Lipid index quality control product as well as preparation method and application thereof

Abstract

The invention provides a lipid index quality control product as well as a preparation method and application thereof. The lipid index quality control product comprises serum, a fat emulsion injection and a preservative; the serum, the fat emulsion injection and the preservative are mixed according to different preset proportions to obtain quality control products with different lipid indexes; the preset proportion is that the volume ratio of the serum to the fat emulsion injection to the preservative is 100: (2-10): (0.05-0.1); the preservative is 0.05% of sodium azide. By applying the provided lipid index quality control product, the accuracy and stability of a detection system on a lipid blood sample can be evaluated, and the reliability of a detection result in the presence of lipid interference is ensured.

Description

Technical Field

[0001] This invention relates to the field of medical testing and measurement technology, specifically to a lipid index quality control product, its preparation method, and its application. Background Technology

[0002] Serum index quality control products are used to monitor the response and detection capabilities of IVD instruments to hemolyzed, jaundiced, and lipemic samples. These quality control products can assist laboratories in improving pre-analytical errors and interferences affecting biochemical and immunological assays. Some existing quality control products require the simultaneous detection of all three indicators—hemolysis index, lipemia index, and jaundice index—which is inconvenient for laboratories and users, and also wastes economic resources and testing costs. Sometimes, the monitoring of specimen appearance (serum index) may be a single or unilateral requirement for laboratories, without the need for combined testing.

[0003] Further research is needed to address the issue of independently detecting lipid indexes and improving the accuracy of the detection.

[0004] Therefore, this product specifically addresses the accuracy of lipid index detection, and specifically monitors the appearance of lipid index in serum, designing and inventing a quality control product. It achieves the most accurate and effective satisfaction of both economic benefits and specific customer needs. Summary of the Invention

[0005] This invention aims to at least partially address one of the technical problems in related technologies. Existing serum index quality control products are mixed serum index quality control products, including hemolysis index, lipemia index, and jaundice index. These three substances require different wavelengths of light in nanometers during IVD instrument detection. Research has shown that detecting the color of a single serum index is more accurate. When detecting the hemolysis index, the red index is mainly detected (because hemolyzed samples have a red appearance). When detecting the lipemia index quality control product, the white index is mainly detected (because lipemia samples have a white appearance). When detecting the jaundice index, the yellow index is mainly detected (jaundice index has a yellow appearance). However, the mixture of yellow, white, and red colors inherently influences and interferes with each other, leading to inaccurate detection of the color index of each substance. Based on this, we have developed a lipemia index quality control product for the detection of lipemia index.

[0006] Specifically, the present invention provides the following technical solution: A first aspect of the present invention provides a lipid index quality control product, comprising: serum, fat emulsion injection, and preservative; The serum, fat emulsion injection and preservative are mixed in different predetermined proportions to obtain quality control products with different lipid indexes; The predetermined ratio is a volume ratio of the serum, fat emulsion injection, and preservative of 100:(2 to 10):(0.05 to 0.1). The preservative is 0.05% sodium azide.

[0007] According to embodiments of the present invention, the lipid index quality control product provided above may further include the following technical features: According to an embodiment of the present invention, the lipid indexes of the quality control products with different lipid indices are 0≤LI<125, 125≤LI<250, 250≤LI<500, 500≤LI<1000, and 1000≤LI.

[0008] According to an embodiment of the present invention, the fat emulsion injection comprises 40 to 50 g / L soybean oil, 4 to 5 g / L lecithin and 5 to 6.6 g / L glycerol.

[0009] According to an embodiment of the present invention, the serum is derived from the serum of at least two healthy subjects.

[0010] A second aspect of the present invention provides a method for preparing the lipid index quality control product described in the first aspect, comprising: (1) Provide fat emulsion injection excipients, mix the fat emulsion injection excipients and a portion of preservatives, and perform high-pressure homogenization to obtain fat emulsion injection; (2) Obtain serum from healthy subjects, and mix the serum from healthy subjects with the remaining preservative to obtain mixed serum; (3) The fat emulsion injection and the mixed serum are mixed and sterilized to obtain the lipid index quality control product.

[0011] According to an embodiment of the present invention, step (1) further includes: (1-1) Mix soybean oil and lecithin and heat to 60-70℃, stirring until completely dissolved to obtain the oil phase; (1-2) Dissolve glycerol in water for injection, heat to the same temperature, add sodium azide to dissolve, so as to obtain an aqueous phase; (1-3) The oil phase is slowly added to the aqueous phase and subjected to high-speed shearing at 6000-8000 rpm for 10 to 15 minutes to obtain the primary emulsion; (1-4) The colostrum is subjected to high-pressure homogenization at a pressure of 800 bar to 100 bar in order to obtain the fat emulsion injection; (1-5) After homogenization, nitrogen is used for filling to reduce oxidation.

[0012] According to an embodiment of the present invention, after high-pressure homogenization in steps (1-4), the particle size of the fat emulsion injection is ≤300 nm and the PDI is <0.2.

[0013] A third aspect of the present invention provides a kit comprising the lipid index quality control product described in the first aspect, or the lipid index quality control product obtained according to the preparation method described in the second aspect.

[0014] A fourth aspect of the present invention provides a method for determining the lipid index of an isolated test sample, comprising: The lipid index of the test sample and the lipid index quality control sample were detected using an automated biochemical analyzer. The automated biochemical analyzer is calibrated and validated based on the lipid index test results of the lipid index quality control material in order to obtain calibration coefficients; The lipid index of the sample to be tested is determined based on the calibration coefficient and the lipid index of the sample to be tested.

[0015] A fifth aspect of the present invention provides a method for detecting the response capability of an instrument to a lipemia sample, comprising: The instrument is used to test the lipid index quality control sample to obtain a measured value; the hemolysis index quality control sample is the lipid index quality control sample described in any one of the first aspects or the lipid index quality control sample obtained according to the preparation method described in the second aspect. The measured values ​​are compared with the theoretical values ​​of the lipemia index of the lipemia index quality control sample to determine the instrument's responsiveness to lipemia samples.

[0016] The beneficial effects achieved by this invention are at least as follows: (1) The lipid index quality control product provided by the present invention can be used to evaluate the accuracy and stability of the detection system for lipid samples, and ensure the reliability of the detection results when lipid interference is present; (2) This application can obtain high concentrations of lipid interference quality control material to verify the instrument’s anti-interference ability and detect instrument results with higher lipid index. Attached Figure Description

[0017] Figure 1 The results are colorimetric cards of CLSI quality control samples with different lipid indices, provided according to embodiments of the present invention. Detailed Implementation

[0018] The embodiments of the present invention are described in detail below. Examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain the present invention, but should not be construed as limiting the present invention.

[0019] In recent years, many patients treated clinically suffer from diabetes, hyperlipidemia, and high lipoprotein intake, resulting in milky-white serum. Clinical laboratory reports often only note "specimen is severely turbid, results are for reference only" or require patients to "re-test after three days of vegetarianism." This inability to provide accurate test results for rapid clinical diagnosis hinders optimal treatment. Ensuring timely and reliable test data is a constant and significant challenge for laboratory staff. Therefore, understanding serum lipid levels is crucial, requiring a quality control product to measure its accuracy. When only the lipid level result is needed, a lipid level quality control product can be used alone to ensure the accuracy of the lipid level test.

[0020] This invention provides a lipid index quality control product, comprising: serum, fat emulsion injection, and preservative; The serum, fat emulsion injection and preservative are mixed in different predetermined proportions to obtain quality control products with different lipid indexes; The predetermined ratio is a volume ratio of serum, fat emulsion injection, and preservative of 100:(2 to 10):(0.05 to 0.1). The preservative is 0.05% sodium azide.

[0021] According to specific embodiments, the lipid indices of the quality control products with different lipid indices are 0≤LI<125, 125≤LI<250, 250≤LI<500, 500≤LI<1000, and 1000≤LI. According to preferred embodiments, the lipid indices of the quality control products with different lipid indices are 50, 150, 300, 600, and 1200. According to preferred embodiments, the lipid indices of the quality control products with different lipid indices are 0, 125, 250, 500, and 1000. The corresponding lipid severity classifications are described in Table 1 below. Figure 1 The results of the CLSI visual lipid colorimetric chart are shown.

[0022] Table 1. Description of the degree of lipemia corresponding to different lipemia indices.

[0023] According to a specific embodiment, the mentioned fat emulsion injection comprises 40 to 50 g / L soybean oil, 4 to 5 g / L lecithin, and 5 to 6.6 g / L glycerol. According to a preferred embodiment, the fat emulsion injection comprises 45 g / L soybean oil, 4.5 g / L lecithin, and 5.5 g / L glycerol.

[0024] According to an embodiment of the present invention, the serum is serum from at least two healthy subjects, such as serum from three, four, five, six, seven, eight, nine, ten or more healthy subjects.

[0025] This invention also provides a method for preparing the above-mentioned lipid index quality control product, comprising: (1) Provide fat emulsion injection excipients, mix the fat emulsion injection excipients and a portion of preservatives, and perform high-pressure homogenization to obtain fat emulsion injection; (2) Obtain serum from healthy subjects, and mix the serum from healthy subjects with the remaining preservative to obtain mixed serum; (3) The fat emulsion injection and the mixed serum are mixed and sterilized to obtain the lipid index quality control product.

[0026] According to an embodiment of the present invention, step (1) further includes: (1-1) Mix soybean oil and lecithin and heat to 60-70℃, stirring until completely dissolved to obtain the oil phase; (1-2) Dissolve glycerol in water for injection, heat to the same temperature, add sodium azide to dissolve, so as to obtain an aqueous phase; (1-3) The oil phase is slowly added to the aqueous phase and subjected to high-speed shearing at 6000-8000 rpm for 10 to 15 minutes to obtain the primary emulsion; (1-4) The colostrum is subjected to high-pressure homogenization at a pressure of 800 bar to 100 bar in order to obtain the fat emulsion injection.

[0027] (1-5) After homogenization, nitrogen is used for filling to reduce oxidation.

[0028] According to an embodiment of the present invention, after high-pressure homogenization in steps (1-4), the particle size of the fat emulsion injection is ≤300 nm and the PDI is <0.2.

[0029] A third aspect of the present invention provides a kit comprising the lipid index quality control product described in the first aspect, or the lipid index quality control product obtained according to the preparation method described in the second aspect.

[0030] A fourth aspect of the present invention provides a method for determining the lipid index of an isolated test sample, comprising: The lipid index of the test sample and the lipid index quality control sample were detected using an automated biochemical analyzer. The automated biochemical analyzer is calibrated and validated based on the lipid index test results of the lipid index quality control material in order to obtain calibration coefficients; The lipid index of the sample to be tested is determined based on the calibration coefficient and the lipid index of the sample to be tested.

[0031] The methods provided can be used for non-disease diagnosis and treatment purposes, such as scientific research, assisting clinical judgment, etc.

[0032] A fifth aspect of the present invention provides a method for detecting the response capability of an instrument to a lipemia sample, comprising: The instrument is used to test the lipid index quality control sample to obtain a measured value; the hemolysis index quality control sample is the lipid index quality control sample described in any one of the first aspects or the lipid index quality control sample obtained according to the preparation method described in the second aspect. The measured values ​​are compared with the theoretical values ​​of the lipemia index of the lipemia index quality control sample to determine the instrument's responsiveness to lipemia samples.

[0033] Example 1 Example 1 prepared a lipid index quality control product using the following method: The remaining serum from healthy subjects after laboratory testing is human matrix serum (the same as the human samples tested in the laboratory; from the perspective of matrix effect, the quality control product of this invention is closer to the actual testing scenario and has no difference in matrix effect) (from the perspective of raw materials, the quality control product of our invention has no matrix effect because it is a homologous substance); then the obtained serum from healthy subjects is mixed with a portion of sodium azide for later use to obtain mixed serum.

[0034] Fat emulsion injections were prepared according to the following method: Oil phase preparation: Mix soybean oil and lecithin and heat to 60-70℃, stirring until completely dissolved.

[0035] Aqueous phase preparation: Dissolve glycerol in water for injection, heat to the same temperature, and add sodium azide to dissolve.

[0036] Colostrum formation: The oil phase is slowly added to the aqueous phase, and the mixture is subjected to high-speed shearing (6000-8000 rpm, 10 minutes).

[0037] High-pressure homogenization: using 800 bar pressure, cycling more than 6 times, controlling the particle size ≤300 nm, PDI <0.2.

[0038] Nitrogen purging protection: After homogenization, nitrogen purging is used for filling to reduce oxidation and obtain fat emulsion injections.

[0039] 30% fat emulsion injection and mixed serum were mixed and then mixed with human serum in proportion to prepare quality control products with different lipid indexes (0, 125, 250, 500, and 1000, respectively). After being dispensed, the products were sterilized at 121°C for 15 minutes.

[0040] Example 2 Example 2 investigated the stability of the lipid index quality control product prepared in Example 1, including: (1) Short-term stability A quality control product with uniform emulsion droplets and qualified sodium azide concentration was successfully prepared. The emulsion should not show obvious stratification within 24 hours after mixing, and the simulated error of the lipemia index should be <5%.

[0041] The physicochemical stability was verified by accelerated testing (40℃, 75% humidity, 30 days). The results showed that the prepared lipid index quality control sample was stable during storage.

[0042] (2) Long-term stability The prepared lipemia index quality control sample was aliquoted and stored at -20°C for 3 months. The lipemia index deviation was <10% and the particle size change was <10%. This can achieve 2-3 years of storage stability, which meets the requirements of ISO guidelines for quality control samples.

[0043] Example 3 Example 3 validated the clinical application of the lipid index quality control product prepared in Example 1. The total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were measured using a fully automated biochemical analyzer (such as the CS-1200 fully automated biochemical analyzer) via continuous monitoring kinetics. Eight ml of peripheral venous blood was collected from the patient using a disposable sterile venous blood collection needle and stored in two disposable sterile vacuum blood collection tubes.

[0044] Centrifuge the blood sample at 3000 r / min for 10 minutes to separate the serum and store it in a -70℃ freezer for later use.

[0045] After reconstituted serum, it was diluted with distilled water to an appropriate concentration before testing.

[0046] The samples are sent into a fully automated biochemical analyzer, where different tests are performed in multiple small chambers.

[0047] Table 2 Different test results

[0048] The test results above show that the quality control samples with different lipid indices exhibit good stability on the biochemical analyzer, and all results are within the quality control range, meeting the quality requirements for clinical testing.

[0049] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A lipid index quality control product, characterized in that, include: Serum, fat emulsion injections, and preservatives; The serum, fat emulsion injection and preservative are mixed in different predetermined proportions to obtain quality control products with different lipid indexes; The predetermined ratio is a volume ratio of the serum, fat emulsion injection, and preservative of 100:(2~10):(0.05~0.1). The preservative is 0.05% sodium azide.

2. The lipid index quality control product according to claim 1, characterized in that, The lipid indexes of the quality control products with different lipid indices are 0≤LI<125, 125≤LI<250, 250≤LI<500, 500≤LI<1000, and 1000≤LI.

3. The lipid index quality control product according to claim 1, characterized in that, The fat emulsion injection comprises 40 to 50 g / L soybean oil, 4 to 5 g / L lecithin, and 5 to 6.6 g / L glycerol.

4. The lipid index quality control product according to claim 1, characterized in that, The serum was derived from at least two healthy subjects.

5. The method for preparing the lipid index quality control product according to any one of claims 1 to 4, characterized in that, include: (1) Provide fat emulsion injection excipients, mix the fat emulsion injection excipients and a portion of preservatives, and perform high-pressure homogenization to obtain fat emulsion injection; (2) Obtain serum from healthy subjects, and mix the serum from healthy subjects with the remaining preservative to obtain mixed serum; (3) The fat emulsion injection and the mixed serum are mixed and sterilized to obtain the lipid index quality control product.

6. The preparation method according to claim 5, characterized in that, Step (1) further includes: (1-1) Mix soybean oil and lecithin and heat to 60-70℃, stirring until completely dissolved to obtain the oil phase; (1-2) Dissolve glycerol in water for injection, heat to the same temperature, add sodium azide to dissolve, so as to obtain an aqueous phase; (1-3) The oil phase is slowly added to the aqueous phase and subjected to high-speed shearing at 6000-8000 rpm for 10 to 15 minutes to obtain the primary emulsion; (1-4) The colostrum is subjected to high-pressure homogenization at a pressure of 800 bar to 100 bar in order to obtain the fat emulsion injection; (1-5) After homogenization, nitrogen is used for filling to reduce oxidation.

7. The preparation method according to claim 6, characterized in that, After high-pressure homogenization in steps (1-4), the particle size of the fat emulsion injection is ≤300 nm and the PDI is <0.

2.

8. A reagent kit, characterized in that, It includes the lipid index quality control product according to any one of claims 1 to 4, or the lipid index quality control product obtained by the preparation method according to any one of claims 5 to 7.

9. A method for determining the lipid index of an isolated test sample, characterized in that, include: The lipid index of the test sample and the lipid index quality control sample were detected using an automated biochemical analyzer. The automated biochemical analyzer is calibrated and validated based on the lipid index test results of the lipid index quality control material in order to obtain calibration coefficients; The lipid index of the sample to be tested is determined based on the calibration coefficient and the lipid index of the sample to be tested. The lipid index quality control product is the lipid index quality control product according to any one of claims 1 to 4 or the lipid index quality control product obtained by the preparation method according to any one of claims 5 to 7.

10. A method for detecting the response capability of an instrument to a lipemia sample, characterized in that, include: The instrument is used to test the lipid index quality control sample to obtain a measured value; the hemolysis index quality control sample is the lipid index quality control sample according to any one of claims 1 to 4 or the lipid index quality control sample obtained by the preparation method according to any one of claims 5 to 7. The measured values ​​are compared with the theoretical values ​​of the lipemia index of the lipemia index quality control sample to determine the instrument's responsiveness to lipemia samples.

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