Natural porphine salts and their use as plant growth regulators and immune inducers

Abstract

The present invention discloses natural porphine salts and their use as plant growth regulators and immune inducers. [Solution] The porphine salt includes a porphine salt or a chlorin salt. Compared with the prior art, the porphine salt of the present invention has good stability, stable and reliable quality under long-term storage, a long shelf life, and is easy to carry and transport. The product has good water solubility, is easy to prepare an aqueous solution, has a stable content, good reproducibility in field use, low application concentration, strong activity, high pest control effect, is easy to use, the raw materials are natural and of abundant origin, the preparation is free of emissions and pollution, and is energy-saving and environmentally friendly. In particular, as an immune inducer, it has a high pest control effect and is significantly more effective in use than existing products.

Description

[Technical Field]

[0001] The present invention relates to a natural porphine salt and its use as a plant growth regulator and immune inducer, and belongs to the technical field of plant growth regulators. [Background technology]

[0002] Chlorophyll and heme are types of natural porphine structures that are produced and exist in plants and animals, and are the material basis for life-sustaining activities such as photosynthesis and oxygen respiration in plants and animals. Among them, heme has a typical porphine structure, while the chlorophyll structure is the parent nucleus of chlorin. Chlorin is significantly asymmetric compared to the porphine structure, and its existing forms and states are more complex and diverse. Currently, various porphine-based products obtained by natural extraction and processing are beginning to be widely used in many fields, including the chemical industry, medicine, food, and agriculture. For example, chlorin / iron (chlorophyllin / iron chloride, CN102285992B) and hemin (heme, CN10048884C) are used as novel plant growth regulators for various crops, chlorophyllin / iron sodium salt and chlorophyllin / copper sodium / potassium salt are used as food additives (food colorings), and protoporphyrin sodium is used as a drug to treat liver cirrhosis. In addition to commercially available products, in recent years, Chinese patents such as CN101045730, CN102351867, CN102775416, and CN102796108 have also been developed to treat liver cirrhosis. The preparation and properties of divalent salts such as lead salt (chlorophyllin / iron zinc salt), chlorophyll / iron calcium salt (chlorophyllin / iron calcium salt), chlorophyll / iron magnesium salt (chlorophyllin / iron magnesium salt), and chlorophyll / iron manganese salt (chlorophyllin / iron manganese salt) have been disclosed. In a paper (Reaction rate, equilibrium constant and structure study of iron chlorophyllin metal salt, Master's thesis, North China University of Science and Technology, March 2017), the preparation of a series of chlorophyllin / iron metal salts was studied, and according to the salt reaction, product structure, and physical and chemical properties, the reaction of chlorophyllin / iron with divalent Mg was investigated. 2+ , Ca 2+ , Mn 2+ and Zn 2+The ions were mixed in a 1:1 molar ratio of sparingly soluble / sparely soluble salts (K sp ≒0 -16 ~10 -18 M 2 ), which has been found to have a fast salification reaction (activation energy is as low as 20 kJ / mol), and due to differences in water solubility, divalent metal salts of these porphine chelates can be prepared by precipitation from aqueous solution, among which the water-soluble sodium salt of porphine is the basic raw material for the preparation of these products. Water-soluble porphine salts reported include sodium / potassium salts of divalent transition metal porphine chelates, such as chlorophyll / zinc sodium (chlorophyllin / zinc sodium), chlorophyll / zinc potassium (chlorophyllin / zinc potassium), chlorophyll / manganese sodium (chlorophyllin / manganese sodium), and chlorophyllin / copper sodium. These sodium / potassium salts are highly soluble, and their aqueous solutions are generally highly alkaline. However, there have been few reports on sodium / potassium salts of trivalent transition metal porphine chelates. Hemin is a trivalent iron porphine chelate, and the chloride axially bonded to the central iron is easily replaced by a hydroxyl group with stronger coordination ability. When hemin is added to an aqueous solution, it becomes hematin sodium (Properties and Detection Methods of Hemin, Chinese Journal of Biochemical Drugs, 1993.66(4):58-59). Experiments have shown that precipitation from alkaline aqueous solutions produces only hematin sodium, not hemin sodium. When sodium iron salt and sodium copper salt of chlorophyllin are used, the product exhibits good water solubility, the stability of the solid sample is greatly improved, and the product is easy to transport and carry. However, at present, there are no reported cases of research and application of natural porphyrin salts as plant growth regulators.

[0003] Plant immunity inducers (also known as plant vaccines) are a new type of biological pesticide developed in recent years based on vaccine engineering technology. They activate the plant's immune system by regulating plant metabolism and growth, and provide effective prevention and control, prevention of crop diseases (quarantine), improvement of crop resistance, increased yield, and improved quality. In addition, they are harmless to humans and animals and do not pollute the environment, making them a popular variety in current research and development of biological pesticides.

[0004] The plant immune inducers discovered so far are mainly small molecules derived from plants or microorganisms, such as salicylic acid and matrine, large molecules such as humic acid and lentinan, and some chemically synthesized fungicides. However, research and application of natural porphyrin salts as plant immune inducers (plant vaccines) has not yet been reported.

[0005] Naturally occurring porphines have a planar parent ring with high electron density, which easily chelates with metal ions to form chelates of chlorophyll, heme, etc. The outer porphine ring is bonded to alkyl or alkenyl groups, as well as groups such as carboxyalkyl groups, resulting in a complex structure with many isomers. For example, acidic chlorin / iron is basically poorly soluble in water and has poor photothermal stability, while planar chlorin / iron molecules have strong association and aggregation properties, high lipophilicity of the entire molecule, and large lattice energy. Research has shown that these solids are difficult to diffuse and disperse, and are difficult to dissolve (K sp As the storage time increases, the association and aggregation become stronger, and the solubility decreases further (K sp(The value gradually decreases), and in solution, the stability of the compound also decreased significantly (Preliminary Study on the Basic Properties of Chlorin / Iron Solutions, Bulletin of Nanjing University of Science (Natural Science Edition), 43:1, 2020, 3:143-148). Many acid-type porphines have low molecular polarity, strong lipid solubility, but low water solubility, and are unstable to light and heat under normal conditions. While storing the product in the solid state at low temperatures provides a sufficient shelf life, in solution, especially in aqueous solution, they are prone to decomposition when exposed to light. This requires strict handling to achieve good results, reducing the convenience of product use and increasing uncertainty about the effects of product use. Therefore, it is necessary to develop stable, easy-to-use, and highly active products tailored to actual usage needs. Summary of the Invention [Problem to be solved by the invention]

[0006] The present invention aims to solve the problems in the prior art by providing a natural porphine salt and its use as a plant growth regulator and immune inducer. The natural porphine salt has good photothermal stability, strong activity, good pest control effect, and its aqueous solution can be quickly prepared, making it convenient for use in fields. [Means for solving the problem]

[0007] To achieve the above objectives, the present invention adopts the following technical solutions: The present invention prepares a series of naturally occurring porphine salts, including salts of porphine-based compounds or salts of chlorin-based compounds.

[0008] In the present invention, naturally occurring porphine compounds or chlorin compounds are used, among which the porphine compounds include protoporphyrin and a series of protoporphyrin chelates, and the chlorin compounds include pheophorbide and a series of pheophorbide chelates.

[0009] In the present invention, the salts of the porphine compounds can be prepared by salifying acid-type protoporphyrin or a series of chelates thereof with metal ions necessary for plant nutrition. The acid-type protoporphyrin and a series of chelates thereof include protoporphyrin, hemin (heme), and hematin. The metal ions necessary for plant nutrition include common monovalent, divalent, or trivalent ions of sodium, potassium, ammonium, magnesium, calcium, iron, zinc, manganese, and copper.

[0010] Protoporphyrin is a naturally occurring porphyrin structure, a product obtained by removing chelated iron from heme extracted from animal blood.

[0011] In the present invention, the salts of the above-mentioned chlorin compounds can be prepared by salifying acid-type pheophorbide (chlorin) and its series of chelates with metal ions necessary for plant nutrition. The acid-type pheophorbide and its series of acid-type chelates include pheophorbide (chlorin), chlorophyllin / iron chloride (chlorin / iron), hydroxychlorophyllin / iron (hydroxychlorin / iron), chlorophyllin / iron (chlorin / ferrous, chlorophyllin / ferrous or chlorophyllin / iron(II)), chlorophyllin / zinc (chlorin / zinc), and chlorophyllin / copper (chlorin / copper). The metal ions necessary for plant nutrition include common monovalent, divalent, or trivalent ions of sodium, potassium, ammonium, magnesium, calcium, iron, zinc, manganese, and copper.

[0012] Pheophorbides, also known as chlorins, are products obtained by hydrolyzing chlorophyll extracted from plants or silkworm feces and removing magnesium. They are a mixture of various isomers of the chlorin nucleophilic structure, mainly including various isomers such as pheophorbide a, pyropheophorbide a, chlorin e6, chlorin e4, chlorin f, chlorin p6, and purpurin 18.

[0013] The porphine salts of the present invention can be prepared using commercially available chlorophyllin-based products such as pheophorbide (sodium salt), chlorophyllin / copper sodium salt, chlorophyllin / iron sodium salt, chlorophyllin / zinc sodium salt, chlorin / iron, protoporphyrin sodium, or heme as raw materials, and generally yield porphine salt products with a content of more than 95% (measured photometrically).

[0014] In preparing the salt of the porphine compound or chlorin compound of the present invention, the monovalent metal ion salt can be prepared by preparing a solution of the acid-form porphine compound or chlorin compound in an alcohol solvent, mixing the solution with an alcohol solution of a monovalent metal hydroxide to cause precipitation, or by salifying the acid-form porphine compound or chlorin compound in an aqueous solution of a monovalent metal hydroxide and precipitating the salt using acetone as a solvent, or by preparing a solution of the acid-form porphine compound or chlorin compound and a monovalent metal hydroxide in acetone as a solvent, passing dry ammonia gas through the solution, and finally filtering, washing, and drying the solution.

[0015] The hydroxides of the monovalent metals include sodium hydroxide, potassium hydroxide, aqueous ammonia, and the like.

[0016] Examples of monovalent metal ion salts of the porphine compound or chlorin compound of the present invention include protoporphyrin potassium, protoporphyrin ammonium, hemin sodium (heme sodium, the same applies hereinafter), hemin potassium, hemin ammonium, hematin potassium, hematin ammonium, chlorin / iron sodium salt (chlorophyllin / iron chloride sodium salt, the same applies hereinafter), chlorin / iron potassium salt, chlorin / iron ammonium salt, hydroxychlorophyllin / iron potassium salt, hydroxychlorophyllin / iron sodium salt, hydroxychlorophyllin / iron ammonium salt, chlorophyllin / iron potassium salt, chlorophyllin / iron ammonium salt, chlorin sodium salt (pheophorbide sodium salt, the same applies hereinafter), chlorin potassium salt, chlorin ammonium salt, chlorophyllin / zinc ammonium salt, chlorophyllin / copper ammonium salt, and the like.

[0017] In preparing the salt of the porphine compound or chlorin compound of the present invention, the divalent or trivalent metal ion salt can be obtained by mixing an aqueous solution of a sodium / potassium salt of the porphine compound or chlorin compound with an aqueous solution of a soluble divalent or trivalent metal ion salt to cause precipitation, followed by filtration, washing, and drying.

[0018] The soluble divalent or trivalent metal ion salts include sulfates, hydrochlorides, nitrates, etc. of divalent or trivalent metal ions.

[0019] The divalent metal ion salts of the porphine compound or chlorin compound of the present invention include magnesium protoporphyrin, calcium protoporphyrin, ferrous protoporphyrin, manganese protoporphyrin, zinc protoporphyrin, copper protoporphyrin, magnesium hemin, calcium hemin, ferrous hemin, manganese hemin, zinc hemin, copper hemin, magnesium hematin, calcium hematin, ferrous hematin, manganese hematin, zinc hematin, copper hematin, magnesium salt of chlorin / iron, calcium salt of chlorin / iron, ferrous salt of chlorin / iron, manganese salt of chlorin / iron, zinc salt of chlorin / iron, copper salt of chlorin / iron, magnesium salt of hydroxychlorophyllin / iron, calcium salt of hydroxychlorophyllin / iron, ferrous salt of hydroxychlorophyllin / iron, manganese salt of hydroxychlorophyllin, and hydroxychlorophyllin. hydroxychlorophyllin / iron zinc salt, hydroxychlorophyllin / iron copper salt, chlorophyllin / iron magnesium salt, chlorophyllin / iron calcium salt, chlorophyllin / iron ferrous salt, chlorophyllin / iron manganese salt, chlorophyllin / iron zinc salt, chlorophyllin / iron copper salt, chlorin magnesium salt, chlorin ferrous salt, chlorin manganese salt, chlorin zinc salt, chlorin copper salt, chlorophyllin / zinc magnesium salt, chlorophyllin / zinc calcium salt, chlorophyllin / zinc ferrous salt, chlorophyllin / zinc manganese salt, chlorophyllin / zinc zinc salt, chlorophyllin / zinc copper salt, chlorophyllin / zinc magnesium salt, chlorophyllin / zinc calcium salt, chlorophyllin / zinc ferrous salt, chlorophyllin / zinc manganese salt, chlorophyllin / zinc zinc salt, chlorophyllin / zinc copper salt, chlorophyllin / zinc magnesium salt, chlorophyllin / copper calcium salt, chlorophyllin / copper ferrous salt, chlorophyllin / copper manganese salt, chlorophyllin / copper zinc salt, chlorophyllin / copper copper salt, etc.

[0020] Trivalent metal ion salts of the porphine compound or chlorin compound of the present invention include protoporphyrin iron salt, hemin iron salt, hematin iron salt, chlorin / iron iron salt (chlorophyllin / iron chloride iron salt), hydroxychlorophyllin / iron iron salt, chlorophyllin / iron iron salt, chlorin iron salt, chlorophyllin / zinc iron salt, chlorophyllin / copper iron salt, etc.

[0021] The porphine salt of the present invention is used as a plant growth regulator or a plant immunity inducer, and the porphine salt is sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of protoporphyrin; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of hemin; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of hematin; Sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorin / iron; Hydroxychlorophyllin / sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of iron, Sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorophyllin / iron; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorins; Sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorophyllin / zinc; Includes sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorophyllin / copper.

[0022] The porphine salt of the present invention maintains the macrocyclic structure of porphine and still exhibits characteristic absorption in the Soret band and Q band of the ultraviolet and visible spectrum. The maximum absorption peak (λ ) of each band is lowered due to the influence of the metal chloride ion. max) is shifted to some extent compared to acid-form porphine. Although the Q band in the visible light region has relatively weak absorption, porphine salt-based products maintain a dark color. In aqueous solution, the Soret band spectrum (360-420 nm) exhibits strong absorption, making the difference easily observable. This can be used to identify and verify salt products (UV-Visible Spectra of Porphyrin and Its Derivatives, Spectral Laboratory, 28:3, 2011, 5:1165-1169). It also serves as a basic standard for photometrically measuring the content of porphine-based products (China National Food Safety Standards, Food Additives Chlorophyll / Copper Potassium Salt gB1886.307-2020). Compared to acid-form porphine, porphine salts have a higher electron density in the conjugated structure of the porphine ring due to the electron-donating properties of the metal, resulting in a higher Soret band λ. max The stronger the metallicity, the wider the red shift. For example, λ of heme (20% methanol solution) max = 368 nm, and in its alkali metal and alkaline earth metal salts (in aqueous solution), the λ of heme sodium, heme potassium, heme magnesium, and heme calcium max The wavelengths of all transition metal salts are red-shifted to 393 nm to 395 nm. The d orbitals of transition metal salts are involved in conjugation, which may affect the electron density of porphines depending on factors such as the valence state of the ions, the electronegativity of the elements, and the number of d electrons. max The red shift varies in width; for example, the heme / manganese salt has a red shift of 395 nm, and the λ max The red shift width is the same as that of alkali metal salts, and the valence layer d 5 This may be related to the electronic structure and the small electronegativity of manganese (1.5). The heme / zinc salt is 377 nm, and the valence layer d 10 The electronegativity of zinc (1.6) is slightly greater than that of manganese, resulting in a significant reduction in the red shift. The heme ferrous salt and heme iron(III) salt have a λ max are 371 nm and 373 nm, respectively, which correspond to the large number of electrons provided by the high-valence state of iron, and ferrous protoporphyrin and iron protoporphyrin have λ maxare 362 nm and 356 nm, respectively, and although there is no chelated metal in the protoporphyrin ring, the iron(III) salt is blue-shifted, which is due to the d 5 The structure of heme copper salts is likely to accept feedback electrons, which may be related to the reduction of the conjugated electron density of porphins. max shows a blue shift of 367 nm, which is due to the d 9 This may be related to its structural properties and its large electronegativity of 1.9.

[0023] The porphine salt of the present invention exhibits a clear endothermic dehydration process (88-120°C) according to thermal analysis (DTA / TG), and thermogravimetry (TG) measurements show that monovalent metal salts usually dehydrate 2 moles of water and divalent metal salts dehydrate 3 moles of water, whereas acid-type porphine usually does not show any clear weight loss before decomposition, or the weight loss is significantly lower than that of the corresponding salt. This indicates that the acid-type sample contains no or almost no water inside, and the internal structure of the solid is dense. Differential thermal analysis (DTA) shows that the porphine salt exhibits a clear endothermic dehydration process (88-120°C). Porphyrin salts show obvious or significant endothermic and exothermic peaks. When the sample is dehydrated, the specific heat of the system gradually increases with increasing temperature, but there is no peak characteristic of phase transition. The decomposition temperature is often higher than that of the corresponding acid-type porphyrin. The high dehydration temperature and large endothermic range (large energy required for dehydration) mean that a small amount of solid water is adsorbed mainly in the form of hydrated ions after the salification of porphyrin. The differential thermal curve of acid-type porphyrin is relatively flat, with no obvious endothermic or exothermic peaks.

[0024] In the porphine salts of the present invention, the monovalent metal ion salts have good water solubility, and the divalent / trivalent metal ion salts are soluble or slightly soluble in water, with their solubility being significantly higher than that of the corresponding acid-type porphines. The aqueous solutions of the porphine salts are neutral to weakly acidic (pH=6.5-7.5) at the dilute concentrations (10 ppm-0.001 ppm) used in the present invention, which are suitable for the physiological requirements of plants, and have good photothermal stability. Accelerated testing has shown that the content of the porphine salts of the present invention remains unchanged even after storage in solid form for more than two years, and there is no obvious decomposition of the aqueous solution even under shading. Even after 2-3 hours of exposure under normal light conditions (not direct, strong light), there is no significant decrease in the content, making them suitable for use in fields and convenient for application.

[0025] Tests have proven that when the aqueous solution of the porphine salt of the present invention is directly irradiated with high-intensity light (illuminance 30,000 lux) for 1 to 3 hours, the content is significantly higher than that of hemin or chlorin / iron in existing acid-type products.

[0026] The porphine salt of the present invention can be diluted by directly adding water and stirring to prepare a solution having a porphine salt content of 0.01 ppm to 10 ppm (approximately 0.014 μmol / L to 14.0 μmol / L), and can be used as a plant growth regulator or a plant immunity inducer.

[0027] A 0.001 ppm to 0.1 ppm solution of the porphine salt of the present invention can be used for soaking seeds and irrigating crop fields, and a 0.01 ppm to 10.0 ppm solution can be used for foliar spraying of crops, thereby promoting seed germination, increasing the germination rate, increasing root length and growth, strengthening plant immunity and stress resistance, promoting seedling growth, increasing chlorophyll content, delaying premature plant aging, and improving yield and quality.

[0028] The porphine salt of the present invention still maintains the chlorophyllase inhibitory effect of acid chlorin, and in vitro tests showed that the chlorophyllase inhibition rate increased with increasing salt sample concentration, proving that it is a typical competitive inhibitor. Tests also showed that the porphine salt of the present invention has the effect of regulating multiple plant signals, such as regulating the NO concentration in crop roots to promote root growth, inducing increased activity of SOD, CAT and POD in plant leaves under salt stress, and promoting proline release in plants. It also reduces oxidative damage caused by salt stress, strengthens the stress tolerance of crops, significantly reduces phytotoxicity caused by herbicides, and significantly increases crop yield.

[0029] In vitro disease resistance tests have proven that the porphyrin salt of the present invention has excellent plant immunity-inducing activity and can significantly weaken infection by Phytophthora capsici, and has a significant control effect against diseases in pepper plants artificially inoculated with Phytophthora spores. The control effect when sprayed with a 2.0 ppm solution reached 71.89%. In field tests using a 0.2 ppm solution of chlorin / iron sodium salt to control tobacco diseases, the control effect reached 71.00%, demonstrating a significant increase in plant stress tolerance and increased crop yield. [Effects of the Invention]

[0030] Compared with the prior art, the porphine salt of the present invention has good stability, stable and reliable quality under long-term storage, a long shelf life, and is easy to carry and transport; the product has good water solubility, is easy to prepare an aqueous solution, has a stable content, has good reproducibility in field use, requires a low use concentration, has strong activity, has a high pest control effect, is easy to use, is made from natural raw materials of abundant origin, is prepared without emissions or pollution, is energy-saving and environmentally friendly; and, as an immunity inducer in particular, has a high pest control effect and is significantly more effective in use than existing products. [Brief explanation of the drawings]

[0031] [Figure 1]Figure 1 shows a comparison of the DTA / TG (differential thermal / thermogravimetry) analysis of chlorophyllin / iron sodium salt and chlorophyllin / iron, with the top being DTA (differential thermal) and the bottom being TG (thermogravimetry). [Figure 2] FIG. 2 shows UV spectrophotometric scans of hemin and magnesium hemin solutions exposed to strong light for 3 hours in a stability study. DETAILED DESCRIPTION OF THE INVENTION

[0032] The present invention will be further described below with reference to specific examples. Example 1: 5.0 grams of protoporphyrin was weighed and dissolved in 1000 mL of ethanol. 12.0 mL of 10% potassium hydroxide ethanol solution was slowly added dropwise while stirring. The mixture was left in a refrigerator overnight, filtered, washed with cold ethanol, and dried under reduced pressure to obtain approximately 4.35 grams of protoporphyrin potassium salt, which was a purple-brown solid that decomposed at >302°C (DTA / TG) and was easily soluble in water. max = 372 nm (aqueous solution).

[0033] Example 2: 5.0 grams of protoporphyrin was weighed and dissolved in 1000 mL of acetone. Dry ammonia gas was slowly passed through the solution while stirring until saturated. The solution was left overnight, filtered, washed with a small amount of cold acetone, and dried under reduced pressure to obtain approximately 3.11 grams of protoporphyrin ammonium salt, which was a purple-brown solid that decomposed at >296°C (DTA / TG), was easily soluble in water, and had a λ max = 371 nm (aqueous solution).

[0034] Example 3: 5.0 grams of protoporphyrin sodium salt was weighed and dissolved in 100 mL of water with stirring. 5.0 grams of magnesium sulfate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 3.50 grams of protoporphyrin magnesium salt, which was a purple-brown solid that decomposed at >361°C (DTA / TG), was soluble in water, and had a λ max= 355 nm (aqueous solution).

[0035] Example 4: 5.0 grams of protoporphyrin sodium salt was weighed and dissolved in 100 mL of water with stirring. 4.5 grams of calcium sulfate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 3.93 grams of protoporphyrin calcium salt, which was a purple-brown solid that decomposed at >365°C (DTA / TG), was soluble in water, and had a λ max = 356 nm (aqueous solution).

[0036] Example 5: 5.0 grams of protoporphyrin sodium salt was weighed and dissolved in 100 mL of water with stirring. 6.0 grams of manganese sulfate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 3.83 grams of protoporphyrin manganese salt, which was a purple-brown solid that decomposed at >367°C (DTA / TG), was soluble in water, and had a λ max = 357 nm (aqueous solution).

[0037] Example 6: 5.0 grams of protoporphyrin sodium salt was weighed and dissolved in 100 mL of water with stirring. 6.5 grams of ferrous sulfate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 4.51 grams of ferrous protoporphyrin salt, which was a purple-brown solid that decomposed at >359°C (DTA / TG), was soluble in water, and had a λ max = 362 nm (aqueous solution).

[0038] Example 7: 5.0 grams of protoporphyrin sodium salt was weighed and dissolved in 100 mL of water with stirring. 4.5 grams of iron trichloride was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then allowed to stand for 1 hour. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 4.63 grams of protoporphyrin iron salt, which was a purple-brown solid that decomposed at >367°C (DTA / TG) and was poorly soluble in water. max = 356 nm (aqueous solution).

[0039] Example 8: 5.0 grams of protoporphyrin sodium salt was weighed and dissolved in 100 mL of water with stirring. 5.0 grams of zinc sulfate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 4.23 grams of protoporphyrin zinc salt, which was a purple-brown solid that decomposed at >361°C (DTA / TG) and was poorly soluble in water. max = 358 nm (aqueous solution).

[0040] Example 9: 5.0 grams of protoporphyrin sodium salt was weighed and dissolved in 100 mL of water with stirring. 6.0 grams of copper sulfate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 4.72 grams of protoporphyrin copper salt, which was a purple-brown solid that decomposed at >363°C (DTA / TG) and was poorly soluble in water. max = 382 nm (aqueous solution).

[0041] Example 10: 10.0 g of hemin was weighed and dissolved in 1000 mL of ethanol. 25.0 mL of 10% potassium hydroxide ethanol solution was slowly added dropwise while stirring. The mixture was left overnight, filtered, washed with cold ethanol, and dried under reduced pressure to obtain approximately 7.14 g of hemin potassium salt, which was a black solid powder that decomposed at >251°C (DTA / TG), was easily soluble in water, and had a λ max = 394 nm (aqueous solution).

[0042] Example 11: 10.0 g of hemin was weighed and dissolved in 1000 mL of ethanol. 25.0 mL of 10% sodium hydroxide ethanol solution was slowly added dropwise while stirring. The mixture was left overnight, filtered, washed with cold ethanol, and dried under reduced pressure to obtain approximately 9.33 g of hemin sodium salt, which was a black solid powder that decomposed at >264°C (DTA / TG), was easily soluble in water, and had a λ max = 393 nm (aqueous solution).

[0043] Control sample: Preparation of hematin sodium salt According to the method described in the literature (Preparation of Hemin Sodium, Chemistry World, 2004.10:540-541), 10.0 g of hemin was dissolved in 120 mL of 0.2 mol / L NaOH solution, stirred for 6 hours, filtered, and 1000 mL of acetone (more than 8 times the volume) was slowly added to the filtrate, resulting in the deposition of a black precipitate. The solution was left overnight, filtered, washed with a small amount of acetone, and dried under reduced pressure to obtain 8.7 g of black solid, which was easily soluble in water and had a λ max = 384 nm (aqueous solution).

[0044] Comparative test 1: Cl - Ion detection Reagents: HNO3 solution (4 mol / L), AgNO3 test solution (0.1 mol / L) Method: After preparing the precipitate as described above and filtering the product, collect the filtrate (reaction mother liquor), completely recover the solvent ethanol or acetone by rotary evaporation, cool to obtain the residual solid, add about 20 mL of pure water to dissolve, and adjust to neutral (pH ≒ 7) with HNO3 to obtain the test solution, take a small amount of test solution (0.5-1 mL) in a test tube, add a few drops of AgNO3 test solution, observe whether there is precipitation / turbidity, if turbidity occurs, continue to add small amounts of HNO3 slowly and observe the changes.

[0045] Results: The test solution recovered from the ethanol filtrate was clear and contained Cl. - No ions were detected. The test solution recovered from the acetone filtrate produced a precipitate (AgCl↓), and when HNO3 was added, the precipitate did not change, and Cl - ions were detected. Description: The product prepared in ethanol (in this example) is hemin sodium. In the process of preparing a control sample by adding acetone to an alkaline solution of hemin to precipitate it (literature method), chloride ions are replaced by hydroxyl ions in the reaction solution (producing NaCl). The resulting product is chloride-free (substituted by hydroxyl) and is not hemin sodium, but hematin sodium.

[0046] Comparison test 2: UV absorption maximum wavelength (λ max ) measurement Sample: Hemin sodium, hematin sodium, hemin Reagents: Pure water, NaOH solution 0.1 mol / L Measuring instrument: Shimadzu UV2600 ultraviolet spectrophotometer Method: Weigh an appropriate amount of sample, dissolve it in pure water or NaOH solution, dilute it to a solution with a concentration of about 20.0 ppm, and measure the maximum absorption wavelength λ of the sample with an ultraviolet instrument. max (Structural characteristics) were scanned and the results are shown in the table below. [Table 1]

[0047] Results: In NaOH solution, the characteristic absorption of all three samples was at 384 nm, which was due to the high OH axial chlorine group attached to the central iron of hemin. - This indicates that the OH group was substituted with hydroxyl group under the ionic concentration to produce the product sodium hematin. - The concentration of ions is not high (20 ppm sodium hemin solution, pH = 7.5), and the chloride groups are retained. The sample in this example exhibits the characteristic absorption of sodium hemin at 393 nm, while the control sample prepared by the literature method exhibits the same characteristic value as the NaOH solution, indicating that the sample is sodium hematin.

[0048] The chlorine atoms in the axial coordination group of the central iron of heme possess electron-donating / withdrawing abilities different from those of hydroxyl groups. The effect of these electrons is related to the electronegativity, lone pair electrons, and valence electron structure of the element. Chlorine atoms have an electronegativity of 3.0 and a 3p valence electron structure, and when axially bound to the central iron, they have five available lone electron pairs. The oxygen atoms in hydroxyl groups have an electronegativity of 3.5 and a 2p valence electron structure, and when axially bound to the central iron, they have four available lone electron pairs. Compared to chlorine, hydroxyl groups have stronger electron-withdrawing abilities and weaker electron feedback capabilities. Therefore, when hydroxyl groups are coordinated, the electron density of the porphyrin conjugated structure decreases, resulting in a blue shift of the characteristic Soret band from 393 nm to 384 nm in sodium hematin.

[0049] Example 12: 5.0 grams of hemin was weighed and dissolved in 1000 mL of acetone. Dry ammonia gas was slowly passed through the solution with stirring until saturated. The solution was left overnight, filtered, washed with a small amount of cold acetone, and dried under reduced pressure to obtain about 3.60 grams of hemin ammonium salt, which was a brown solid that decomposed at >243°C (DTA / TG), was easily soluble in water, and had a λ max = 386 nm (aqueous solution).

[0050] Example 13: 10.0 grams of hemin sodium salt was weighed and dissolved in 200 mL of water with stirring. 11.0 grams of magnesium sulfate was weighed and dissolved in 100 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 7.19 grams of hemin magnesium salt, which was a dark brown solid that decomposed at >292°C (DTA / TG), was soluble in water, and had a λ max = 395 nm (aqueous solution).

[0051] Example 14: 10.0 grams of hemin sodium salt was weighed and dissolved in 200 mL of water with stirring. 9.0 grams of calcium chloride was weighed and dissolved in 100 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain about 7.37 grams of hemin calcium salt, which was a dark brown solid that decomposed at >311°C (DTA / TG), was soluble in water, and had a λ max = 395 nm (aqueous solution).

[0052] Example 15: 10.0 grams of hemin sodium salt was weighed and dissolved in 200 mL of water with stirring. 11.0 grams of manganese sulfate was weighed and dissolved in 150 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain about 8.57 grams of hemin manganese salt, which was a dark brown solid that decomposed at >329°C (DTA / TG), was poorly soluble in water, and had a λ max = 395 nm (aqueous solution).

[0053] Example 16: 10.0 grams of hemin sodium salt was weighed and dissolved in 200 mL of water with stirring. 14.0 grams of ferrous sulfate was weighed and dissolved in 150 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 8.63 grams of hemin ferrous salt, which was a dark brown solid that decomposed at >327°C (DTA / TG), was poorly soluble in water, and had a λ max = 373 nm (aqueous solution).

[0054] Example 17: 10.0 grams of hemin sodium salt was weighed and dissolved in 200 mL of water with stirring. 11.0 grams of iron chloride was weighed and dissolved in 120 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 9.06 grams of hemin iron salt, which was a dark brown solid that decomposed at >334°C (DTA / TG) and was poorly soluble in water. max = 371 nm (aqueous solution).

[0055] Example 18: 10.0 grams of hemin sodium salt was weighed and dissolved in 200 mL of water with stirring. 14.0 grams of zinc sulfate was weighed and dissolved in 150 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain about 9.13 grams of hemin zinc salt, which was a dark brown solid that decomposed at >274°C (DTA / TG), was poorly soluble in water, and had a λ max = 377 nm (aqueous solution).

[0056] Example 19: 10.0 grams of hemin sodium salt was weighed and dissolved in 200 mL of water with stirring. 15.0 grams of copper sulfate was weighed and dissolved in 150 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain about 9.06 grams of hemin copper salt, which was a dark brown solid that decomposed at >248°C (DTA / TG) and was poorly soluble in water. max = 367 nm (aqueous solution).

[0057] Example 20: 1000.0 g of chlorine / iron parent compound (manufactured by Nanjing Baite Biotechnology Co., Ltd., the same applies below) was weighed and mixed with 15 L of industrial alcohol. Stirring was continued until fully dissolved, and 2.5 L of 15% potassium hydroxide ethanol solution was slowly added dropwise. The precipitate was left overnight, filtered, washed with cold ethanol, and dried under reduced pressure to obtain 896.0 g of chlorine / iron potassium salt, which was a dark green powder that decomposed at >313°C (DTA / TG) and was easily soluble in water. max = 400 nm (aqueous solution).

[0058] Example 21: 1000.0 grams of chlorine / iron base compound was weighed and mixed with 15 L of industrial alcohol to dissolve thoroughly. 2.3 L of 15% sodium hydroxide ethanol solution was slowly added dropwise. The precipitate was left overnight, filtered, washed with cold ethanol, and dried under reduced pressure to obtain 907 grams of chlorine / iron sodium salt, which was a dark green powder that decomposed at >301°C (DTA / TG) and was easily soluble in water. max = 399 nm (aqueous solution).

[0059] Example 22: 100.0 grams of chlorine / iron parent compound was weighed and dissolved in 1.5 L of acetone. Dry ammonia gas was slowly passed through to precipitate the compound. The precipitate was left overnight, filtered, washed with cold acetone, and dried under reduced pressure to obtain approximately 51.2 grams of ammonium salt of chlorine / iron, which was a dark green powder that decomposed at >270°C (DTA / TG), was easily soluble in water, and had a λ max = 401 nm (aqueous solution).

[0060] Example 23: 100.0 grams of sodium salt of chlorine / iron was weighed and dissolved in 1 L of water with stirring. 110.0 grams of magnesium chloride hexahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed twice with cold water, filtered under pressure and dried under reduced pressure to obtain 73.1 grams of magnesium salt of chlorine / iron, which was a dark green powder that decomposed at >302°C (DTA / TG), was soluble in water, and had a λ max = 400 nm (aqueous solution).

[0061] Example 24: 100.0 grams of sodium salt of chlorine / iron was weighed and dissolved in 1 L of water with stirring. 100.0 grams of calcium chloride dihydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to precipitate, and the mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 80.2 grams of calcium salt of chlorine / iron, which was a dark green powder that decomposed at >377°C (DTA / TG), was soluble in water, and had a λ max= 399 nm (aqueous solution).

[0062] Example 25: 100.0 grams of sodium salt of chlorin / iron was weighed and dissolved in 1 L of water with stirring. 110.0 grams of ferrous chloride tetrahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed with cold water, filtered under pressure and dried under reduced pressure to obtain 91.1 grams of ferrous salt of chlorin / iron, which was a dark green powder, decomposed at >303°C (DTA / TG), poorly soluble in water, and had a λ max = 401 nm (aqueous solution).

[0063] Example 26: 100.0 grams of sodium salt of chlorine / iron was weighed and dissolved in 1 L of water with stirring. 110.0 grams of iron trichloride hexahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed with cold water, filtered under pressure and dried under reduced pressure to obtain 89.2 grams of iron salt of chlorine / iron, which was a dark green powder, decomposed at >329°C (DTA / TG), poorly soluble in water, and had a λ max = 405 nm (aqueous solution).

[0064] Example 27: 100.0 grams of sodium salt of chlorine / iron was weighed and dissolved in 1 L of water with stirring. 100.0 grams of manganese chloride tetrahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed with cold water, filtered under pressure and dried under reduced pressure to obtain 87.8 grams of manganese salt of chlorine / iron, which was a dark green powder that decomposed at >299°C (DTA / TG), was poorly soluble in water, and had a λ max = 400 nm (aqueous solution).

[0065] Example 28: 100.0 grams of sodium salt of chlorine / iron was weighed and dissolved in 1 L of water with stirring. 100.0 grams of zinc chloride was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed with cold water, filtered under pressure and dried under reduced pressure to obtain 91.4 grams of zinc salt of chlorine / iron, which was a dark green powder that decomposed at >300°C (DTA / TG), was poorly soluble in water, and had a λ max = 399 nm (aqueous solution).

[0066] Example 29: 100.0 grams of sodium salt of chlorine / iron was weighed and dissolved in 1 L of water with stirring. 95.0 grams of copper chloride dihydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed with cold water, filtered under pressure and dried under reduced pressure to obtain 87.2 grams of copper salt of chlorine / iron, which was a dark green powder, decomposed at >222°C (DTA / TG), poorly soluble in water, and had a λ max = 398 nm (aqueous solution).

[0067] Example 30: 1000.0 g of commercially available sodium chlorophyllin / iron salt was weighed and dissolved in 10 L of water with stirring. 15% diluted sulfuric acid was slowly added dropwise at room temperature with stirring until the pH reached 3-4. The mixture was left overnight in the dark, filtered, washed twice with cold water, filtered under pressure, and dried under reduced pressure. The resulting chlorophyllin / iron was mixed with 15 L of industrial alcohol and dissolved by stirring. 2.5 L of 15% potassium hydroxide ethanol solution was slowly added dropwise. The precipitate was left overnight, filtered, washed with cold ethanol, and dried under reduced pressure to obtain approximately 783.0 g of potassium chlorophyllin / iron salt, which was a dark green powder that decomposed at >309°C (DTA / TG) and was easily soluble in water. max = 397 nm (aqueous solution).

[0068] Example 31: 100.0 g of commercially available sodium chlorophyllin / iron salt was weighed and dissolved in 1 L of water with stirring. 15% diluted sulfuric acid was slowly added dropwise at room temperature with stirring until the pH reached 3-4. The solution was left overnight in the dark, filtered, washed twice with cold water, filtered under pressure, and dried under reduced pressure. The resulting chlorophyllin / iron was dissolved in 1.5 L of acetone, and precipitated by slowly passing dry ammonia gas through the solution with stirring. The solution was left overnight, filtered, washed with cold acetone, and dried under reduced pressure to obtain approximately 37.2 g of ammonium chlorophyllin / iron salt. The powder was a dark green color and decomposed at >224°C (DTA / TG). It was easily soluble in water and had a λ max = 398 nm (aqueous solution).

[0069] Example 32: 100.0 grams of commercially available sodium chlorophyllin / iron salt was weighed and dissolved in 1 L of water with stirring. 120.0 grams of ferrous sulfate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 90.9 grams of ferrous chlorophyllin / iron salt, which was a dark green powder that decomposed at >278°C (DTA / TG), was poorly soluble in water, and had a λ max = 400 nm (aqueous solution).

[0070] Example 33: 100.0 grams of commercially available sodium salt of chlorophyllin / iron was weighed and dissolved in 1 L of water with stirring. 90.0 grams of iron chloride was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation. The mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 91.6 grams of iron salt of chlorophyllin / iron, which was a dark green powder that decomposed at >343°C (DTA / TG), was poorly soluble in water, and had a λ max = 399 nm (aqueous solution).

[0071] Example 34: 100.0 grams of commercially available sodium chlorophyllin / iron salt was weighed and dissolved in 1 L of water with stirring. 110.0 grams of copper sulfate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation. The mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 91.2 grams of copper chlorophyllin / iron salt, which was a dark green powder that decomposed at >208°C (DTA / TG), was poorly soluble in water, and had a λ max = 401 nm (aqueous solution).

[0072] Example 35: 500.0 grams of commercially available sodium chlorophyllin / zinc salt was weighed and dissolved in 5 L of pure water. The pH was adjusted to 4-5 with dilute sulfuric acid while stirring, resulting in a precipitate. The precipitate was filtered, washed with water, and dried to obtain chlorophyllin / zinc. 100.0 grams of chlorophyllin / zinc was weighed and dissolved in 1.5 L of acetone. Dry ammonia gas was gently passed through the precipitate, and the mixture was left overnight. The precipitate was filtered, washed with cold acetone, and dried under reduced pressure to obtain approximately 40.2 grams of ammonium chlorophyllin / zinc salt. This black powder decomposed at >251°C (DTA / TG), was easily soluble in water, and had a λ max = 404 nm (aqueous solution).

[0073] Example 36: 100.0 grams of commercially available sodium chlorophyllin / zinc salt was weighed and dissolved in 1 L of water with stirring. 100.0 grams of magnesium chloride hexahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation. The mixture was left overnight in the dark, filtered, washed twice with cold water, filtered under pressure, and dried under reduced pressure to obtain 87.6 grams of commercially available magnesium chlorophyllin / zinc salt, which was a black powder that decomposed at >333°C (DTA / TG), was soluble in water, and had a λ max = 405 nm (aqueous solution).

[0074] Example 37: 100.0 grams of commercially available sodium salt of chlorophyllin / zinc was weighed and dissolved in 1 L of water with stirring. 100.0 grams of calcium chloride dihydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to precipitate, and the precipitate was left overnight in the dark. The precipitate was filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 83.7 grams of calcium salt of chlorophyllin / zinc, which was a black powder that decomposed at >365°C (DTA / TG), was soluble in water, and had a λ max = 405 nm (aqueous solution).

[0075] Example 38: 100.0 grams of commercially available sodium salt of chlorophyllin / zinc was weighed and dissolved in 1 L of water with stirring. 120.0 grams of ferrous chloride tetrahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 90.2 grams of ferrous salt of chlorophyllin / zinc, which was a black powder that decomposed at >371°C (DTA / TG), was poorly soluble in water, and had a λ max = 406 nm (aqueous solution).

[0076] Example 39: 100.0 grams of commercially available sodium salt of chlorophyllin / zinc was weighed and dissolved in 1 L of water with stirring. 100.0 grams of iron trichloride hexahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation. The mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 89.8 grams of iron salt of chlorophyllin / zinc, which was a black powder that decomposed at >341°C (DTA / TG), was poorly soluble in water, and had a λ max = 403 nm (aqueous solution).

[0077] Example 40: 100.0 grams of commercially available sodium salt of chlorophyllin / zinc was weighed and dissolved in 1 L of water with stirring. 100.0 grams of manganese chloride tetrahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation. The precipitate was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 90.7 grams of manganese salt of chlorophyllin / zinc, which was a black powder that decomposed at >336°C (DTA / TG), was poorly soluble in water, and had a λ max = 404 nm (aqueous solution).

[0078] Example 41: 100.0 grams of commercially available sodium salt of chlorophyllin / zinc was weighed and dissolved in 1 L of water with stirring. 100.0 grams of zinc chloride was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation. The mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 92.5 grams of zinc salt of chlorophyllin / zinc. The zinc salt was a black powder that decomposed at >320°C (DTA / TG), was poorly soluble in water, and had a λ max = 409 nm (aqueous solution).

[0079] Example 42: 100.0 grams of commercially available sodium salt of chlorophyllin / zinc was weighed and dissolved in 1 L of water with stirring. 95.0 grams of copper chloride dihydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation. The mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 90.3 grams of copper salt of chlorophyllin / zinc, which was a black powder that decomposed at >271°C (DTA / TG), was poorly soluble in water, and had a λ max = 409 nm (aqueous solution).

[0080] Example 43: 100.0 g of commercially available sodium chlorophyllin / copper salt was weighed and dissolved in 2 L of water with stirring. 15% diluted sulfuric acid was slowly added dropwise at room temperature with stirring until the pH reached 3-5. The solution was left overnight in the dark, filtered, washed twice with cold water, filtered under pressure, and dried under reduced pressure to obtain chlorophyllin / copper. The solution was mixed with 2 L of industrial alcohol and stirred to dissolve. 250 mL of 15% potassium hydroxide ethanol solution was slowly added dropwise. The precipitate was left overnight, filtered, washed with cold ethanol, and dried under reduced pressure to obtain approximately 79.8 g of potassium chlorophyllin / copper salt. The powder was a dark green color and decomposed at >296°C (DTA / TG). It was easily soluble in water and had a λ max = 405 nm (aqueous solution).

[0081] Example 44: 100.0 grams of commercially available sodium chlorophyllin / copper salt was weighed and dissolved in 2 L of water with stirring. 15% diluted sulfuric acid was slowly added dropwise at room temperature with stirring until the pH reached 3-5. The solution was left overnight in the dark, filtered, washed twice with cold water, filtered under pressure, and dried under reduced pressure to obtain chlorophyllin / copper. The solution was mixed with 2 L of acetone and stirred to dissolve. Dry ammonia gas was slowly passed through the solution until saturated, and the precipitate was left overnight. The precipitate was filtered, washed with cold acetone, and dried under reduced pressure to obtain approximately 44.6 grams of ammonium chlorophyllin / copper salt. The powder was a dark green color and decomposed at >277°C (DTA / TG), easily soluble in water, and had a λ max = 404 nm (aqueous solution).

[0082] Example 45: 100.0 grams of commercially available sodium chlorophyllin / copper salt was weighed and dissolved in 1 L of water with stirring. 100.0 grams of magnesium sulfate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to precipitate the precipitate. The precipitate was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 74.1 grams of magnesium chlorophyllin / copper salt, which was a dark green powder that decomposed at >280°C (DTA / TG), was soluble in water, and had a λ max = 406 nm (aqueous solution).

[0083] Example 46: 100.0 grams of commercially available sodium chlorophyllin / copper salt was weighed and dissolved in 1 L of water with stirring. 90.0 grams of calcium chloride was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to precipitate, and the precipitate was left overnight in the dark. The precipitate was filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 87.4 grams of calcium chlorophyllin / copper salt, which was a dark green powder that decomposed at >305°C (DTA / TG), was soluble in water, and had a λ max = 405 nm (aqueous solution).

[0084] Example 47: 100.0 grams of commercially available sodium chlorophyllin / copper salt was weighed and dissolved in 1 L of water with stirring. 90.0 grams of manganese chloride tetrahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to precipitate, and the precipitate was left overnight in the dark. The precipitate was filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 70.3 grams of manganese chlorophyllin / copper salt, which was a dark green powder that decomposed at >311°C (DTA / TG), was soluble in water, and had a λ max = 405 nm (aqueous solution).

[0085] Example 48: 100.0 grams of commercially available sodium chlorophyllin / copper salt was weighed and dissolved in 1 L of water with stirring. 120.0 grams of ferrous sulfate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation. The mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 90.4 grams of ferrous chlorophyllin / copper salt, which was a dark green powder that decomposed at >266°C (DTA / TG), was poorly soluble in water, and had a λ max = 403 nm (aqueous solution).

[0086] Example 49: 100.0 grams of commercially available sodium salt of chlorophyllin / copper was weighed and dissolved in 1 L of water with stirring. 90.0 grams of iron chloride was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation. The mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 89.7 grams of iron salt of chlorophyllin / copper, which was a dark green powder that decomposed at >357°C (DTA / TG), was poorly soluble in water, and had a λ max = 406 nm (aqueous solution).

[0087] Example 50: 100.0 grams of commercially available sodium chlorophyllin / copper salt was weighed and dissolved in 1 L of water with stirring. 100.0 grams of zinc chloride was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation. The precipitate was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 90.5 grams of zinc chlorophyllin / copper salt, which was a dark green powder that decomposed at >263°C (DTA / TG), was poorly soluble in water, and had a λ max = 404 nm (aqueous solution).

[0088] Example 51: 100.0 grams of commercially available sodium salt of chlorophyllin / copper was weighed and dissolved in 1 L of water with stirring. 110.0 grams of copper sulfate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation. The mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 91.7 grams of copper salt of chlorophyllin / copper, which was a dark green powder that decomposed at >242°C (DTA / TG), was poorly soluble in water, and had a λ max = 404 nm (aqueous solution).

[0089] Example 52: 1000 grams of commercially available sodium chlorophyllin / magnesium salt was weighed and dissolved in 20 L of pure water preheated to 60°C. Heating was stopped, and dilute sulfuric acid was slowly added dropwise until the pH reached 3-4. The mixture was left overnight in the dark, filtered, washed twice with cold water, filtered under pressure, and dried under reduced pressure to obtain 886.3 grams of pheophorbide (chlorin). 100.0 grams of dried chlorin was dissolved in 1.6 L of industrial alcohol and stirred until fully dissolved. 300 mL of 15% potassium hydroxide in ethanol was slowly added dropwise to precipitate the precipitate. The precipitate was left overnight, filtered, washed with cold ethanol, and dried under reduced pressure to obtain 69.2 grams of potassium chlorin, a black powder that decomposed at >283°C (DTA / TG) and was easily soluble in water. max = 405 nm (aqueous solution).

[0090] Example 53: Weigh 1000 grams of dry chlorin (prepared according to the method of Example 52), mix with 15 L of industrial alcohol, dissolve thoroughly, add 2.3 L of 15% sodium hydroxide ethanol solution dropwise, leave the precipitate overnight, filter, wash with cold ethanol, and dry under reduced pressure to obtain 723.0 grams of chlorin sodium salt, a black powder that decomposes at >277°C (DTA / TG), is easily soluble in water, and has a λ max = 404 nm (aqueous solution).

[0091] Example 54: Weigh out 100.0 grams of dry chlorin (prepared according to the method of Example 52), mix with 1.5 L of acetone to dissolve, and precipitate by slowly passing dry ammonia gas through it. Allow to stand overnight, filter, wash with cold acetone, and dry under reduced pressure to obtain about 37.6 grams of chlorin ammonium salt, which is a brown powder that decomposes at >281°C (DTA / TG), is easily soluble in water, and has a λ max = 403 nm (aqueous solution).

[0092] Example 55: 100.0 grams of chlorin sodium salt (prepared according to the method of Example 53) was weighed and dissolved in 1 L of water with stirring. 110.0 grams of magnesium chloride hexahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, and the mixture was left overnight in the dark, filtered, washed twice with cold water, filtered under pressure, and dried under reduced pressure to obtain 80.7 grams of chlorin magnesium salt, which was a dark brown powder that decomposed at >312°C (DTA / TG), was soluble in water, and had a λ max = 404 nm (aqueous solution).

[0093] Example 56: 100.0 grams of chlorin sodium salt (prepared according to the method of Example 53) was weighed and dissolved in 1 L of water with stirring. 100.0 grams of calcium chloride dihydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to precipitate, and the mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 83.1 grams of chlorin calcium salt, which was a black powder that decomposed at >373°C (DTA / TG), was soluble in water, and had a λ max = 404 nm (aqueous solution).

[0094] Example 57: 100.0 grams of chlorin sodium salt (prepared according to the method of Example 53) was weighed and dissolved in 1 L of water with stirring. 110.0 grams of ferrous chloride tetrahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 83.9 grams of ferrous chlorin salt, which was a black powder that decomposed at >292°C (DTA / TG), was poorly soluble in water, and had a λ max = 405 nm (aqueous solution).

[0095] Example 58: 100.0 grams of chlorin sodium salt (prepared according to the method of Example 53) was weighed and dissolved in 1 L of water with stirring. 110.0 grams of iron trichloride hexahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, and the mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 84.5 grams of chlorin iron salt, which was a black powder that decomposed at >303°C (DTA / TG), was poorly soluble in water, and had a λ max = 403 nm (aqueous solution).

[0096] Example 59: 100.0 grams of chlorin sodium salt (prepared according to the method of Example 53) was weighed and dissolved in 1 L of water with stirring. 100.0 grams of manganese chloride tetrahydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, and the mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 81.4 grams of chlorin manganese salt, which was a black powder that decomposed at >284°C (DTA / TG), was poorly soluble in water, and had a λ max = 404 nm (aqueous solution).

[0097] Example 60: 100.0 grams of chlorin sodium salt (prepared according to the method of Example 53) was weighed and dissolved in 1 L of water with stirring. 100.0 grams of zinc chloride was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to precipitate, and the mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 86.0 grams of chlorin zinc salt, which was a black powder that decomposed at >349°C (DTA / TG), was poorly soluble in water, and had a λ max = 409 nm (aqueous solution).

[0098] Example 61: 100.0 grams of chlorin sodium salt (prepared according to the method of Example 53) was weighed and dissolved in 1 L of water with stirring. 95.0 grams of copper chloride dihydrate was weighed and dissolved in 500 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, and the mixture was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 84.6 grams of chlorin copper salt, which was a black powder that decomposed at >245°C (DTA / TG), was poorly soluble in water, and had a λ max = 405 nm (aqueous solution).

[0099] Example 62: 10.0 g of hemin was weighed and dissolved in 1000 mL of 6.0% potassium hydroxide aqueous solution, and the mixture was stirred for 5 hours in the dark. 8000 mL of acetone was slowly added, and the mixture was stirred for 1 hour in the dark and then left overnight. The mixture was filtered, washed with cold acetone, and dried under reduced pressure to obtain approximately 8.37 g of hematin potassium salt, which was a black solid powder that decomposed at >303°C (DTA / TG) and was easily soluble in water. max = 388 nm (aqueous solution).

[0100] Example 63: 5.0 grams of hematin potassium was weighed and dissolved in 1000 mL of water. 5% sulfuric acid was slowly added dropwise to the mixture while stirring in the dark until the pH was approximately 4. The mixture was left standing overnight, filtered, and the filter cake was washed with water until the filter cake was neutral and dried under reduced pressure. The resulting solid was dissolved in 1000 mL of acetone, and dry ammonia gas was slowly passed through the filter cake while stirring until the cake was saturated. The mixture was left standing overnight, filtered, washed with a small amount of cold acetone, and dried under reduced pressure to obtain approximately 3.17 grams of hematin ammonium salt, which was a brown solid that decomposed at >287°C (DTA / TG) and was easily soluble in water. max = 383 nm (aqueous solution).

[0101] Example 64: 10.0 grams of hematin potassium salt was weighed and dissolved in 200 mL of water with stirring. 12.0 grams of magnesium sulfate was weighed and dissolved in 100 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with a small amount of cold water, and dried under reduced pressure to obtain about 7.25 grams of hematin magnesium salt, which was a dark brown solid that decomposed at >293°C (DTA / TG), was soluble in water, and had a λ max = 387 nm (aqueous solution).

[0102] Example 65: 10.0 grams of hematin potassium salt was weighed and dissolved in 200 mL of water with stirring. 10.0 grams of calcium chloride was weighed and dissolved in 100 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain about 7.46 grams of hematin calcium salt, which was a dark brown solid that decomposed at >307°C (DTA / TG), was soluble in water, and had a λ max = 392 nm (aqueous solution).

[0103] Example 66: 10.0 grams of hematin potassium salt was weighed and dissolved in 200 mL of water with stirring. 11.0 grams of manganese sulfate was weighed and dissolved in 150 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain about 8.12 grams of hematin manganese salt, which was a dark brown solid that decomposed at >322°C (DTA / TG), was poorly soluble in water, and had a λ max = 390 nm (aqueous solution).

[0104] Example 67: 10.0 grams of hematin potassium salt was weighed and dissolved in 200 mL of water with stirring. 14.0 grams of ferrous sulfate was weighed and dissolved in 150 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 7.71 grams of hematin ferrous salt, which was a dark brown solid that decomposed at >331°C (DTA / TG) and was poorly soluble in water. max= 371 nm (aqueous solution).

[0105] Example 68: 10.0 grams of hematin potassium salt was weighed and dissolved in 200 mL of water with stirring. 11.0 grams of iron chloride was weighed and dissolved in 120 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 8.43 grams of hematin iron salt, which was a dark brown solid that decomposed at >328°C (DTA / TG) and was poorly soluble in water. max = 367 nm (aqueous solution).

[0106] Example 69: 10.0 grams of hematin potassium salt was weighed and dissolved in 200 mL of water with stirring. 14.0 grams of zinc sulfate was weighed and dissolved in 150 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain about 8.94 grams of hematin zinc salt, which was a dark brown solid that decomposed at >303°C (DTA / TG), was poorly soluble in water, and had a λ max = 375 nm (aqueous solution).

[0107] Example 70: 10.0 grams of hematin potassium salt was weighed and dissolved in 200 mL of water with stirring. 15.0 grams of copper sulfate was weighed and dissolved in 150 mL of water with stirring. The two solutions were gently mixed with stirring, stirred for 1 hour, and then left overnight. The mixture was filtered, washed with cold water, and dried under reduced pressure to obtain approximately 8.22 grams of hematin copper salt, which was a dark brown solid that decomposed at >266°C (DTA / TG) and was poorly soluble in water. max = 363 nm (aqueous solution).

[0108] Example 71: 100.0 g of chlorin / iron parent compound (manufactured by Nanjing Baite Biotechnology Co., Ltd.) was weighed and dissolved in 1000 mL of 6% potassium hydroxide aqueous solution under stirring. The mixture was stirred for 5 hours in the dark, and then 8000 mL of acetone was slowly added. The mixture was stirred for 1 hour in the dark and then left overnight. The mixture was filtered, washed with cold acetone, and dried under reduced pressure to obtain 9.02 g of potassium salt of hydroxychlorophyllin / iron, which was a dark green powder that decomposed at >320°C (DTA / TG) and was easily soluble in water. max = 395 nm (aqueous solution).

[0109] Example 72: Hydroxychlorophyllin / iron sodium salt (8.46 g) was obtained in the same manner as in Example 71, except that potassium hydroxide solution was used instead of sodium hydroxide solution. The obtained product was a dark green powder that decomposed at >311°C (DTA / TG), was easily soluble in water, and had a λ max = 396 nm (aqueous solution).

[0110] Example 73: 5.0 grams of sodium salt of hydroxychlorophyllin / iron was weighed and dissolved in 1000 mL of water. 5% sulfuric acid was slowly added dropwise to the solution while stirring in the dark until the pH was approximately 4. The solution was left overnight, filtered, and the filter cake was washed with water until the filter cake was neutral and dried under reduced pressure. The resulting solid was dissolved in 1000 mL of acetone, and dry ammonia gas was slowly passed through the solution while stirring until the solution was saturated. The solution was left overnight, filtered, washed with a small amount of cold acetone, and dried under reduced pressure to obtain 3.34 grams of ammonium salt of hydroxychlorophyllin / iron. The powder was a dark green color and decomposed at >255°C (DTA / TG). It was easily soluble in water and had a λ max = 404 nm (aqueous solution).

[0111] Example 74: 10.0 grams of sodium salt of hydroxychlorophyllin / iron was weighed and dissolved in 100 mL of water with stirring. 11.0 grams of magnesium chloride hexahydrate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed twice with cold water, filtered under pressure, and dried under reduced pressure to obtain 7.27 grams of magnesium salt of hydroxychlorophyllin / iron, a dark green powder that decomposed at >317°C (DTA / TG), was soluble in water, and had a λ max = 401 nm (aqueous solution).

[0112] Example 75: 10.0 grams of sodium salt of hydroxychlorophyllin / iron was weighed and dissolved in 100 mL of water with stirring. 10.0 grams of calcium chloride dihydrate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to precipitate, and the precipitate was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 7.71 grams of calcium salt of hydroxychlorophyllin / iron, which was a dark green powder that decomposed at >332°C (DTA / TG), was soluble in water, and had a λ max = 402 nm (aqueous solution).

[0113] Example 76: 10.0 grams of sodium salt of hydroxychlorophyllin / iron was weighed and dissolved in 110 mL of water with stirring. 10.0 grams of ferrous chloride tetrahydrate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 9.26 grams of ferrous salt of hydroxychlorophyllin / iron, which was a dark green powder that decomposed at >315°C (DTA / TG), was poorly soluble in water, and had a λ max = 398 nm (aqueous solution).

[0114] Example 77: 10.0 grams of sodium salt of hydroxychlorophyllin / iron was weighed and dissolved in 110 mL of water with stirring. 10.0 grams of iron trichloride hexahydrate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 8.11 grams of iron salt of hydroxychlorophyllin / iron, which was a dark green powder that decomposed at >304°C (DTA / TG), was poorly soluble in water, and had a λ max = 401 nm (aqueous solution).

[0115] Example 78: 10.0 grams of sodium salt of hydroxychlorophyllin / iron was weighed and dissolved in 100 mL of water with stirring. 10.0 grams of manganese chloride tetrahydrate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 87.8 grams of manganese salt of hydroxychlorophyllin / iron, which was a dark green powder that decomposed at >331°C (DTA / TG), was poorly soluble in water, and had a λ max = 403 nm (aqueous solution).

[0116] Example 79: 10.0 grams of sodium salt of hydroxychlorophyllin / iron was weighed and dissolved in 100 mL of water with stirring. 10.0 grams of zinc chloride was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to precipitate, and the precipitate was left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 8.67 grams of zinc salt of hydroxychlorophyllin / iron, which was a dark green powder that decomposed at >327°C (DTA / TG), was poorly soluble in water, and had a λ max = 403 nm (aqueous solution).

[0117] Example 80: 10.0 grams of sodium salt of hydroxychlorophyllin / iron was weighed and dissolved in 100 mL of water with stirring. 9.0 grams of copper chloride dihydrate was weighed and dissolved in 50 mL of water with stirring. The two solutions were gently mixed at room temperature with stirring to cause precipitation, which was then left overnight in the dark, filtered, washed with cold water, filtered under pressure, and dried under reduced pressure to obtain 8.42 grams of copper salt of hydroxychlorophyllin / iron, which was a dark green powder that decomposed at >264°C (DTA / TG), was poorly soluble in water, and had a λ max = 395 nm (aqueous solution).

[0118] Example 81: Differential Thermal / Thermogravimetric Analysis (DTA / TG) of Sodium Chlorin / Iron and Chlorin / Iron Test equipment: Shimadzu DTG-60 differential thermal and thermogravimetric simultaneous analyzer Test sample: sodium salt of chlorine / iron, chlorine / iron Reference: Aluminum trioxide Test gas: Nitrogen gas Heating rate: 10°C / min Temperature range: 25℃~500℃ Sample weight: 4.3 to 4.5 milligrams Analysis software: ta60WS Ver. 2.11 Analysis of the results: The DTA / TG spectra of the sodium salt of chlorine / iron and chlorine / iron were measured, and the spectra of the two samples were combined using analysis software. The results are shown in Figure 1, which shows the DTA comparison chart (upper panel) and TG comparison chart (lower panel) of the samples, respectively.

[0119] The DTA / TG spectra of the samples after converting chlorin / iron into sodium salts show that there is no melting phase transition peak (no melting point) in the two samples, and the difference in thermal response is very obvious. Within the same measurement range (uV), the DTA curve of the acid-form chlorin / iron (upper panel of Figure 1) is stable with little fluctuation, and the endothermic and exothermic peaks are almost flat, whereas the curve of the sodium salt of chlorin / iron shows large fluctuations, with two large exothermic peaks at 271.67°C and 453.24°C. The endothermic peak at 90.46°C is the dehydration or decomposition of low-boiling-point small molecules in the chlorin / iron sample, and the heat release peak at 219.16°C is the heat release peak for the chlorin / iron sample. The change in specific heat of the samples within the measurement range is clear. The chlorine / iron sodium salt, which is not white (almost horizontal), shows obvious endothermic heat due to dehydration. The endothermic peak shifts to 71.7°C, and the endothermic heat increases significantly. The specific heat of the sample increases up to 110°C, and the curve rises. From 180°C, decomposition begins, with intense heat release, resulting in a peak at 271.67°C. A second peak appears at 453.24°C, and after further decomposition, the specific heat begins to decrease. The difference in the thermal reactions of the two samples is very obvious, and the decomposition point of the salt sample is significantly higher than that of the acid sample. Comparing the TG spectra (bottom panel of Figure 1), the TG curve of chlorine / iron salt showed a slow change before reaching 140°C, with a small weight loss of 3.256%. The TG curve of the sodium salt of chlorine / iron salt showed a rapid decline, with a dehydration weight loss of 7.511%, corresponding to the bimolecular hydration (salt) content. The sodium salt of chlorine / iron salt showed a gradual weight loss (decomposition) after reaching 180°C, then a rapid weight loss at 390°C (increased decomposition). The sodium salt of chlorine / iron salt showed a decrease in weight loss above 200°C, a decrease in weight loss at 295°C, and a rapid increase above 405°C (increased decomposition). This indicates that the decomposition temperature of the salt sample was significantly higher than that of the acid sample. The results show that the two samples only have decomposition points when heated, and the thermal stability of the chlorine iron after salification is obviously stronger than that of the acid form sample.

[0120] Example 82: Dilution of sample solutions such as chlorine / iron sodium salts and pH measurement Samples: chlorophyllin / iron sodium salt (commercially available food additive, food class, content 96%), protoporphyrin sodium (commercially available pharmaceutical raw material, pharmaceutical class, content 95.0%), chlorin / iron sodium salt (content measured by photometry was 95.5%, same below), heme potassium (content 96.0%), chlorophyllin / iron magnesium salt (content 97.3%), heme magnesium (content 95.7%), pure water (pH=7). Measuring equipment: FA1604N electronic balance, PHS-3C benchtop acidity meter, ultrasonic meter, volumetric flask. Method: Accurately weigh 0.2500 grams of sample into a 25 mL volumetric flask, dissolve in pure water, and dilute to the mark to prepare a 1.00% (10,000.0 ppm) stock solution. The stock solution was then diluted with pure water at different concentrations to prepare a series of solutions ranging from 1,000.0 ppm to 0.1 ppm. Each sample was measured three times and its pH was determined. For poorly soluble salts, a solution with a concentration below the saturated solubility was prepared. An appropriate amount of sample was weighed, dissolved, and diluted in the same manner. The corresponding mol / L concentrations and the pH values ​​of the solutions are shown in the table below.

[0121] [Table 2]

[0122] As shown in the table, the measured samples showed that when the concentration of readily soluble sodium / potassium salts was 1% (10,000.0 ppm), the solution pH was 10.21-10.95, indicating strong alkalinity. Even when diluted 10 times, it remained strongly alkaline. As the concentration was further diluted, the pH decreased and the alkalinity decreased significantly. When soluble magnesium salts (100 ppm) were used, the pH was similar to that of sodium / potassium salt solutions. At 30.0 ppm, the solution pH was <8.0 (pH = 7.50-7.63), which is within the physiologically appropriate pH range for plants. When diluted to 10.0 ppm, the solution was close to neutral. Furthermore, at concentrations of 1.0 ppm to 0.1 ppm (~10 ppm), which are the concentrations typically used for the samples of this invention, the solution pH was close to neutral. -6m When diluted to the 0.05% (0.05% mol / L) range, the sample solutions unexpectedly exhibited weak acidity. In the samples tested, chlorophyllin / iron sodium salt (a food additive) and protoporphyrin sodium (a drug for treating liver disease) were all commercially available products, and the dilute solutions unexpectedly exhibited weak acidity, as did the samples such as chlorin / iron sodium salt and heme sodium salt newly prepared according to the present invention (no literature reports).

[0123] Generally, acid or alkaline solutions are dissolved in water at a concentration of 10 -7 When diluted to a concentration close to or even lower than 1000 mol / L, the pH of the solution approaches or is approximately equal to 7.0, but does not exceed 7.0 (going from acidic to alkaline, or from alkaline to acidic). A salt solution of a strong base or a weak acid is a strong base under normal concentration conditions (concentration>>10 -7 mol / L), the aqueous solution of salt becomes obviously alkaline due to the hydrolysis of weak acid radicals. After diluting the solution, as the concentration decreases, the hydrolysis effect of salt gradually weakens, reaching 10 -6 ~10 -7When the solution was diluted to 0.1 mol / L (the sample concentration in this example was 1.0 ppm to 0.1 ppm), hydrogen ions generated by the dissociation of water gradually became dominant, and the pH of the solution should be close to or approximately equal to 7.0. However, in this measurement, the pH was <7.0, and the solution unexpectedly changed from weakly alkaline to weakly acidic. This result suggests that the acidic components present in the system, i.e., the measured porphine salt, may coexist with small or trace amounts of acid-form porphine in the sample, and these residual acid-form porphines are the reason why the solution exhibits a weak acidity after dilution.

[0124] Acid-form porphines are polybasic acids. For example, protoporphyrin sodium and heme are both dibasic acids, while chlorine and chelates are mixtures of monobasic, dibasic, and tribasic acids. The soluble sodium, potassium, or ammonium salt samples used in the present invention are all prepared by precipitation, in which acid-form porphines are neutralized with alkali in an organic solvent. Divalent and trivalent metal salts, such as magnesium salts, are prepared by metathesis precipitation. During the salification reaction, various process factors, such as the concentration of the organism, the rate of alkali addition, the uniformity of stirring, and the reaction temperature, can limit the formation of precipitates and the composition of the products. Experiments have shown that incomplete salification during the preparation of porphine salts or a small amount of unreacted acid-form molecules are simultaneously surrounded by the precipitate, resulting in a coprecipitation phenomenon. This proves that incomplete salification during preparation is unavoidable because the planar porphine molecules have a tendency to aggregate with each other, which is an inherent factor that makes them prone to coprecipitation. Such unexpected pH changes enhance the utility of the products of the present invention, making them more suited to the physiological conditions of the crops and more useful for use on field crops.

[0125] Example 83: Measurement of sample solution stability in dark / light Samples: A total of 90 samples of porphyrin salts and chlorin salts (all contents were ≥ 95.0% and were measured by photometry) Control: chlorine / iron, hemin Measuring instruments: Shimadzu UV2600 ultraviolet spectrophotometer, Smart Sensor AR813A illuminance meter Measurement: An appropriate amount of salt sample was taken, dissolved and diluted in water to prepare a test solution with a concentration of approximately 10 ppm. For the acid-type control product (which is very poorly soluble in water), a test solution with a concentration of approximately 10 ppm was prepared using a 30% to 50% aqueous ethanol solution. After preparing each test solution, the absorbance (at time 0) was scanned using an ultraviolet spectrophotometer in the range of 200 nm to 800 nm. Each sample test solution was divided into two parts, one part of the test solution was left in the dark at room temperature of 25°C, and the other part was placed under a light intensity condition of 30,000 lux. Solutions were sampled at 1, 2, and 3 hours, and the absorbance of the test solution at each time point was repeatedly measured by scanning.

[0126] The absorbance at the maximum absorption peak of each test solution at time 0 (initial time) was taken as 100% relative content, and the absorbance value at each time point was compared with the value at time 0 to calculate the relative content %. The results showed that under dark conditions, the contents of the test salt samples and acid-type control samples remained essentially unchanged or even slightly decreased in some cases (the relative contents were still >96.5%).

[0127] In comparative studies under light conditions, the stability of the salt sample solutions was significantly different from that of the control sample. Taking the light stability of hemin and hemin magnesium salt solutions as an example, a comparison of their absorbance scans is shown in Figure 2. The hemin solution content dropped to less than 30% after 1 hour of light exposure, and after 3 hours, the content was only 20.21%. The relative content of the corresponding hemin magnesium salt after 3 hours was 75.61%, significantly higher than that of the acid-form control. The same results were obtained when comparing chlorin / iron and its sodium salt. The changes in relative content of each control sample are shown in the table below.

[0128] [Table 3]

[0129] Furthermore, the time it takes for the relative content of each sample solution to decrease by 50% under strong light conditions (half-life t 1 / 2 ) were measured and compared, and the calculated results are shown in the table below. [Table 4] The results were as follows: Under strong light, the half-life (t 1 / 2 The half-lives of the magnesium and sodium salts were 0.69 and 0.73 hours, respectively, and the half-lives of the magnesium and sodium salts increased to 6.0 and 3.41 hours, respectively, an increase of 8.69 and 4.67 times, indicating that the stability of the salted compound solutions under light exposure was significantly enhanced compared to the acid-form control. Under dark conditions, the t 1 / 2 The stability of the salt type compound is better than that of the acid type compound solution, which is more suitable for the complex application environment of the field and has more practical value.

[0130] Aqueous solutions of 90 salt-type compounds (measured concentrations were 12 ppm to 25 ppm) were exposed to strong light for 3 hours to detect their stability, and the results are shown in the table below. [Table 5] TIFF2025532772000007.tif226170

[0131] The results showed that the series of salt-type compounds of the present invention all have strong stability, and even after 3 hours of irradiation, the relative content of most sample solutions can maintain more than 50%. This shows that the stability is significantly superior to that of currently used acid-type products, making them capable of meeting a wide range of practical needs and of greater practical value.

[0132] Example 84: Effects of chlorin / iron sodium salt, heme potassium, and protoporphyrin potassium on wheat germination and seedling growth Test variety: Huaimu 35 Sample: sodium salt of chlorine / iron, potassium heme, potassium protoporphyrin, purified water control (CK) Methods: Each treatment was replicated three times. 50 whole, consistently sized seeds were selected for each sample. They were sterilized with sodium hypochlorite for 5 minutes, rinsed with purified water, and placed in a plastic box covered with 100g of sand. 25ml of the prepared sample solution was poured over each seed. The seeds were then neatly arranged, capped, and placed in a 25°C incubator for cultivation. Germination was defined as the length of the wheat seed sprout exceeding half the seed diameter (long axis). Seed whiteness and number of germinated seeds were recorded daily according to the wheat growth conditions. On the sixth day, 10 wheat seedlings were randomly selected and measured for root length, plant length, and root number. On the seventh day, root weight and aboveground weight were measured. All measurements were repeated three times.

[0133] The wheat germination rates of the three samples at different concentrations are shown in Table 1 below. [Table 6] The results showed that compared to the control (CK = 0 ppm), all samples in the concentration range of 0.2500 ppm to 0.0025 ppm showed a clear promotion effect on wheat germination, with 0.2500 ppm heme potassium showing the best promotion effect on germination rate, and the other samples showing a preferred concentration of 0.0250 ppm.

[0134] The effect of different concentrations of chlorin / iron sodium salt solution on the roots / seedlings of wheat seedlings is shown in Table 2 below. [Table 7] The results showed that chlorine / iron sodium salt had different effects on the growth of early wheat seedlings, with a clear promoting effect on root growth and a relatively weak effect on the above-ground seedling parts, and that 0.0250 ppm was a suitable concentration for use.

[0135] The effect of different concentrations of heme potassium solution on wheat seedling roots / seedlings is shown in Table 3 below. [Table 8] The results showed that low concentrations of heme potassium (0.0250 ppm or 0.0025 ppm) could significantly promote the growth of wheat seedlings above ground and the increase in the number of root systems, but had a weak effect on promoting root length.

[0136] The effect of different concentrations of potassium protoporphyrin solution on the roots / seedlings of wheat seedlings is shown in Table 4 below. [Table 9] The results showed that low concentrations of potassium protoporphyrin (0.0250 ppm or 0.0025 ppm) had some effect on promoting root growth of wheat, but had little effect on promoting shoot growth at the seedling stage.

[0137] Example 85: In vitro chlorophyllase inhibitory activity test According to a literature method (Northwestern Botanical Journal, 2003, 23(5):750-754), 2.00 g of acetone powder prepared from mung bean leaves was accurately weighed and extracted with 50 mL of phosphate buffer (pH 7.0-7.3) for 4.0 hours, then centrifuged at 4-5°C for 10 minutes. The supernatant was a chlorophyll solution with an active enzyme concentration of 0.13 mmol / L. Chlorin salt samples were added to the enzyme solution at concentrations of 0.3 ppm to 30.0 ppm, and the substrate chlorophyll (120 ppm) was added. The chlorophyllase inhibitory activity (IC) of the salt samples was measured at each concentration. 50 The values ​​were calculated and the results are shown in the table below. [Table 10] Chlorin salts are extremely important and valuable for practical use because they maintain the strong chlorophyllase inhibitory activity of acid chlorins.

[0138] Example 86: Adventitious root induction test of mung bean hypocotyls To prepare the test solution, 8 mL of commercially available STANLEY general-purpose nutrient solution was diluted with tap water to 3200 mL (400-fold dilution) to obtain the diluted nutrient solution. Salt samples, such as sodium protoporphyrin and potassium hemin, were weighed appropriately and used to prepare 0.010 ppm and 0.1 ppm test solutions. 150 mL of each was taken. Another 150 mL of the diluted nutrient solution served as a blank control. The test and control solutions were placed in brown bottles, the tops of which were covered with black film. Mung bean seedlings with uniform growth and height of 7-8 cm were selected and cut 3 cm below the first node of the true leaves. Eight mung bean seedlings (roots excluded) were placed in each bottle, with the lower radicle immersed in the solution. The solution was changed every two days at room temperature (20-25°C) for a total of 7 days. A clear difference was observed on the 2nd or 3rd day after treatment, and observations and measurements were made on the 7th day, and the number of adventitious roots per cutting and the average total length of the adventitious roots were calculated. The results are shown in the table below. [Table 11] The results showed that compared with the blank control, each test sample had a significant effect of promoting the rooting range and number of roots in mung bean hypocotyls, and the effects were all superior to those of hemin, an existing plant growth regulator product.

[0139] Example 87: Experiment to increase rice yield by controlling sheath blight (use of plant immunity inducer) Location: Zhonghua Family Farm, Guangming Village, Dazhong Road, Dafeng District, Jiangsu Province, China. Six acres of flat, fertile clay loam rice paddy were selected. The previous crop was wheat. Rice variety tested: Nanjing 9108 The seeds were sown on May 16, 2021, and transplanted on June 12, 2021. The plant spacing for machine planting was 18 cm, and the row spacing was 25 cm. Two treatments were established in accordance with GB / T 17980.20-2000 Guidelines for Field Efficacy Trials. Treatment 1 was chlorine / iron sodium salt at 3.0 mg / row, and treatment 2 was purified water (control). Taking into account plant growth characteristics and pest control needs, the chemical was sprayed once at the end of tillering (July 22, 2021), internode elongation / booting stage (August 16, 2021), and full heading stage (August 30, 2021). The water requirements per row were 30 L, 45 L, and 45 L, respectively (concentrations of 0.100 ppm, 0.067 ppm, and 0.067 ppm). The test samples were dissolved, diluted, and mixed uniformly before spraying. Each treatment area was set at 3 rows.

[0140] Before harvest, five sampling points per treatment (20 m per sampling point) were collected. 2 ) and the pathological condition of rice sheath blight was investigated in detail, and the corrected control effect obtained is shown in the table below. [Table 12]

[0141] The harvest date was November 8, 2021. Prior to harvest, the yield composition and structure were investigated and the yield per row was measured. The results are shown below. [Table 13] The experiments showed that chlorin / iron sodium salt is a plant immunity inducer that is clearly effective in controlling rice sheath blight and also has excellent effects in increasing rice yield.

[0142] Example 88: Experiment to increase yield by controlling rice leaf blast (use of plant immunity inducer) Location: Zhonghua Family Farm, Guangming Village, Dazhong Road, Dafeng District, Jiangsu Province, China. The previous crop was oilseed rape. Test field: Flat terrain, clay loam soil, good fertility. Rice variety: Nanjing 9108. The seeds were sown on May 16, 2021, and transplanted on June 14, 2021, with a plant spacing of 18 cm and row spacing of 25 cm when planted mechanically.

[0143] The experiment was conducted in accordance with the experimental standard GB / T17980.19-2000. Two treatments were used: Treatment 1, which was chlorine / iron sodium salt, and Treatment 2, which was a blank control (water). The treatments were thoroughly diluted and dissolved before spraying. Based on the characteristics of pest control, the treatments were sprayed once at the end of tillering (July 22, 2021), the internode elongation / booting stage (August 16, 2021), and the full-heading stage (August 30, 2021). The chlorine / iron sodium salt dosage was 3 mg / row, and the water requirements per row were 30 L, 45 L, and 45 L, respectively (sample concentrations were 0.1 ppm, 0.067 ppm, and 0.067 ppm). The area of ​​each treatment was 3 acres, and five points were sampled for each treatment using a five-point sampling method (20 m per sampling point). 2 Before harvest, the occurrence of rice leaf blast was investigated in the paddy fields, and the corrected control effect is shown in the table below. [Table 14] The test results showed that chlorine / iron sodium salt significantly improved rice's resistance to rice leaf blast disease, demonstrating the effectiveness of using a plant immunity inducer.

[0144] The harvest took place on November 8, 2020. The yield, composition, and structure were investigated before harvesting, and the yield per row was measured. The yield-increasing effect of the sample was clear, and the specific results are shown in the table below. [Table 15]

[0145] Example 89: Yield increase test of peppers sprayed with heme potassium salt solution Experimental location: Litangfang Village, Wucheng Town, Wucheng County, Dezhou City, China. Test site: medium loam soil, medium to high fertility, previous crop was sorghum. Test variety: Hebei Jieze chili pepper, planting density is 3,500 plants per row. Planting method: mechanical transplantation. Planting time: May 8, 2020. Test method: The test and control groups consisted of 10 acres each. The treatment group was sprayed with a heme potassium salt solution, the control group was sprayed with heme (at the same dose and concentration as the treatment group), and the blank group was sprayed with purified water. The treatment and control groups were sprayed on May 19 (after transplanting) and July 2 (at full bloom), respectively. The dose was 1.5 mg / acre, and the water requirements per acre were 15 L and 30 L, respectively (sample concentrations were 0.1 ppm and 0.05 ppm). On September 13, a sampling survey was conducted for the treatment and control groups by selecting 10 chili pepper plants with uniform growth. The number of fruits and individual fruit weights were recorded. The entire field was harvested three times: August 30, September 13, and September 25. Yields were also statistically compared, and the results are shown below. [Table 16] The test showed that spraying heme potassium salt on chili peppers significantly increased the fruit set rate and fruit weight, resulting in a significant yield increase. The heme in the control group also showed a good yield increase effect, but the increase in yield was not as great as that of heme potassium salt, and the difference was clear.

[0146] Example 90: Test to increase yield of tomatoes by spraying with chlorophyllin / sodium iron salt Experimental site: The 3rd Regiment of the 22nd Regiment of the Xinjiang Construction Corps, China. The original crop was a seedbed, and the soil quality was light. Variety: Processed Tomato 1615. The cultivation method employed flat planting, plastic covering, seedling transplanting, and pressurized drip irrigation. Seedlings were transplanted at 2,600 plants per row on April 29th. The control and treatment areas were 20 rows each, and the treatments were sprayed twice, once at the beginning of flowering (May 22, 2021) and once at the fruit-setting stage (June 5, 2021). The chlorophyllin / iron sodium salt solution was sprayed at a concentration of 0.05 ppm.

[0147] Harvest was on August 3. During harvest, five consecutive plantings from each treatment were randomly selected for sampling statistics, the data of which are presented below. [Table 17] The results showed that chlorophyllin / iron sodium salt could increase the fruit set rate and single fruit weight of tomatoes, and increase the tomato yield per row by more than 27%. At the same time, after spraying chlorophyllin / iron sodium salt, the red / green ratio of tomatoes reached 3.36 (compared to the control's red / green ratio of 3.17), an increase of 5.91%, indicating that chlorophyllin / iron sodium salt not only increases yield, but also has the obvious effect of promoting early coloring of tomatoes and improving the quality of fruits and vegetables. The test proved that chlorophyllin / iron sodium salt has high practical value and potential for wide application.

[0148] Example 91: Comparison of spraying chlorine / iron and its potassium salt to increase wheat yield Experimental site: Wheat experimental field at Huanghai Farm Agricultural Science Station, Jiangsu Province, China, previous crop Sowing date: December 10, 2020, sowing amount: 30kg / row, rice. Crops: Huaimai No. 35, chlorine / iron (powder) treated fields, potassium salt treated fields and control fields (sprayed purified water) each cover 7 acres. Method: The chlorin / iron potassium salt solution was sprayed once each at the isshin-sanba stage (March 14, 2021), the booting / panicle splitting stage (April 14, 2021), and the flowering stage (May 1, 2021). The amount of chlorin / iron potassium salt used per spray was 3 mg / row, and the amount of water used per row was 15 L, 30 L, and 30 L, respectively (concentrations of 0.2 ppm, 0.1 ppm, and 0.1 ppm).

[0149] The entire field was harvested on June 11, 2021. The actual measured data is shown below. [Table 18] The amount of chlorine / iron and its potassium salt used was 3 mg / row, and the yield-increasing effect of spraying was obvious, but the yield-increasing effect of the potassium salt was even greater.

[0150] Example 92: Control test of tobacco diseases by spraying sodium salt of chlorin / iron Location: Wujiang Village, Tianwen Town, Lian'an County, Guizhou Province, China. Variety: Unen 87, transplanted on May 13th, with 100cm spacing between rows and 50cm spacing between plants. Samples: chlorine / iron sodium salt, chlorine / iron powder (positive control), lentinan solution (positive control), purified water (blank control) Methods: Six treatments were set up, treatments 1-3 were three different concentrations of chlorine / iron sodium salt solutions, control 1 was a chlorine / iron powder solution, control 2 was 0.5% lentinan solution, and control 3 was a blank control. For each treatment, four relatively dispersed plots were selected in the field, each 50 square meters in size. The pesticide was sprayed twice on May 28th and June 4th during the root elongation period after transplanting the tobacco leaves (when there are 12 to 14 leaves). The spray volume for each treatment was the same (30 liters per furrow) to ensure that the leaves were sprayed evenly without leaving any blind spots during the work.

[0151] On July 15th, all tobacco plants in each treatment area were inspected, and the disease classification was conducted according to the Chinese National Standard for Tobacco Disease Classification GB / T23222-2008. The disease incidence rate and disease index for each treatment area were statistically calculated, and the results are shown below. [Table 19] The results showed that chlorin / iron sodium salt significantly enhanced tobacco immunity, resisted tobacco mosaic virus, and reduced the incidence of the disease, and its effects were superior to those of the positive controls chlorin / iron and lentinan.

[0152] Example 93: Test to promote tobacco yield by spraying sodium salt of chlorine / iron Test site: Yutang Village, Huanghua Town, Changsha County, Hunan Province, China. Variety: Unen 87, planting density 16,500 plants / hectare, spacing 50cm between plants, and row spacing 110-120cm. Method: Four spray concentrations were set, with four plots per treatment, each 20 square meters. The drug was sprayed twice on May 8, 2019, during the root elongation period after transplanting tobacco leaves (when there were 12 to 14 leaves), and on May 15, 2019, with a water usage of 45 L per row (spray volume was 1.35 L per plot).

[0153] At harvest, the height of the plant, the length and width of the leaves, and the weight of each leaf were sampled and statistically recorded. All leaves in the plot were then hot-air-cured into flue-cured tobacco, and the concentration of each sample (chlorine / iron sodium salt), the growth status of the flue-cured tobacco after spraying, and the yield were statistically recorded. The results are shown in the table below. [Table 20] There was a certain correlation between the effect of spraying sodium chlorinate / iron on tobacco leaves and the concentration. At the tested concentrations, sodium chlorinate / iron significantly increased tobacco leaf weight, increased the width and length of the leaf blades, and increased tobacco yield per row. However, 0.1 ppm was more suitable and contributed to increasing tobacco leaf yield.

[0154] Example 94: Cotton wilt control test by spraying chlorophyllin / sodium salt of iron Experimental location: East Red No. 1 Unit, Wutubulage Town, Bole City, Xinjiang Uygur Autonomous Region, China. The previous crop was cotton. Cotton variety: Shinrikuchu 66, sown on April 21, 2020, with 4 rows per mulch film, 15,000 plants per ridge, and each treatment area was 1 ridge. Samples: chlorophyllin / iron sodium salt, VitaCat (positive control), purified water (blank control) Method: Three concentrations of samples were set, along with a positive control and a blank control, for a total of five treatments. Each was sprayed once at the cotton seedling stage (May 28th) and the bud stage (June 28th), with the concentration adjusted each time. The concentration changes for each treatment of chlorophyllin / iron sodium salt and the control (treatments 1 to 5) are shown in the table below. [Table 21]

[0155] The incidence of wilt disease was measured 20 days after each sample application using a three-point survey method for each treatment, with 100 plants selected at each point. The control effect was investigated, and the results are shown below. [Table 22]

[0156] The harvesting period was November 15th, 2020. At the time of harvesting, three sampling points were selected from each treatment plot, with an area of ​​0.01 mu per sampling point. The cotton growth status was recorded, and the cotton was harvested and the yield increase was calculated. The data is shown below. [Table 23] The results showed that chlorophyllin / iron sodium salt could significantly reduce the incidence of cotton wilt and increase cotton yield. As the spray concentration of seedling samples increased, the control effect of cotton wilt improved, and the overall effect was superior to that of the positive control.

[0157] Example 95: Effect of potassium chlorin / iron salt on germination of salt-stressed rice In this experiment, three NaCl concentrations of 0, 2.5, and 4.5 g / L were set, and 0.2 ppm chlorine / iron potassium salt was added. One salt stress blank control (CK: 4.5 g / L NaCl solution) was also added. There were four treatments in total: (1) 4.5 g / L NaCl solution (CK), (2) 0 g / L NaCl solution-0.2 ppm chlorine / iron potassium salt solution, (3) 2.5 g / L NaCl solution-0.2 ppm chlorine / iron potassium salt solution, and (4) 4.5 g / L NaCl solution-0.2 ppm chlorine / iron potassium salt solution.

[0158] Selected plump rice seeds (cultivar Nanjing 9108) were sterilized by immersion in a sodium hypochlorite solution for 2–3 minutes and then rinsed 2–3 times with purified water. After washing, 50 plump seeds were selected and placed in the prepared seed soaking solution. The seeds were completely soaked at 25°C for 48 hours. After soaking, the seeds were transferred to a plastic box lined with two layers of paper towels. The germination beds were moistened with a solution of the appropriate salt concentration for each treatment and grown in a constant-temperature incubator at 25°C under a 12-hour light / 12-hour dark cycle with 70% humidity. The germination beds were kept moist throughout the cultivation period. Rice seed germination and seedling growth were observed daily. The germination rate for each treatment was recorded on day 7. On day 14, 10 seeds from each treatment were randomly selected and measured for seedling length, root length, root number, root weight, and seedling weight. The germination vigor was measured on the 5th day of germination and the germination percentage was measured on the 14th day of germination. The results are shown in the table below. [Table 24]

[0159] [Table 25]

[0160] [Table 26]

[0161] [Table 27] The results showed that chlorine / iron potassium salt had good stress tolerance properties and could promote the germination of salt-stressed rice seeds, improve germination rate and vigor, promote rooting of seedlings and above-ground growth of plantlets, increase root length, and promote increases in seedling and root weight, thereby significantly reducing the damage caused by salt stress to plantlets.

[0162] Example 96: Testing flooded and submerged rice with sprays of chlorophyllin / sodium iron Location: Irrigation area along the river in Xiaosijiao Village, Guzu Town, Dangtu County, Ma'anshan City, Anhui Province, China. Rice variety: Nannen 9108 Waterlogging: After the water subsided, the rice plants became weak, soft, and yellowed and drooped. On July 10, 2021, an area of ​​20 mu was flooded, and the water subsided on July 20. Over the course of 10 days, the entire rice plant was submerged for nearly four days, and most of the rice plant was submerged for nearly six days. After the water subsided, the rice plants became weak, and the leaves turned yellow and drooped. Method and dosage: A 10.0 ppm chlorophyllin / iron sodium salt solution was sprayed by aerial spraying, spraying treatment on 10 acres, with an average dosage of 1.2 L / acre (the amount used was 12.0 mg / acre), and the blank control field was 10 acres (0 mg / acre). Results: The harvest took place on October 20, 2021, and 320 jin / mu (moisture content 16.5%) was harvested in the control field, while 720 jin / mu (moisture content 16.5%) was harvested in the treated field. The spraying of chlorophyllin / iron sodium salt had a significantly significant effect.

[0163] Example 97: Testing the spraying of potassium chlorine / iron salts on drought-stricken rice seedlings Sample: chlorine / iron potassium salt, purified water (control) Variety: Huiliangyou 473 rice seeds Method: Plump rice seeds were selected and sterilized in 6% sodium hypochlorite for 2-3 minutes, then rinsed thoroughly with water. The seeds were then soaked in purified water at room temperature (30±1°C) until they turned white. They were then transferred to seedling trays for seedling cultivation. After approximately 15 days, rice seedlings with uniform growth conditions were selected and transferred to buckets for further cultivation. Each bucket had three holes, with two seedlings planted per hole. The soil was clay loam, and was crushed, sieved, and mixed uniformly before transplanting. The barrels were 30 cm high and had an inner diameter of 30 cm. Water, fertilizer, and pest control were managed centrally during the potting period. Two concentrations of chlorine / iron potassium salt samples were prepared and sprayed on the transplanted rice seedlings.

[0164] Drought model: Transplanting took place on August 18, 2020, and natural drought and sample spray treatment of rice seedlings began on the second day (September 19). Soil moisture content was measured according to the method of NY / T 52-1987. The specific method and drought model results are shown in Table 1 below. [Table 28] As shown in the table, after natural drought, the soil moisture content changed from normal (moisture content was 26.0%) on day 0 to mild drought (moisture content was 6.0%-10.0%) on day 3, and then to moderate drought (moisture content was 6.0%-8.0%) on day 6.

[0165] After the drought stress and sample spraying started at the rice tillering stage (September 19th), the chlorophyll content (mg / g) of the leaves was measured twice on the third and sixth days (September 21st and September 25th), respectively, and the results are shown in Table 2 below. [Table 29] The results showed that under drought conditions, the chlorophyll content of rice leaves gradually decreased over time (blank treatment), but the effect of drought on the change of chlorophyll content in rice leaves was obvious; with the treatment of chlorin / iron potassium salt solution, the chlorophyll content of rice leaves did not decrease but increased over time, proving that the samples could maintain chlorophyll content even under drought stress, delaying the decomposition of chlorophyll and possessing obvious stress tolerance.

[0166] Example 98: In vitro study of chlorin / iron sodium salts inducing plant resistance to pepper diseases Experimental method: Leaves of Nicotiana benthamiana were injected with sterilized ultrapure water (DDW), 20.0 ppm chlorine / iron sodium salt, and 2.0 ppm chlorine / iron sodium salt. After 7 and 10 days, the leaves were inoculated with a fungal seed cake of Phytophthora capsici LT263. The results were observed 36 hours after inoculation. Results: Compared with DDW treatment, treatment of N. benthamiana with 20.0 ppm and 2.0 ppm of chlorin / iron sodium salt could obviously reduce the lesion area inoculated with LT263 after 7 and 10 days. Conclusion: Treating Nicotiana benthamiana with a chlorin / iron sodium salt solution can effectively attenuate and reduce Phytophthora capsici infection after 7 or 10 days.

[0167] Example 99: Potted experiment of sodium chlorin / iron salt to induce resistance to late blight in pepper Test medium V8 medium: 1 g of CaCO3 was added to 100 mL of V8 vegetable and fruit juice medium, and the mixture was centrifuged at 6000 r / min for 10 minutes. The supernatant was collected and diluted 10 times. 15% agar powder was added and the mixture was sterilized at 121°C for 20 minutes. Test variety: Sujiao No. 5, developed by the Vegetable Research Institute of Jiangsu Academy of Agricultural Sciences. Preparation of Phytophthora capsici zoospores: A fungal dish was punched from the edge of a plate of freshly activated Phytophthora capsici. A 6-mm fungal dish was then transferred, fungal side up, to an empty 9-cm-diameter culture dish with a needle, with 10 fungal dishes placed per Petri dish. 15 mL of sterile water was added to the Petri dish, which was then placed on an ultraclean work surface under light. The water was changed every 30 minutes for a total of three changes. The remaining wastewater was removed with a straw, and 10 mL of V8 liquid medium was added. The mixture was incubated in a dark incubator at 25°C for 24 hours to produce a large number of cystospores. The mixture was then placed in a refrigerator at 4°C for 30 minutes, followed by another 30 minutes of incubation at 25°C, during which a large number of zoospores were released. 10 μL of the mixture was then taken and observed under a microscope using a hemocytometer. The remaining sample was collected for later use.

[0168] Determining the efficacy of sodium chlorin / iron salts in controlling late blight of pepper in pots. The experiment consisted of two treatments. Control group (CK): purified water was sprayed. Treatment group: sprayed with a 2.0 ppm solution of chlorin / iron sodium salt. Pepper seedlings at the 4-6 leaf stage were transplanted and treated with a solution of chlorine / sodium iron per potted plant. Three days later, spores of Phytophthora capsicum were inoculated into the seedlings. The inoculum amount was approximately 1 x 10 5 Three replicates were set for each treatment, with 18 pots per replicate. After treatment, the pepper plants were observed once a day to record the disease incidence rate, and the disease index and control efficiency were calculated.

[0169] According to the method in the Standards for the Investigation and Forecasting of Pepper Phytophthora blight (NY / T2060.1-2011), the situation of plant phytophthora blight was investigated, and the incidence rate, pathological index, and control effect were calculated (Ye Minshuo et al., 2019). Disease incidence rate = number of diseased plants / total number of plants x 100% Pathological index = [(number of diseased plants at each level × corresponding disease grade) / (total number of plants surveyed × 5) × 100% Control effect = [(control disease index - treatment disease index) / control disease index] x 100%

[0170] Extraction of plant total RNA For RNA extraction, 0.2 g of pepper leaves (samples) were collected after treatment with chlorin / iron sodium salt for 12, 24, 36, 48, 60, and 72 hours. RNA was extracted using the TRIzol method. A 2 mL sample was transferred to a centrifuge tube, thoroughly polished with liquid nitrogen, and then 1 mL of TRIzol reagent (Thermo Fisher Scientific, USA) was added. The tube was mixed thoroughly and allowed to stand for 5 minutes. 200 μL of chloroform was added, shaken to mix, and allowed to stand for 5 minutes. The tube was then centrifuged at 12,000 g for 15 minutes at 4°C. The supernatant was transferred to a 1.5 mL centrifuge tube, an equal volume of pre-chilled isopropanol was added, shaken to mix, allowed to stand for 30 minutes at -80°C, and centrifuged at 12,000 g for 10 minutes at 4°C. The supernatant was discarded, and 75% alcohol was added. The tube was centrifuged at 12,000 g for 5 minutes at 4°C. The tube was washed twice. The supernatant was discarded, and the precipitate was dissolved in 30 μL of 0.1% DEPC-treated water and stored at -80°C.

[0171] cDNA first-strand synthesis and fluorescent quantitative PCR First-strand cDNA synthesis was performed using the EasyScript® One-Step gDNA Removal and cDNA Synthesis SuperMix Kit (Transgene, Beijing), while simultaneously removing genomic DNA remaining in the RNA template. The total volume was 20 μL. Total RNA was 1000 ng, RNase-free water was 7 μL, and random primer (N9) was 1 μL, for a total of 8 μL. After mixing, the mixture was incubated at 65°C for 5 minutes and placed on ice for 2 minutes. Then, 10 μL of 2xES reaction mix, 1 μL of EasyScript® RT / RI Enzyme Mix, and 1 μL of gDNA remover were added. The mixture was gently mixed, incubated at 25°C for 10 minutes, then at 42°C for 15 minutes. The mixture was inactivated by heating at 85°C for 5 seconds and stored at 4°C. The mixture was diluted 40-fold with cDNA and stored at -20°C for future use.

[0172] The expression of the disease resistance-related genes PR1, WRKY40, WRKY53, ACCO, and GST in pepper leaves treated with chlorin / iron sodium salt for different times was detected by real-time PCR. Actin was used as an internal control. The sequences of the specific primers are shown in the table below. [Table 30]

[0173] cDNA was used as a template and amplification was performed using a ROCHE LightCycler® 96 real-time fluorescent quantitative PCR system. The amplification system consisted of 10 μL of 2x PerfectStart™ greengreen qPCR SuperMix (Transgene, Beijing), 0.4 μL (10 μmol / L) upstream and downstream primers, 1 μL of cDNA, and 8.2 μL of nuclease-free water for a total volume of 20 μL. The reaction process consisted of pre-denaturation at 95°C for 180 seconds, followed by 40 cycles of 95°C for 10 seconds, 60°C for 30 seconds, 95°C for 10 seconds, 65°C for 10 seconds, 97°C for 1 second, and finally cooling to 37°C for 30 seconds. The specificity of the reaction system was determined from the melting curve and fluorescence curve. The experiment was repeated three times, and the relative expression level of the gene was calculated using 2-ΔΔCt. The results are shown in the table below. [Table 31] After treatment with chlorin / iron sodium salt, the incidence of pepper late blight was significantly reduced, and the control effect reached 71.89%, indicating that chlorin / iron sodium salt at a concentration of 2.0 ppm immediately possessed very strong plant immune-inducing activity.

[0174] Example 100: Reducing herbicide damage and significantly increasing rice yield Location: Huaian City, Jiangsu Province, China ▲ Shu ▼ ▲ Yi ▼ Mahu Village, Rice variety: Kinja 818 rice, Sample: Sodium chlorine / iron salt (0.25 ppm aqueous solution) Period: June-November 2021 Field conditions: The previous crop was wheat, the soil fertility is the same, and the field management is the same. Method: The treatment field was set to 1.5 mu and the control field to 1.5 mu. The herbicide was sprayed on the rice plants, and after the herbicide damage appeared, the sample was sprayed twice. The purified water blank was used as the control. The specific procedures were as follows:

[0175] On June 20th, when the weeds had three leaves and the rice had 3-3.5 leaves, the treated and control fields were treated with 4% imazamox 200ml / row + 10% pyribenzoxim 40ml / row by foliage spraying, using 30kg of water per row. After one week, the weed control effect was good, but obvious damage to the rice plants appeared. Ten days after using the herbicide (June 30th), the treated fields were sprayed with 7.5mg of the sample per row, using 30L of solution per row. The control fields were sprayed with purified water, and the same spraying method was used again at the rice panicle stage (August 25th). Results: Harvested on November 10th, and yield data is shown below. [Table 32]

[0176] The actual field tests showed that the herbicidal effect on rice fields was clear, with no clear difference between the treated and control fields. However, in the fields sprayed with the chlorin / iron sodium salt solution, there was some phytotoxicity that affected crop growth. However, the number of panicles per row and the number of kernels per panicle were better than in the blank control, and the yield per 1,000 kernels increased significantly, reaching an increase of more than 22% in yield per row. It was found that spraying the chlorin / iron sodium salt solution not only did not affect the herbicidal effect, but also significantly reduced the adverse effects of the herbicide and increased yield per row.

Claims

1. Porphine salts include salts of porphine compounds or salts of chlorin compounds.

2. The porphine salt according to claim 1, characterized in that the salt of the porphine compound includes a salt of protoporphyrin or a salt of protoporphyrin chelate, and the salt of the chlorin compound includes a salt of pheophorbide or a salt of pheophorbide chelate.

3. 2. The porphin salt according to claim 1, wherein the porphin compound is selected from protoporphyrin, hemin, or hematin, and the chlorin compound is selected from pheophorbide, chlorin / iron (chlorophyllin / iron chloride), hydroxychlorophyllin / iron, chlorophyllin / iron, chlorophyllin / zinc, or chlorophyllin / copper.

4. The porphine salt according to claim 1, characterized in that the salt of the porphine compound is prepared by salifying an acid form of protoporphyrin or its chelate with a metal ion, and the salt of the chlorin compound is prepared by salifying an acid form of pheophorbide or its chelate with a metal ion.

5. The porphine salt according to claim 1, wherein the salt of the porphine compound includes a monovalent, divalent, or trivalent metal ion salt of the porphine compound, and the salt of the chlorin compound includes a monovalent, divalent, or trivalent metal ion salt of the chlorin compound.

6. 2. The porphine salt according to claim 1, wherein the salt of the porphine compound includes a sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, or copper salt of the porphine compound, and the salt of the chlorin compound includes a sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, or copper salt of the chlorin compound.

7. The porphine salt is potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of protoporphyrin; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of hemin; potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of hematin; Sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorin / iron (chlorophyllin / iron chloride); Hydroxychlorophyllin / sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of iron; Chlorophyllin / ammonium, ferrous, iron, and copper salts of iron; potassium, ammonium, magnesium, ferrous, iron, zinc, manganese, and copper salts of chlorins (pheophorbides); potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorophyllin / zinc; Ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorophyllin / copper 2. The porphine salt of claim 1, comprising:

8. 8. A method for preparing the porphine salt according to claim 1, comprising reacting an acid-form porphine compound or chlorin compound with a hydroxide of a monovalent metal to form a precipitate, which is then separated to obtain a monovalent metal ion salt of the porphine compound or chlorin compound; or reacting a monovalent metal salt of the porphine compound or chlorin compound with a soluble polyvalent metal ion salt to form a precipitate, which is then separated to obtain a polyvalent metal ion salt of the porphine compound or chlorin compound.

9. the metal hydroxide is selected from sodium hydroxide, potassium hydroxide, or aqueous ammonia; 9. The method for preparing a polyvalent metal ion salt of a porphine compound or a chlorin compound according to claim 8, wherein the monovalent metal salt of the porphine compound or the chlorin compound is selected from sodium salts or potassium salts of the porphine compound or the chlorin compound, and the soluble polyvalent metal ion salt includes sulfate, hydrochloride, or nitrate of a divalent or trivalent metal ion.

10. The method for preparing the monovalent metal ion salt of the porphine compound or chlorin compound includes the steps of: the step of preparing a solution of an acid-form porphine compound or a chlorin compound in an organic solvent, mixing the solution with an organic solvent of a monovalent metal ion salt to cause precipitation, the step of salifying the acid-form porphine compound or the chlorin compound and the monovalent metal ion salt in an aqueous solution, and causing precipitation with an organic solvent, or the step of preparing a solution of an acid-form porphine compound or a chlorin compound in an organic solvent, introducing dry ammonia gas to cause precipitation, and filtering, washing, and drying the solution, The method for preparing the polyvalent metal ion salt of the porphine compound or chlorin compound includes the steps of: The method includes steps of mixing an aqueous solution of a monovalent metal salt of a porphine compound or a chlorin compound with an aqueous solution of a soluble polyvalent metal ion salt, followed by precipitation, filtration, washing, and drying.

9. A method for preparing the porphine salt according to claim 8.

11. Use of the porphine salt according to any one of claims 1 to 6 as a plant growth regulator or a plant immunity inducer.

12. The porphine salt is sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of protoporphyrin; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of hemin; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of hematin; Sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorin / iron; Hydroxychlorophyllin / sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of iron; Sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorophyllin / iron; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorins; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorophyllin / zinc; Sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorophyllin / copper The use according to claim 11, characterized in that it comprises

13. The use according to claim 11, characterized in that it includes spraying, smearing, seed soaking, aerial application, spike soaking, irrigation or dusting of plants or the environment in which the plants are growing, in order to achieve the purpose of regulating the growth of the plants or enhancing the stress tolerance of the plants in need of such growth or enhancement of the stress tolerance.

14. Use according to claim 11, characterized in that during use the concentration / content of the applied porphine salt is between 0.001 ppm and 10 ppm.

15. The use according to claim 11, characterized in that when used for soaking seeds and irrigating crop fields, the concentration / content of the applied porphine salt is 0.001 ppm to 0.1 ppm, and when used for foliar spraying of crops, the concentration / content of the applied porphine salt is 0.01 ppm to 10 ppm.

16. The use according to claim 11, characterized in that the functions of the plant growth regulator include promoting seed germination, increasing germination rate, increasing root length, promoting root growth, strengthening plant immunity and stress resistance, promoting seedling growth, increasing chlorophyll content, delaying premature plant aging, and improving yield and quality.

17. The use according to claim 11, characterized in that the functions of the plant immunity inducer include increasing stress tolerance of plants and increasing crop yield.

18. Use of a composition comprising the porphine salt according to any one of claims 1 to 6 as a plant growth regulator or a plant immunity inducer.

19. The porphine salt is sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of protoporphyrin; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of hemin; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of hematin; Sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorin / iron; Hydroxychlorophyllin / sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of iron; Sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorophyllin / iron; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorins; sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorophyllin / zinc; Sodium, potassium, ammonium, magnesium, calcium, ferrous, iron, zinc, manganese, and copper salts of chlorophyllin / copper 19. The use according to claim 18, characterized in that it comprises

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