Method for controlling impurities in L-arabinose and its use

Abstract

The present invention relates to a method for controlling impurities in L-arabinose and its use. The method includes removing impurities from L-arabinose starting materials using an adsorption resin. The method and use provided by the present invention can effectively control the content of impurities such as 5-hydroxymethylfurfural and furfural in L-arabinose products, particularly by removing impurities from the source and upstream processes, thereby reducing the risk of toxicity and side effects of impurities, thereby improving the quality and safety of L-arabinose products.

Description

[Technical Field]

[0001] This application claims priority to a prior patent application bearing patent application number 202310055013.8 and entitled "Method for controlling impurities in L-arabinose and use thereof," filed with the State Intellectual Property Administration of China on February 3, 2023, which is incorporated herein by reference in its entirety. The present invention relates to the field of sugar production processes, and in particular to a method for controlling impurities in L-arabinose and its use. [Background technology]

[0002] L-arabinose (C5H 10 O5) is an aldopentose, also known as L(+)-aldo gum. It was originally a monosaccharide isolated from gum arabic. Research has shown that L-arabinose has good laxative properties, as well as beneficial blood sugar lowering and weight loss effects. L-arabinose is primarily derived from lignocellulosic biomass, such as corn, sugarcane, and beets. Literature sources reveal that L-arabinose undergoes dehydration under acidic and high-temperature conditions to produce the mutagenic impurity furfural, the degradation pathway of which is shown in the diagram below. [ka]

[0003] According to the preliminary classification of carcinogens list published by the World Health Organization's International Agency for Research on Cancer on October 27, 2017, furfural is a mutagenic impurity and is classified as a Group 3 carcinogen.

[0004] 5-Hydroxymethylfurfural (5-HMF) is nephrotoxic, mutagenic, and potentially genotoxic. Pharmacopoeias clearly specify that 5-HMF is the limiting component in glucose injections. However, no extensive research has been conducted on the 5-HMF-related content and control of L-arabinose preparations.

[0005] In order to improve the quality and safety of various L-arabinose products, it is necessary to further effectively control the content of impurities such as 5-hydroxymethylfurfural and furfural from the source and upstream production process, and reduce the risk of toxicity and side effects of impurities, which will be beneficial for the industrialized large-scale production of L-arabinose products and their further use in the food and pharmaceutical fields. Summary of the Invention

[0006] To solve the problems existing in the prior art, in a first aspect, the present invention provides a method for controlling impurities, said method comprising: The method includes step (Ia) of removing impurities in the L-arabinose starting material using an adsorption resin. According to an embodiment of the present invention, the impurities are at least one selected from furfural and 5-hydroxymethylfurfural.

[0007] According to an embodiment of the present invention, the adsorption resin is at least one selected from a macroporous adsorption resin with a high specific surface area, a styrene-based macroporous resin, and a styrene-divinylbenzene copolymer skeleton macroporous resin.

[0008] In some embodiments, the adsorption resin is at least one selected from a styrene-based macroporous strongly acidic cation exchange resin or a styrene-based macroporous weakly basic anion exchange resin.

[0009] In some embodiments, the adsorption resin is selected from sulfonic acid group cation exchange resins, for example, cation exchange resins having sulfonic acid groups (-SO3H) in a styrene-divinylbenzene copolymer.

[0010] According to an embodiment of the present invention, the styrene-divinylbenzene copolymer skeleton macroporous resin is selected from those having weak polarity and those having medium polarity.

[0011] According to an embodiment of the present invention, the adsorption resin is at least one selected from LSA-220, D309, SQD-96, D301-M, D301, D319, LSA-20, LX-10B, LX-100B, LX-20B, LX-360, LX-150, LSA-10, LSA-12, LSA-10B, LSA-100, LSA-210, HPD296, HPD500, HPD300L, D-900, and 001x7. Preferably, the resin is at least one selected from LSA-220, SQD-96, D319, LSA-20, and LX-10B, and more preferably, the resin is at least one selected from LSA-220 and SQD-96. More preferably, the adsorption resin is selected from the combination of D301 and 001x7.

[0012] In some embodiments, the L-arabinose starting material is first adsorbed onto a D301 resin, then onto a 001x7 resin, and finally onto a mixed resin of D301 and 001x7; or first onto a 001x7 resin, then onto a D301 resin, and finally onto a mixed resin of D301 and 001x7, wherein the volume ratio of D301 to 001x7 in the mixed resin is 1:1 to 3:1, e.g., 2:1. In some embodiments, the L-arabinose starting material is first adsorbed onto D301 resin, then onto 001x7 resin, then onto a mixed resin of D301 and 001x7, and finally onto LSA-220; or first onto 001x7 resin, then onto D301 resin, then onto a mixed resin of D301 and 001x7, and finally onto LSA-220, where the volume ratio of D301 to 001x7 in the mixed resin is 1:1 to 3:1, e.g., 2:1, and the volume ratio of D301:001x7:mixed resin is (1-3):1:(1-3), e.g., 2:1:2.

[0013] According to an embodiment of the present invention, the adsorption resin is purchased from Lanxiao or Suqing.

[0014] According to an embodiment of the present invention, the resin is at least one selected from Lanxiao LSA-220, Suqing D309, Suqing SQD-96, Suqing D301-M, Suqing D301, Suqing D319, Lanxiao LSA-20, Lanxiao LX-10B, and Suqing 001x7. Preferably, the resin is at least one selected from Lanxiao LSA-220, Suqing SQD-96, Suqing D319, Lanxiao LSA-20, Lanxiao LX-10B, and Suqing 001x7, more preferably, the resin is at least one selected from Lanxiao LSA-220 and Suqing SQD-96.

[0015] In some embodiments, the L-arabinose starting material is first adsorbed onto a Suoqing D301 resin, then onto a Suoqing 001x7 resin, and finally onto a mixed resin of Suoqing D301 and Suoqing 001x7; or first onto a Suoqing 001x7 resin, then onto a Suoqing D301 resin, and finally onto a mixed resin of Suoqing D301 and Suoqing 001x7, where the volume ratio of Suoqing D301 to Suoqing 001x7 in the mixed resin is 1:1 to 3:1, e.g., 2:1. The volume ratio of Suoqing D301 resin:Suoqing 001x7:mixed resin is (1-3):1:(1-3), e.g., 2:1:2.

[0016] In some embodiments, the L-arabinose starting material is first adsorbed with Sosei D301 resin, then with Sosei 001x7 resin, then with a mixed resin of Sosei D301 and Sosei 001x7, and finally with LSA-220; or first with Sosei 001x7 resin, then with Sosei D301 resin, then with a mixed resin of Sosei D301 and Sosei 001x7, and finally with LSA-220, wherein the volume ratio of Sosei D301 to Sosei 001x7 in the mixed resin is 1:1 to 3:1, e.g., 2:1. The volume ratio of the Sosei D301 resin:Sosei 001×7:mixed resin is (1 to 3):1:(1 to 3), for example, 2:1:2.

[0017] According to an embodiment of the present invention, in step (Ia), the L-arabinose starting material can be selected from an L-arabinose solid material, an L-arabinose-containing syrup, or an L-arabinose-containing preparation.

[0018] In some embodiments, the L-arabinose starting material is selected from commercially available or synthetic L-arabinose solids, e.g., the purity of the L-arabinose in the solids is ≧90%.

[0019] In some embodiments, the L-arabinose starting material is selected from L-arabinose-containing crude liquid products extracted from natural products, and crystallization mother liquor in an L-arabinose-containing syrup purification process.

[0020] In some embodiments, the L-arabinose starting material is selected from an L-arabinose-containing syrup, which can be selected from commercially available products.

[0021] In some embodiments, the sugar content of the L-arabinose-containing syrup is 20 to 90% Brix, for example, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90% Brix.

[0022] In some embodiments, the L-arabinose starting material is selected from an L-arabinose-containing aqueous solution having a sugar content of 20 to 90% Brix, e.g., 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90% Brix, or a mass concentration of L-arabinose in the L-arabinose-containing aqueous solution of ≥ 10%, e.g., ≥ 10%, ≥ 15%, ≥ 20%, ≥ 25%, ≥ 30%, ≥ 35%, ≥ 40%, ≥ 45%, ≥ 50%, ≥ 55%, ≥ 60%, ≥ 65%, ≥ 70%, ≥ 75%, ≥ 80%, ≥ 85%, or ≥ 90% Brix.

[0023] More specifically, in some embodiments, the mass concentration of L-arabinose in the L-arabinose-containing syrup is ≧10%, for example, the mass concentration of L-arabinose is ≧10%, ≧15%, ≧20%, ≧25%, ≧30%, ≧35%, ≧40%, ≧45%, ≧50%, ≧55%, ≧60%, ≧65%, ≧70%, ≧75%, ≧80%, ≧85%, ≧90%.

[0024] In some embodiments, the L-arabinose starting material is selected from any L-arabinose-containing formulation, for example, the formulation may be a liquid formulation, and the mass concentration of L-arabinose in the liquid formulation is ≧10%, ≧15%, ≧20%, ≧25%, ≧30%, ≧35%, ≧40%, ≧45%, ≧50%, ≧55%, ≧60%, ≧65%, ≧70%, ≧75%, ≧80%, ≧85%, or ≧90%. The formulation may contain L-arabinose as the only active ingredient or may contain other active ingredients.

[0025] According to an embodiment of the present invention, in step (Ia), the furfural content in the L-arabinose starting material is greater than 10,000 ng / ml, e.g., greater than 10,000, 20,000, 30,000, 40,000, 50,000, 60,000, 70,000, 80,000, 90,000, 100,000, 110,000, 120,000, 130,000, 140,000, or 150,000 ng / ml, and the 5-hydroxymethylfurfural content is greater than 5,000 ng / ml, e.g., greater than 5,000, 10,000, 20,000, 30,000, 40,000, or 50,000 ng / ml. In some embodiments, the furfural content in the L-arabinose starting material is greater than 50,000 ng / ml. In some embodiments, the L-arabinose starting material contains 5-hydroxymethylfurfural at a concentration greater than 10,000 ng / ml.

[0026] According to an embodiment of the present invention, in step (Ia), the L-arabinose starting material has a Brix of ≦60% Brix upon passing through the column, for example, 30, 35, 40, 45, 50, or 55% Brix, preferably ≦50% Brix. More preferably, the Brix is ​​≦40% Brix. Optionally, step (Ia) includes dissolving or diluting the L-arabinose starting material with water to a Brix suitable for passing through the column.

[0027] According to an embodiment of the present invention, in step (Ia), the column flow rate of the material is 0.5 BV / h to 4 BV / h, for example, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, or 4.0 BV / h, the column flow temperature is 10 to 50°C, preferably room temperature (25±5°C), and the elution solvent used is water.

[0028] In some embodiments, step (Ia) is a step of passing the raw syrup through a Suqing D301 resin, a Suqing 001x7 resin, and a mixed resin (the volume ratio of D301 to 001x7 is 2:1) at a rate of 1 to 3 BV / h, and then passing it through a Lanxiao LSA-220, where the volume ratio of the Suqing D301 resin:Suqing 001x7 resin:mixed resin is (1 to 3):1:(1 to 3), for example, 2:1:2.

[0029] According to an embodiment of the present invention, the method may further comprise a step (Ib) of treating with activated carbon, which may be performed before or after step (Ia).

[0030] According to an embodiment of the present invention, in step (Ib), the activated carbon is selected from powdered activated carbon and granular activated carbon. The adsorption temperature of the activated carbon is 20 to 80°C, for example, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, or 75°C. The adsorption time of the selected activated carbon is 10 to 100 min, preferably 20 to 90 min, for example, 30, 40, 50, 60, 70, 80, or 90 min. The amount of activated carbon used is 0.0005 to 0.010 g per mL of material, for example, 0.0005, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, or 0.010 g.

[0031] In some embodiments, step (Ib) comprises heating the syrup obtained in step (Ia) to 40 to 60°C, adding activated carbon for adsorption, and then filtering by suction to remove the activated carbon. The activated carbon adsorption time is 20 to 40 minutes, and 0.002 to 0.006 g of activated carbon is used for adsorption per mL of material.

[0032] In a second aspect, the present invention provides the use of the impurity control method in a process for producing an L-arabinose product.

[0033] In some embodiments, the present invention provides a process for producing an L-arabinose product, the process comprising steps (Ia) and (Ib) as defined above.

[0034] According to an embodiment of the present invention, the manufacturing process comprises: (I) subjecting the L-arabinose starting material to activated carbon treatment in step (Ia) and, optionally, step (Ib) to obtain a clarified sugar solution; and (II) concentrating, crystallizing, centrifuging and drying the resulting clean sugar solution.

[0035] According to an embodiment of the present invention, said step (Ia) and step (Ib) are as defined above.

[0036] In some embodiments, step (II) is selected from step (II-a) of placing a portion of the clear syrup obtained in step (I) (e.g., about half of the clear syrup) in an evaporative crystallizer, concentrating the sugar solution until the sugar content reaches 55 to 65% Brix, adding an appropriate amount of L-arabinose as seed crystals, and then starting to feed the remaining clear syrup, and continuing concentration after the feeding is completed.

[0037] According to an embodiment of the present invention, the evaporative crystallizer can be selected from common crystallization reactors such as an evaporative crystallizer.

[0038] According to an embodiment of the present invention, in the step (II-a), the sugar solution is concentrated preferably until the sugar content reaches 55 to 60% Brix, more preferably 58 to 60% Brix, and after the end of feeding, concentration is continued until the concentration of the solid-liquid mixture reaches 75 to 85%.

[0039] According to an embodiment of the present invention, in the step (II-a), the degree of supersaturation of the sugar solution is controlled to 1.01 to 1.19, preferably 1.01 to 1.10, and more preferably 1.03 to 1.08 during the fed-batch process.

[0040] In some embodiments, step (II) is selected from step (II-b), which comprises adding an appropriate amount of L-arabinose raw material as seed crystals and lowering the temperature at a rate of 0.5 to 3°C / hour when the clear syrup obtained in step (I) is added to an evaporative crystallizer to concentrate the syrup until the sugar content reaches 55 to 65% Brix.

[0041] According to an embodiment of the present invention, in step (II-b), the sugar solution is preferably concentrated until the sugar content reaches 58 to 60% Brix, and the temperature is preferably lowered at a rate of 0.5 to 2.5°C / hour, for example, 0.5, 1.0, 1.5, or 2.0°C / hour. In some embodiments, the temperature is lowered at a rate of 0.5 to 2°C / hour, and in some embodiments, the temperature is lowered at a rate of 0.5 to 1.5°C / hour.

[0042] In some embodiments, step (II-b) is selected from step (II-b1), which comprises filtering off the activated carbon, adding the resulting material to an evaporative crystallizer for concentration, and adding an appropriate amount of L-arabinose raw material as seed crystals and lowering the temperature at a rate of 0.5 to 3°C / hour when concentrating until the sugar content reaches 55 to 65% Brix.

[0043] According to an embodiment of the present invention, in step (II-b1), the sugar solution is preferably concentrated until the sugar content reaches 58 to 60% Brix, and the temperature is preferably lowered at a rate of 0.5 to 2.5°C / hour, for example, 0.5, 1.0, 1.5, or 2.0°C / hour. In some embodiments, the temperature is lowered at a rate of 0.5 to 2°C / hour, and in some embodiments, the temperature is lowered at a rate of 0.5 to 1.5°C / hour.

[0044] According to an embodiment of the present invention, step (II) may further include a step of adding activated carbon, which may be selected from powdered activated carbon and granular activated carbon. The adsorption temperature of the activated carbon is 20 to 80°C, e.g., 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, or 75°C. The adsorption time of the selected activated carbon is 10 to 100 min, preferably 20 to 90 min, e.g., 30, 40, 50, 60, 70, 80, or 90 min. Regarding the amount of activated carbon used, 0.0005 to 0.010 g of activated carbon is used for adsorption per 1 mL of material, for example, 0.0005, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, or 0.010 g of activated carbon is used.

[0045] According to an embodiment of the present invention, in step (II), after the concentration is completed, the material can be discharged and centrifuged. After the centrifugation, the obtained L-arabinose crystals can be further dried. Preferably, the drying temperature for the crystals is 50 to 60°C, the drying time is 1.5 to 2.5 hours, for example, 2 hours, and the vacuum degree is -0.01 to -0.1 MPa, for example, -0.05 to -0.1 MPa.

[0046] In some embodiments, step (II) further comprises evaporating the cleaned sugar solution obtained in step (I) until the sugar content of the sugar solution reaches 55-65% Brix, followed by evaporative crystallization at 50-75°C and a vacuum of -0.05 to -0.1 MPa, adding an appropriate amount of L-arabinose as seed crystals, and then starting to feed the remaining cleaned sugar solution. After the feeding is completed, the solid-liquid mixture is concentrated until the concentration reaches 75-85%, and then the material is discharged and centrifuged. The degree of supersaturation of the sugar solution is controlled to 1.01 to 1.19 throughout the entire fed-batch crystallization process. Furthermore, the L-arabinose crystals obtained by centrifugation are dried at 50-60°C for 2 hours at a vacuum of -0.05 to -0.1 MPa. According to an embodiment of the present invention, the furfural content in the obtained L-arabinose is ≦5 ppm, for example, ≦4.5 ppm, ≦4.0 ppm, and the 5-hydroxymethylfurfural content is ≦3.0 ppm, for example, ≦2.5 ppm, ≦2.0 ppm, ≦1.5 ppm, ≦1.0 ppm.

[0047] According to an embodiment of the present invention, a person skilled in the art will understand that in step (II) (step II-a or step II-b), the seed crystals can be selected from commonly obtained solid L-arabinose raw materials (e.g., commercially available L-arabinose products with a purity of ≥98%), and the raw materials can be further pulverized by passing them through a sieve, such as a No. 5-6 Pharmacopoeia standard sieve.

[0048] According to an embodiment of the present invention, the manufacturing process comprises: (1) adsorbing about 1-3 L of the L-arabinose starting material at a rate of about 2 BV / h first with D301 resin, then with 001x7 resin, then with a mixed resin of D301 and 001x7, and finally with LSA-220, in which the mixed resin volume ratio of D301 and 001x7 is 1:1-3:1, for example, 2:1; then heating the material to 40-60°C, adding activated carbon, adsorbing for 20-40 minutes, and filtering by suction to remove the activated carbon, to obtain a clear syrup; and (2) evaporating the clear syrup obtained in step (1) until the sugar content reaches 55-65% Brix, followed by evaporative crystallization at 50-75°C and a vacuum of -0.05 to -0.1 MPa. After adding an appropriate amount of L-arabinose raw material as seed crystals, the remaining clear syrup is started to be fed as a feed. After the feeding is completed, the solid-liquid mixture is concentrated until the concentration reaches 75-85%, and then the material is discharged and centrifuged. The degree of supersaturation of the sugar solution is controlled to 1.01-1.19 throughout the entire crystallization process by feeding.

[0049] The present invention further provides a process for producing an L-arabinose product. The process can perform only step (II-b) on an L-arabinose starting material without performing step (I). The process can remove impurities from the L-arabinose starting material. The impurities are at least one selected from furfural and 5-hydroxymethylfurfural. Specifically, the manufacturing process comprises: The method includes the steps of adding an L-arabinose starting material to an evaporative crystallizer and concentrating it until the sugar content reaches 55 to 65% Brix, adding an appropriate amount of L-arabinose starting material as seed crystals, and lowering the temperature at a rate of 0.5 to 3°C / hour.

[0050] According to an embodiment of the present invention, in the above step, the sugar solution is preferably concentrated until the sugar content reaches 58 to 60% Brix, and the temperature is preferably lowered at a rate of 0.5 to 2.5°C / hour, for example, 0.5, 1.0, 1.5, or 2.0°C / hour. In some embodiments, the temperature is lowered at a rate of 0.5 to 2°C / hour, and in some embodiments, the temperature is lowered at a rate of 0.5 to 1.5°C / hour.

[0051] According to an embodiment of the present invention, the L-arabinose starting material is the same as the "L-arabinose starting material" used in step (Ia). The seed crystals, similar to those defined in step (II), can be selected from commonly available solid L-arabinose materials (e.g., commercially available L-arabinose products with a purity of 98% or higher), and the material may be further pulverized by passing it through a sieve such as a No. 5-6 Pharmacopoeia standard sieve.

[0052] According to embodiments of the present invention, the L-arabinose starting material is preferably selected from L-arabinose-containing syrups having a sugar content of 20 to 90% Brix, such as 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90% Brix, and in some embodiments, the sugar content of the L-arabinose-containing syrup is 40 to 50% Brix.

[0053] According to an embodiment of the present invention, the manufacturing process may further include a step of adding activated carbon, which may be selected from powdered activated carbon and granular activated carbon. The adsorption temperature of the activated carbon is 20 to 80°C, for example, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, or 75°C. The adsorption time of the selected activated carbon is 10 to 100 min, preferably 20 to 90 min, for example, 30, 40, 50, 60, 70, 80, or 90 min. Regarding the amount of activated carbon used, 0.0005 to 0.010 g of activated carbon is used for adsorption per 1 mL of material, for example, 0.0005, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, or 0.010 g of activated carbon is used.

[0054] According to an embodiment of the present invention, the manufacturing process comprises: The method includes the steps of evaporating and concentrating the starting syrup (with a sugar content in the range of 40-50% Brix) at 50-75°C and a vacuum of -0.05 to -0.1 MPa until the sugar content reaches 58-60% Brix, adding an appropriate amount of L-arabinose as seed crystals, and then initiating crystallization while decreasing the temperature at a rate of approximately 2°C / h.

[0055] According to an embodiment of the present invention, in the production process, after the temperature is lowered and crystallization is completed, the material can be discharged and centrifuged. After the centrifugation, the obtained L-arabinose crystals can be further dried. Preferably, the drying temperature for the crystals is 50 to 60°C, the drying time is 1.5 to 2.5 hours, for example, 2 hours, and the vacuum degree is -0.01 to -0.1 MPa, for example, -0.05 to -0.1 MPa.

[0056] According to an embodiment of the present invention, the furfural content in the obtained L-arabinose is ≦5 ppm, for example, ≦4.5 ppm, ≦4.0 ppm, and the 5-hydroxymethylfurfural content is ≦3.0 ppm, for example, ≦2.5 ppm, ≦2.0 ppm, ≦1.5 ppm, ≦1.0 ppm.

[0057] In a third aspect, the present invention provides a product obtained according to the impurity control method of the first aspect.

[0058] According to an embodiment of the present invention, the content of furfural in the product is ≦5 ppm, for example, ≦4.5 ppm, ≦4.0 ppm, and the content of 5-hydroxymethylfurfural is ≦3.0 ppm, for example, ≦2.5 ppm, ≦2.0 ppm, ≦1.5 ppm, ≦1.0 ppm.

[0059] In a fourth aspect, the present invention provides an L-arabinose product obtained according to the manufacturing process of the second aspect.

[0060] According to an embodiment of the present invention, the furfural content in the obtained L-arabinose is ≦5 ppm, for example, ≦4.5 ppm, ≦4.0 ppm, and the 5-hydroxymethylfurfural content is ≦3.0 ppm, for example, ≦2.5 ppm, ≦2.0 ppm, ≦1.5 ppm, ≦1.0 ppm.

[0061] In a fifth aspect, the present invention provides L-arabinose, wherein the furfural content in the L-arabinose is ≦5.0 ppm, for example, ≦4.5 ppm, ≦4.0 ppm, and the 5-hydroxymethylfurfural content in the L-arabinose is ≦3.0 ppm, for example, ≦2.5 ppm, ≦2.0 ppm, ≦1.5 ppm, ≦1.0 ppm. [Effects of the Invention]

[0062] The method and use provided by the present invention can effectively control the content of impurities such as 5-hydroxymethylfurfural and furfural in L-arabinose products, especially by removing impurities from the source and upstream processes, and reducing the risk of toxicity and side effects of impurities, thereby helping to improve the quality and dosage safety of L-arabinose products. DETAILED DESCRIPTION OF THE INVENTION

[0063] The technical means of the present invention will be described in more detail below with reference to specific examples. Note that the following examples are merely for illustrative purposes and are not intended to limit the scope of protection of the present invention. Any technology realized based on the above content of the present invention is included in the scope of protection of the present invention.

[0064] Unless otherwise specified, raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0065] In the following examples, "syrup" or "raw syrup" refers to L-arabinose-containing syrup. All of the raw materials can be purchased commercially.

[0066] Example 1: Effect of activated carbon treatment on impurity removal rate Experimental materials and equipment: syrup (Zhejiang Huakang Pharmaceutical Co., Ltd., 50-70% Brix), activated carbon (Nanping Yuanli Co., Ltd.), sugar content meter ATAGO PAL-α, furfural and 5-hydroxymethylfurfural detection (HPLC), microporous filtration membrane (Shanghai Xingya Purification Materials Factory).

[0067] 1.1 The raw syrup was diluted with purified water to a sugar content of 32.2% Brix, and 200ml of the syrup was dispensed into four Erlenmeyer flasks. 0.8g of activated carbon (4‰ of activated carbon per syrup volume) was added to each Erlenmeyer flask, and the flasks were incubated in a water bath at temperatures of 30°C, 50°C, 60°C, and 75°C for 30 minutes each. The flasks were then filtered through 0.45μm and 0.22μm micropore filtration membranes, respectively, and then sampled for detection. The experimental results are shown in Table 1.

[0068] [Table 1]

[0069] 1.2 The raw syrup was diluted with purified water to a sugar content of 32.1% Brix, and 600ml of the syrup was dispensed into four Erlenmeyer flasks. 0.3g, 1.2g, 2.4g, and 4.8g of activated carbon (0.5‰, 2‰, 4‰, and 8‰ of activated carbon per syrup volume) were added to each Erlenmeyer flask. The flasks were then incubated in a 60°C water bath for 60 minutes, after which the syrup was filtered through 0.45um and 0.22um micropore filtration membranes, respectively, and then sampled for detection. The experimental results are shown in Table 2.

[0070] [Table 2]

[0071] 1.3 The raw syrup was adjusted to a sugar content of 32.1% Brix, and 200 ml of the syrup was dispensed into three Erlenmeyer flasks. 0.8 g of activated carbon (4‰ of activated carbon relative to the volume of the syrup) was added to each Erlenmeyer flask, and the flasks were incubated in a 60°C water bath for 30, 60, and 90 minutes, respectively. The flasks were then filtered through 0.45 μm and 0.22 μm micropore filtration membranes, and samples were taken for detection. The experimental results are shown in Table 3.

[0072] [Table 3]

[0073] As can be seen from Table 3, once the treatment time with activated carbon reaches 90 minutes, the amount of adsorption of impurities does not increase any further, and there is no point in extending the adsorption time.

[0074] As can be seen from Tables 1 to 3, the removal rates of furfural and 5-hydroxymethylfurfural were all less than 50% for different adsorption times, adsorption temperatures, and amounts of activated carbon used. Even when the amount of activated carbon used was 8‰ (equivalent to approximately 2.5% of the dry matter), the removal rates of furfural and 5-hydroxymethylfurfural were 32.97% and 46.49%, respectively, when the decolorization rate reached 87%.

[0075] Example 2: Effect of adsorption resin on impurity removal rate Experimental materials and equipment: syrup (Tang Chuan Biotechnology (Xiamen) Co., Ltd., 30%-50% Brix), sugar content meter ATAGO PAL-α, furfural and 5-hydroxymethylfurfural detection (HPLC), resin (Xi'an Lanxiao Co., Ltd. and Jiangsu Suqing Co., Ltd.).

[0076] 2.1 Preliminary selection of adsorption resin Experimental procedure: 1.0 g of resin was taken, and 25 g of 32.4% Brix syrup was added. The mixture was shaken at room temperature at 1000 rpm for 1.5 hours. After filtering the resin, a sample of the syrup was taken and analyzed. The results are shown in Table 4.

[0077] [Table 4]

[0078] As can be seen from the detection results, the furfural removal effect of Suqing D301-M and Suqing D301 when used alone is obviously low, whereas Lanxiao LSA-220, Suqing SQD-96, D319, Lanxiao LSA-20, Lanxiao Lx-10B, etc. have good furfural removal effect, and in particular, Lanxiao LSA-220 has the best removal effect on furfural and 5-hydroxymethylfurfural.

[0079] 2.2 Removal of impurities by resin adsorption Experimental materials: Raw syrup (Tangchuan Biotechnology (Xiamen) Co., Ltd., 30%~50% Brix).

[0080] Resin type: LSA-220 manufactured by Lanxiao, macroporous adsorption resin manufactured by Xi'an Lanxiao, SQD-96 manufactured by Suqing, macroporous weakly basic styrene-based resin manufactured by Jiangsu Suqing. The column was packed by a wet method, the column packing volume was 130 ml (ie, the volume of 1 BV), the feed flow rate was about 2 BV / h, and the sampling sequence was as shown in the table.

[0081] [Table 5]

[0082] [Table 6]

[0083] 2.3 Effect of different sugar content on resin adsorption Experimental equipment and materials: syrup (Zhejiang Huakang Pharmaceutical Co., Ltd., 50-70% Brix), sugar content meter ATAGO PAL-α, furfural and 5-hydroxymethylfurfural detection (HPLC), resin (Xi'an Lanxiao Co., Ltd.).

[0084] Experimental procedure: 30g of syrup with different sugar contents (sugar contents can be further adjusted by adding purified water to the raw syrup) and 2g of resin were placed on a shaker (1000 rpm) at room temperature for 1.5 hours to adsorb, and then sampled and detected.

[0085] [Table 7]

[0086] As can be seen from Table 7, when the sugar content is controlled within 50% brix, the adsorption rates for furfural and 5-hydroxymethylfurfural are both over 75%, and when the sugar content is controlled within 40% brix, the resin adsorption effect is most favorable.When the sugar content is controlled at 60% brix, the resin adsorption rate for 5-hydroxymethylfurfural is less than 60%.

[0087] 2.4 Impurity removal effect of LSA-220 Experimental materials: Raw syrup (Tangchuan Biotechnology (Xiamen) Co., Ltd., 30%~50% Brix).

[0088] Resin type: LSA-220 made by Lanxiao.

[0089] The column was packed by the wet method. 500 ml of Lanxiao LSA-220 resin was packed into a 2.5 cm diameter chromatography column by the wet method at room temperature. The column flow rate was 2 BV / h. The sampling sequence is shown in Table 8.

[0090] [Table 8]

[0091] 2.5 Impurity removal effect by combining resins Experimental materials: Raw syrup (Tangchuan Biotechnology (Xiamen) Co., Ltd., 30%~50% Brix).

[0092] Resin type: D-301 manufactured by Jiangsu Suqingsha, 001×7 manufactured by Jiangsu Suqingsha.

[0093] The columns were packed using a wet method. Three 2.5 cm diameter columns were used. The starting syrup was passed through 150 ml of Suoshin D301 resin, 75 ml of Suoshin 001x7 resin, and 150 ml of the mixed resin (the volume ratio of D301 to 001x7 was 2:1), packed into the columns using a wet method, in that order, at a column passing rate of 2 BV / h.

[0094] [Table 9]

[0095] Example 3: Study of impurity content during the process 3.1 Experimental materials and equipment: syrup (manufactured by Tangchuan Biotechnology (Xiamen) Co., Ltd.), sugar content meter ATAGO PAL-α, furfural and 5-hydroxymethylfurfural detection (HPLC), L-arabinose seed crystals (i.e., L-arabinose raw material was ground and then passed through No. 5-6 standard sieves of the Pharmaceutical Codex; the raw material was purchased from Tangchuan Biotechnology (Xiamen) Co., Ltd.).

[0096] 3.2 Experimental process: The starting syrup for crystallization batch number 1213 had furfural at 180,856 ng / ml, 5-hydroxymethylfurfural at 21,056 ng / ml, and a sugar content of 42.9% Brix. This starting syrup was concentrated to approximately 800 ml, and approximately 1,200 ml was added as a feed. Approximately 1,055 g of dry matter was added.

[0097] The starting syrup for crystallization batch number 1215 had furfural 24074 ng / ml, 5-hydroxymethylfurfural 885 ng / ml, and a sugar content of 43.9% Brix. This starting syrup was concentrated to approximately 800 ml, and approximately 1200 ml was added as a feed. Approximately 1073 g of dry matter was added.

[0098] The starting syrup was evaporated to 55-65% Brix, then subjected to evaporation crystallization at 50-75°C and a vacuum of -0.05--0.1 MPa. After adding 1.5 g of L-arabinose as seed crystals, the remaining starting syrup was added. After the addition, the mixture was concentrated until the solid-liquid mixture reached a concentration of 75-85%, after which the material was discharged and centrifuged. The degree of supersaturation of the sugar solution was controlled at 1.01-1.19 throughout the entire crystallization process.

[0099] The arabinose crystals obtained by centrifugation were dried at a drying temperature of 50 to 60°C for 2 hours at a vacuum of -0.05 to -0.1 MPa.

[0100] [Table 10]

[0101] As can be seen from Table 10-1, when not treated with resin, the contents of 5-hydroxymethylfurfural in the two batches were 885 ng / ml and 21056 ng / ml, respectively.

[0102] 3.3 Experimental process: Approximately 1000 mL of starting syrup (sugar content in the range of 40-50% Brix) was evaporated to 60% Brix at 50-75°C and a vacuum of -0.05 to -0.1 MPa. The temperature was lowered and crystallization was initiated in a 65°C water bath. 0.75 g of L-arabinose was added as seed crystals, and crystallization was initiated by lowering the temperature at a rate of approximately 2°C / h. When the temperature reached 31°C, the material was discharged and centrifuged. The arabinose crystals obtained after centrifugation were dried at 50-60°C for 2 hours at a vacuum of -0.05 to -0.1 MPa.

[0103] Activated carbon can be used for adsorption, and the adsorption conditions were as follows: about 0.8 g of activated carbon was used, and the adsorption was carried out at 50° C. for 30 minutes.

[0104] During the crystallization process, the impurity content in each sample was measured and is shown in Table 10-2 below.

[0105] [Table 11]

[0106] As can be seen from Table 10-2 above, using the temperature-reducing and crystallization process of 3.3 has a certain impurity removal effect on furfural and 5-HMF.

[0107] Example 4: Effect of resin adsorption on impurities in the manufacturing process The starting syrup for crystal batch numbers 1221 and 1229 had furfural of 180856ng / ml, 5-hydroxymethylfurfural of 21056ng / ml, and a sugar content of 42.9% brix.

[0108] 4.1 Experimental materials and equipment: syrup (Tangchuan Biotechnology (Xiamen) Co., Ltd.), ATAGO PAL-α sugar content meter, furfural and 5-hydroxymethylfurfural detection (HPLC), resins: Lanxiao LSA-220, Xi'an Lanxiao, Jiangsu Suqing D-301, Jiangsu Suqing 001x7, activated carbon (Nanping Yuanli). L-arabinose seed crystals (i.e., powder obtained by grinding L-arabinose raw material and passing it through No. 5-6 standard sieves; the raw material was purchased from Tangchuan Biotechnology (Xiamen) Co., Ltd.).

[0109] 4.2 Experimental process: (1) Approximately 2 L of raw syrup was passed through a column packed with 150 ml of D301 resin (manufactured by Suqing), 75 ml of 001x7 resin, and 150 ml of mixed resin (the volume ratio of D301 to 001x7 was 2:1) at a rate of approximately 2 BV / h, and then through 150 ml of LSA-220 (manufactured by Lanxiao). The syrup was then heated to 50°C, and 8 g of activated carbon was added and adsorbed for 30 minutes. The activated carbon was removed by suction filtration, and a purified syrup was obtained. (2) The purified sugar solution obtained in step (1) was evaporated until the sugar content of the sugar solution reached 55-65% Brix, and then subjected to evaporation crystallization at 50-75°C and a vacuum of -0.05 to -0.1 MPa. 1.5 g of L-arabinose was added as seed crystals, and the remaining purified sugar solution was then fed. After the feeding was completed, the sugar solution was concentrated until the solid-liquid mixture reached a concentration of 75-85%, after which the material was discharged and centrifuged. The degree of supersaturation of the sugar solution was controlled at 1.01-1.19 throughout the entire fed-batch crystallization process. (3) The arabinose crystals obtained by centrifugation were dried at a drying temperature of 50 to 60°C for 2 hours at a vacuum of -0.05 to -0.1 MPa.

[0110] [Table 12]

[0111] As can be seen from Table 11, when the 5-hydroxymethylfurfural in the purified syrup was controlled to 458 ng / ml or less, neither 5-hydroxymethylfurfural nor furfural was detected in the arabinose crystal sample, which indicates that the adsorption resin used in the present invention can effectively control furfural and 5-hydroxymethylfurfural, and reduce the risk of their possible toxicity and side effects to a very low level.

[0112] Example 5 Bowel cleansing effect of L-arabinose as a bowel preparation drug Experimental materials: L-arabinose selected from the L-arabinose crystal sample of batch number 1221 in the example was dissolved in purified water to a concentration of 0.4 g / ml. PEG (Shu Taiqing) was dissolved in purified water to a concentration of 0.521 g / ml. SPF rats (from Zhejiang Litonglihua) were used.

[0113] 5.1 Experimental animals and their care: Specific pathogen-free (SPF) rats (200±10g) aged 6 weeks were used. They were fed a standard diet (D12450-B) containing 67.35wt% carbohydrate, 19.2wt% crude protein, and 4.3wt% fat. The room temperature was maintained at 24-26°C and humidity at 40-60%.

[0114] 5.2 Experimental methods and procedures: SPF rats were randomly divided into a blank control group, a positive control group, and a test group, with 10 rats in each group, half of which were male and half of which were female. The doses for each group are shown in Table 12 below. Administration method: The animals in each group were fasted from 10 PM the night before the test, but not from water. On the morning of the test, half of the calculated dose was administered (intragastric administration) at 7 AM and 9 AM, and 2.0 mL of pure water was administered intragastrically at 12 PM and 1 PM, respectively. The animals were dissected 3.5 hours after the last administration.

[0115] [Table 13]

[0116] 5.3 Experimental results: (1) In both the positive control group and the test group, watery and loose stools were observed before autopsy. (2) After dissection, no residue was found in the rectum of the positive control group and the test group, demonstrating a good intestinal cleansing effect. Apparently, the dose of L-arabinose used in the test group was only 41% of that used in the positive control group, achieving the same effect, demonstrating the superiority of the dosage.

[0117] Example 6: Therapeutic effect of L-arabinose on constipation model rats Experimental materials: L-arabinose selected from arabinose crystal samples of batch number 1229 in the examples.

[0118] 6.1 Experimental preparation Modeling process: Modeling with diphenoxylate. Experimental chemicals and reagents: Activated carbon (5%), arabinose chemicals were prepared in 0.4 g / ml aqueous solution. Positive control: PEG (Shu Taiqing). SPF rats (derived from Zhejiang Litonglihua).

[0119] 6.2 Experimental process: In addition to the normal group, mice that successfully modeled were randomly divided into five groups, including a model group, a low-dose group, an intermediate-dose group, a high-dose group, and a positive control group, each consisting of eight rats. Each rat was housed in an independent cage and operated on daily between 10:00 and 12:00 PM. The normal and model groups received 0.2 mL of pure water, the positive control group received 0.133 mL of mosapride solution, and the low-dose, intermediate-dose, and high-dose groups received 0.01 mL, 0.02 mL, and 0.04 mL of the test drug, respectively. Any amount less than 0.2 mL was supplemented with pure water. Animals were allowed free access to food and water. The administration was continued for seven consecutive days.

[0120] At the first administration, the test group received the drug in the form of 5% activated charcoal (0.2g x 5%) and administered intragastrically, while the normal and model groups received the drug in the form of 0.2ml of purified water. The time of first black stool passage was recorded.

[0121] The test substance was administered intragastrically once daily, and on the seventh day after administration, feces from that day were collected in a sterile centrifuge tube and stored at -100°C. The collected feces were dried at 105°C until a constant weight was reached, and the fecal moisture content was calculated as follows: fecal moisture content = (1 - mass after drying / mass before drying) × 100%.

[0122] The rats were administered intragastrically once daily for 14 days, followed by a 12-hour fast. One hour after the last dose, each animal in each group received 0.15 ml of 5% activated charcoal. Thirty minutes later, rats were anesthetized with an intraperitoneal injection of 10% chloral hydrate. Immediately after anesthesia, the laparotomy was completed, and the intestinal tract from the pylorus to the ileocecal junction was dissected and placed on a glass plate. The length of the intestinal tract through which the ink passed and the total length of the intestinal tract were measured. The intestinal propulsion rate was calculated as the ratio of the two. The total length of the step from the pylorus to the ileocecal junction and the distance from the pylorus to the leading edge of the carbon ink were measured, and the carbon ink propulsion percentage was calculated. Intestinal propulsion rate = distance from the tip of the carbon ink to the pylorus / total length of the small intestine × 100%.

[0123] [Table 14]

[0124] The results of the t-test showed that the small intestinal propulsion rate, fecal water content, and first black stool defecation time of the three dosage groups were all significantly different from the model group ( * p<0.01). There was a tendency for small intestinal propulsion rate to increase from the low-dose group to the high-dose group, but there was no significant difference between the groups, demonstrating the clear effect of drug intervention, with a clear increase in the mouse's water content and smooth defecation. There was no significant difference compared to the positive control drug, indicating that the small intestinal propulsion rate and fecal water content were equivalent to those of the positive control drug.

[0125] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention should be included within the protection scope of the present invention.

Claims

1. (Ia) removing impurities in the L-arabinose starting material using an adsorption resin; the impurity is at least one selected from furfural and 5-hydroxymethylfurfural; The method for controlling impurities, wherein the adsorption resin is at least one selected from a macroporous adsorption resin with a high specific surface area, a styrene-based macroporous resin, and a styrene-divinylbenzene copolymer skeleton macroporous resin.

2. The adsorption resin is at least one selected from a styrene-based macroporous strongly acidic cation exchange resin and a styrene-based macroporous weakly basic anion exchange resin, or the adsorption resin is selected from a sulfonic acid group cation exchange resin; Preferably, the sulfonic acid group cation exchange resin is a styrene-divinylbenzene copolymer having a sulfonic acid group (—SO ３ H), Preferably, the adsorption resin is at least one selected from LSA-220, D309, SQD-96, D301-M, D301, D319, LSA-20, Lx-10B, LX-100B, LX-20B, LSA-10, LSA-12, LSA-10B, LSA-100, LSA-210, HPD500, HPD300L, D-900, 001x7; More preferably, the resin is at least one selected from LSA-220, SQD-96, D319, LSA-20, and Lx-10B; More preferably, the resin is at least one selected from LSA-220 and SQD-96; More preferably, the adsorption resin is selected from the combination of D301 and 001x7.

3. 3. The method according to claim 1, wherein the L-arabinose starting material is first adsorbed onto a D301 resin, then onto a 001x7 resin, and finally onto a mixed resin of D301 and 001x7, wherein the volume ratio of D301 to 001x7 in the mixed resin is 1:1 to 3:1, for example 2:

1.

4. 4. The method according to claim 1, wherein the L-arabinose starting material is selected from an L-arabinose-containing syrup, and the sugar content of the L-arabinose-containing syrup is 20 to 70% Brix.

5. 4. The method according to claim 1, wherein the L-arabinose starting material has a Brix of ≦60% Brix, preferably ≦50% Brix, more preferably ≦40% Brix, upon passing through the column; and wherein, in step (I-a), the material is passed through the column at a rate of 0.5 BV / h to 4 BV / h, the temperature of the material passing through the column is 10 to 50°C, and the elution solvent used is water.

6. further comprising a step (Ib) of treating with activated carbon, The method according to any one of claims 1 to 3, characterized in that in step (Ib), the adsorption temperature of the activated carbon is 20 to 80°C, the adsorption time of the selected activated carbon is 10 to 100 min, and the amount of activated carbon used for adsorption is 0.0005 to 0.010 g per mL of L-arabinose material.

7. Use of the method for controlling impurities according to any one of claims 1 to 6 in a process for producing an L-arabinose product, the process comprising: (I) subjecting the L-arabinose starting material to activated carbon treatment in step (I-a) and, optionally, step (I-b) to obtain a clarified sugar solution; (II) concentrating, crystallizing, centrifuging and drying the resulting cleaned sugar solution; 7. Use, characterized in that the steps (Ia) and (Ib) are defined according to any one of claims 1 to 6.

8. The step (II) a step (II-a) of placing a part of the clarified syrup obtained in step (I) (for example, about half of the clarified syrup) into an evaporative crystallizer, concentrating the sugar solution until the sugar content reaches 55 to 65% Brix, adding an appropriate amount of L-arabinose as seed crystals, and then starting to feed the remaining clarified syrup, and continuing to concentrate after the feeding is completed; In the step (II-a), the sugar solution is concentrated preferably until the sugar content reaches 55 to 60% Brix, more preferably 58 to 60% Brix, and after the feeding is completed, the concentration is continued until the concentration of the solid-liquid mixture reaches 75 to 85%; Preferably, in the step (II-a), the degree of supersaturation of the sugar solution is controlled to 1.01 to 1.19 during the feeding process; Alternatively, the step (II) is selected from step (II-b), which comprises adding an appropriate amount of L-arabinose raw material as seed crystals and lowering the temperature at a rate of 0.5 to 3°C / hour when the purified syrup obtained in step (I) is added to an evaporative crystallizer to concentrate the purified syrup until the sugar content reaches 55 to 65% Brix; In the step (II-b), the sugar solution is preferably concentrated until the sugar content reaches 58 to 60% Brix, and the temperature is preferably lowered at a rate of 0.5 to 1.5°C / hour. In the step (II), after the concentration is completed, the material can be discharged and centrifuged. More preferably, after the centrifugation, the obtained L-arabinose crystals are further dried. More preferably, the crystals are dried at a temperature of 50 to 60°C for a period of 1.5 to 2.5 hours, and at a vacuum of -0.05 to -0.1 MPa.

9. 1. A process for producing an L-arabinose product, comprising the steps of adding an L-arabinose starting material to an evaporative crystallizer to concentrate it, and adding an appropriate amount of L-arabinose starting material as seed crystals and lowering the temperature at a rate of 0.5 to 3°C / hour when the L-arabinose starting material is concentrated to a sugar content of 55 to 65% Brix, wherein the L-arabinose starting material is selected from L-arabinose-containing syrup, and the sugar content of the L-arabinose-containing syrup is 40 to 50% Brix.

10. An L-arabinose product obtained by the production process according to claims 7 to 9.

11. 11. The product of claim 10, wherein the furfural content is ≦5.0 ppm and the 5-hydroxymethylfurfural content is ≦3.0 ppm.

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