Microscopic identification method for American cockroach powder

Abstract

The invention relates to the technical field of periplaneta americana powder microscopic flaking, and discloses a periplaneta americana powder microscopic identification method which comprises the following steps: S1, sample degreasing: dividing a periplaneta americana adult into five parts, namely a head part, a chest back plate, wings, an abdomen part and chest feet by using tweezers, respectively grinding into powder, taking 100g of powder of each part of the periplaneta americana, and uniformly mixing the powder with the powder of each part of the periplaneta americana; respectively filling the materials into a filter paper tube, putting the filter paper tube into an extraction tube of a Soxhlet extractor, soaking the American cockroach powder with an extracting solution, and removing grease; s2, sample tabletting: taking the degreased powder, respectively adding distilled water, chloral hydrate and dilute glycerol for tabletting, and preparing a water-loaded tablet, a chloral hydrate cold-loaded tablet and a chloral hydrate permeabilized tablet; s3, microscopic observation: placing the prepared loading sheet under an optical microscope, and carrying out image acquisition and analysis on powder characteristics by using image processing software equipped for the microscope; and S4, identification standard. Compared with the prior art, the method has the advantages that grease is convenient to remove, the observation definition and accuracy are improved, a microscopic identification standard is established, and an identification system is perfected.

Description

Technical Field

[0001] The invention relates to the technical field of microscopic identification of American cockroach powder, in particular to a microscopic identification method of American cockroach powder. Background Art

[0002] The American cockroach, a traditional Chinese medicinal herb, possesses diverse pharmacological activities, including promoting blood circulation and dispersing blood stasis, detoxifying and relieving malnutrition, promoting tissue repair, and modulating immunity. However, due to its propensity to feed on oily substances, its body fat content is significantly elevated (adult cockroaches typically contain over 8% fat). This results in directly pulverized powders with significant oil interference, prone to clumping, and blurred microscopic features, seriously compromising the accuracy and stability of microscopic observations.

[0003] Currently, the treatment of American cockroaches primarily focuses on optimizing the degreasing process to reduce oil toxicity and improve medicinal safety. However, existing patents and standards fail to address the microscopic identification of American cockroach powder: there are no national or local microscopic identification standards, nor is there systematic research on the characteristics of different anatomical parts (such as the head, wings, and abdomen).

[0004] Therefore, it is urgent to establish a microscopic identification method based on fine processing of anatomical parts to overcome the observation obstacles caused by high oil content and provide a reliable basis for the quality control of this medicinal material. Summary of the Invention

[0005] The technical problem to be solved by the present invention is that in the traditional preparation of American cockroach slices, due to the oil in the slices, particles stick together and the visual field is blurred during microscopic structure observation, which affects the clarity and accuracy of observation. In addition, the local standard for American cockroaches only has trait identification and thin layer identification methods, and lacks standards for microscopic identification.

[0006] To solve the above technical problems, the present invention provides a technical solution: a method for microscopic identification of American cockroach powder, comprising the following steps:

[0007] S1. Sample defatting: Use tweezers to separate the American cockroach adult into five parts: head, pronotum, wings, abdomen, and thoracic legs. Powder each part separately. Take 100 g of the powder from each part of the American cockroach, load it into a filter paper tube and place it in the extraction tube of a Soxhlet extractor. Use the extracting solution to soak the American cockroach powder and dissolve the oil. When the extract in the extraction tube reaches the height of the siphon, it automatically flows back to the flask with the oil. Reflux and defatting for 3-4 hours until the extract is colorless. Stop heating, remove the powder, and evaporate the extract.

[0008] S2. Sample preparation: Take the above-mentioned defatted powder that has passed through a No. 5 sieve, add distilled water, chloral hydrate, and dilute glycerin, respectively, and prepare water-mounted slides, chloral hydrate cold-mounted slides, and chloral hydrate permeabilized slides;

[0009] S3. Microscopic observation: Place the prepared slide under an optical microscope, use an objective lens to observe the microstructure from low to high, and use the image processing software equipped with the microscope to collect and analyze the powder characteristics;

[0010] S4. Identification criteria.

[0011] Furthermore, the extract in S1 is obtained by adding 300 ml of petroleum ether to a round-bottom flask, heating the flask in a water bath at 50-60° C. to boil the petroleum ether, and condensing the steam through a condenser to obtain the extract.

[0012] Furthermore, the reflux method used in S1 is Soxhlet extraction reflux.

[0013] Furthermore, the identification criteria observed in S4 are as follows:

[0014] Setae: There are two types: thick and thin. The thick setae are oblong-conical, straight or slightly curved, dark brown or tan, often broken, with a sub-rounded base, slightly swollen, and blunt-rounded tips. The thin setae are lanceolate or needle-shaped, straight or slightly curved, yellow or yellow-brown, with sharp tips, smooth surface, and no texture.

[0015] Wing fragments: rare, light yellow, yellow-brown or yellow-green, with dense scaly or wavy texture on the surface;

[0016] Striated muscle fibers: common, brownish yellow or colorless and transparent, with fine and dense horizontal stripes, sometimes scattered in cross, vertical, or spiral patterns;

[0017] Body wall fragments: colorless, yellow, light yellow or yellow-brown, with reticular, hexagonal or fish-scale textures, sometimes with circular pores, the edges of which are darker in color; yellow-brown or yellow-brown body wall fragments, with a smooth surface and dotted white dot-like textures, and transparent dot-like hair pits;

[0018] Compound eye fragments: occasionally seen, irregular blocks, with typical ommatidia structure, ommatidia are circular or polygonal, arranged closely, like a honeycomb.

[0019] The advantages of the present invention compared with the prior art are:

[0020] This application utilizes the principle of reflux extraction of fat-soluble components by organic solvents, adopts a Soxhlet extractor, and uses petroleum ether as a solvent to fully remove the oil from American cockroach powder through reflux degreasing, thereby reducing the obstruction of microstructure by oil, improving the clarity of microscopic observation, and providing standardized technical support for its microscopic research and quality analysis.

[0021] This application, through its innovative "partial fine powdering and observation" strategy, breaks through the bottleneck of traditional microscopic identification of whole powders and establishes a highly accurate, systematic, exclusive, and reproducible standard system for microscopic identification of American cockroaches. This not only greatly enhances the ability and reliability of authenticity and adulteration detection, providing a powerful tool for quality control and standardization, but also establishes significant technical barriers and has significant commercial value and application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The present invention provides microscopic images of defatted and non-defatted parts of American cockroach powder according to a microscopic identification method of the present invention.

[0023] Figure 2 The present invention provides a microscopic image of fine bristles in a method for microscopic identification of American cockroach powder according to the present invention.

[0024] Figure 3 The present invention provides a microscopic image of coarse setae in a method for microscopic identification of American cockroach powder according to the present invention.

[0025] Figure 4 The present invention provides a microscopic image of wing fragments used in a method for microscopic identification of American cockroach powder.

[0026] Figure 5 The present invention provides a microscopic image of striated muscle fibers in a method for microscopic identification of American cockroach powder according to the present invention.

[0027] Figure 6 The present invention provides a microscopic image of body wall fragments used in a method for microscopic identification of American cockroach powder.

[0028] Figure 7 The present invention provides a microscopic image of compound eye fragments used in a method for microscopic identification of American cockroach powder. DETAILED DESCRIPTION

[0029] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0030] A method for microscopic identification of American cockroach powder comprises the following steps:

[0031] S1. Sample defatting: Use tweezers to separate the American cockroach adult into five parts: head, pronotum, wings, abdomen, and thoracic legs. Powder each part separately. Take 100g of each powder of the American cockroach part, load it into filter paper tubes and place it in the extraction tube of a Soxhlet extractor; add 300ml of petroleum ether to a round-bottom flask, heat it in a water bath at 50-60°C, boil the petroleum ether, and condense the steam through a condenser to obtain an extract. Use the extract to soak the American cockroach powder and dissolve the oil. When the extract in the extraction tube reaches the height of the siphon, it automatically refluxes the oil back to the flask through the Soxhlet extraction reflux mode. Reflux and defatting for 3-4 hours until the extract is colorless. Stop heating, remove the powder, and evaporate the extract;

[0032] S2. Sample preparation: Take the defatted powder that has passed through a No. 5 sieve, add 1-2 drops of distilled water, chloral hydrate, and dilute glycerol, respectively, to prepare water-mounted slides, chloral hydrate cold-mounted slides, and chloral hydrate permeabilized slides;

[0033] S3. Microscopic observation: Place the prepared slide under an optical microscope, use the objective lens to observe the microstructure from low to high, and use the image processing software equipped with the microscope to collect and analyze the powder characteristics. Figure 1 As shown in the figure, after degreasing, the interference of oil is eliminated, the powder is prevented from clumping and sticking, the dispersion is improved, and the key features such as body wall patterns, hair pits, punctures and glands are made clearer; at the same time, the influence of oil oxidation and deformation is reduced, ensuring that the microscopic identification results are repeatable and accurate.

[0034] S4. Identification criteria:

[0035] As attached Figure 2 and attached Figure 3 As shown, the bristles are of two thicknesses: the coarse setae are oblong-conical, straight or slightly curved, dark brown or tan, mostly broken, with a sub-circular base, slightly swollen, and blunt tip; the fine setae are lanceolate or needle-shaped, straight or slightly curved, yellow or yellow-brown, with a sharp tip, a smooth surface, and no texture.

[0036] As attached Figure 4 As shown, wing fragments: rare, light yellow, yellow-brown or yellow-green, with dense scaly or wavy textures on the surface.

[0037] As attached Figure 5 As shown, striated muscle fibers are common, brown-yellow or colorless and transparent, with densely arranged horizontal stripes, sometimes scattered in a cross, vertical, or spiral pattern.

[0038] As attached Figure 6 As shown, body wall fragments are: colorless, yellow, light yellow or yellow-brown, with reticular, hexagonal or fish-scale textures, sometimes with circular wall pores, and the edges of the wall pores are darker in color; yellow-brown or yellowish-brown body wall fragments have a smooth surface with dotted white dot-like textures, and transparent dot-like hair pits can be seen.

[0039] As attached Figure 7 As shown, compound eye fragments: occasionally seen, irregular blocks, with typical eyelet structure, the eyelets are circular or polygonal, arranged closely, like a honeycomb.

[0040] It should be noted that, in this document, relational terms such as first and second, etc., are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprises," "comprising," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a list of elements includes not only those elements but also other elements not explicitly listed, or elements inherent to such process, method, article, or apparatus.

[0041] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

[0042] The present invention and its embodiments are described above. This description is not restrictive. The drawings show only one embodiment of the present invention, and the actual structure is not limited thereto. In short, if a person skilled in the art is inspired by this and, without departing from the purpose of the present invention, designs structures and embodiments similar to this technical solution without inventiveness, they shall fall within the scope of protection of the present invention.

Claims

1. A method for microscopic identification of American cockroach powder, characterized in that: The following steps are involved: S1. Sample defatting: Use tweezers to separate the American cockroach adult into five parts: head, pronotum, wings, abdomen, and thoracic legs. Powder each part separately. Take 100 g of the powder from each part of the American cockroach, load it into a filter paper tube and place it in the extraction tube of a Soxhlet extractor. Use the extracting solution to soak the American cockroach powder and dissolve the oil. When the extract in the extraction tube reaches the height of the siphon, it automatically flows back to the flask with the oil. Reflux and defatting for 3-4 hours until the extract is colorless. Stop heating, remove the powder, and evaporate the extract. S2. Sample preparation: Take the above-mentioned defatted powder that has passed through a No. 5 sieve, add distilled water, chloral hydrate, and dilute glycerol, respectively, and prepare water-mounted slides, chloral hydrate cold-mounted slides, and chloral hydrate permeabilized slides; S3. Microscopic observation: Place the prepared slide under an optical microscope, use an objective lens to observe the microstructure from low to high, and use the image processing software equipped with the microscope to collect and analyze the powder characteristics; S4. Identification criteria.

2. The method for microscopic identification of American cockroach powder according to claim 1, wherein: The extract in S1 is obtained by adding 300 ml of petroleum ether into a round-bottom flask, heating it in a water bath at 50-60° C. to boil the petroleum ether, and condensing the steam through a condenser to obtain the extract.

3. The method for microscopic identification of American cockroach powder according to claim 1, wherein: The reflux method used in S1 is Soxhlet extraction reflux.

4. The method for microscopic identification of American cockroach powder according to claim 1, wherein: The identification criteria observed in S4 are as follows: Setae: There are two types: thick and thin. The thick setae are oblong-conical, straight or slightly curved, dark brown or tan, often broken, with a sub-rounded base, slightly swollen, and blunt-rounded tips. The thin setae are lanceolate or needle-shaped, straight or slightly curved, yellow or yellow-brown, with sharp tips, smooth surface, and no texture. Wing fragments: rare, light yellow, yellow-brown or yellow-green, with dense scaly or wavy texture on the surface; Striated muscle fibers: common, brownish yellow or colorless and transparent, with fine and dense horizontal stripes, sometimes scattered in cross, vertical, or spiral patterns; Body wall fragments: colorless, yellow, light yellow or yellow-brown, with reticular, hexagonal or fish-scale textures, sometimes with circular pores, the edges of which are darker in color; yellow-brown or yellow-brown body wall fragments, with a smooth surface and dotted white dot-like textures, and transparent dot-like hair pits; Compound eye fragments: occasionally seen, irregular blocks, with typical ommatidia structure, ommatidia are circular or polygonal, arranged closely, like a honeycomb.

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