A composition for distinguishing three kinds of head oil of aquatic shrimps and application thereof

Abstract

The application discloses a kind of compositions for distinguishing three kinds of aquatic shrimp head oils and application thereof, belong to aquatic product and its product quality detection technical field.The composition is specifically: PC (24:0 / 20:4), PE (16:0 / 16:0), PS (18:0 / 18:0), PS (18:0 / 16:0), LPC (22:6), TG (16:1 / 18:2 / 18:2), MG (20:4).The application extracts and lipidomic detection to the oil of three kinds of aquatic shrimps head, and determines the composition capable of distinguishing the difference of three kinds of shrimp head oils by multivariate statistical analysis.The application takes the composition as core component, and realizes the rapid and accurate identification of three kinds of aquatic shrimp head oils by combining with lipidomic detection method.The application method is simple in operation, and the discrimination effect is remarkable, and can be applied to aquatic oil traceability, quality monitoring and true and false identification field.

Description

Technical Field

[0001] This invention belongs to the field of quality testing technology for aquatic products and their products, specifically relating to a composition for distinguishing the head oil of whiteleg shrimp, giant freshwater prawn, and red swamp crayfish, and its application. Background Technology

[0002] Litopenaeus vannamei, Macrobrachium rosenbergii, and Procambarus clarkii are important economic shrimp species in my country, with their aquaculture and processing scale continuously growing. Shrimp heads are a major by-product of shrimp processing, and realizing their resource utilization is a key link in promoting the sustainable development of the industry. Extracting shrimp oil is a feasible solution. Shrimp head oil is rich in various bioactive lipids, possessing good nutritional value and market potential. However, shrimp head oil from different shrimp species differs in function and characteristics, and its application should also vary. During the distribution process, the rapid and accurate differentiation and identification of shrimp head oil from different shrimp species is of great significance for ensuring product quality, preventing adulteration, and promoting its differentiated and high-value utilization.

[0003] Traditional chemical analysis methods are cumbersome to operate and lack specificity, making it difficult to effectively identify shrimp oil species and hindering the high-value development and product traceability management of shrimp oil. Summary of the Invention

[0004] One objective of this invention is to provide a lipid molecular composition, and another objective is to provide its application in order to effectively distinguish the head oil of whiteleg shrimp, giant freshwater prawn and red swamp crayfish, thereby overcoming the shortcomings of the prior art.

[0005] Based on lipidomics data, this invention identifies the lipid composition of shrimp head oil from Litopenaeus vannamei, Macrobrachium rosenbergii, and Procambarus clarkii at the molecular and species level, screens and determines characteristic lipid biomarkers, and establishes a rapid and accurate method for identifying shrimp head oil species.

[0006] To achieve the above objectives, the technical solution provided by the present invention is as follows:

[0007] A composition for distinguishing the head oil of three types of aquatic shrimp, the composition comprising icosicocarbonate-eicosatraenoic acid-phosphatidylcholine (PC(24:0 / 20:4)), hexadecanoic acid-hexadecanoic acid-phosphatidylethanolamine (PE(16:0 / 16:0)), octadecanoic acid-octadecanoic acid-phosphatidylserine (PS(18:0 / 18:0)), octadecanoic acid-hexadecanoic acid-phosphatidylserine (PS(18:0 / 16:0)), docosahexaenoic acid-lysophosphatidylcholine (LPC(22:6)), hexadecanoic acid-octadecadienoic acid-octadecadienoic acid-triglyceride (TG(16:1 / 18:2 / 18:2)), and icosicocarbonate-monoglyceride (MG(20:4)).

[0008] The composition is used in the traceability, quality control, and adulteration identification of the head oil of Litopenaeus vannamei, Macrobrachium rosenbergii, and Procambarus clarkii.

[0009] The method for detecting the head oil of Litopenaeus vannamei, Macrobrachium rosenbergii, and Procambarus clarkii, using the composition, includes the following steps: (1) Take three kinds of fresh shrimp head samples; (2) Extract oil from three types of shrimp head samples to obtain oil extracts; (3) Pretreatment of the oil extract; (4) Analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS / MS); (5) Principal component analysis was performed to distinguish different shrimp head oils based on the relative abundance (RA) of the seven lipid molecules contained in the composition.

[0010] Preferably, in step (1), the shrimp head samples of the three types of shrimp are homogenized to prepare uniform tissue samples, with each sample weighing 50g to 5000g.

[0011] Preferably, in step (2), the tissue sample is added to 12 to 36 times the volume of chloroform-methanol solution (2:1, v / v), extracted at room temperature for 6 to 24 hours, filtered, and the filtrate is collected. Then, 1 / 4 volume of 0.88% potassium chloride solution is added, mixed thoroughly, and allowed to stand at room temperature overnight. The lower phase is collected, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate is collected and desolventized by rotary evaporation to obtain the oil extract.

[0012] Preferably, in step (3), 50 mg of the oil extract is accurately weighed into a 2 mL centrifuge tube, and a 6 mm diameter grinding bead is added. Then, 280 µL of 30% methanol solution and 400 µL of methyl tert-butyl ether are added to the sample in sequence, mixed, and ground in a cryogenic grinder for 6 min (-10℃, 50 Hz). After low-temperature ultrasonic extraction for 30 min (5℃, 40 kHz), the sample is allowed to stand at -20℃ for 30 min, centrifuged at high speed for 15 min (13000 g, 4℃), and 350 µL of the supernatant is transferred to a centrifuge tube, dried under nitrogen, and reconstituted with 100 µL of isopropanol-acetonitrile solution (1:1, v / v). After vortexing for 30 s, the sample is sonicated for 5 min (ice bath, 40 kHz), centrifuged at high speed for 10 min (13000 g, 4℃), and the supernatant is transferred to a brown sample vial for LC-MS / MS analysis.

[0013] Preferably, the LC-MS / MS method in step (4) is as follows: Chromatographic conditions: injection volume is 2 µL, column is Accucore C30 column (100 mm × 2.1 mm id, 2.6 µm), column temperature is 40℃, mobile phase A is 50% acetonitrile aqueous solution (containing 10 mM ammonium acetate, 0.1% formic acid), mobile phase B is 2 mM ammonium acetate acetonitrile / isopropanol / water (10 / 88 / 2, v / v / v, containing 0.02% formic acid), flow rate is 0.40 mL / min, gradient elution program is: 0 min A to B volume ratio 65:35, 3 min A to B volume ratio 40:60, 10 min A to B volume ratio 15:85, 12.5 min A to B volume ratio 0:100, 13.9 min A to B volume ratio 0:100, 14 min A to B volume ratio 65:35, 16 ... The elution program ended at min. Mass spectrometry conditions: positive and negative ion scanning modes, scan range m / z 200~2000, positive mode ion spray voltage 3000 V, negative mode ion spray voltage -3000 V, sheath gas 60 arb, auxiliary heating gas 20 arb, ion source heating temperature 370℃, cyclic collision energy 20 V-40 V-60 V, data acquisition using DDA mode.

[0014] Preferably, in step (5), the relative abundances of seven lipid molecules—PC (24:0 / 20:4), PE (16:0 / 16:0), PS (18:0 / 18:0), PS (18:0 / 16:0), LPC (22:6), TG (16:1 / 18:2 / 18:2), and MG (20:4)—are used as the basis for the principal component formula: PC1 = 0.042 × RA PC(24:0 / 20:4) +0.000×RA PE(16:0 / 16:0)+0.236×RA PS(18:0 / 18:0) +0.215×RA PS(18:0 / 16:0) +0.241×RA LPC(22:6) -0.215×RA TG(16:1 / 18:2 / 18:2) +0.219×RA MG(20:4) PC2 = 0.354 × RA PC(24:0 / 20:4) +0.353×RA PE(16:0 / 16:0) -0.115×RA PS(18:0 / 18:0) +0.165×RA PS(18:0 / 16:0) -0.109×RA LPC(22:6) -0.181×RA TG(16:1 / 18:2 / 18:2) -0.163×RA MG(20:4) Principal component analysis was performed to distinguish the oils in different shrimp heads;

[0015] Among them, the oil samples from the heads of Litopenaeus vannamei were distributed in the region of 1.0 < PC1 < 3.0 and 0.5 < PC2 < 2.0; the oil samples from the heads of Macrobrachium rosenbergii were distributed in the region of -1.0 < PC1 < 1.0 and -3.0 < PC2 < -1.5; and the oil samples from the heads of Procambarus clarkii were distributed in the region of -3.0 < PC1 < -1.5 and 0 < PC2 < 2.0.

[0016] Compared with the prior art, the beneficial effects of the present invention are:

[0017] This invention, for the first time, screened seven key biomarkers at the lipid molecule level that can accurately and stably distinguish the head oils of Litopenaeus vannamei, Macrobrachium rosenbergii, and Procambarus clarkii. The limited number of biomarkers and their high specificity avoid the high cost and data processing redundancy of full-spectrum analysis. Principal component analysis based on the composition of these seven lipid molecules enables clear and accurate separation of the head oils from these three shrimp species. This invention can be applied to quality control, authenticity verification, and origin traceability of shrimp by-product oils. It can also support the refined development and high-value utilization of these oils in the fields of general food, health food, and biopharmaceuticals, improving the economic benefits and resource utilization of the shrimp processing industry. Furthermore, this invention has strong scalability and can provide technical reference for the identification and quality control of other aquatic oils. Attached Figure Description

[0018] Figure 1 This is a diagram showing the main lipid subclass composition of the head oil from the three types of aquatic shrimp in Example 1.

[0019] Figure 2This is a volcano diagram of the differential lipid molecules in the head oil of three types of aquatic shrimp in Example 1; where (a) is the head oil of Litopenaeus vannamei vs. head oil of Macrobrachium rosenbergii, (b) is the head oil of Litopenaeus vannamei vs. head oil of Procambarus clarkii, and (c) is the head oil of Macrobrachium rosenbergii vs. head oil of Procambarus clarkii.

[0020] Figure 3 The diagram shows the KEGG metabolic pathway topology analysis of the head oil of three types of aquatic shrimp in Example 1; where (a) is the KEGG topology analysis (head oil of Litopenaeus vannamei vs. head oil of Macrobrachium rosenbergii), (b) is the KEGG topology analysis (head oil of Litopenaeus vannamei vs. head oil of Procambarus clarkii), and (c) is the KEGG topology analysis (head oil of Macrobrachium rosenbergii vs. head oil of Procambarus clarkii).

[0021] Figure 4 This is a heatmap comparing the relative abundance of the composition in the head oil of three types of aquatic shrimp in Example 1.

[0022] Figure 5 This is a principal component analysis diagram of the shrimp head oil of three types of aquatic shrimp in Example 1, based on the composition.

[0023] Figure 6 This is a heatmap comparing the relative abundance of the composition in the head oil of three types of aquatic shrimp in Example 2.

[0024] Figure 7 This is a principal component analysis diagram of shrimp head oil in Example 2, which distinguishes three types of aquatic shrimp based on the composition. Detailed Implementation

[0025] The technical solution of the present invention will be further described and illustrated below with reference to the embodiments.

[0026] Example 1: In this embodiment, 5000 g each of fresh whiteleg shrimp, giant freshwater prawn, and red swamp crayfish were collected from the Qingdao Aquatic Products Market in Shandong Province, China.

[0027] The three types of fresh shrimp samples were separated into cephalothoraxes according to anatomical location and homogenized to prepare uniform shrimp head tissue samples. Each group was divided into six parallel experimental groups (i.e., each experimental group was no less than 200 g).

[0028] Oil extraction was performed on three types of shrimp head samples. 12 times the volume of chloroform-methanol solution (2:1, v / v) was added, and the samples were extracted at room temperature for 6 h. After filtration, 1 / 4 volume of 0.88% potassium chloride solution was added to the filtrate. The mixture was thoroughly mixed and allowed to stand at room temperature overnight. The lower phase was collected, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was collected and desolvated by rotary evaporation to obtain the oil extract.

[0029] Accurately weigh 50 mg of the oil extract into a 2 mL centrifuge tube, add a 6 mm diameter grinding bead; then add 280 µL of 30% methanol solution and 400 µL of methyl tert-butyl ether to the sample, mix well, grind in a cryo-tissue homogenizer for 6 min (-10℃, 50 Hz), extract by low-temperature sonication for 30 min (5℃, 40 kHz), let stand at -20℃ for 30 min, centrifuge at high speed for 15 min (13000 g, 4℃), transfer 350 µL of supernatant into a centrifuge tube, dry under nitrogen, add 100 µL of isopropanol-acetonitrile solution (1:1, v / v) to reconstitute, vortex mix for 30 s, sonicate for 5 min (ice bath, 40 kHz), centrifuge at high speed for 10 min (13000 g, 4℃), transfer the supernatant into a brown sample vial for LC-MS / MS analysis.

[0030] LC-MS / MS Method: Chromatographic conditions: injection volume 2 µL, column Accucore C30 column (100 mm × 2.1 mm id, 2.6 µm), column temperature 40℃, mobile phase A 50% acetonitrile aqueous solution (containing 10 mM ammonium acetate, 0.1% formic acid), mobile phase B 2 mM ammonium acetate acetonitrile / isopropanol / water (10 / 88 / 2, v / v / v, containing 0.02% formic acid), flow rate 0.40 mL / min, gradient elution program as shown in Table 1, specifically: 0 min A:B volume ratio 65:35, 3 min A:B volume ratio 40:60, 10 min A:B volume ratio 15:85, 12.5 min A:B volume ratio 0:100, 13.9 min A:B volume ratio 0:100, 14 min A:B volume ratio 65:35, elution program ends at 16 min. Mass spectrometry conditions: positive and negative ion scanning modes, scanning range of m / z 200~2000, positive mode ion spray voltage of 3000 V, negative mode ion spray voltage of -3000 V, sheath gas of 60 arb, auxiliary heating gas of 20 arb, ion source heating temperature of 370℃, cyclic collision energy of 20 V-40 V-60 V, data acquisition using DDA mode.

[0031] Table 1. Gradient elution program for mobile phase

[0032] Results analysis: Lipidomics analysis was performed on the head oil of Litopenaeus vannamei, Macrobrachium rosenbergii, and Procambarus clarkii. Results are shown below. Figure 1A total of 4,588 lipid molecules were identified, mainly divided into 5 categories: 2,487 glycerophospholipids (GP), 1,377 glycerol esters (GL), 588 sphingolipids (SP), 104 fatty acyls (FA), and 32 sterol esters (ST).

[0033] Based on the lipid molecular species composition data of the head oil of Litopenaeus vannamei, Macrobrachium rosenbergii, and Procambarus clarkii, multivariate statistical analysis was performed. Using P < 0.05 and VIP PLS-DA > 1 as criteria, molecules with significant differences in lipid molecular species composition among different shrimp heads were screened. Volcano plots were obtained through pairwise comparisons. (See figure) Figure 2 In the comparison of the head oil of Litopenaeus vannamei and Macrobrachium rosenbergii, a total of 1704 differential lipid molecules were identified; in the comparison of the head oil of Litopenaeus vannamei and Procambarus clarkii, a total of 1717 differential lipid molecules were identified; and in the comparison of the head oil of Macrobrachium rosenbergii and Procambarus clarkii, a total of 1684 differential lipid molecules were identified.

[0034] KEGG pathway enrichment analysis was performed on significantly different lipid molecules in the head oil of three types of shrimp. The identified differentially expressed lipid molecules were uploaded to the KEGG database to collect relevant information, and topological analysis of the metabolic pathways was conducted. The results are shown in [Figure 1]. Figure 3 The main KEGG pathways involved in the differentially expressed lipid molecules in the head oil of the three shrimp species include glycerophospholipid metabolism, glycerol ester metabolism, linoleic acid metabolism, sphingolipid metabolism, and α-linolenic acid metabolism. Further screening of key differentially expressed lipid molecules involved in multiple important metabolic pathways yielded key differential markers that can be used to distinguish the head oil of the three aquatic shrimp species, including: PC (24:0 / 20:4) (tetracosuccinic acid-eicosatetraenoic acid-phosphatidylcholine) and PE (16:0 / 16:0) (hexadecanoic acid-hexadecanoic acid-phosphatidylethanolamine). PS(18:0 / 18:0) (octadecanoic acid-octadecanoic acid-phosphatidylserine), PS(18:0 / 16:0) (octadecanoic acid-hexadecanoic acid-phosphatidylserine), LPC(22:6) (docosahexaenoic acid-lysophosphatidylcholine), TG(16:1 / 18:2 / 18:2) (hexadecanoic acid-octadecadienoic acid-octadecadienoic acid-triglycerides), MG(20:4) (eicosatraenoic acid-monoglycerides). A heatmap comparing the relative abundance of these seven lipid molecules in the head oil of three types of shrimp is shown below. Figure 4 Principal component analysis was performed based on the relative abundance of the selected compositions, and the results are shown in [Figure number missing]. Figure 5The head oil of the three types of shrimp was clearly separated and distributed in different regions of the two-dimensional spectrum. Specifically, the head oil sample of Litopenaeus vannamei was distributed in the region of 1.0 < PC1 < 3.0 and 0.5 < PC2 < 2.0; the head oil sample of Macrobrachium rosenbergii was distributed in the region of -1.0 < PC1 < 1.0 and -3.0 < PC2 < -1.5; and the head oil sample of Procambarus clarkii was distributed in the region of -3.0 < PC1 < -1.5 and 0 < PC2 < 2.0. Using a composition containing the seven key markers, the source of head oil in the three types of shrimp can be accurately distinguished.

[0035] Example 2: In this embodiment, 3000 g each of live whiteleg shrimp, giant freshwater prawn, and red swamp crayfish were purchased from the seafood market.

[0036] The three types of fresh shrimp samples were separated into cephalothoraxes according to anatomical location and homogenized to prepare uniform shrimp head tissue samples. Each group was divided into six parallel experimental groups (i.e., each experimental group was no less than 100 g).

[0037] Oil extraction was performed on three types of shrimp head samples. 12 times the volume of chloroform-methanol solution (2:1, v / v) was added, and the samples were extracted at room temperature for 6 h. After filtration, 1 / 4 volume of 0.88% potassium chloride solution was added to the filtrate. The mixture was thoroughly mixed and allowed to stand at room temperature overnight. The lower phase was collected, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was collected and desolvated by rotary evaporation to obtain the oil extract.

[0038] Accurately weigh 50 mg of the oil extract into a 2 mL centrifuge tube, add a 6 mm diameter grinding bead; then add 280 µL of 30% methanol solution and 400 µL of methyl tert-butyl ether to the sample, mix well, grind in a cryo-tissue homogenizer for 6 min (-10℃, 50 Hz), extract by low-temperature sonication for 30 min (5℃, 40 kHz), let stand at -20℃ for 30 min, centrifuge at high speed for 15 min (13000 g, 4℃), transfer 350 µL of supernatant into a centrifuge tube, dry under nitrogen, add 100 µL of isopropanol-acetonitrile solution (1:1, v / v) to reconstitute, vortex mix for 30 s, sonicate for 5 min (ice bath, 40 kHz), centrifuge at high speed for 10 min (13000 g, 4℃), transfer the supernatant into a brown sample vial for LC-MS / MS analysis.

[0039] LC-MS / MS Method: Chromatographic conditions: injection volume 2 µL, column Accucore C30 column (100 mm × 2.1 mm id, 2.6 µm), column temperature 40℃, mobile phase A 50% acetonitrile aqueous solution (containing 10 mM ammonium acetate, 0.1% formic acid), mobile phase B 2 mM ammonium acetate acetonitrile / isopropanol / water (10 / 88 / 2, v / v / v, containing 0.02% formic acid), flow rate 0.40 mL / min, gradient elution program as shown in Table 1, specifically: 0 min A:B volume ratio 65:35, 3 min A:B volume ratio 40:60, 10 min A:B volume ratio 15:85, 12.5 min A:B volume ratio 0:100, 13.9 min A:B volume ratio 0:100, 14 min A:B volume ratio 65:35, elution program ends at 16 min. Mass spectrometry conditions: positive and negative ion scanning modes, scanning range of m / z 200~2000, positive mode ion spray voltage of 3000 V, negative mode ion spray voltage of -3000 V, sheath gas of 60 arb, auxiliary heating gas of 20 arb, ion source heating temperature of 370℃, cyclic collision energy of 20 V-40 V-60 V, data acquisition using DDA mode.

[0040] Results analysis:

[0041] The relative abundances of the following lipid molecules in the head oil of Litopenaeus vannamei, Macrobrachium rosenbergii, and Procambarus clarkii were determined: PC (24:0 / 20:4) (tetracosanoic acid-eicosatraenoic acid-phosphatidylcholine), PE (16:0 / 16:0) (hexadecanoic acid-hexadecanoic acid-phosphatidylethanolamine), PS (18:0 / 18:0) (octadecanoic acid-octadecanoic acid-phosphatidylserine), PS (18:0 / 16:0) (octadecanoic acid-hexadecanoic acid-phosphatidylserine), LPC (22:6) (docosahexaenoic acid-lysophosphatidylcholine), TG (16:1 / 18:2 / 18:2) (hexadecanoic acid-octadecadienoic acid-octadecadienoic acid-triglyceride), and MG (20:4) (eicosatraenoic acid-monoglyceride). A heatmap comparing the relative abundances of these seven lipid molecules in the head oil of the three shrimp species is shown below. Figure 6 The relative abundance results of the seven lipid molecules were substituted into the principal component formula (PC1 = 0.042 × RA). PC(24:0 / 20:4) +0.000×RA PE(16:0 / 16:0) +0.236×RA PS(18:0 / 18:0) +0.215×RA PS(18:0 / 16:0) +0.241×RA LPC(22:6) -0.215×RA TG(16:1 / 18:2 / 18:2) +0.219×RA MG(20:4) PC2 = 0.354 × RAPC(24:0 / 20:4) +0.353×RA PE(16:0 / 16:0) -0.115×RA PS(18:0 / 18:0) +0.165×RA PS(18:0 / 16:0) -0.109×RA LPC(22:6) -0.181×RA TG(16:1 / 18:2 / 18:2) -0.163×RA MG(20:4) The analysis was performed, and the results are shown below. Figure 7 The head oil of the three types of shrimp was clearly separated and distributed in different regions of the two-dimensional spectrum. Specifically, the head oil sample of Litopenaeus vannamei was distributed in the region of 1.0 < PC1 < 3.0 and 0.5 < PC2 < 2.0; the head oil sample of Macrobrachium rosenbergii was distributed in the region of -1.0 < PC1 < 1.0 and -3.0 < PC2 < -1.5; and the head oil sample of Procambarus clarkii was distributed in the region of -3.0 < PC1 < -1.5 and 0 < PC2 < 2.0. Using a composition containing the seven key markers, the source of head oil in the three types of shrimp can be accurately distinguished.

[0042] Example 3: The application of the composition of the seven key biomarkers obtained in Example 1, specifically PC (24:0 / 20:4) (tetracosuccinic acid-eicosapentaenoic acid-phosphatidylcholine), PE (16:0 / 16:0) (hexadecanoic acid-hexadecanoic acid-phosphatidylethanolamine), PS (18:0 / 18:0) (octadecanoic acid-octadecanoic acid-phosphatidylserine), PS (18:0 / 16:0) (octadecanoic acid-hexadecanoic acid-phosphatidylserine), LPC (22:6) (docosahexaenoic acid-lysophosphatidylcholine), TG (16:1 / 18:2 / 18:2) (hexadecanoic acid-octadecadienoic acid-octadecadienoic acid-triglyceride), and MG (20:4) (eicosapentaenoic acid-monoglyceride), specifically includes:

[0043] The composition can be applied to the traceability identification, quality monitoring, and adulteration detection of oil from the heads of whiteleg shrimp, giant freshwater prawn, and red swamp crayfish.

[0044] This invention not only provides a reliable scientific basis for quality control, authenticity identification, and origin traceability of oils derived from shrimp by-products, but also promotes their refined development and high-value utilization in the fields of general food, health food, and biopharmaceutical products, thereby improving the economic benefits and resource utilization rate of the shrimp processing industry. Furthermore, this invention has strong scalability and can provide technical reference for the identification and quality control of other aquatic oils.

[0045] The embodiments described above are merely preferred embodiments of the present invention and do not constitute a limitation on the scope of the claims. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be within the protection scope of the present invention.

[0046] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the scope of the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A composition for distinguishing the head oil of three types of aquatic shrimp, characterized in that, The composition comprises: icosicocarbonate-eicosatraenoic acid-phosphatidylcholine, hexadecanoate-hexadecanoate-phosphatidylethanolamine, octadecanoate-octadecanoate-phosphatidylserine, octadecanoate-hexadecanoate-phosphatidylserine, docosahexaenoic acid-lysophosphatidylcholine, hexadecanoic acid-octadecadienoic acid-octadecadienoic acid-triglyceride, and icosicocarbonate-monoglyceride; the three aquatic shrimp species are Litopenaeus vannamei, Macrobrachium rosenbergii, and Procambarus clarkii.

2. The application of the composition according to claim 1 in the traceability identification, quality control and adulteration identification of the head oil of Litopenaeus vannamei, Macrobrachium rosenbergii and Procambarus clarkii.

3. The method for detecting the head oil of Litopenaeus vannamei, Macrobrachium rosenbergii, and Procambarus clarkii using the composition of claim 1, characterized in that, Includes the following steps: (1) Take three kinds of fresh shrimp head samples; (2) Extract oil from three types of shrimp head samples to obtain oil extracts; (3) Pretreatment of the oil extract; (4) Analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS / MS); (5) Principal component analysis was performed on the relative abundance (RA) of the seven lipid molecules contained in the composition to distinguish different shrimp head oils.

4. The detection method as described in claim 3, characterized in that, In step (1), the shrimp head samples of the three types of shrimp were homogenized.

5. The detection method as described in claim 3, characterized in that, In step (2), the tissue sample is added to 12 to 36 times the volume of chloroform-methanol solution, and extracted at room temperature for 6 to 24 hours. After filtration, the filtrate is collected and potassium chloride solution is added. After thorough mixing, the mixture is left to stand at room temperature overnight. The lower phase is collected, dehydrated by anhydrous sodium sulfate, filtered, and the filtrate is collected and desolventized by rotary evaporation to obtain the oil extract.

6. The detection method as described in claim 3, characterized in that, In step (3), the oil extract is ground, subjected to low-temperature ultrasonic extraction and allowed to stand, centrifuged at high speed, the supernatant is dried, isopropanol-acetonitrile solution is added to redissolve, mixed, ultrasonicated, centrifuged at high speed, and the supernatant is taken for LC-MS / MS analysis.

7. The detection method as described in claim 3, characterized in that, In step (4), the LC-MS / MS method used was as follows: Chromatographic conditions: injection volume was 2... µ L, the chromatographic column was an Accucore C30 column, the column temperature was 40℃, mobile phase A was 50% acetonitrile aqueous solution containing 10 mM ammonium acetate and 0.1% formic acid; mobile phase B was a mixed solution of 2 mM ammonium acetate acetonitrile, isopropanol and water, with a volume ratio of 10:88:2, containing 0.02% formic acid; the flow rate was 0.40 mL / min; the gradient elution program was as follows: 0 min A:B volume ratio 65:35, 3 min A:B volume ratio 40:60, 10 min A:B volume ratio 15:85, 12.5 min A:B volume ratio 0:100, 13.9 min A:B volume ratio 0:100, 14 min A:B volume ratio 65:35, and the elution program ended at 16 min.

8. The detection method as described in claim 3, characterized in that, In step (5), based on the relative abundance of seven lipid molecules—PC (24:0 / 20:4), PE (16:0 / 16:0), PS (18:0 / 18:0), PS (18:0 / 16:0), LPC (22:6), TG (16:1 / 18:2 / 18:2), and MG (20:4)—the formula is substituted into the principal component equation: PC1 = 0.042 × RA PC(24:0 / 20:4) +0.000×RA PE(16:0 / 16:0) +0.236×RA PS(18:0 / 18:0) +0.215×RA PS(18:0 / 16:0) +0.241×RA LPC(22:6) -0.215×RA TG(16:1 / 18:2 / 18:2) +0.219×RA MG(20:4) ; PC2 = 0.354 × RA PC(24:0 / 20:4) +0.353×RA PE(16:0 / 16:0) -0.115×RA PS(18:0 / 18:0) +0.165×RA PS(18:0 / 16:0) -0.109×RA LPC(22:6) -0.181×RA TG(16:1 / 18:2 / 18:2) -0.163×RA MG(20:4) Principal component analysis was performed to distinguish the oil from different shrimp heads: the oil samples from Litopenaeus vannamei were distributed in the regions of 1.0 < PC1 < 3.0 and 0.5 < PC2 < 2.0; the oil samples from Macrobrachium rosenbergii were distributed in the regions of -1.0 < PC1 < 1.0 and -3.0 < PC2 < -1.5; and the oil samples from Procambarus clarkii were distributed in the regions of -3.0 < PC1 < -1.5 and 0 < PC2 < 2.0.

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