Analysis Method of Donkey Bone Components in Donkey Hide Glue

Abstract

The present invention discloses an analysis method for donkey bone-derived components in donkey hide glue. The analysis method comprises the following steps: (1) analyzing the P element content and the Cu element content in the donkey hide glue to be tested, and calculating the ratio of the P element content to the Cu element content; (2) judging whether the donkey hide glue to be tested contains donkey bone-derived components according to the ratio of the P element content to the Cu element content, or further judging the content of the donkey bone-derived components in the donkey hide glue to be tested. The present invention provides a new analysis method for identifying whether donkey hide glue is doped with donkey bone-derived components.

Description

Technical Field

[0001] The invention relates to an analytical method for identifying donkey bone-derived components in donkey hide glue, belonging to the technical field of medicine and food detection. Background Art

[0002] Donkey hide glue, also known as donkey-hide glue, is a glue-like substance made from donkey hide. It is a traditional Chinese tonic that can be used as both medicine and food. All editions of the "Chinese Pharmacopoeia" have stipulated that the raw material of donkey hide glue is donkey hide.

[0003] The characteristic peptide method can be applied to the identification of non-donkey-derived raw materials such as cattle, pigs, and horses in donkey hide gelatin. For example, CN107525873A discloses a method for qualitatively analyzing the source of gelatin in donkey hide gelatin cake. The method comprises the following steps: (1) extracting and purifying the gelatin component in donkey hide gelatin cake; (2) enzymatically hydrolyzing the extracted and purified gelatin to obtain an enzymatic hydrolysis product; (3) separating and detecting the enzymatic hydrolysis product using a high performance liquid chromatograph-linear ion trap orbital trap high resolution combined mass spectrometer coupling device, and recording the obtained mass spectrum; (4) determining the characteristic peptides of donkey hide gelatin, pig hide gelatin, and cow hide gelatin, and comparing the characteristic peaks in the obtained mass spectrum with the characteristic peptides of donkey hide gelatin, pig hide gelatin, and cow hide gelatin. In theory, since the cow bone, pig bone, and horse bone-derived components contain the same characteristic peptides as the corresponding skin-derived components, the characteristic peptide method can also be applied to the identification of cow bone, pig bone, and horse bone-derived components.

[0004] Regarding the identification of donkey bone-derived components in donkey hide glue, CN110824083A discloses a method for detecting donkey bone glue components in animal hide glue and its products: the animal hide glue product to be tested is subjected to enzymatic digestion after protein purification, the enzymatic solution is injected into a liquid-mass spectrometer, and MRM detection is performed in a secondary mass spectrometry mode. The reference substance is a pure donkey bone glue or a negative sample to be tested as a matrix, and is prepared by adding a characteristic polypeptide of donkey bone glue. However, since the polypeptides of donkey bones and donkey skins have a high similarity, and the content of the characteristic polypeptide is easily affected by the glue making process, the characteristic polypeptide method has the disadvantages of low accuracy and difficulty in quantification for the donkey bone-derived components added to donkey hide glue.

[0005] In addition, the fatty acid detection method is currently used to detect and analyze the donkey bone-derived components in donkey skin glue. The theoretical basis of the fatty acid detection method is that there are differences in the fatty acid composition or content of donkey skin and donkey bones. The fatty acids are detected by gas chromatography, and donkey skin glue and donkey bone glue are distinguished according to the different fatty acid components. This method requires the steps of oil extraction, oil hydrolysis, and methyl esterification of the sample. The process is very complicated and requires the use of a large amount of organic reagents, which is costly and polluting. Furthermore, the composition of fatty acids is easily affected by the preparation process, resulting in the inability of this method to accurately conduct quantitative analysis.

[0006] Therefore, there is a need for a method that can accurately and quickly identify whether donkey hide glue contains donkey bone-derived components or the content of donkey bone-derived components, so as to efficiently judge the quality of donkey hide glue. Summary of the invention

[0007] In view of this, the object of the present invention is to provide a new analytical method for identifying whether donkey hide glue is mixed with donkey bone-derived components. Further, the analytical method can accurately determine whether donkey hide glue is mixed with donkey bone-derived components. Further, the analytical method can determine the amount of donkey bone-derived components mixed in donkey hide glue. Further, the analytical method is simple, easy to operate, consumes less resources, has low cost, and is convenient for practical application.

[0008] The present invention provides a method for analyzing donkey bone-derived components in donkey hide glue, comprising the following steps:

[0009] (1) analyzing the P element content and the Cu element content in the donkey hide glue to be tested, and calculating the ratio of the P element content to the Cu element content;

[0010] (2) judging whether the donkey hide glue to be tested contains donkey bone-derived components based on the ratio of the P content to the Cu content, or further judging the content of the donkey bone-derived components in the donkey hide glue to be tested.

[0011] According to the analysis method of the present invention, preferably, in step (2), when the ratio of the P element content to the Cu element content is higher than 20, the donkey skin glue to be tested is mixed with donkey bone-derived components.

[0012] According to the analysis method of the present invention, preferably, when the ratio of the P element content to the Cu element content is higher than 25, the donkey bone-derived components mixed in the donkey hide glue are not less than 10%; or

[0013] When the ratio of P content to Cu content is higher than 50, the donkey bone-derived ingredients mixed in the donkey hide glue are not less than 20%; or

[0014] When the ratio of P content to Cu content is higher than 80, the donkey bone-derived ingredients in the donkey hide glue are not less than 30%; or

[0015] When the ratio of P content to Cu content is higher than 120, the donkey bone-derived components mixed in the donkey hide glue are not less than 40%; or

[0016] When the ratio of P content to Cu content is higher than 180, the donkey bone-derived ingredients mixed in the donkey hide glue are not less than 50%; or

[0017] When the ratio of P element content to Cu element content is higher than 250, the donkey bone-derived components mixed in the donkey hide glue are not less than 60%.

[0018] According to the analysis method of the present invention, preferably, in step (1), an inductively coupled plasma mass spectrometer is used to determine the P element content and the Cu element content in the donkey hide glue.

[0019] According to the analysis method of the present invention, preferably, the parameters of the inductively coupled plasma mass spectrometer are:

[0020] The plasma gas flow rate is 10-20 L / min; the auxiliary gas flow rate is 0.5-2 L / min; the carrier gas flow rate is 0.5-2 L / min; the argon tank pressure is 400-800 KPa; the atomizing gas pressure is 300-350 KPa; the radio frequency power is 1200-1700 W; and the atomizing chamber temperature is 0.5-4°C.

[0021] According to the analysis method of the present invention, preferably, in step (1), when determining the Cu element content, the internal standard substance used is Ge; when determining the P element content, the internal standard substance used is Se.

[0022] According to the analysis method of the present invention, preferably, the donkey hide glue to be tested is dried and then crushed to form donkey hide glue powder to be tested; the donkey hide glue powder to be tested is subjected to microwave digestion to obtain a digestion product; the digestion product is fixed to volume with water to obtain a donkey hide glue test solution to be tested; and then an inductively coupled plasma mass spectrometer is used to detect the P element content and the Cu element content of the donkey hide glue test solution to be tested;

[0023] The conditions for microwave digestion are as follows: heating from the initial temperature to the T1 temperature, and maintaining the temperature at the T1 temperature for 3 to 10 minutes; then heating from the T1 temperature to the T2 temperature, and maintaining the temperature at the T2 temperature for 10 to 30 minutes; wherein the T1 temperature is 100 to 150°C, the T2 temperature is 160 to 250°C, the time for heating from the initial temperature to the T1 temperature is 5 to 20 minutes, and the time for heating from the T1 temperature to the T2 temperature is 5 to 20 minutes.

[0024] The applicant unexpectedly discovered that the ratio of the P element content to the Cu element content in donkey hide glue and donkey bone glue has a great difference, and this difference is very stable in donkey hide glue and donkey bone glue of various sources and batches. Therefore, it is possible to accurately determine whether the donkey hide glue to be tested is mixed with donkey bone-derived components by determining the characteristic of the ratio of the P element content to the Cu element content in the donkey hide glue to be tested, and further, it is possible to accurately determine the content of the donkey bone-derived components mixed in the donkey hide glue to be tested. Based on this, the present invention provides a new analysis method for donkey bone-derived components in donkey hide glue, which can accurately identify whether the donkey hide glue to be tested is mixed with donkey bone glue; further, it is possible to accurately determine the content of donkey bone glue mixed in the donkey hide glue to be tested. Compared with the existing fatty acid detection method, the method of the present invention uses fewer reagents and instruments, has a simple sample pretreatment process, is easy to operate, has a fast detection speed, a low detection cost, consumes less resources, and has determined parameters for each process, which is easy to control and suitable for practical application. DETAILED DESCRIPTION

[0025] The present invention is further described below in conjunction with specific embodiments, but the protection scope of the present invention is not limited thereto.

[0026] In the present invention, the "donkey hide glue to be tested" refers to the donkey hide glue that needs to be tested, which may not contain donkey bone-derived components (or donkey bone glue), may contain donkey bone-derived components, or may even be entirely counterfeit donkey bone-derived components.

[0027] In the present invention, the "pure donkey hide glue" refers to donkey hide glue made only from donkey hide, which does not contain any non-donkey-derived components or donkey bone-derived components.

[0028] In the present invention, the "P element content" and the "Cu element content" in the donkey hide glue to be tested both represent mass content.

[0029] Donkey hide glue and donkey bone glue are both derived from donkeys, so for donkey hide glue only mixed with donkey bone glue, the conventional characteristic polypeptide method is difficult to distinguish. The inventor unexpectedly found that the ratio of the P element content to the Cu element content in donkey hide glue and donkey bone glue is very different and very stable. This feature can be used to easily determine whether donkey hide glue is mixed with donkey bone glue, or further, to determine the content of donkey bone glue mixed in the donkey hide glue. Therefore, the method of the present invention can be used in conjunction with the characteristic polypeptide method to jointly judge the quality of donkey hide glue. When the characteristic polypeptide method is used to detect that the donkey hide glue to be tested does not contain non-donkey-derived components, the method of the present invention is then used to detect whether it contains donkey bone-derived components, or to further determine the content of donkey bone-derived components.

[0030] The method for analyzing donkey bone-derived components in donkey hide glue of the present invention comprises the following steps: (1) determining the ratio of the P element content to the Cu element content in the donkey hide glue to be tested; (2) determining whether the donkey hide glue to be tested contains the content of donkey bone-derived components, or further, determining the content of donkey bone-derived components contained in the donkey hide glue to be tested. The following is a detailed introduction.

[0031] <Determine the ratio of P content to Cu content in the donkey hide glue to be tested>

[0032] The P and Cu contents in the donkey hide glue to be tested were analyzed, and the ratio of the P content to the Cu content was calculated.

[0033] The P element content and the Cu element content in the donkey hide glue to be tested can be measured by any known method, preferably, by inductively coupled plasma mass spectrometry. According to one embodiment of the present invention, the internal standard method is used for determination. When measuring the P element content, the internal standard substance used is Ge. When measuring the Cu element content, the internal standard substance used is Se. The donkey hide to be tested and the internal standard substance are both formed into liquid samples, which are detected by inductively coupled plasma mass spectrometry.

[0034] Specifically, the donkey hide glue test solution to be tested and the blank sample are respectively injected into the inductively coupled plasma mass spectrometer; Ge in the mixed internal standard solution is used as the internal standard for the determination of the Cu element content, and Se in the mixed internal standard solution is used as the internal standard for the determination of the P element content, and the signal corresponding values ​​of the Cu element, Ge element, P element and Se element are determined. The Cu element content and the P element content in the donkey hide glue to be tested are obtained according to the standard curve.

[0035] The Cu element content and the P element content in the donkey hide glue to be tested can be measured multiple times, and the average value of the multiple measurements is taken as the final Cu element content and the P element content.

[0036] The mixed internal standard solution is a nitric acid aqueous solution of Ge and Se. The concentration of Ge in the mixed internal standard solution can be 1 μg / ml. The concentration of Se in the mixed internal standard solution can be 1 μg / ml. The blank sample is a nitric acid aqueous solution.

[0037] The parameters of the inductively coupled plasma mass spectrometer can be: plasma gas flow rate of 10 to 20 L / min; auxiliary gas flow rate of 0.5 to 2 L / min; carrier gas flow rate of 0.5 to 2 L / min; argon tank pressure of 400 to 800 KPa; atomizing gas pressure of 300 to 350 KPa; radio frequency power of 1200 to 1700 W; atomizing chamber temperature of 0.5 to 4° C. The carrier gas can be argon.

[0038] According to one embodiment of the present invention, the parameters of the inductively coupled plasma mass spectrometer are: plasma gas flow rate is 15L / min; auxiliary gas flow rate is 1L / min; carrier gas flow rate is 1 L / min; argon tank pressure is 500-700KPa; nebulizing gas pressure is 330KPa; RF power is 1550W; and nebulizing chamber temperature is 2°C.

[0039] The donkey hide glue test solution to be tested can be obtained by microwave digestion, which can include the following steps: drying the donkey hide glue to be tested, and then crushing it to form donkey hide glue powder to be tested; microwave digesting the donkey hide glue powder to be tested to obtain a digested product; and diluting the digested product with water to obtain the donkey hide glue test solution to be tested.

[0040] The microwave digestion conditions can be: heating from the initial temperature to T1 temperature, maintaining at T1 temperature for 3 to 10 minutes; then heating from T1 temperature to T2 temperature, maintaining at T2 temperature for 10 to 30 minutes. The T1 temperature is 100 to 150°C, the T2 temperature is 160 to 250°C, the time from the initial temperature to T1 temperature is 5 to 20 minutes, and the time from T1 temperature to T2 temperature is 5 to 20 minutes.

[0041] According to one embodiment of the present invention, the conditions for microwave digestion are: heating from the initial temperature to 120°C, maintaining at 120°C for 5 minutes, and the time used to heat from the initial temperature to 120°C is 10 minutes; then heating from 120°C to 190°C, maintaining at 190°C for 20 minutes, and the time used to heat from 120°C to 190°C is 10 minutes.

[0042] In the present invention, the content of donkey bone-derived components in the donkey hide glue to be tested is 0-100wt%, preferably 0-80wt%, and more preferably 0-60wt%. The doping amount of donkey bone-derived components in donkey hide glue on the market is mostly below 60wt%.

[0043] In the present invention, the Cu element and the P element can be measured by drawing a standard curve by the internal standard method. The following method can be used to obtain the standard curve: the standard solution of the Cu element and the standard solution of the P element of different concentrations are injected into the inductively coupled plasma mass spectrometer successively, Ge in the mixed internal standard solution is used as the internal standard for the determination of the Cu element content, and Se in the mixed internal standard solution is used as the internal standard for the determination of the P element content, and the signal corresponding values ​​of the Cu element, the Ge element, the P element and the Se element are measured. The concentration of the Cu element in the standard solution is used as the abscissa, and the ratio of the signal response value of the Cu element and the signal corresponding value of the Ge element is used as the ordinate to draw the standard curve of the Cu element. The concentration of the P element in the standard solution is used as the abscissa, and the ratio of the signal response value of the P element and the signal corresponding value of the Se element is used as the ordinate to draw the standard curve of the P element. The mixed internal standard solution is as described above. The parameters of the inductively coupled plasma mass spectrometer are the same as the parameters for measuring the P element content and the Cu element content in the donkey skin glue to be measured.

[0044] <Determination of the content of donkey bone-derived components in the donkey hide glue to be tested>

[0045] According to the ratio of the P element content to the Cu element content, it is judged whether the donkey skin glue to be tested contains donkey bone-derived components, or further judged the content of the donkey bone-derived components.

[0046] In the present invention, when the ratio of the P element content to the Cu element content is higher than 20, the donkey-bone derived components are mixed in the donkey-skin glue to be tested.

[0047] In the present invention, when the ratio of the P element content to the Cu element content is higher than 25, the donkey bone-derived component mixed in the donkey hide glue is not less than 10%; or

[0048] When the ratio of P content to Cu content is higher than 50, the donkey bone-derived ingredients mixed in the donkey hide glue are not less than 20%; or

[0049] When the ratio of P content to Cu content is higher than 80, the donkey bone-derived ingredients in the donkey hide glue are not less than 30%; or

[0050] When the ratio of P content to Cu content is higher than 120, the donkey bone-derived components mixed in the donkey hide glue are not less than 40%; or

[0051] When the ratio of P content to Cu content is higher than 180, the donkey bone-derived ingredients mixed in the donkey hide glue are not less than 50%; or

[0052] When the ratio of P element content to Cu element content is higher than 250, the donkey bone-derived components mixed in the donkey hide glue are not less than 60%.

[0053] In the present invention, the percentages mentioned are all percentages by mass.

[0054] In the present invention, the relationship between the ratio of the P element content to the Cu element content and the content of the donkey bone-derived component in the donkey hide glue can be obtained by one of the following methods:

[0055] Method 1: Determine the P and Cu contents in pure donkey hide glue and donkey hide glue containing different contents of donkey bone-derived components, and calculate the ratio of the P content to the Cu content, thereby forming a relationship between the ratio of the P content to the Cu content and the ratio of the donkey bone-derived components in donkey hide glue.

[0056] Method 2: Determine the P and Cu contents in pure donkey hide glue, donkey hide glue containing different contents of donkey bone-derived components and pure donkey bone glue, and calculate the ratio of the P content to the Cu content, forming a relationship between the ratio of the P content to the Cu content and the ratio of the donkey bone-derived components in donkey hide glue.

[0057] The method for determining the P element content and the Cu element content in pure donkey hide glue, donkey hide glue containing different donkey bone-derived components and pure donkey bone glue is the same as the method for determining the P element content and the Cu element content in the donkey hide glue to be tested, and will not be repeated here.

[0058] The relationship between the ratio of P element content to Cu element content and the content of donkey bone-derived components in donkey hide glue can be expressed in the form of a functional equation, and the range of the ratio of P element content to Cu element content and the corresponding range of the content of donkey bone-derived components in donkey hide glue can also be summarized.

[0059] Example 1 - Sample preparation

[0060] 1.1 Preparation of standard solution and mixed internal standard solution

[0061] (1) Preparation of standard solution

[0062] 5 wt% nitric acid aqueous solution was used to prepare standard solutions of Cu element with concentrations of 1 μg / L, 10 μg / L, 50 μg / L, 100 μg / L, 150 μg / L, and 200 μg / L. 5 wt% nitric acid aqueous solution was used to prepare standard solutions of P element with concentrations of 10 μg / L, 100 μg / L, 500 μg / L, 1000 μg / L, 1500 μg / L, and 3000 μg / L.

[0063] (2) Preparation of mixed internal standard solution

[0064] A mixed internal standard solution with Ge and Se concentrations of 1 μg / ml was prepared using 5 wt % nitric acid aqueous solution.

[0065] 1.2 Preparation of pure donkey hide glue, pure donkey bone glue, donkey hide glue containing different donkey bone-derived components (mixed glue) and blank samples

[0066] (1) Preparation of pure donkey hide glue samples

[0067] Eight donkey hide samples from different origins on the market were used to prepare pure donkey hide glue samples (numbered EJ1-EJ8) using the following method: the donkey hide samples were boiled to obtain pure donkey hide glue. The pure donkey hide glue was dried and then crushed into powder. 0.5g of pure donkey hide glue powder and 5ml of 68wt% nitric acid were placed in a microwave digestion tank and digested according to the procedure in Table 1. The digested product was diluted to 50ml with ultrapure water and mixed to obtain a pure donkey hide glue sample. Two portions of each donkey hide sample were digested in parallel to prepare two pure donkey hide glue samples.

[0068] (2) Preparation of pure donkey bone glue samples

[0069] Seven donkey bone samples from different origins on the market were used to prepare pure donkey bone glue samples (numbered LGJ1-LGJ7) using the following method: the donkey bone samples were boiled to obtain pure donkey bone glue. The pure donkey bone glue was dried and then crushed into powder. 0.5g of pure donkey bone glue powder and 5ml of 68wt% nitric acid were placed in a microwave digestion tank and digested according to the procedure in Table 1. The digested product was diluted to 50ml with ultrapure water and mixed to obtain a pure donkey bone glue sample. Two portions of each donkey bone sample were digested in parallel to prepare two portions of pure donkey bone glue samples.

[0070] (3) Preparation of donkey hide glue (mixed glue) samples containing different donkey bone-derived components

[0071] According to the composition shown in Table 2, pure donkey hide glue numbered EJ-1 and pure donkey bone glue numbered LGJ-1 were mixed to obtain donkey hide glue containing different donkey bone-derived components (numbered respectively as mixed glue 1 to mixed glue 6).

[0072] Prepare mixed glue samples from mixed glue 1-6: Dry the mixed glue and then crush it into powder. Place 0.5g of mixed glue powder and 5ml of concentrated nitric acid in a microwave digestion tank and digest according to the procedure in Table 1. Dilute the digested product to 50ml with ultrapure water and mix well to obtain a mixed glue sample. Digest 2 portions of each mixed glue sample in parallel to prepare two mixed glue samples.

[0073] (4) Preparation of blank samples

[0074] 5 ml of concentrated nitric acid was placed in a microwave digestion tank and treated according to the procedure in Table 1. The treated product was diluted to 50 ml with ultrapure water and mixed to obtain a blank sample.

[0075] Table 1

[0076] step temperature Time (min) 1 From the initial temperature to 120℃ 10 2 120℃ 5 3 Heating from 120℃ to 190℃ 10 4 190℃ 20

[0077] Table 2

[0078] serial number Donkey bone glue content (wt%) Donkey hide glue content (wt%) Mixed glue 1 10 90 Mixed glue 2 20 80 Mixed glue 3 30 70 Mixed glue 4 40 60 Mixed glue 5 50 50 Mixed glue 6 60 40

[0079] Example 2 - Elemental Analysis

[0080] 2.1 Drawing the standard curve

[0081] Ge in the mixed internal standard solution prepared in Example 1 is used as the internal standard for the determination of Cu element content, and Se in the mixed internal standard solution prepared in Example 1 is used as the internal standard for the determination of P element content. The standard solutions of different concentrations prepared in Example 1 are injected into an inductively coupled plasma mass spectrometer (ICP-MS) successively, and the signal response values ​​of the elements to be measured (Cu, P) and the internal standard elements (Ge and Se) are measured according to the following parameters. The concentration of the element to be measured in the standard solution is used as the abscissa, and the ratio of the signal response value of the element to be measured and the signal response value of the internal standard element corresponding thereto is used as the ordinate to draw a standard curve, and the standard curve equation is shown in the table below.

[0082] ICP-MS parameters: plasma gas flow rate is 15L / min; auxiliary gas flow rate is 1 L / min; carrier gas (argon) flow rate is 1L / min; argon tank pressure is 500-700KPa; nebulizing gas pressure is 330KPa; RF power is 1550W; and nebulizing chamber temperature is 2°C.

[0083] Table Standard Curve Equation

[0084] element Standard curve equation r Cu <![CDATA[Y=7.209×10 -2 X+6.448×10 -3 ]]> 0.9997 P <![CDATA[Y=2.228×10 -1 X+9.174×10 -4 ]]> 0.9999

[0085] 2.2 Determination of Cu and P contents in pure donkey hide glue samples

[0086] The blank sample and the pure donkey hide glue sample prepared in Example 1 were respectively injected into an inductively coupled plasma mass spectrometer (ICP-MS), and Ge in the mixed internal standard solution prepared in Example 1 was used as the internal standard for the determination of the Cu element content, and Se in the mixed internal standard solution prepared in Example 1 was used as the internal standard for the determination of the P element content. The signal response values ​​of the elements to be measured (Cu, P) and the internal standard elements (Ge and Se) were determined according to the ICP-MS parameters in 2.1. The contents of Cu and P in the pure donkey hide glue sample were obtained according to the standard curve, and the ratio of the P content to the Cu content (P / Cu) was calculated. The results are shown in Table 3.

[0087] Table 3

[0088] sample P content (mg / kg) Cu content (mg / kg) P / Cu EJ1 130.26 24.38 5.34 EJ2 163.05 11.04 14.77 EJ3 122.88 10.77 11.40 EJ4 144.40 8.33 17.34 EJ5 268.97 28.05 9.59 EJ6 113.00 27.00 4.19 EJ7 79.80 16.60 4.81 EJ8 149.00 25.90 5.75

[0089] Note: The P content and Cu content in the above table are the average values ​​of the results of two measurements for each sample.

[0090] 2.3 Determination of Cu and P contents in pure donkey bone glue samples

[0091] The blank sample and pure donkey bone glue sample prepared in Example 1 were injected into an inductively coupled plasma mass spectrometer (ICP-MS) respectively, Ge in the mixed internal standard solution prepared in Example 1 was used as the internal standard for the determination of Cu element content, Se in the mixed internal standard solution prepared in Example 1 was used as the internal standard for the determination of P element content, and the signal response values ​​of the elements to be measured (Cu, P) and the internal standard elements (Ge and Se) were determined according to the ICP-MS parameter in 2.1. The content of Cu and P in the pure donkey bone glue sample was obtained according to the standard curve, and the ratio (P / Cu) of P content to Cu content was calculated. The results obtained are shown in Table 4.

[0092] Table 4

[0093] sample P content (mg / kg) Cu content (mg / kg) P / Cu LGJ1 2719.91 1.02 2666.58 LGJ2 3041.68 1.31 2321.89 LGJ3 609.56 0.43 1417.58 LGJ4 2618.18 0.99 2644.63 LGJ5 576.00 0.34 1694.12 LGJ6 333.00 0.37 900.00 LGJ7 263.00 0.41 641.46

[0094] Note: The P content and Cu content in the above table are the average values ​​of the results of two measurements for each sample.

[0095] 2.4 Determination of Cu and P contents in donkey hide glue (mixed glue) containing different donkey bone-derived components

[0096] The blank sample and the mixed gel sample prepared in Example 1 were respectively injected into an inductively coupled plasma mass spectrometer (ICP-MS), and Ge in the mixed internal standard solution prepared in Example 1 was used as the internal standard for the determination of Cu content, and Se in the mixed internal standard solution prepared in Example 1 was used as the internal standard for the determination of P content, and the signal response values ​​of the elements to be measured (Cu, P) and the internal standard elements (Ge and Se) were determined according to the ICP-MS parameters in 2.1. The contents of Cu and P in the mixed gel sample were obtained according to the standard curve, and the ratio of P content to Cu content (P / Cu) was calculated. The results are shown in Table 5.

[0097] Table 5

[0098] sample Donkey bone glue content (wt%) P content (mg / kg) Cu content (mg / kg) P / Cu Mixed glue 1 10 630.237 25.26 24.95 Mixed glue 2 20 1065.556 21.47 49.63 Mixed glue 3 30 1410.5 17.5 80.60 Mixed glue 4 40 1718.55 14.25 120.60 Mixed glue 5 50 1897.474 10.89 174.24 Mixed glue 5 60 1647.236 6.59 249.96

[0099] Note: The P content and Cu content in the above table are the average values ​​of the results of two measurements for each sample.

[0100] From Tables 3-5, we can summarize the relationship between the ratio of P content to Cu content (P / Cu) shown in Table 6 and the amount of donkey bone-derived components doped in the donkey hide glue to be tested.

[0101] Table 6

[0102]

[0103] Example 3-Determination of P and Cu content in donkey hide glue and analysis of donkey bone source in donkey hide glue Content of sexual ingredients

[0104] The following methods were used to determine the P and Cu contents in three batches of donkey hide glue to be tested (donkey hide glue 1 to be tested, donkey hide glue 2 to be tested, and donkey hide glue 3 to be tested, all of which were prepared by boiling donkey hide with a certain amount of donkey bones, i.e., doped with a specific content of donkey bone-derived components):

[0105] The donkey hide glue to be tested is dried and then crushed into powder. 0.5g of the donkey hide glue powder to be tested and 5ml of concentrated nitric acid are placed in a microwave digestion tank and digested according to the procedure of Table 2 in Example 1. The digested product is diluted to 50ml with ultrapure water and mixed to obtain the donkey hide glue test solution to be tested. Two portions of each donkey hide glue to be tested are digested in parallel to prepare two portions of donkey hide glue test solutions to be tested.

[0106] The donkey hide glue test solution to be tested and the blank sample prepared in Example 1 were respectively injected into an inductively coupled plasma mass spectrometer (ICP-MS), Ge in the mixed internal standard solution prepared in Example 1 was used as the internal standard for the determination of Cu element content, Se in the mixed internal standard solution prepared in Example 1 was used as the internal standard for the determination of P element content, and the signal response values ​​of the elements to be tested (Cu, P) and the internal standard elements (Ge and Se) were determined according to the ICP-MS parameters in 2.1. The contents of Cu and P in the donkey hide glue to be tested were obtained according to the standard curve, and the ratio of P content to Cu content (P / Cu) was calculated. The content of donkey bone-derived components in the donkey hide glue to be tested was analyzed according to the ratio of P content to Cu content (P / Cu) and the data of Table 6. The results are shown in Table 7.

[0107] Table 7

[0108]

[0109] Note: The P content and Cu content in the above table are the average values ​​of the results of two measurements for each sample.

[0110] The results showed that the judgment result was consistent with the actual amount of donkey bone-derived components impregnated in the donkey hide glue to be tested.

[0111] The present invention is not limited to the above-mentioned embodiments. Without departing from the essential content of the present invention, any deformation, improvement and substitution that can be conceived by those skilled in the art shall fall within the scope of the present invention.

Claims

1. A method for analyzing donkey bone-derived components in donkey hide glue, characterized in that: The steps include: (1) analyzing the P element content and the Cu element content in the donkey hide glue to be measured, and calculating the ratio of the P element content to the Cu element content; (2) judging whether the donkey hide glue to be tested contains donkey bone-derived components according to the ratio of the P element content to the Cu element content, or further judging the content of the donkey bone-derived components in the donkey hide glue to be tested; When the ratio of P element content to Cu element content is higher than 20, the donkey hide glue to be tested contains donkey bone-derived components; or When the ratio of P element content to Cu element content is higher than 25, the donkey bone-derived components mixed in the donkey hide glue to be tested are not less than 10%; or When the ratio of P element content to Cu element content is higher than 50, the donkey bone-derived components mixed in the donkey hide glue to be tested are not less than 20%; or When the ratio of P element content to Cu element content is higher than 80, the donkey bone-derived components mixed in the donkey hide glue to be tested are not less than 30%; or When the ratio of P element content to Cu element content is higher than 120, the donkey bone-derived components mixed in the donkey hide glue to be tested are not less than 40%; or When the ratio of P element content to Cu element content is higher than 180, the donkey bone-derived components mixed in the donkey hide glue to be tested are not less than 50%; or When the ratio of the P element content to the Cu element content is higher than 250, the donkey bone-derived components mixed in the tested donkey hide glue are not less than 60%.

2. The analysis method according to claim 1, characterized in that: In step (1), the P element content and the Cu element content in the donkey hide glue are determined by inductively coupled plasma mass spectrometry.

3. The analysis method according to claim 2, characterized in that The parameters of the inductively coupled plasma mass spectrometer are: The plasma gas flow rate is 10-20 L / min; the auxiliary gas flow rate is 0.5-2 L / min; the carrier gas flow rate is 0.5-2 L / min; the argon tank pressure is 400-800 KPa; the atomizing gas pressure is 300-350 KPa; the radio frequency power is 1200-1700 W; and the atomizing chamber temperature is 0.5-4°C.

4. The analysis method according to claim 2, characterized in that In step (1), when determining the Cu element content, the internal standard substance used is Ge; when determining the P element content, the internal standard substance used is Se.

5. The analysis method according to claim 2, characterized in that The donkey hide glue to be tested is dried and then crushed to form donkey hide glue powder to be tested; the donkey hide glue powder to be tested is subjected to microwave digestion to obtain a digestion product; the digestion product is diluted to volume with water to obtain a donkey hide glue test solution to be tested; and then an inductively coupled plasma mass spectrometer is used to detect the P element content and the Cu element content of the donkey hide glue test solution to be tested; The conditions for microwave digestion are as follows: heating from the initial temperature to the T1 temperature, and maintaining the T1 temperature for 3 to 10 minutes; then heating from the T1 temperature to the T2 temperature, and maintaining the T2 temperature for 10 to 30 minutes; wherein the T1 temperature is 100 to 150°C, the T2 temperature is 160 to 250°C, the time for heating from the initial temperature to the T1 temperature is 5 to 20 minutes, and the time for heating from the T1 temperature to the T2 temperature is 5 to 20 minutes.

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