CN114002206A - Method and device for rapidly detecting bioluminescence intensity of traditional Chinese medicine decoction - Google Patents
Method and device for rapidly detecting bioluminescence intensity of traditional Chinese medicine decoction Download PDFInfo
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- CN114002206A CN114002206A CN202110753721.XA CN202110753721A CN114002206A CN 114002206 A CN114002206 A CN 114002206A CN 202110753721 A CN202110753721 A CN 202110753721A CN 114002206 A CN114002206 A CN 114002206A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
- G01N21/763—Bioluminescence
Abstract
The invention relates to quality detection of a traditional Chinese medicine decoction, and belongs to the field of medicine analysis. A method for detecting the biological luminous intensity of the decoction of Chinese traditional medicine rapidly, said method places the decoction obtained by decocting Chinese traditional medicine into the darkroom and measures the fluorescence intensity of the decoction quantitatively, and under the condition of different concentration decoctions and different time decoction of standing, measure the fluorescence intensity repeatedly in order to set up the standard curve; and finally, putting the decoction to be detected in a dark room to quantitatively detect the fluorescence intensity of the decoction, and determining the concentration and the standing time of the decoction according to a standard curve. According to the technical scheme, the condition that the internal effective medicinal components of the Chinese medicinal decoction change gradually along with the time lapse can be accurately identified through UPE analysis, the content of the effective medicinal components of the Chinese medicinal decoction can be effectively, quickly and accurately obtained, and the quality condition of the Chinese medicinal decoction can be conveniently obtained.
Description
Technical Field
The invention relates to detection of traditional Chinese medicine quality, in particular to a method and a device for rapidly detecting bioluminescence intensity of a traditional Chinese medicine decoction.
Background
The traditional Chinese medicine decoction is used as the traditional preparation form of the traditional Chinese medicine, and plays an important role in guaranteeing the health and health care of human beings. In view of the inconvenience in decocting and carrying traditional Chinese medicine decoction, more and more hospitals or drug stores provide the traditional Chinese medicine decoction-replacing service, and after the decoction is decocted once, the decoction is stored in a packaging bag in a sealed mode in a single dose, so that the decoction is convenient to carry and store. The cold storage of the refrigerator at ordinary times, warm water heating when taking, it is convenient to take and use, and is deeply welcomed by patients. However, the content of the total extract and the change of the effective components during the storage process due to the complex compatibility of the traditional Chinese medicine components are not reported, and the effect of treating diseases is similar to that of the traditional Chinese medicine which is decocted at present.
The popular method for detecting the content of the active ingredients of the traditional Chinese medicine is to measure the content of the active ingredients according to the method for measuring the active ingredients in Chinese pharmacopoeia. The high performance liquid chromatograph is adopted, and the operation process is generally as follows: the chromatographic condition is groped → the solution of the reference substance is prepared → the solution of the test substance is prepared → the precision and the repeatability are tested → the content of a certain active ingredient. The content of the effective components can be detected by utilizing the conventional universal methods such as high performance liquid chromatography and the like, but the method has the defects of complicated operation process, time consumption, high detection cost, high technical condition requirement and the like, is inconvenient for real-time rapid detection, and is not beneficial to popularization.
Therefore, the method for detecting the quality of the traditional Chinese medicine has high universality, has a universal detection effect on the quality of the traditional Chinese medicine, is quick, efficient and low in cost, and is one of important means for ensuring the quality of the traditional Chinese medicine.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method and a device for rapidly detecting the bioluminescence intensity of a traditional Chinese medicine decoction, and solving the problems of complex process, high cost and inconvenience in comprehensive popularization of the traditional Chinese medicine decoction processing quality detection.
Technical scheme
A method for rapidly detecting bioluminescence intensity of a traditional Chinese medicine decoction is characterized by comprising the following steps:
a. decocting the Chinese medicinal materials to obtain decoction;
b. quantitatively detecting the fluorescence intensity of the decoction in a dark room;
c. repeating the steps a and b under the conditions of decoction liquids with different concentrations and decoction liquids with different standing times, and establishing a standard curve;
d. and (3) putting the decoction to be detected in a dark room, quantitatively detecting the fluorescence intensity of the decoction, and determining the concentration and the standing time of the decoction according to a standard curve.
Further, before the decoction is placed in a dark room for detection in the step b, dark cooling is carried out in the dark room for 20-50 min in advance, so as to eliminate the influence of natural light on the electronic state activation of organic luminescent groups in the decoction;
further, in the process of establishing the standard curve in the step c, after the fluorescence intensity of the standard solution is measured, HPLC quantitative analysis is carried out for digital quantification of the standard curve.
The device for rapidly detecting the bioluminescence intensity of the traditional Chinese medicine decoction is characterized by comprising a darkroom, wherein a sample table is arranged in the darkroom, photomultiplier tubes are arranged around the sample table, and the photomultiplier tubes are electrically connected with a photon counting unit and a PLC (programmable logic controller).
Furthermore, the inner wall and the periphery of the darkroom are subjected to blackening treatment, and the spontaneous light emitting effect of the material of the inner wall of the darkroom is subjected to substrate correction in the PLC.
Further, the photon counting unit comprises a photon counter and a photon counter card, wherein the photon counter is used for discriminating and counting the received electric signals; the photon counter card is used for collecting, analyzing and processing signals.
Furthermore, a shutter is arranged in front of the photomultiplier probe and is electrically connected with the PLC.
Further, the PLC controller realizes the opening and closing of the photomultiplier shutter through a computer program, records the sample luminescence information obtained by the photomultiplier, feeds back the sample luminescence information to a computer loaded with data processing software, and outputs a result.
Further, the photomultiplier is disposed directly above the sample.
Advantageous effects
By adopting the method and the device for rapidly detecting the bioluminescence intensity of the traditional Chinese medicine decoction, the UPE and the effective medicinal components are obviously related through the correlation analysis of the UPE change trend of different traditional Chinese medicine decoction liquids and the content change trend of the effective chemical components, the condition that the effective medicinal components in the decoction liquid are gradually changed along with the lapse of time can be accurately identified through the UPE analysis, and the condition that the concentration of the effective medicinal components is gradually reduced along with the lapse of time can be accurately detected. The UPE detection can effectively, quickly and accurately obtain the content of the effective medicinal components of the traditional Chinese medicine decoction, and conveniently obtain the quality condition of the traditional Chinese medicine decoction.
Drawings
FIG. 1 is a schematic view of a bioluminescence intensity measurement system of a decoction of Chinese medicinal materials according to the present invention;
FIG. 2 is a flow chart of measurement according to the present invention;
FIG. 3 is a graph showing that the bioluminescence intensity of the decoction of the traditional Chinese medicine of the present invention increases linearly with the increase of the volume of the medicine added;
FIG. 4 is a UPE intensity variation curve of the decoction of Panax notoginseng with prolonged standing time;
FIG. 5 is a UPE intensity variation curve of pummelo peel with prolonged standing time according to the present invention;
fig. 6 is a graph showing the variation of the total amount of ginsenosides Rb, Rg and R of panax notoginseng according to the invention along with the prolonging of the standing time.
FIG. 7 is a graph showing the change of naringin content of pummelo peel with the time of storage.
Wherein: 1-darkroom, 2-sample stage, 3-photomultiplier, 4-high voltage power supply, 5-photon counting unit, 6-PLC controller, 7-computer.
Detailed Description
The invention will be further elucidated with reference to the specific embodiments and the accompanying drawings.
This technical scheme firstly provides a device of short-term test traditional chinese medicine decoction bioluminescence intensity, including darkroom 1, darkroom inside sets up sample platform 2, sets up photomultiplier 3 around the sample platform, and photon counting unit 5 and PLC controller 6 are connected to the photomultiplier electricity, and PLC controller electricity connects computer 7. In the device, the inner wall and the periphery of the darkroom are subjected to blackening treatment, and the spontaneous light emitting effect of the material of the inner wall of the darkroom is subjected to substrate correction in the PLC. The photon counting unit comprises a photon counter and a photon counter card, wherein the photon counter is used for discriminating and counting received electric signals, and the photon counter card is used for collecting, analyzing and processing the signals.
The shutter is arranged in front of the photomultiplier probe in the device and is electrically connected with the PLC controller. The PLC controller realizes the opening and closing of the photomultiplier shutter through a computer program, records the sample luminescence information obtained by the photomultiplier, feeds back the sample luminescence information to a computer loaded with data processing software, and outputs a result.
In this embodiment, the configuration of the apparatus is as follows:
a photomultiplier tube: ET Enterprises,9235QA (sensitivity to low light reaches 10-17W/cm)2Background noise of 6-7 photons/second at 20 DEG C
A high-voltage power supply: Sens-Tech, PM20 SP;
photon counter: HAMAMATSU, C9744;
a counter card: national Instruments, NI PCIe-6320;
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
Firstly, decocting traditional Chinese medicines:
weighing 40g of traditional Chinese medicines, placing the traditional Chinese medicines in a marmite, adding 800mL of tap water for soaking for 30min, heating the traditional Chinese medicines by using an electric ceramic stove, adjusting the firepower to 2000, boiling the traditional Chinese medicines, adjusting the firepower to 800, decocting the traditional Chinese medicines for 20min, filtering liquid medicines by using 6 layers of gauze, repeating the decoction process on dregs of a decoction, combining and concentrating the liquid medicines decocted for 2 times, cooling the liquid medicines to room temperature, and finally obtaining 200mL of traditional Chinese medicine decoction of 0.2 g/mL.
And (3) equally dividing 20 parts of the decoction, putting into a sterile centrifuge tube, and cooling to room temperature in a dark place for later use.
Secondly, setting an instrument:
(1) the laboratory air conditioner is started 2 hours ahead of time, the temperature is adjusted to 20 ℃, and the environmental humidity is controlled to be between 40 and 50 percent.
(2) Setting of test parameters
Running the test software PC-device-DH, starting the self-test of the test system, and operating when the self-test of the system is finished
The interface completes the setting of corresponding parameters:
photomultiplier tube high pressure: 906V
Measurement mode: SE measurement
Measuring time interval: 1s
Measuring time: for 10min
Initial state of the filter plate: no filter
Third, measurement process
1) And after the power-on self-test process is finished and before the sample is tested, testing the background noise of the system, and recording as background.
2) All light sources in the room where the test system is located are turned off, 1 part of 5mL of the traditional Chinese medicine decoction is sucked and placed in a quartz plate, and the quartz plate is placed on a sample table in a dark room. The whole process needs to be operated in a dark environment, so that the traditional Chinese medicine decoction is prevented from being influenced by external light again, and the photon intensity of the traditional Chinese medicine decoction obtained by testing is ensured;
3) the height of the sample stage is adjusted to enable the upper edge of the sample stage to be close to the lower edge of the photomultiplier, so that the distance between the sample and the photomultiplier probe is shortened as far as possible, and photons radiated by the sample are collected to the greatest extent;
4) confirming that the measurement parameters are correct, clicking 'Measure' on the operation software, and starting to count;
5) and after 10min, finishing the measurement, clicking built-in data processing software, analyzing the data and exporting a result.
6) Marking the decoction of the Chinese medicinal materials decocted in the same day as No. 1, and performing the above detection on the 3 rd day, the 5 th day, the 7 th day, the 11 th day, the 14 th day, the 21 st day and the 30 th day after the decoction of the Chinese medicinal materials of the rest No. 2-8 days; on the day of measuring the bioluminescence intensity of the traditional Chinese medicine decoction, the content of the active ingredients is detected by HPLC, so that the correlation analysis of the bioluminescence intensity and the content of the active ingredients of the traditional Chinese medicine decoction is carried out.
Fourth, data analysis
Pearson correlation analysis.
Fifth, test results
FIG. 3 shows that the bioluminescence intensity of the decoction increases linearly with the increase of the volume of the drug, indicating that the bioluminescence intensity is linearly related to the volume of the drug or the concentration of the drug in the sample cell.
FIG. 4 shows UPE strength change of Notoginseng radix decoction with prolonged standing time; FIG. 5 shows the UPE intensity change of exocarpium Citri Grandis with prolonged standing time, which indicates that with the lapse of time, the content of chemiluminescent group in the decoction of Chinese medicinal materials is reduced, the amount and type of organic molecules in the Chinese medicinal materials are changed, and the change of molecular structure is consistent with the change of drug effect.
The embodiment also provides the total amount of ginsenoside Rb, Rg and R in the attached figure 6, and the naringin content in the attached figure 7, wherein the content of effective components of ginsenoside and naringin in the decoction changes along with the prolonged standing time, and the change trend of the content of the effective components of ginsenoside and naringin in the decoction is consistent with the change trend of UPE in the attached figures 4 and 5, which further shows that the quantity and the types of organic molecules in the traditional Chinese medicine are changed along with the lapse of time, and the drug effect is damaged.
Pearson correlation analysis of UPE Strength and substance content
The UPE strength of the panax notoginseng water decoction liquid with different standing days and the corresponding total amount of ginsenoside Rb, Rg and R, the UPE strength of the pummelo peel water decoction liquid with different standing days and the corresponding naringin content are respectively subjected to Pearson correlation analysis, the result shows that the UPE strength change of the traditional Chinese medicine water decoction liquid and the content of the effective components are in positive correlation along with the prolonging of the standing time, and the result is shown in Table 1.
TABLE 1 Pearson correlation analysis results for each index
Note: indicates significant correlation at the 0.05 level and indicates significant correlation at the 0.001 level.
Therefore, in this embodiment, it can be seen from the correlation analysis of the variation trend of the UPE of different traditional Chinese medicine decoction solutions and the content variation trend of the active chemical ingredients, that the UPE is significantly related to the active pharmaceutical ingredients, and the UPE analysis can accurately identify that the internal active pharmaceutical ingredients of the traditional Chinese medicine decoction solutions gradually change and the concentrations of the active pharmaceutical ingredients gradually decrease with the passage of time, and the change of the active pharmaceutical ingredients has significant feasibility through the UPE detection.
Claims (9)
1. A method for rapidly detecting bioluminescence intensity of a traditional Chinese medicine decoction is characterized by comprising the following steps:
a. decocting the Chinese medicinal materials to obtain decoction;
b. quantitatively detecting the fluorescence intensity of the decoction in a dark room;
c. repeating the steps a and b under the conditions of decoction liquids with different concentrations and decoction liquids with different standing times, and establishing a standard curve;
d. and (3) putting the decoction to be detected in a dark room, quantitatively detecting the fluorescence intensity of the decoction, and determining the concentration and the standing time of the decoction according to a standard curve.
2. The method for rapidly detecting bioluminescence intensity of a Chinese medicinal decoction according to claim 1, wherein in the step c of establishing the standard curve, after the standard solution is subjected to fluorescence intensity measurement, HPLC quantitative analysis is performed for digital quantification of the standard curve.
3. The method for rapidly detecting the bioluminescence intensity of a traditional Chinese medicine decoction according to claim 1, wherein the decoction is subjected to dark cooling in a dark room for 20-50 min before being placed in the dark room for detection in the step b.
4. The device for rapidly detecting the bioluminescence intensity of the traditional Chinese medicine decoction is characterized by comprising a darkroom, wherein a sample table is arranged in the darkroom, photomultiplier tubes are arranged around the sample table, and the photomultiplier tubes are electrically connected with a photon counting unit and a PLC (programmable logic controller).
5. The device of claim 4, wherein the inner wall and the outer periphery of the darkroom are blackened, and the spontaneous emission effect of the material of the inner wall of the darkroom is corrected by the PLC.
6. The device for rapidly detecting the bioluminescence intensity of a traditional Chinese medicine decoction according to claim 4, wherein the photon counting unit comprises a photon counter and a photon counter card, wherein the photon counter is used for screening and counting received electric signals; the photon counter card is used for collecting, analyzing and processing signals.
7. The device of claim 4, wherein the shutter is disposed in front of the photomultiplier probe and electrically connected to the PLC controller.
8. The device for rapidly detecting the bioluminescence intensity of a traditional Chinese medicine decoction according to claim 4, wherein the PLC controller realizes the opening and closing of a photomultiplier shutter through a computer program, records the sample luminescence information obtained by the photomultiplier, feeds back the sample luminescence information to a computer loaded with data processing software, and outputs the result.
9. The device of claim 4, wherein the photomultiplier is disposed directly above the sample.
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Citations (5)
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| CN109187464A (en) * | 2018-09-10 | 2019-01-11 | 中国农业大学 | A method of quick, Accurate Determining Chinese medicine Se content |
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- 2021-07-02 CN CN202110753721.XA patent/CN114002206A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102004089A (en) * | 2010-11-19 | 2011-04-06 | 山东省医药生物技术研究中心 | Device and method for detecting property of traditional Chinese medicine |
| CN103645164A (en) * | 2013-09-22 | 2014-03-19 | 吉林化工学院 | Fluorescent measuring method for inorganic arsenic in traditional Chinese medicinal material under-forest ginseng |
| CN203688443U (en) * | 2013-12-23 | 2014-07-02 | 西安理工大学 | Biological ultraweak photon radiation spectroscopic detector |
| CN106525821A (en) * | 2016-10-31 | 2017-03-22 | 山东省医药生物技术研究中心 | Method and device for detecting medicinal property of traditional Chinese medicine |
| CN109187464A (en) * | 2018-09-10 | 2019-01-11 | 中国农业大学 | A method of quick, Accurate Determining Chinese medicine Se content |
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