Cuttlefish ink composition as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly discloses a cuttlefish ink composition and a preparation method and application thereof.The cuttlefish ink composition comprises an active ingredient and auxiliary materials with effective treatment dose, the active ingredient is cuttlefish ink, and the cuttlefish ink is a dry ink bag of sepia esculenta or sepiella maindroni of cuttlefish animals; the auxiliary materials comprise a flavoring agent, an excipient and a hollow capsule, and the nitrogen content in every 1g of the cuttlefish ink composition is not less than 65mg. The cuttlefish ink composition and the preparation method and application thereof are used for systemic hemostasis, and the occurrence rate of side effects is obviously lower than that of existing medicines.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine preparation technology, and in particular to a cuttlefish ink composition, its preparation method, and its application. Background Technology

[0002] Cuttlefish ink is the dried ink sac of the golden cuttlefish (Sepia esculenta Hoyle) or the needleless cuttlefish (Sepiella maindroni de Rochebrune). It can be collected year-round. The processing method involves harvesting the cuttlefish, removing the ink sac, washing, and drying. Cuttlefish ink is named for its color and origin. The ink content varies among different species, ranging from approximately 1-10g in dry weight, accounting for about 1.28% of the cuttlefish's total body weight. Studies have found that cuttlefish ink contains various active ingredients. The ink is a viscous suspension composed of black particles invisible to the naked eye, with approximately 200mg of particles per mL (20% w / v). Cuttlefish ink particles are spherical with a diameter varying considerably, between 90-250nm. The black particles have a two-layer structure: a high-density core and a low-density shell. The core is the so-called cuttlefish ink melanin, while the shell may be a proteoglycan complex attached to the melanin. Cuttlefish ink melanin is a polymer of indolequinone, which may or may not be bound to proteins.

[0003] Existing research indicates that cuttlefish ink has high medicinal value. Ink is extracted from fresh cuttlefish ink sacs, homogenized, filtered, and freeze-dried to obtain cuttlefish ink powder. 5% cuttlefish ink can significantly shorten clotting and bleeding times in mice and enhance the function of clotting factors, platelets, and capillaries. In existing technology, cuttlefish ink hemostatic capsules are made by adding maltose and corn starch, followed by drying and pulverization to form capsules. Each capsule (0.33g) contains no less than 12.5mg of nitrogen (N), possessing astringent and hemostatic effects. This drug is mainly used to treat functional uterine bleeding and excessive menstrual bleeding. However, systematic research on cuttlefish ink is still lacking, and its medicinal value remains in the exploratory stage. Summary of the Invention

[0004] The purpose of this invention is to provide a cuttlefish ink composition, its preparation method, and its application for systemic hemostasis, with a significantly lower incidence of side effects than existing drugs.

[0005] To achieve the above objectives, the present invention provides a cuttlefish ink composition comprising a therapeutically effective amount of an active ingredient and excipients. The active ingredient is cuttlefish ink, which is the dried ink sac of the cuttlefish species Cuttlefish or Cuttlefish scaberne. The excipients include flavoring agents, excipients, and empty capsules. The cuttlefish ink composition contains not less than 65 mg of nitrogen per 1 g.

[0006] Preferably, the cuttlefish ink composition is one of capsules, tablets, or pills.

[0007] Preferably, the flavoring agent is an oligosaccharide, specifically xylooligosaccharide, and the excipient is croscarmellose sodium.

[0008] The xylooligosaccharides used in this invention, also known as xylooligosaccharides, are low-polymer sugars composed of 2-10 xylose molecules linked by -1,4-glycosidic bonds. They belong to the category of functional oligosaccharides. They are obtained by directional hydrolysis, separation, and purification of xylan-rich raw materials. Compared with commonly used soybean oligosaccharides, fructooligosaccharides, and isomaltooligosaccharides, they have unique advantages. They can selectively promote the proliferation of intestinal bifidobacteria. Their bifidus factor function is 10-20 times that of other polymeric sugars.

[0009] Xylooligosaccharides are difficult for human digestive enzymes to break down. Digestion experiments using saliva, gastric juice, pancreatic juice, and small intestinal enzymes have shown that these digestive juices can hardly break down xylooligosaccharides. Its energy value is almost zero; it neither affects blood glucose concentration nor increases insulin levels, and it does not form fat deposits. Therefore, it can function effectively in low-energy foods, maximizing the satisfaction of those who love sweets but are concerned about diabetes and obesity. Thus, it is safe for diabetics, obese patients, and those with hypoglycemia to consume. Furthermore, it exhibits good acid and heat stability and is difficult to ferment. Compared to other oligosaccharides, xylooligosaccharides are characterized by their excellent stability. Even under acidic conditions (pH=2.5-7) and heated to 100℃, it remains essentially undecomposed, while some functional oligosaccharides decompose easily under acidic conditions, thus reducing their activity in promoting bifidobacteria growth.

[0010] This invention also provides a method for preparing a cuttlefish ink composition, comprising the following steps: S1. Ethanol spraying: Select squid ink, remove impurities, spray in 10-30% ethanol by weight of squid ink, and let it soak for 12-20 hours. S2. Drying: Place in a vacuum dryer and dry under vacuum at 60~80℃ for 6~8 hours; S3. Grinding: Grind into 80-100 mesh to obtain cuttlefish ink powder; S4. One-step granulation: The cuttlefish ink powder obtained in S3 is mixed with croscarmellose sodium, and then an aqueous solution of xylooligosaccharide is added. The mixture is then granulated in one step to obtain granules. S5. Granule crushing and capsule filling: Crush the granules to 40-60 mesh and fill them into gelatin hollow capsules to obtain squid ink capsules.

[0011] Preferably, the mass ratio of the cuttlefish ink powder, the cross-linked sodium carboxymethyl cellulose, and the xylooligosaccharide is 1:0.02:0.64.

[0012] Preferably, the aqueous solution of the xylooligosaccharide has a specific concentration of 25-26%.

[0013] The present invention also provides an application of the cuttlefish ink composition in systemic hemostasis.

[0014] Therefore, the present invention employs the above-mentioned cuttlefish ink composition, its preparation method, and its application, and the beneficial effects are as follows: This invention is used for systemic hemostasis and is remarkably effective for various types of bleeding, including gynecological, surgical, and internal medicine bleeding. It is particularly effective for bleeding caused by uterine bleeding, hemoptysis due to pulmonary tuberculosis, and gastrointestinal ulcers and inflammation. It also shows varying degrees of efficacy for uterine bleeding, hematuria, hemoptysis due to lung cancer, and cervical cancer bleeding. The incidence of side effects is significantly lower than that of existing drugs.

[0015] This invention utilizes cuttlefish ink (containing nitrogenous active ingredients) to promote platelet aggregation and enhance the activity of coagulation factors, achieving biological hemostasis; it utilizes the high water absorption and swelling properties of cross-linked carboxymethyl cellulose sodium (compression hemostasis) and its porous structure (reducing rebleeding caused by inflammatory stimulation) to achieve physical hemostasis; and it utilizes xylooligosaccharides, which are difficult to be broken down by human digestive enzymes and have good acid and heat stability, to selectively promote the proliferation activity of intestinal bifidobacteria (indirectly assisting mucosal repair), reducing edema and secondary bleeding at the bleeding site. The three components work synergistically to achieve biological, physical, and repair-based hemostasis, resulting in shorter hemostasis time, less bleeding, and a reduced risk of rebleeding compared to using cuttlefish ink alone.

[0016] The xylooligosaccharides of this invention effectively inhibit the oxidative denaturation of nitrogen-containing components, ensuring the retention rate of nitrogen-containing active ingredients and making the nitrogen content of the finished product more stable.

[0017] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0018] Figure 1 This is a thin-layer chromatogram of a cuttlefish ink composition, its preparation method, and its application examples according to the present invention. Detailed Implementation

[0019] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.

[0020] Unless otherwise defined, the technical or scientific terms used in this invention shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0021] Example 1: A cuttlefish ink composition, specifically cuttlefish ink capsules, comprising the following ingredients: 100g squid ink, 2g croscarmellose sodium, 64g xylooligosaccharides.

[0022] Its preparation method is as follows: 1. Ethanol spraying: Select squid ink, remove impurities, spray in 30% of the squid ink's mass of 75% ethanol, and let it soak for 12 hours.

[0023] 2. Drying: Place the squid ink that has been moistened with 75% ethanol into a vacuum dryer and dry it under vacuum at 60°C for 8 hours to obtain the dried product.

[0024] 3. Pulverization: The dried product is pulverized at 80 mesh to obtain cuttlefish ink powder.

[0025] 4. One-step granulation: Squid ink powder is mixed with croscarmellose sodium, and then an aqueous solution of xylooligosaccharide is added. The mixture is then granulated in one step to obtain granules.

[0026] 5. Granule crushing: Crush the granules to a 40-mesh size to obtain medicinal powder.

[0027] 6. Capsule filling: Fill gelatin hollow capsules with 40-mesh medicinal powder to obtain squid ink capsules.

[0028] Example 2 A cuttlefish ink composition, specifically cuttlefish ink capsules, comprising the following ingredients: 100g squid ink, 2g croscarmellose sodium, 64g xylooligosaccharides.

[0029] Its preparation method is as follows: 1. Ethanol spraying: Select squid ink, remove impurities, spray in 10% of the squid ink's mass of 75% ethanol, and let it soak for 20 hours.

[0030] 2. Drying: Place the squid ink that has been moistened with 75% ethanol into a vacuum dryer and dry it under vacuum at 80°C for 8 hours to obtain the dried product.

[0031] 3. Grinding: Grind the dried product into 100-mesh powder to obtain cuttlefish ink powder.

[0032] 4. One-step granulation: Squid ink powder is mixed with croscarmellose sodium, and then an aqueous solution of xylooligosaccharide is added. The mixture is then granulated in one step to obtain granules.

[0033] 5. Granule crushing: Crush the granules to a 60-mesh size to obtain medicinal powder.

[0034] 6. Capsule filling: Fill gelatin hollow capsules with 60-mesh medicinal powder to obtain squid ink capsules.

[0035] Comparative Example 1: A squid ink capsule comprising the following ingredients: 100g squid ink, 5g maltose, 60g cornstarch.

[0036] Its preparation method is as follows: 1. Drying: Take cuttlefish ink, rinse with water, rinse with clean water, place in a stainless steel pan and dry at 120℃ for 10 hours. After drying, remove and let cool.

[0037] 2. Grinding: Take the above dried product, grind it into fine powder, and pass it through a 100-mesh sieve.

[0038] 3. Granulation: Take starch and cuttlefish ink powder, add maltose and an appropriate amount of purified water, mix them to form a soft material, granulate it under a 12-mesh sieve, place it in a stainless steel pan, and dry it at 70℃.

[0039] 4. Grinding: Take the above dried product, grind it, and pass it through a 40-mesh sieve.

[0040] 5. Capsule filling: Fill the gelatin hollow capsules with the above 40-mesh powder to obtain squid ink capsules.

[0041] Experimental testing: 1. Qualitative identification of Cuttlefish Ink Capsules was performed using thin-layer chromatography (Chinese Pharmacopoeia 2025 Edition, Part IV, 0502). The test method is as follows: Take 1.0 g of the product from Example 1, grind it finely, add 30 mL of 70% ethanol, sonicate for 20 min, filter, evaporate the filtrate to dryness, add 1 mL of 70% ethanol to the residue, dissolve, and use as the test solution. Take 1.0 g of cuttlefish ink, add 30 mL of 70% ethanol, sonicate for 20 min, filter, evaporate the filtrate to dryness, add 1 mL of 70% ethanol to the residue, dissolve, and use as the reference drug solution. Take tyrosine reference standard, add 0.5 mol / L hydrochloric acid solution to prepare a solution containing 0.25 mg tyrosine per 1 mL, and use as the reference solution. Take 3 μL each of the above test solution and reference drug solution, and 1 μL of the reference solution, and spot them separately on the same silica gel G thin-layer plate, two parallel spots. Use n-butanol-glacial acetic acid-ethanol-water (volume ratio 4:1:1:2) as the developing solvent, develop, remove, air dry, spray with 0.5% ninhydrin ethanol solution, and heat at 105℃ until the spots are clearly visible. The test results are as follows. Figure 1 As shown.

[0042] Depend on Figure 1 As can be seen, from left to right, the chromatograms are of the cuttlefish ink reference material solution, the tyrosine reference solution, the test solution, and parallel spots of the test solution. At the corresponding positions as the reference material and reference standard chromatograms, the test solution shows spots of the same color. This indicates that the cuttlefish ink capsules conform to the characteristics of cuttlefish ink (containing cuttlefish ink-specific components), contain tyrosine, have good quality consistency, and show no impurities (no additional amino impurities).

[0043] 2. The content of cuttlefish ink capsules was determined by nitrogen determination method (Chinese Pharmacopoeia 2025 Edition, Part IV, Method 0704, Method I). The test method is as follows: Ten grains of the product from Example 1 were ground into a fine powder, and an appropriate amount (equivalent to 0.5g of cuttlefish ink) was accurately weighed. The test results showed that each 1g of this product contained 65mg of nitrogen (N).

[0044] Take 10 grains of the product from Comparative Example 1, grind them into a fine powder, and accurately weigh an appropriate amount (equivalent to 0.5g of cuttlefish ink). The test results show that each 1g of this product contains 61mg of nitrogen (N).

[0045] This demonstrates that the contribution of squid ink in this embodiment is more substantial, the retention rate of nitrogen-containing components in the process is further optimized, providing a material basis for improved hemostatic effect, while also reflecting production controllability.

[0046] 3. Modern clinical verification of hemostatic effect: The usual dosage is 1-1.5g of cuttlefish ink per day per person, but the specific dosage can be determined by a physician based on the patient's condition and other specific circumstances. Forty patients (aged 25-45 years, with a disease duration of 1-6 months) meeting the diagnostic criteria for uterine bleeding in *Obstetrics and Gynecology* were selected and observed. After oral administration of the cuttlefish ink capsules described in Example 1, 67.5% of patients achieved short-term control (bleeding stopped 5-7 days after starting medication), with no significant adverse reactions (such as gastrointestinal irritation or allergies). 42.1% of patients were cured after one or several cycles of treatment (bleeding stopped 5-7 days after medication, and no recurrence occurred 3 months after discontinuation). This indicates that cuttlefish ink not only stops bleeding but may also improve overall bodily function by effectively controlling bleeding, mobilizing the body's self-regulating functions, and restoring normal function.

[0047] Ten patients with bleeding due to peptic ulcer disease or gastritis were observed and treated with cuttlefish ink capsules (Example 1), achieving a 100% cure rate. Simultaneously, ten patients with bleeding due to peptic ulcer disease or gastritis were recruited and treated with intramuscular or intravenous injections of tranexamic acid, vitamin K, and para-aminomethylbenzoic acid (PAMBA) as a control group. Both groups of patients met the diagnostic criteria for peptic ulcer (gastric ulcer / duodenal ulcer) or acute gastritis in *Internal Medicine*, exhibited clear bleeding symptoms (such as hematemesis, melena, and a positive fecal occult blood test), were aged 18-65 years, had a disease duration of 1-14 days, and had no history of coagulation disorders, liver or kidney dysfunction, drug allergies, or malignant tumors. Although all patients in the control group were cured, the average hemostasis time with cuttlefish ink was 4.2 days, compared to 8.6 days in the control group, indicating that cuttlefish ink was more effective.

[0048] Eighteen patients with pulmonary tuberculosis and hemoptysis meeting the diagnostic criteria of "Clinical Tuberculosis" were selected for observation. These 18 patients with hemoptysis due to pulmonary tuberculosis included 13 cases of infiltrative pulmonary tuberculosis and 5 cases of cavitary pulmonary tuberculosis. The patients' ages ranged from 22 to 68 years, and the duration of illness ranged from 3 months to 5 years. The severity of hemoptysis was graded as follows: 14 cases of long-term low-volume hemoptysis (daily hemoptysis <100ml, lasting ≥7 days), and 4 cases of massive hemoptysis (daily hemoptysis ≥300ml or a single hemoptysis ≥100ml). Oral administration of the cuttlefish ink capsules (Example 1) resulted in a total effective rate of 72.2%, with an average hemostasis time of 6.5 days. The observation showed a significant effect on patients with long-term low-volume hemoptysis, but no significant effect on patients with massive hemoptysis.

[0049] During the treatment of the aforementioned patients, no allergic reactions (skin itching, rash, difficulty breathing), no gastrointestinal discomfort (nausea, diarrhea, abdominal distension), and no abnormal liver or kidney function were observed. Furthermore, the acute oral LD50 of the cuttlefish ink capsules from Examples 1 and 2 was investigated in rats. 50 No liver or kidney damage was observed after long-term administration of >5000mg / kg for 6 months. No serious irreversible toxicity was observed within the range of 10 to 100 times the planned clinical dose. This indicates that the side effects of the cuttlefish ink capsules in this embodiment are controllable, have a wide safety window, and a reasonable risk-benefit ratio.

[0050] Therefore, the present invention employs the above-mentioned cuttlefish ink composition, its preparation method, and its application for systemic hemostasis, with a significantly lower incidence of side effects than existing drugs.

[0051] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A squid ink composition, characterized in that: The composition includes a therapeutically effective amount of active ingredient and excipients. The active ingredient is cuttlefish ink, which is the dried ink sac of the cuttlefish species Cuttlefish or Cuttlefish scaberne. The excipients include flavoring agents, excipients, and empty capsules. The nitrogen content in the cuttlefish ink composition shall not be less than 65 mg per 1 g.

2. The cuttlefish ink composition according to claim 1, characterized in that: The flavoring agent is an oligosaccharide, specifically xylooligosaccharide, and the excipient is croscarmellose sodium.

3. A method for preparing a cuttlefish ink composition, characterized in that, Includes the following steps: S1. Ethanol spraying: Select squid ink, remove impurities, spray in 10-30% ethanol by weight of squid ink, and let it soak for 12-20 hours. S2. Drying: Place in a vacuum dryer and dry under vacuum at 60~80℃ for 6~8 hours; S3. Grinding: Grind into 80-100 mesh to obtain cuttlefish ink powder; S4. One-step granulation: The cuttlefish ink powder obtained in S3 is mixed with croscarmellose sodium, and then an aqueous solution of xylooligosaccharide is added. The mixture is then granulated in one step to obtain granules. S5. Granule crushing and capsule filling: Crush the granules to 40-60 mesh and fill them into gelatin hollow capsules to obtain squid ink capsules.

4. The method for preparing a cuttlefish ink composition according to claim 3, characterized in that: The mass ratio of the cuttlefish ink powder, the cross-linked sodium carboxymethyl cellulose, and the xylooligosaccharide is 1:0.02:0.

64.

5. The method for preparing a cuttlefish ink composition according to claim 3, characterized in that: The specific concentration of the aqueous solution of the xylooligosaccharide is 25-26%.

6. The use of a cuttlefish ink capsule prepared by the method of preparing a cuttlefish ink composition according to any one of claims 1 to 2 or a cuttlefish ink composition according to any one of claims 3 to 5 in systemic hemostasis.

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