Tegillarca inflata reeve, tegillarca subcrenata and tegillarca granosa species

Abstract

The invention discloses feature peptides of arca inflata reeve, arca subcrenata and arca granosa and application thereof. The feature peptide of the scapharca inflata reeve species comprises a feature peptide of scapharca inflata reeve: PSVQGAAAQLTADVK, a feature peptide of scapharca subcrenata: VAELANAVVSNADQK and a feature peptide of scapharca subcrenata: DSWAALGSDK, and can be used for identifying the species of scapharca inflata reeve, scapharca subcrenata and scapharca subcrenata and evaluating the quality of medicinal materials. The invention also provides a detection method based on the characteristic peptide of the tegillarca granosa species and a liquid chromatography-triple quadrupole mass spectrometry technology, and the detection method is used for detecting components in tegillarca granosa medicinal materials or other related products, so that efficient and accurate identification of the tegillarca broughtonii, the tegillarca subcrenata and the tegillarca granosa is realized. The method is simple and convenient to operate, strong in specificity, high in sensitivity and good in stability, provides a reliable quality detection method for safe and effective use of the scapharca granosa in the fields of food and medicines, and has a good application prospect in quality control of the scapharca granosa medicinal materials and related products.

Description

Technical Field

[0001] The present invention belongs to the technical field of clam species identification, and specifically relates to clam species characteristic peptides of Scapharca broughtonii, Tegillarca granosa and Arca subcrenata and their applications. Background Art

[0002] The traditional Chinese medicine clam (Scapharcae et tegillarcae musculus) is the meat of animals in the family Arcidae, such as Scapharca broughtonii Scapharca broughtonii (Schrenck), Tegillarca granosa Tegillarca granosa (Linnaeus), and Arca subcrenata Scapharca subcrenata etc., which has the effects of invigorating qi and nourishing blood, strengthening the stomach and warming the middle, promoting digestion and resolving food stagnation. Modern pharmacological research further shows that it has significant biological activities such as antioxidant, anti-inflammatory, antibacterial and anti-tumor. At the same time, as a high-protein seafood, clam meat is rich in hemoglobin, iron element and various essential amino acids, and is a natural iron supplement, with both food therapy value.

[0003] As a food and medicine with economic value, the quality control of clams still faces challenges at present. The high similarity of morphological characteristics among species makes it difficult to identify the original species. The identification methods based on phenotypic differences such as the morphology of the original animal shell and the number of radial ribs are easily affected by the individual development stage and environmental factors, and are prone to misjudgment. Although DNA barcoding technology (such as COI gene sequencing) has strong specificity, there are limitations such as the easy degradation of DNA in high-temperature processed samples, resulting in inaccurate determination, and it is difficult to meet the needs of rapid detection at the grass-roots level. In recent years, the phenomenon of adulteration with related species has been found in commercially available clam products, which directly affects the material basis of its medicinal efficacy and food safety. Summary of the Invention

[0004] The purpose of the present invention is to provide clam species characteristic peptides of Scapharca broughtonii, Arca subcrenata and Tegillarca granosa, and to provide specific applications of the characteristic peptides to make up for the deficiencies of the existing technology.

[0005] Clams are very few organisms in the class Bivalvia with a hemoglobin oxygen-carrying system. It is speculated that using the characteristic peptides in clam hemoglobin for species identification can quickly and accurately identify different species of clams and overcome the limitations of traditional methods. The present invention screens and identifies the characteristic peptides in the hemoglobin of Scapharca broughtonii, Tegillarca granosa and Arca subcrenata, provides an effective supplement to the existing quality control system, and provides an effective solution for the identification and quality control of clams.

[0006] To achieve the above purpose, the present invention adopts the following technical solutions: The clam species characteristic peptides of Scapharca broughtonii, Arca subcrenata and Tegillarca granosa include: The sequence of the Scapharca broughtonii characteristic peptide is: PSVQGAAAQLTADVK (PSV); The sequence of the Scapharca subcrenata characteristic peptide is: VAELANAVVSNADQK (VAE); The sequence of the Tegillarca granosa characteristic peptide is: DSWAALGSDK (DSW).

[0007] Application of the characteristic peptides of the species of Scapharca broughtonii, Scapharca subcrenata and Tegillarca granosa in the identification of the three original species of Scapharca medicinal materials.

[0008] A method for identifying the three original species of Scapharca medicinal materials is to detect whether the sample contains the characteristic peptides of the species of Scapharca broughtonii, Scapharca subcrenata and Tegillarca granosa, namely PSV, VAE and DSW; the judgment principle is: If PSV is detected in the sample and VAE and DSW are not detected, it is determined that the sample to be tested is Scapharca broughtonii; If VAE is detected in the sample and DSW and PSV are not detected, it is determined that the sample to be tested is Scapharca subcrenata If DSW is detected in the sample and VAE and PSV are not detected, it is determined that the sample to be tested is Tegillarca granosa; If PSV, VAE and DSW are not detected in the sample, it is determined that the sample to be tested is not any one of Scapharca broughtonii, Scapharca subcrenata and Tegillarca granosa.

[0009] A method for identifying the original species of Scapharca (Scapharca broughtonii, Scapharca subcrenata and Tegillarca granosa) based on the characteristic peptides of the Scapharca species includes the following steps: (1) Preparation of the test solution: Take the powder of the Scapharca medicinal material sample for enzymatic hydrolysis, centrifuge, collect the supernatant, freeze-dry to obtain the freeze-dried powder of the Scapharca enzymatic hydrolysate; take the freeze-dried powder of the Scapharca enzymatic hydrolysate, prepare a Scapharca enzymatic hydrolysate solution, perform desalting treatment, and then re-dissolve to obtain the test solution; (2) Preparation of the reference solution: Using the three characteristic peptides of Scapharca species, PSV, VAE and DSW, as reference substances, dissolve them in 0.1% formic acid water to prepare the reference solution; (3) Detection and analysis: Use liquid chromatography-triple quadrupole mass spectrometry (LC-MS / MS) method for detection and analysis; (4) If PSV is detected in the sample to be tested and VAE and DSW are not detected, it is determined that the sample to be tested is Scapharca broughtonii; if VAE is detected in the sample to be tested and DSW and PSV are not detected, it is determined that the sample to be tested is Scapharca subcrenata; if DSW is detected in the sample to be tested and VAE and PSV are not detected, it is determined that the sample to be tested is Tegillarca granosa; if PSV, VAE and DSW are not detected in the sample to be tested, it is determined that the sample to be tested is not Scapharca broughtonii, Scapharca subcrenata and Tegillarca granosa.

[0010] Further, in the step (1): the enzymatic hydrolysis conditions are as follows: take the powder of the ark shellfish medicinal material sample, add PBS buffer according to the solid-liquid ratio of 1:10 (w / v, g / mL), add trypsin (final concentration 800 U / mL), and perform enzymatic hydrolysis at 50 °C and pH 8.0 for 5 hours; the desalting treatment is to desalt the sample using a desalting column; the reconstitution is to reconstitute with 50 μL of 0.1% formic acid aqueous solution.

[0011] Further, the chromatographic conditions in the step (3) are as follows: mobile phase A is 0.1% formic acid aqueous solution, mobile phase B is 0.1% formic acid acetonitrile solution, and the flow rate is set at 0.3 mL / min. The elution gradient is as follows: 0 - 1 min, 95% A; 1 - 2 min, 95 - 80% A; 2 - 8 min, 80 - 60% A; 8 - 10 min, 60 - 10% A; 10 - 11.5 min, 10% A; 11.5 - 11.6 min, 10 - 95% A; 11.6 - 14 min, 95% A. The monitored ion pairs include VAE as m / z 765.3 → 171.1; DSW as m / z 525.5 → 661.4; PSV as m / z 728.9 → 185.1.

[0012] Compared with the prior art, the present invention has at least the following beneficial effects: The characteristic peptides provided by the present invention have excellent specificity and stability for ark shellfish of three origins, with strong specificity, can be used for the identification of the origin of ark shellfish medicinal materials, and have good application prospects. The present invention can be widely applied to the component detection of ark shellfish medicinal materials or other related products, can accurately distinguish whether the product contains ark shellfish components, and is particularly suitable for the identification of three ark shellfish origins - Scapharca broughtonii, Arca subcrenata, and Tegillarca granosa; it can effectively prevent forgery and adulteration phenomena, and provides a reliable technical means for the quality control of ark shellfish products.

[0013] The present invention also provides a discrimination method based on three ark shellfish species characteristic peptides. This method has remarkable specificity, sensitivity and stability, and can efficiently and accurately identify three ark shellfish origins - Scapharca broughtonii, Arca subcrenata, and Tegillarca granosa. Compared with the prior art, the present invention effectively overcomes the common problems of cross-contamination and misjudgment in traditional methods, ensuring the reliability and accuracy of the discrimination results. In addition, the method of the present invention is easy to operate, the sample treatment process is simplified, and it can complete efficient discrimination in a short time, with strong practicability. This method effectively fills the blank in the prior art that cannot accurately identify the origin of ark shellfish, and its wide application potential makes it have important application value in the research and industrialization of pharmaceuticals, foods and related fields. Description of the Drawings

[0014] Figure 1 are the secondary fragment mass spectra of PSV, VAE, and DSW.

[0015] Figure 2 It is the ion current chromatogram of the standard products (50 ng / mL) of the characteristic peptides of Scapharca broughtonii (PSV), Scapharca subcrenata (VAE), and Tegillarca granosa (DSW).

[0016] Figure 3 It is the extracted ion current chromatogram of PSV, VAE, and DSW in Scapharca broughtonii.

[0017] Figure 4 It is the extracted ion current chromatogram of PSV, VAE, and DSW in Scapharca subcrenata.

[0018] Figure 5 It is the extracted ion current chromatogram of PSV, VAE, and DSW in Tegillarca granosa.

[0019] Figure 6 It is the extracted ion current chromatogram of PSV, VAE, and DSW in the HWZ05 sample.

[0020] Figure 7 It is the extracted ion current chromatogram of PSV, VAE, and DSW in the HLYG06 sample.

[0021] Figure 8 It is the extracted ion current chromatogram of PSV, VAE, and DSW in the HLYG11 sample.

[0022] Figure 9 It is the extracted ion current chromatogram of PSV, VAE, and DSW in the HYT20 sample. Detailed implementation manners

[0023] The invention will be further explained and illustrated below through specific examples in combination with the accompanying drawings.

[0024] Example 1: Screening of characteristic peptides of three species of ark shellfish, namely Scapharca broughtonii, Scapharca subcrenata, and Tegillarca granosa Preparation of test sample solutions Respectively take 0.1 g of the powder samples of the three ark shellfish medicinal materials, Scapharca broughtonii, Scapharca subcrenata, and Tegillarca granosa, add 1 mL of PBS buffer solution, add 800 U of trypsin, enzymatically hydrolyze at 50 °C and pH 8.0 for 5 hours, centrifuge at 8000 rpm for 10 minutes at 4 °C, collect the supernatant, freeze-dry to obtain the freeze-dried powder of the ark shellfish hydrolysate. Take the prepared freeze-dried powder of the ark shellfish hydrolysate, dissolve it in water, and prepare a 10 mg / mL ark shellfish hydrolysate solution. Use the ZIPTIP C18 desalting column produced by Millipore Corporation to desalt the sample, and redissolve it with 50 μL of 0.1% formic acid aqueous solution to obtain the test sample solution. The ark shellfish samples of the three species, Scapharca broughtonii, Scapharca subcrenata, and Tegillarca granosa, are all subjected to 3 batches of repeated experiments.

[0025] (2) Selection of characteristic ions and sequence speculation LC-MS / MS parameters: Analysis was performed using an EASY-nLC1000 nano ultra-high performance liquid chromatography system equipped with a nano peptide trapping column Acclaim PepMap C18 (100 μm × 20 mm, 3 μm) and a nano peptide analysis column Acclaim PepMap C18 (75 μm × 250 mm, 3 μm). Column temperature: 50 °C; Mobile phase A was an aqueous solution of 0.1% formic acid, and mobile phase B was an acetonitrile solution of 0.1% formic acid, with gradient elution; The injection volume was 2 μL. A Q-Exactive mass spectrometer with an electrospray ionization (ESI) source was used to collect mass spectrometry information in the positive ion mode under the data-dependent acquisition (DDA) mode.

[0026] Database search and screening of characteristic peptides: The mass spectrometry data analysis and retrieval software MaxQuant 2.4.3.0 was used, and the database used was the Uniprot protein database of ark shell ( Arcidae ), including three species of ark shell, Scapharca broughtonii ( Scapharca broughtonii ), Scapharca subcrenata ( Scapharca subcrenata ), Tegillarca granosa ( Tegillarca granosa ), and other species of ark shell.

[0027] Combined with the analysis results of MaxQuant and protein-protein BLAST analysis, and chemometric methods, characteristic peptides of Scapharca broughtonii, Scapharca subcrenata, and Tegillarca granosa were screened respectively. The specific requirements are as follows: (1) Proteins / peptides with a missing value greater than 2 in one species were excluded; (2) The peptide must be stably detected and have a relatively high response in at least one of the three species; (3) The peptide is derived from the hemoglobin of the ark shell genus; (4) VIP > 1 in the OPLS-DA model; (5) The fold change of Fold change is at least 2 (Ratio > 2), and P < 0.01 is satisfied. Finally, the characteristic peptides of Scapharca broughtonii were obtained: as Figure 1 shown, PSVQGAAAQLTADVK (PSV), the characteristic peptide of Scapharca subcrenata: VAELANAVVSNADQK (VAE), and the characteristic peptide of Tegillarca granosa: DSWAALGSDK (DSW).

[0028] According to the deduced amino acid sequence, a reference substance of the characteristic peptide was commissioned for synthesis. The reference substance and the test solutions of Scapharca subcrenata, Scapharca broughtonii, and Tegillarca granosa were detected simultaneously. The retention times and the secondary mass spectrometry information of both were consistent, thus confirming the correctness of the sequence.

[0029] Example 2: Detection method for characteristic peptides of three species of ark shell, Scapharca broughtonii, Scapharca subcrenata, and Tegillarca granosa Three species-specific peptides in the clam hydrolysate solution were detected by LC-MS / MS.

[0030] (1) Sample preparation Preparation of test solution: Take 0.1 g of powder samples of three clam medicinal materials, namely Scapharca broughtonii, Arca subcrenata, and Tegillarca granosa, add 1 mL of PBS buffer solution, add 800 U of trypsin, enzymatically hydrolyze at 50 °C and pH 8.0 for 5 hours, centrifuge at 8000 rpm for 10 minutes at 4 °C, collect the supernatant, freeze-dry to obtain the freeze-dried powder of clam hydrolysate. Take the freeze-dried powder of clam hydrolysate, dissolve it in water, and prepare a 10 mg / mL clam hydrolysate solution. Desalt the sample using a ZIPTIP C18 desalting column produced by Millipore Corporation, and redissolve it with 50 μL of 0.1% formic acid aqueous solution to obtain the test solution.

[0031] Preparation of reference solution: Weigh accurately three species-specific peptide reference substances of clams, dissolve them in 0.1% formic acid in water to obtain a stock solution with a concentration of 1 mg / mL, store it in a -20 °C refrigerator for standby. Accurately measure the stock solution of the species-specific peptide reference substance of clams, dilute it step by step with 0.1% formic acid aqueous solution, and add an internal standard to obtain a series of reference solutions with concentrations of 5 ng / mL, 10 ng / mL, 25 ng / mL, 50 ng / mL, 100 ng / mL, 200 ng / mL, 400 ng / mL, 500 ng / mL, and 600 ng / mL. The concentration of the internal standard is 250 ng / mL.

[0032] Spiked matrix sample: Take the species-specific peptide reference substance of clams, add the enzymatic hydrolysis system matrix and internal standard, and prepare spiked matrix samples of the species-specific peptide of clams with concentrations of 10 ng / mL, 300 ng / mL, and 500 ng / mL. The concentration of the internal standard is 250 ng / mL.

[0033] Quality control sample: Take the stock solution of the species-specific peptide reference substance of clams, dilute it step by step with 0.1% formic acid aqueous solution, add an internal standard, and prepare quality control solutions of the species-specific peptide of clams with three concentrations, namely high, medium, and low, with concentrations of 10 ng / mL (LQC), 300 ng / mL (MQC), and 500 ng / mL (HQC). The concentration of the internal standard is 250 ng / mL.

[0034] (2) LC-MS / MS detection method An UPLC H-Class PLUS ultra-high performance liquid chromatography system equipped with an Xevo TQ-XS mass spectrometer detector was used. The chromatographic column was an ACQUITY UPLC BEH C18 column (2.1 × 100 mm, 1.7 μm), the column temperature was 40°C, mobile phase A was an aqueous solution of 0.1% formic acid, mobile phase B was an acetonitrile solution of 0.1% formic acid, and gradient elution was carried out according to Table 1; the injection volume was 4 μL. The mass spectrometer detector used ESI and multi-reaction monitoring (MRM) was carried out in the positive ion mode, the capillary voltage was 3.0 kV, and the desolvation gas temperature was 500°C. Nitrogen was used as the desolvation gas (1000 L / Hr) and the cone gas (150 L / Hr), and the collision gas was argon (0.15 mL / min). The monitored ion pairs are shown in Table 2.

[0035] Table 1 Liquid phase conditions for the detection method of scallop species characteristic peptides

[0036] Table 2 Mass spectrometry MRM conditions for the detection method of scallop species characteristic peptides

[0037] Note: TTLLEDDIAK ( 13 C6, 15 N2) is an isotope-labeled peptide and is used as an internal standard.

[0038] (3) Specificity In the extracted ion current chromatogram of the ark shell sample ( Figure 3 ), at the retention time position corresponding to the ark shell characteristic peptide PSV reference solution (as Figure 2 shown), chromatographic peaks with consistent retention times were all presented, and there were no chromatographic peaks corresponding to the mud ark shell characteristic peptide DSW and the blood ark shell characteristic peptide VAE reference solutions; in the extracted ion current chromatogram of the blood ark shell sample ( Figure 4 ), at the retention time position corresponding to the blood ark shell characteristic peptide VAE reference solution (as Figure 2 shown), chromatographic peaks with consistent retention times were all presented, and there were no chromatographic peaks corresponding to the mud ark shell characteristic peptide DSW and the ark shell characteristic peptide PSV reference solutions; in the extracted ion current chromatogram of the mud ark shell sample ( Figure 5 ), at the retention time position corresponding to the mud ark shell characteristic peptide DSW reference solution (as Figure 2 shown), chromatographic peaks with consistent retention times were all presented, and there were no chromatographic peaks corresponding to the blood ark shell characteristic peptide VAE and the ark shell characteristic peptide PSV reference solutions. It shows that the three scallop species characteristic peptides were only detected in their respective species, and the specificity is strong.

[0039] (4) Linear relationship, detection limit and quantitation limit The standard curves were plotted with the ratios of the peak areas of the quantitative ion pairs of VAE, DSW, and PSV to the peak area of the internal standard and the corresponding mass concentrations. The calculated lowest detection limits (LOD, S / N≥3), lowest quantification limits (LOQ, S / N≥10), and regression equations are shown in Table 3. The three characteristic peptides of the clam species had good linear relationships within the range of 5 - 600 ng / mL.

[0040] Table 3. Sensitivity and linearity of the detection method for characteristic peptides of clam species

[0041] (5) Matrix effect By comparing the peak areas of the reference substance solutions of the three characteristic peptides of the clam species with those of the spiked matrix samples, the matrix effects of the samples at each concentration were calculated. The results showed that the matrix effects of VAE, DSW, and PSV were 100.4% - 109.1%, 94.8% - 106.5%, and 96.1% - 99.9% respectively, indicating that there was no obvious matrix ion suppression or enhancement effect interfering with the determination of the three characteristic peptides of the clam species in the samples.

[0042] (6) Recovery rate By comparing the peak areas of LQC, MQC, and HQC and the standard substances after being prepared and processed according to the test solution, the recovery rates were calculated. As shown in Table 4, the recovery rates of VAE, DSW, and PSV were 107.5% - 110.3%, 88.1% - 104.7%, and 95.0% - 102.9% respectively, indicating that the method had good accuracy and was not affected by the sample treatment process.

[0043] Table 4. Matrix effect and recovery rate of the detection method for characteristic peptides of clam species (n = 6)

[0044] (7) Accuracy and precision The analysis of accuracy and precision was carried out through intra-day and inter-day experiments at four concentrations: the lowest quantification limit (LLOQ), LQC, MQC, and HQC concentrations. As shown in Table 5, the intra-day and inter-day precisions (RSD) of the determination results of VAE, DSW, and PSV were 2.1% - 9.3%, 2.7% - 9.3%, and 1.4% - 9.3% respectively; the accuracies (RE) were -5.7% - 11.3%, -2.9% - 13.3%, and -5.5% - 3.0% respectively, indicating that the detection method had reliable accuracy and good reproducibility.

[0045] Table 5. Precision and accuracy of the detection method for characteristic peptides of clam species (n = 6)

[0046] (8) Stability Placed at the autosampler temperature for 72 h, subjected to 3 cycles of repeated freezing and thawing, and stored at -40 °C for 7 days, the accuracy (RE) of the detection results of VAE, DSW, and PSV was -7.4% to 7.9%, -1.8% to 7.1%, and -9.5% to 10.4% (Table 6), indicating that the three species-specific peptides of blood clams were stable under storage and detection conditions.

[0047] Table 6. Stability of the detection method for blood clam species-specific peptides (n = 6)

[0048] Example 3: Detection and application of blood clam species-specific peptides Twenty-seven batches of blood clam samples were collected and identified by DNA barcoding for their origins. There were 6 batches of Scapharca subcrenata, 12 batches of Scapharca broughtonii, 5 batches of Tegillarca granosa, and 4 batches with failed detection. Samples were prepared according to the method described in item (1) Preparation of test solution in Example 2 and analyzed by the LC-MS / MS method in Example 2.

[0049] As shown in Tables 7, 8, and 9, only the chromatographic peak of the Scapharca subcrenata species-specific peptide VAE appeared in all 6 batches of Scapharca subcrenata, with an average content of 0.42 μg / g; only the chromatographic peak of the Tegillarca granosa species-specific peptide DSW appeared in all 5 batches of Tegillarca granosa, with an average content of 0.10 μg / g; only the chromatographic peak of the Scapharca broughtonii species-specific peptide PSV appeared in all 12 batches of Scapharca broughtonii, with an average content of 0.27 μg / g. As Figure 6 、 Figure 7 、 Figure 8 、Figure 9 shows, among the 4 batches of samples with failed DNA barcoding detection, the Tegillarca granosa species-specific peptide DSW was detected in HWZ05 and HLYG06, the Scapharca broughtonii species-specific peptide PSV was detected in HLYG11, while neither DSW, PSV, nor VAE was detected in HYT20, indicating that HWZ05 and HLYG06 are Tegillarca granosa, HLYG11 is Scapharca broughtonii, and HYT20 is neither Tegillarca granosa, Scapharca broughtonii, nor Scapharca subcrenata.

[0050] Table 7 Contents of VAE, PSV, and DSW in Scapharca subcrenata samples (n = 3)

[0051] Table 8 Contents of VAE, PSV, and DSW in Tegillarca granosa samples (n = 3)

[0052] Table 9 Contents of VAE, PSV, and DSW in Tegillarca granosa samples (n = 3)

[0053] The above results indicate that the VAE, DSW, and PSV characteristic peptides screened in the present invention can be used as characteristic index components to accurately identify the three origins of Scapharca subcrenata, Scapharca broughtonii, and Tegillarca granosa.

[0054] The specific embodiments described above further elaborate on the object, technical solution, and beneficial effects of the present disclosure. It should be understood that the above are only specific embodiments of the present disclosure and are not intended to limit the present disclosure. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present disclosure shall be included within the protection scope of the present disclosure.

Claims

1. A clam species characteristic peptide of giant clam, hairy clam and mud clam, characterized in that: The characteristic peptides of this clam species include: The sequence of the characteristic peptide PSV of Kuihan is: PSVQGAAAQLTADVK; The sequence of the characteristic peptide VAE of C. serrata is: VAELANAVVSNADQK; The sequence of the characteristic peptide DSW of mud clam is: DSWAALGSDK.

2. Use of the clam species characteristic peptides of giant clam, hairy clam and mud clam as described in claim 1 in identifying the original species of clam medicinal materials.

3. The use according to claim 2, characterized in that The application is a method for identifying the original species of the clam medicinal material. The identification method is to detect whether the sample contains the clam species characteristic peptides of the giant clam, hairy clam and mud clam, namely PSV, VAE and DSW; the judgment principle is: If PSV is detected in the sample, and VAE and DSW are not detected, the sample to be tested is determined to be Clam; If VAE is detected in the sample, but DSW and PSV are not detected, the sample is judged to be a hairy clam. If DSW is detected in the sample, and VAE and PSV are not detected, the sample to be tested is determined to be mud cockle; If PSV, VAE and DSW are not detected in the sample, it is determined that the sample to be tested is not any of the giant clams, hairy clams and mud clams.

4. A method for identifying the original species of clams based on the clams species characteristic peptide of claim 1, characterized in that: The following steps are involved: (1) Preparation of the test solution: hydrolyze the powder of the clam medicinal material sample, centrifuge, collect the supernatant, freeze-dry, and obtain a clam enzymatic hydrolyzate freeze-dried powder; prepare a clam enzymatic hydrolyzate solution from the clam enzymatic hydrolyzate freeze-dried powder, desalt the powder, and then re-dissolve the powder to obtain a test solution; (2) Preparation of reference solution: PSV, VAE and DSW, three clam species characteristic peptides, were used as reference substances, and 0.1% formic acid water was added to dissolve them to prepare reference solution; (3) Detection and analysis: Detection and analysis are performed using liquid chromatography-triple quadrupole mass spectrometry; (4) If PSV is detected in the sample, but VAE and DSW are not detected, the sample is judged to be a giant clam; if VAE is detected in the sample, but DSW and PSV are not detected, the sample is judged to be a hairy clam; if DSW is detected in the sample, but VAE and PSV are not detected, the sample is judged to be a mud clam; if PSV, VAE and DSW are not detected in the sample, the sample is judged not to be a giant clam, hairy clam or mud clam.

5. The method according to claim 4, characterized in that In the step (1), the enzymatic hydrolysis conditions are as follows: taking the powder of the clam medicinal material sample, adding PBS buffer at a solid-liquid ratio of 1:10, adding trypsin at a final concentration of 800 U / mL, and enzymatic hydrolysis at 50°C and pH 8.0 for 5 hours; the desalting treatment is to desalt the sample using a desalting column; and the re-dissolution is to re-dissolve with 50 μL of a 0.1% formic acid aqueous solution.

6. The method according to claim 4, characterized in that The chromatographic conditions in step (3) are as follows: mobile phase A is 0.1% formic acid in water, mobile phase B is 0.1% formic acid in acetonitrile, and the flow rate is set to 0.3 mL / min; the elution gradient is as follows: 0-1 min, 95% A, 1-2 min, 95-80% A, 2-8 min, 80-60% A, 8-10 min, 60-10% A, 10-11.5 min, 10% A, 11.5-11.6 min, 10-95% A, 11.6-14 min, 95% A; the monitoring ion pairs include VAE m / z 765.3→171.1; DSW is m / z 525.5→661.4; PSV is m / z 728.9→185.1.

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