CN108147976A - A kind of extracting method Ledermycined - Google Patents
A kind of extracting method Ledermycined Download PDFInfo
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- CN108147976A CN108147976A CN201611110609.XA CN201611110609A CN108147976A CN 108147976 A CN108147976 A CN 108147976A CN 201611110609 A CN201611110609 A CN 201611110609A CN 108147976 A CN108147976 A CN 108147976A
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Abstract
The present invention provides a kind of extracting method Ledermycined, and includes the following steps：The pH value of zymotic fluid is readjusted the distribution to acidity, zymotic fluid and thalline separation of solid and liquid, obtains liquid 1；The cationic surfactant of gained liquid 1 and 3 4% is 8.0 8.5 in pH value, complex reaction occurs, separation of solid and liquid obtains complex compound 1；Cationic surfactant is tetradecyl trimethyl ammonium chloride and/or hexadecyltrimethylammonium chloride；The percentage accounts for the percentage of the quality of liquid 1 for the quality of cationic surfactant；The pH value of gained complex compound 1 is adjusted, complex reaction occurs with urea, separation of solid and liquid obtains complex compound 2；The pH value of gained complex compound 2 is adjusted, is allowed to dissolve, separation of solid and liquid, obtains liquid 2；The pH value of gained liquid 2 is adjusted with hydrochloric acid, cooling, crystallization obtain DMCTC hydrochlorides.The extracting method of the methyl aureomycin can obtain the DMCTC hydrochlorides of purity >=94%.
Description
Technical field
The present invention relates to a kind of extracting methods Ledermycined.
Background technology
Ledermycining, (English name Demethylchlortetracycline, abridge DMCTC, structure such as formula 1
It is shown) there is wider antimicrobial spectrum, staphylococcus aureus, streptococcus of resistance to tetracycline etc. are included to gram positive bacteria and leather is blue
The resistance to plucked instrument bacterium of gonorrhoea in negative bacterium plays the role of very strong, is production meter Luo Huan elements and tigecycline key precursor, is a kind of heavy
The medical material wanted.
It Ledermycins by streptomyces aureofaciens (Streptomyces aureus) fermenting and producing, then passes through separation
Extraction obtains bulk pharmaceutical chemicals and Ledermycins salt, and the technique for the separation and Extraction mainly reported at present mainly has：(1) zymotic fluid passes through
Acidification, extraction, back extraction, condensation crystallization method (Chinese antibiotic magazine, 2000,25：341-343), the drawback is that using
A large amount of organic solvent, pollution are big；(2) using column chromatography method of purification extraction Ledermycin (Chinese antibiotic magazine,
2013,38：915-917), the drawback is that production cost is high, industrialized production is not suitable for it；(3) zymotic fluid is adjusted to alkaline pH,
Add in the method (CN201310575459) of seeded crystallization.Although easy to operate, due to Ledermycining in alkaline condition
Stability inferior is poor, easily forms DMCTC epimers；(4) zymotic fluid for concentrating decoloration is adjusted into pH in 4.5-8.5, crystallization
Precipitation, is then refined (CN201310380480) using double salt method of Ledermycining.But separation process needs to utilize resin pair
Zymotic fluid decoloration reduces impurity, increases operating cost.
The cheap of DMCTC, dosage are big, there is higher requirement to its purity in, made from existing technique
The purity of DMCTC is unable to reach more than 94%, therefore, can limit its large-scale production and application, thus develop a kind of pollution it is small, into
This low, simple possible production technology so that the purity of DMCTC reaches more than 94%, even up to USP quality requirements (purity >=
95%) it is that need solve the problems, such as at present.
Invention content
The technical problems to be solved by the invention are that organic solvent usage amount is big, production cost in order to overcome in the prior art
The defects of high, and provide a kind of pollution the extracting method Ledermycined small, at low cost.
The present invention provides a kind of extracting methods Ledermycined, and include the following steps：
(1) it adjusts containing the pH value of zymotic fluid Ledermycined to acidity, the zymotic fluid carries out separation of solid and liquid with thalline,
Obtain liquid 1；
(2) cationic surfactant of gained liquid 1 and 3-4% is 8.0-8.5 in pH value, complex reaction occurs, Gu
Liquid detaches, and obtains complex compound 1；The cationic surfactant is tetradecyl trimethyl ammonium chloride and/or cetyl front three
Ammonium chloride；The percentage accounts for the percentage of the quality of the liquid 1 for the quality of the cationic surfactant；
(3) pH value of gained complex compound 1 is adjusted, and complex reaction occurs with urea, separation of solid and liquid obtains complex compound 2；
(4) pH value of gained complex compound 2 is adjusted, gained complex compound 2 dissolves, and separation of solid and liquid obtains liquid 2；
(5) pH value of gained liquid 2 is adjusted with hydrochloric acid to acidity, cooling, crystallization obtain DMCTC hydrochlorides.
In the present invention, described is by streptomyces aureofaciens described in the routine of this field containing the zymotic fluid to Ledermycin
The zymotic fluid that (Streptomyces aureus) fermenting and producing obtains, wherein containing Ledermycining.
In step (1), the operation of the pH value for readjusting the distribution zymotic fluid is conventional for this field, and the pH value is preferably adjusted to 1.0-
2.0, the pH value is more preferably adjusted to 1.5.The substance for adjusting pH value is this field conventional pH regulators, such as aqueous sulfuric acid.
In step (1), the separation of solid and liquid is conventional for this field, preferably micro-filtration, the micro-filtration that the micro-filtration uses
Film is conventional for this field, preferably 0.45 μm of ceramic membrane.
In step (2), the tetradecyl trimethyl ammonium chloride is conventional for this field, preferably PurumTTAC150,
The hexadecyltrimethylammonium chloride is conventional for this field, preferably PurumCTAC150, the PurumTTAC150 and
PurumCTAC150 is purchased from idchem Co., Ltds of South Korea.
In step (2), the substance for adjusting pH value is this field conventional pH regulators, such as sodium hydrate aqueous solution.
In step (2), the separation of solid and liquid is conventional for this field, generally filters.
In step (3), the operation of the pH value of complex compound 1 is conventional for this field obtained by the tune, and the pH value is preferably adjusted
To 1.0-2.0, the pH value is more preferably adjusted to 1.5.The substance of the tune pH value is this field conventional pH regulators, such as sulfuric acid water
Solution.
In step (3), the separation of solid and liquid is conventional for this field, generally filters.
In step (4), the operation of the pH value of complex compound 2 is conventional for this field obtained by the tune, and the pH value is preferably adjusted
To 2.0-3.0, the pH value is more preferably adjusted to 2.5.The substance for adjusting pH value is this field conventional pH regulators, such as hydroxide
Sodium water solution.
In step (4), the separation of solid and liquid is conventional for this field, generally filters.
In step (5), the operation of the pH value of liquid 2 is conventional for this field obtained by the adjusting, and the pH value is preferably adjusted
To 0.2-0.8, the pH value is more preferably adjusted to 0.5.
In step (5), the operation of the cooling is conventional for this field, and the cooling is preferably cooled to 5 DEG C；The crystallization
Operation it is conventional for this field, the time of the crystallization is preferably 6 hours.
On the basis of common knowledge of the art, above-mentioned each optimum condition can be combined arbitrarily to get each preferable reality of the present invention
Example.
The reagents and materials used in the present invention are commercially available.
The positive effect of the present invention is：The extracting method that the present invention is Ledermycined, by cation form
The type of face activating agent, dosage and after adding in cationic surfactant the creative of pH value find, can obtain purity >=
94% DMCTC hydrochlorides, even up to USP quality requirements (purity >=95%).
Description of the drawings
HPLC collection of illustrative plates of the Fig. 1 for the zymotic fluid that Ledermycins in the embodiment of the present invention 1.
Fig. 2 is the HPLC collection of illustrative plates of demethyl aureomycin hydrochloride obtained in the embodiment of the present invention 1.
Specific embodiment
It is further illustrated the present invention below by the mode of embodiment, but does not therefore limit the present invention to the reality
It applies among a range.Test method without specific conditions in the following example, according to conventional methods and conditions or according to quotient
Product specification selects.
Streptomyces aureofaciens (Streptomyces aureus) ATCC 21658 is purchased from American Type Culture collection warehousing
(ATCC)。
The preparation method containing the zymotic fluid to Ledermycin in following embodiments is as follows：By streptomyces aureofaciens
The mycelium inoculation of (Streptomyces aureus) ATCC 21658, in the case where cultivation temperature is 28 DEG C, is cultivated to seed culture medium
After 40 hours, fermentation medium is inoculated by 10% inoculum concentration, at 28 DEG C of cultivation temperature, cultivates 7-8 days, collects containing demethyl
The zymotic fluid of aureomycin, for isolating and purifying for next step.
Seed culture medium (g/L)：Soluble starch 2.0, glucose 1.5, soybean cake powder 1.5, sodium chloride 0.3；
Fermentation medium (g/L):Soluble starch 5.0, maltodextrin 2.0, glucose 1.0, soybean cake powder 3.0, dry ferment
Female powder 5.0, soya-bean oil 2.0, CaCO3 0.3。
Cationic surfactant PurumTTAC150 and PurumCTAC150 in following embodiments are purchased from South Korea
Idchem Co., Ltds.
DMCTC HPLC analysis methods are as follows：
Analyze fixed column：ODSC18 columns, 4.6 × 250mm, 5um；Column temperature：40℃；
Mobile phase A is mutually potassium dihydrogen phosphate (0.1% phosphoric acid) aqueous solution of 0.02mol/L:Acetonitrile=90：10 (V/V), B
It is mutually potassium dihydrogen phosphate (0.1% phosphoric acid) aqueous solution of 0.02mol/L:Acetonitrile=70：30(V/V)；Flow velocity is 1.0mL/min；
Detector wavelength 254nm；Sample size 10uL, run time 20min.
Gradient elution table：
| Time (min) | A phases (%) | B phases (%) |
| 0 | 100 | 0 |
| 5 | 100 | 0 |
| 8 | 0 | 100 |
| 18 | 0 | 100 |
| 23 | 100 | 0 |
| 30 | 100 | 0 |
Embodiment 1
Containing the zymotic fluid to Ledermycin, as shown in Figure 1, fermentation unit is 3250mg/L, volume is collection of illustrative plates
1000mL adjusts pH value to 1.5 with aqueous sulfuric acid, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collect filtrate.Add
Enter 3.0%PurumTTAC150, then adjust pH value to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.By washing
After washing, pH value is adjusted to 1.5 with aqueous sulfuric acid, it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH water
Solution tune pH to 2.5 is stirred, and filtrate is collected in filtering.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, knot
6 hours brilliant, filter cake is collected in filtering, and 60 DEG C of dryings obtain DMCTC hydrochlorides in 8 hours, and collection of illustrative plates is as shown in Fig. 2, 13min or so goes out
Peak, total recovery 74%, purity 95.5%.
Embodiment 2
By containing the zymotic fluid to Ledermycin, fermentation unit 3250mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 3.5%PurumTTAC150,
Then pH value is adjusted to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 74.5%, purity 95.4% in 8 hours.
Embodiment 3
By containing the zymotic fluid to Ledermycin, fermentation unit 3250mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 4.0%PurumTTAC150,
Then pH value is adjusted to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 75.1%, purity 95.2% in 8 hours.
Comparative example 1
By containing the zymotic fluid to Ledermycin, fermentation unit 3250mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 2.5%PurumTTAC150,
Then pH value is adjusted to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 63%, purity 93.2% in 8 hours.
Comparative example 2
By containing the zymotic fluid to Ledermycin, fermentation unit 3250mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 4.5%PurumTTAC150,
Then pH value is adjusted to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 68%, purity 94.3% in 8 hours.
| Project | Total recovery/% of DMCTC | Purity/% of DMCTC |
| Embodiment 1 | 74 | 95.5 |
| Embodiment 2 | 74.5 | 95.4 |
| Embodiment 3 | 75.1 | 95.2 |
| Comparative example 1 | 63 | 93.2 |
| Comparative example 2 | 68 | 94.3 |
Embodiment 4
By containing the zymotic fluid to Ledermycin, fermentation unit 3200mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 3.0%PurumCTAC150,
Then pH value is adjusted to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 60.2%, purity 94.2% in 8 hours.
Embodiment 5
By containing the zymotic fluid to Ledermycin, fermentation unit 3200mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 3.5%PurumCTAC150,
Then pH value is adjusted to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 63%, purity 94.6% in 8 hours.
Embodiment 6
By containing the zymotic fluid to Ledermycin, fermentation unit 3200mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 4.0%PurumCTAC150,
Then pH value is adjusted to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 62%, purity 94.2% in 8 hours.
Comparative example 3
By containing the zymotic fluid to Ledermycin, fermentation unit 3200mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 2.5%PurumCTAC150,
Then pH value is adjusted to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 59%, purity 91.1% in 8 hours.
Comparative example 4
By containing the zymotic fluid to Ledermycin, fermentation unit 3200mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 4.5%PurumCTAC150,
Then pH value is adjusted to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 60%, purity 92.8% in 8 hours.
| Project | Total recovery/% of DMCTC | Purity/% of DMCTC |
| Embodiment 4 | 60.2 | 94.2 |
| Embodiment 5 | 63 | 94.6 |
| Embodiment 6 | 62 | 94.2 |
| Comparative example 3 | 59 | 91.1 |
| Comparative example 4 | 60 | 92.8 |
Embodiment 7
By containing the zymotic fluid to Ledermycin, fermentation unit 3500mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 3.0%PurumTTAC150,
Then pH value is adjusted to 8.0 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 73%, purity 95.8% in 8 hours.
Embodiment 8
By containing the zymotic fluid to Ledermycin, fermentation unit 3500mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 3.0%PurumTTAC150,
Then pH value is adjusted to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 74%, purity 95.5% in 8 hours.
Comparative example 5
By containing the zymotic fluid to Ledermycin, fermentation unit 3500mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 3.0%PurumTTAC150,
Then pH value is adjusted to 7.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 40%, purity 93.3% in 8 hours.
Comparative example 6
By containing the zymotic fluid to Ledermycin, fermentation unit 3500mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 3.0%PurumTTAC150,
Then pH value is adjusted to 9.0 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 61%, purity 92.4% in 8 hours.
| Project | Total recovery/% of DMCTC | Purity/% of DMCTC |
| Embodiment 7 | 73 | 95.8 |
| Embodiment 8 | 74 | 95.5 |
| Comparative example 5 | 40 | 93.3 |
| Comparative example 6 | 61 | 92.4 |
Embodiment 9
By containing the zymotic fluid to Ledermycin, fermentation unit 3230mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 3.0%PurumCTAC150,
Then pH value is adjusted to 8.0 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 58%, purity 94.2% in 8 hours.
Embodiment 10
By containing the zymotic fluid to Ledermycin, fermentation unit 3230mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 3.0%PurumCTAC150,
Then pH value is adjusted to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 60%, purity 94.2% in 8 hours.
Comparative example 7
By containing the zymotic fluid to Ledermycin, fermentation unit 3230mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 3.0%PurumCTAC150,
Then pH value is adjusted to 7.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 52%, purity 93.7% in 8 hours.
Comparative example 8
By containing the zymotic fluid to Ledermycin, fermentation unit 3230mg/L, volume 1000mL use aqueous sulfuric acid
PH value is adjusted to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 3.0%PurumCTAC150,
Then pH value is adjusted to 9.0 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, adjusted with aqueous sulfuric acid
PH value is to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH to 2.5, stirring, mistake
Filtrate is collected in filter.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C, crystallizes 6 hours, and filter cake is collected in filtering,
60 DEG C of dryings obtain DMCTC hydrochlorides, total recovery 59%, purity 93.5% in 8 hours.
| Project | Total recovery/% of DMCTC | Purity/% of DMCTC |
| Embodiment 9 | 58 | 94.2 |
| Embodiment 10 | 60 | 94.2 |
| Comparative example 7 | 52 | 93.7 |
| Comparative example 8 | 59 | 93.5 |
For comparative example 9 by containing the zymotic fluid to Ledermycin, fermentation unit 3200mg/L, volume 1000mL use sulphur
Aqueous acid adjusts pH value to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 4.0%
Then PurumCEP adjusts pH value to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, sulfuric acid is used
Aqueous solution adjusts pH value to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH extremely
2.5, it stirs, filtrate is collected in filtering.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C.Crystallization 6 hours, mistake
Filter, collects filter cake, and 60 DEG C of dryings obtain DMCTC hydrochlorides in 8 hours.
For comparative example 10 by containing the zymotic fluid to Ledermycin, fermentation unit 3200mg/L, volume 1000mL use sulphur
Aqueous acid adjusts pH value to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 4.0%
Then PurumBC150 adjusts pH value to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, sulphur is used
Aqueous acid adjusts pH value to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH
To 2.5, stir, filtrate is collected in filtering.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C.Crystallization 6 hours,
Filtering, collects filter cake, and 60 DEG C of dryings obtain DMCTC hydrochlorides in 8 hours.
For comparative example 11 by containing the zymotic fluid to Ledermycin, fermentation unit 3200mg/L, volume 1000mL use sulphur
Aqueous acid adjusts pH value to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.Add in 4.0%
Then PurumBTAC adjusts pH value to 8.5 with NaOH aqueous solutions, plate-frame filtering collects filter cake.After water washing, sulfuric acid is used
Aqueous solution adjusts pH value to 1.5, and it is 1.5 to add in urea and maintain pH value, and filter cake is collected in filtering.With NaOH aqueous solution tune pH extremely
2.5, it stirs, filtrate is collected in filtering.It is 0.5 to add in hydrochloric acid to pH value to filtrate, and stirring is cooled to 5 DEG C.Crystallization 6 hours, mistake
Filter, collects filter cake, and 60 DEG C of dryings obtain DMCTC hydrochlorides in 8 hours.
| Project | Total recovery/% of DMCTC | Purity/% of DMCTC |
| Comparative example 9 | 20 | 90.5 |
| Comparative example 10 | 35 | 89.8 |
| Comparative example 11 | 38 | 86.2 |
Comparative example 12
According to the method that patent CN201310575459 is recorded, inventor has carried out following experiment, and obtains purity data.
It will be containing the zymotic fluid to Ledermycin, fermentation unit 3200mg/L, volume 1000ml, with aqueous sulfuric acid tune
PH value is saved to 1.5, stirs 30 minutes, by 0.45 μm of ceramic membrane filter, collects filtrate.8 DEG C are cooled to, with NaOH aqueous solution tune
PH value is saved to 9.2, crystal is precipitated, plate-frame filtering collects filter cake.After water washing, with salt acid for adjusting pH value to 2.8, vacuum
It is dry, obtain product 1.7g, total recovery 53.1%, purity 84.5%.
Claims (9)
1. a kind of extracting method Ledermycined, it is characterised in that：It includes the following steps：
(1) it adjusts containing the pH value of zymotic fluid Ledermycined to acidity, the zymotic fluid carries out separation of solid and liquid with thalline, obtains liquid
Body 1；
(2) cationic surfactant of gained liquid 1 and 3-4% is 8.0-8.5 in pH value, complex reaction, solid-liquid point occurs
From obtaining complex compound 1；The cationic surfactant is tetradecyl trimethyl ammonium chloride and/or cetyl trimethyl chlorine
Change ammonium；The percentage accounts for the percentage of the quality of the liquid 1 for the quality of the cationic surfactant；
(3) pH value of gained complex compound 1 is adjusted, and complex reaction occurs with urea, separation of solid and liquid obtains complex compound 2；
(4) pH value of gained complex compound 2 is adjusted, gained complex compound 2 dissolves, and filters, obtains liquid 2；
(5) pH value of gained liquid 2 is adjusted with hydrochloric acid, cooling, crystallization obtain DMCTC hydrochlorides.
2. extracting method as described in claim 1, it is characterised in that：In step (1), the pH value is adjusted to 1.0-2.0, described
PH value is preferably adjusted to 1.5.
3. extracting method as described in claim 1, it is characterised in that：In step (1), the separation of solid and liquid is micro-filtration.
4. extracting method as claimed in claim 3, it is characterised in that：The microfiltration membranes that the micro-filtration uses is 0.45 μm of ceramics
Film.
5. extracting method as described in claim 1, it is characterised in that：In step (2), the tetradecyl trimethyl ammonium chloride
For PurumTTAC150, the hexadecyltrimethylammonium chloride is PurumCTAC150, the PurumTTAC150 and
PurumCTAC150 is purchased from idchem Co., Ltds of South Korea.
6. extracting method as described in claim 1, it is characterised in that：In step (3), the pH value is adjusted to 1.0-2.0, described
PH value is preferably adjusted to 1.5.
7. extracting method as described in claim 1, it is characterised in that：In step (4), the pH value is adjusted to 2.0-3.0, described
PH value is preferably adjusted to 2.5.
8. extracting method as described in claim 1, it is characterised in that：In step (5), the pH value is adjusted to 0.2-0.8, described
PH value is preferably adjusted to 0.5.
9. extracting method as described in claim 1, it is characterised in that：In step (5), the cooling is is cooled to 5 DEG C；It is described
The time of crystallization is 6 hours.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3053893A (en) * | 1960-07-22 | 1962-09-11 | American Cyanamid Co | Recrystallization of demethylchlortetracycline |
| GB917467A (en) * | 1959-12-09 | 1963-02-06 | American Cyanamid Co | Improvements in or relating to antibiotics |
| CN103641739A (en) * | 2013-11-18 | 2014-03-19 | 宁夏泰瑞制药股份有限公司 | Method for producing demeclocycline hydrochloride by utilizing demeclocycline fermentation broth |
| CN104418767A (en) * | 2013-08-28 | 2015-03-18 | 河南天方药业股份有限公司 | Demeclocycline extraction process |
-
2016
- 2016-12-06 CN CN201611110609.XA patent/CN108147976B/en active Active
Patent Citations (4)
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|---|---|---|---|---|
| GB917467A (en) * | 1959-12-09 | 1963-02-06 | American Cyanamid Co | Improvements in or relating to antibiotics |
| US3053893A (en) * | 1960-07-22 | 1962-09-11 | American Cyanamid Co | Recrystallization of demethylchlortetracycline |
| CN104418767A (en) * | 2013-08-28 | 2015-03-18 | 河南天方药业股份有限公司 | Demeclocycline extraction process |
| CN103641739A (en) * | 2013-11-18 | 2014-03-19 | 宁夏泰瑞制药股份有限公司 | Method for producing demeclocycline hydrochloride by utilizing demeclocycline fermentation broth |
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| 廖福荣: "尿素在四环素类抗菌素提纯上的应用", 《医药工业》 * |
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