecting protein content in tobacco leaves

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Step 1: Weigh 0.5-1g of tobacco leaf powder to be tested into a homogenizer, add phosphate buffer solution (containing SDS, pH 6.8) with a concentration of 0.05mol / L, fully grind to obtain a homogenate, and the homogenizer is ultrasonically oscillated Centrifuge after 5-10min, the speed of centrifugation is 4800r / min, and the centrifugation time is 5min. Take the supernatant and filter it with qualitative filter paper and transfer it to a 50-100mL volumetric flask;

[0038] Step 2: add the residue obtained by centrifugation in step 1 to phosphate buffer, and use the same method as step 1 to process, repeat three times, combine the filtrate, use phosphate buffer to make up volume, mix well, and obtain tobacco leaf sample liquid;

[0039] Step 3: Take 1-2 mL of the tobacco leaf sample solution described in Step 2, add 1% volume of 2% sodium deoxycholate, shake for 1 min and mix well, then add 20-40 mL of 15% trichloroacetic acid, 0.07% β-mercaptoethanol acetone solution ...

Embodiment 2

[0046] Take by weighing tobacco leaf sample 0.5g, obtain test solution by the method of embodiment 1, get and measure test solution 2mL, according to standard operation

[0047] a. Precision experiment

[0048] The protein contents of Qingyang 2013 X3L and Longnan 2011 B2L tobacco leaves were determined 7 times by the determination method of the present invention. The results showed that the coefficients of variation of the two samples were 1.16% and 1.41%, respectively, indicating that the method had high precision and good repeatability.

[0049] Table 1 Precision measurement results

[0050] sample protein content% average value% RSD% Qingyang 2013 X 3 L

9.64 9.67 9.85 9.72 9.74 9.90 9.71 9.74 1.16 Longnan 2011 B 2 L

9.28 9.47 9.17 9.31 9.11 9.22 9.10 9.24 1.41

[0051] b. Recovery rate experiment:

[0052] Tobacco leaf sample 0.5g (determined protein content is 9.6%), add different amount of casein standard sample, the additio...

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Abstract

The invention discloses a method for rapidly detecting the content of protein in tobacco leaves, which comprises the following steps: taking a certain amount of tobacco leaf powder to be detected in a homogenizer, adding 0.05 mol/L phosphate buffer solution containing SDS (sodium dodecyl sulfate) with the pH value of 6.8, fully grinding to obtain homogenate, carrying out ultrasonic oscillation on the homogenizer, then carrying out centrifugal separation, then carrying out centrifugal separation at the rotating speed of 4800r/min, taking the supernatant, filtering, and carrying out freeze drying to obtain the protein content in the tobacco leaves. According to the method, cell tissues are fully damaged in the homogenizing process, exudation of protein is facilitated, protein can be efficiently separated out from the screened protein precipitation solution, the influence of non-protein nitrogen is avoided, and the result is more accurate; compared with a Kendall method and a continuous flow analysis method, the method has the advantages that the steps are simplified, the digestion time is obviously shortened, and the total consumed time is less; the ultraviolet and visible spectrophotometer can realize simultaneous detection of a plurality of samples and has the characteristics of high efficiency and large flux, and other devices such as a centrifugal machine and a water bath kettle are standing devices in a laboratory.

Description

technical field [0001] The invention relates to the fields of tobacco leaf processing, alcoholization, cigarette production and the like, in particular to a method for rapidly detecting protein content in tobacco leaves. Background technique [0002] Tobacco is an important economic crop in my country and plays a very important role in the process of national economic and social development. Tobacco leaves contain many chemical substances, and the content of protein is the most abundant. After the protein is burned, with the generation of aza compounds, the smoke shows a bitter, pungent and irritating taste. Excessive protein has an adverse effect on the smoking taste of cigarettes, but if the protein content is too low, the smoke will appear flat, eating Taste and aroma also deteriorate. At the same time, protein can be decomposed into amino acids, and the content of amino acids has an obvious positive correlation with the aroma and fullness of smoke. Therefore, in the p...

Claims

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Application Information

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IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 丁润波花蕾马斌吕长平郭磊杜青霞
Owner GANSU TOBACCO IND
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