A pretreatment method and a raman spectrum detection method for detecting components of urinary calculi

Abstract

The application discloses a pretreatment method for detecting components of urinary calculi, which comprises the following steps: mixing urine containing calculi with a sodium hydroxide solution and performing ultrasonic oscillation; centrifuging, removing the upper liquid and reserving the precipitate; mixing the precipitate with water and a hydrogen peroxide solution, filtering through a filter membrane, and collecting the residual urine on the surface of the filter membrane, so as to obtain the urine to be detected. A Raman spectrum detection method based on the pretreatment is also disclosed, wherein the urine to be detected obtained through the pretreatment is dropped onto a silicon wafer treated with aqua regia, and then Raman spectrum detection is performed, so that efficient detection of the components of urinary calculi in the urine can be realized.

Description

Technical Field

[0001] This invention belongs to the field of urine detection, and particularly relates to a pretreatment method and a Raman spectroscopy detection method for detecting the components of urine stones. Background Technology

[0002] Urinary tract stones are a major disease worldwide, causing severe pain, urinary tract obstruction, and infection, and potentially leading to complete kidney damage. The recurrence rate of kidney stones within 5 years is greater than 50%, and prevention is only possible through appropriate medication. Stone recurrence not only brings physical pain and financial burden but also increases psychological stress for patients. Urinary stones are complex and diverse particulate components, and their formation is influenced by factors such as urine pH, temperature, the components that form the stones, and their concentration and solubility in urine. The presence of stone particles in urine is the basis for the formation of urinary tract stones. Analysis of the composition of urinary stone particles can provide a scientific basis and correct guidance for stone prevention, significantly reducing the recurrence rate of various types of stones.

[0003] Currently, the main method for detecting urinary stones is microscopic examination of urine sediment. This involves taking a well-mixed urine sample, centrifuging it in a centrifuge tube, discarding the supernatant, and leaving a precipitate. The precipitate is then prepared into a slide and observed first under a low-power microscope, followed by observation under a high-power microscope. This entire process is cumbersome, time-consuming, and costly. Furthermore, due to the irregular shape of the crystals and the presence of excessive substances or contaminants in the precipitate that interfere with the field of view, it is often impossible to accurately determine the crystal type.

[0004] Raman spectroscopy can rapidly and effectively identify and classify urinary stones, offering advantages such as independence from sample conditions and real-time monitoring. Applying Raman spectroscopy to the identification and classification of urinary stones allows for molecular-level measurement of samples, providing effective data for identification and classification without damaging the sample. However, urine containing stones often contains foreign matter such as red blood cells, white blood cells, epithelial cells, and bacteria, which severely affect the detection of the target analytes. Therefore, pretreatment methods for urinary stone detection are crucial for detecting the Raman spectral signal of the stones. Summary of the Invention

[0005] To address the aforementioned technical issues, this invention provides a pretreatment method and a Raman spectroscopy detection method for detecting the components of urine stones. The pretreatment effectively removes interfering substances from urine samples, thereby improving the effectiveness of Raman spectroscopy detection.

[0006] The specific solution of this invention is as follows:

[0007] One objective of this invention is to provide a pretreatment method for detecting the components of urine stones, comprising: mixing urine containing stones with sodium hydroxide solution and ultrasonically vibrating; centrifuging to remove the supernatant and retain the precipitate; mixing the precipitate with water and hydrogen peroxide solution, filtering with a filter membrane, and collecting the urine remaining on the surface of the filter membrane to obtain the urine to be tested.

[0008] Preferably, the volume ratio of sodium hydroxide solution to urine containing stones is 0.3-0.5:8-12.

[0009] Preferably, the centrifugation speed is 2000-3000 r / min and the centrifugation time is 5-8 min.

[0010] Preferably, the hydrogen peroxide solution concentration is 20-30%.

[0011] Preferably, the filter membrane pore size is 1-5 μm.

[0012] This invention pretreatment of urine containing stones reduces interference from foreign matter in Raman detection, facilitating in-situ Raman spectroscopy detection of urinary stones. Specifically, sodium hydroxide is first added to alter the pH of the urine, causing protein precipitation, and then ultrasonic vibration separates the protein from the urinary stones. Next, centrifugation removes most interfering substances from the urine, followed by further washing with water to reduce the concentration of uric acid and other substances. Hydrogen peroxide is then added to remove organic impurities. This treatment effectively removes interference from other components in the urine for Raman spectroscopy signal detection, resulting in better spectral detection performance when Raman spectroscopy is applied.

[0013] The second objective of this invention is to provide a Raman spectroscopy detection method, in which the urine sample obtained from the above pretreatment is dropped onto a silicon wafer treated with aqua regia for Raman spectroscopy detection.

[0014] Preferably, the excitation wavelength is 633nm, the laser power is 2.5-6mW, and the acquisition time is 5-10s.

[0015] Preferably, the excitation wavelength is 633nm, the laser power is 5mW, and the acquisition time is 5s.

[0016] The beneficial effects of this invention are:

[0017] This invention provides a pretreatment method for detecting the composition of urinary stones. By treating urine containing urinary stones, interfering impurities can be efficiently removed. The pretreated urine is then subjected to Raman spectroscopy for detection. The detection sensitivity is high, and the composition of urinary stone particles with a size of 5 μm and above can be accurately measured. This method is suitable for rapid on-site detection of urinary stone particles. Attached Figure Description

[0018] Figure 1 The Raman spectrum obtained in Example 1;

[0019] Figure 2 The Raman spectrum obtained in Example 2;

[0020] Figure 3 The Raman spectrum obtained in Comparative Example 1; Detailed Implementation

[0021] The technical solution of the present invention will be described in detail below through specific embodiments. However, it should be clearly stated that these embodiments are for illustrative purposes only and are not intended to limit the scope of the present invention.

[0022] Example 1

[0023] A pretreatment method for detecting the components of urinary stones includes:

[0024] (1) Use a pipette to draw 10 mL of urine containing calcium oxalate stones and put it into a 15 mL centrifuge tube; then draw 0.3 mL of 0.1 M sodium hydroxide solution and add it to the 15 mL centrifuge tube containing the urine containing the stones, and mix thoroughly with an ultrasonic vibrator for 1 min.

[0025] (2) Use a pipette to draw 1 mL of the mixture into a 1.5 mL centrifuge tube. Repeat this process for 10 tubes. Centrifuge at 2500 r / min for 5 min. After centrifugation, remove the supernatant and leave 20 μL of the precipitate in the tube.

[0026] (3) Collect the precipitate from 10 centrifuge tubes and put it into a 15mL centrifuge tube. Add deionized water to 10mL, then add 1mL of 30% hydrogen peroxide solution and mix well. Use a syringe to draw up the treated urine and filter it. After filtration, collect the urine residue on the surface of the filter membrane in the filter, which is the urine to be tested. The syringe head is equipped with a detachable filter with a diameter of 13mm, and a filter membrane with a pore size of 5μm is placed in the filter.

[0027] A Raman spectroscopy detection method involves taking 5 μL of the urine sample obtained through the pretreatment described in this embodiment and adding it onto a silicon wafer treated with aqua regia for Raman spectroscopy detection. The excitation wavelength of the Raman spectrometer is 633 nm, the laser power is 5 mW, and the acquisition time is 5 s.

[0028] The Raman spectrum obtained in this embodiment is as follows: Figure 1 As shown, it can be seen that at 911cm -1 and 1477cm -1 At this location, a peak of calcium oxalate dihydrate appeared, indicating that the urinary stone was composed of calcium oxalate.

[0029] Example 2

[0030] A pretreatment method for detecting the components of urinary stones is the same as in Example 1.

[0031] A Raman spectroscopy detection method involves taking 5 μL of the urine sample obtained through the pretreatment described in this embodiment and dropping it onto a silicon wafer treated with aqua regia for Raman spectroscopy detection. The excitation wavelength of the Raman spectrometer is 633 nm, the laser power is 2.5 mW, and the acquisition time is 5 s.

[0032] The Raman spectrum obtained in this embodiment is as follows: Figure 2 As shown, it can be seen that at 911cm -1 and 1477cm -1 At this location, a peak of calcium oxalate dihydrate appeared, but the peak intensity was weak.

[0033] Comparative Example 1

[0034] A pretreatment method for detecting the composition of urinary stones includes: using a pipette to draw 1 mL of urine from a patient containing calcium oxalate stones into a 1.5 mL centrifuge tube, centrifuging at 2500 r / min for 5 min, extracting the supernatant after centrifugation, and retaining 20 μL of the precipitate in the tube to obtain the urine to be tested.

[0035] A Raman spectroscopy detection method involves taking 5 μL of the urine sample obtained from the pretreatment of the above comparative example and adding it to a silicon wafer treated with aqua regia for Raman spectroscopy detection. The excitation wavelength of the Raman spectrometer is 633 nm, the laser power is 5 mW, and the acquisition time is 5 s.

[0036] The Raman spectrum obtained in this comparative example is as follows: Figure 3 As shown, the detection effect is poor and the signal-to-noise ratio is low.

[0037] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A pretreatment method for detecting the components of urinary stones, characterized in that, include: The urine containing the stones was mixed with a sodium hydroxide solution and then subjected to ultrasonic vibration. Centrifuge to remove the supernatant and retain the precipitate; mix the precipitate with water and hydrogen peroxide solution, filter through a filter membrane, and collect the urine residue on the surface of the filter membrane to obtain the urine sample to be tested.

2. The pretreatment method for detecting the components of urinary stones according to claim 1, characterized in that, The volume ratio of sodium hydroxide solution to urine containing stones is 0.3-0.5: 8-12.

3. The pretreatment method for detecting the components of urinary stones according to claim 1 or 2, characterized in that, The centrifugation speed is 2000-3000 r / min, and the centrifugation time is 5-8 min.

4. The pretreatment method for detecting the components of urinary stones according to claim 1 or 2, characterized in that, The concentration of hydrogen peroxide solution is 20-30%.

5. The pretreatment method for detecting the components of urinary stones according to claim 1 or 2, characterized in that, The filter membrane has a pore size of 1-5 μm.

6. A Raman spectroscopy detection method, characterized in that, The urine sample obtained by any one of the pretreatment methods of claims 1-5 is dropped onto a silicon wafer treated with aqua regia and then subjected to Raman spectroscopy detection.

7. The Raman spectroscopy detection method according to claim 6, characterized in that, The excitation wavelength is 633nm, the laser power is 2.5-6 mW, and the acquisition time is 5-10s.

8. The Raman spectroscopy detection method according to claim 6, characterized in that, The excitation wavelength was 633 nm, the laser power was 5 mW, and the acquisition time was 5 s.

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