CN113583062A - 一种林可霉素的衍生化方法、林可霉素衍生物及衍生化方法的应用 - Google Patents
一种林可霉素的衍生化方法、林可霉素衍生物及衍生化方法的应用 Download PDFInfo
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Abstract
本发明公开了一种林可霉素的衍生化方法、林可霉素衍生物及衍生化方法的应用;衍生化方法为在林可霉素中加入不含活泼氢的反应溶剂和衍生化试剂进行反应;林可霉素衍生物的结构通式为C50H62N6O18S5;林可霉素的衍生化方法的应用,为通过林可霉素的衍生化方法得到林可霉素衍生物,然后将林可霉素衍生物用于测定林可霉素的含量;以该衍生化方法对林可霉素进行处理,可以提高在液相中检测林可霉素含量时的灵敏度。
Description
技术领域
本发明涉及一种林可霉素的衍生化方法、林可霉素衍生物及衍生化方法的应用,属于化合物技术领域。
背景技术
林可霉素是从林可链球菌发酵液中提取出来的一类抗生素,现有技术公开的测定样品中的林可霉素主要有高效液相色谱法、高效液相色谱-质谱联用、气相色谱—质谱测定方法等。上述测定方法中高效液相色谱-质谱联用、气相色谱—质谱测定方法所需仪器价格昂贵,不易普及。高效液相色谱仪检测林可霉素时,因林可霉素紫外吸收弱,上机时的检测限的和定量限都较高,灵敏度较低。
因此,有必要在分析化学的领域中,进一步针对林可霉素的检测效率、灵敏度等提高进行研究。
发明内容
为了克服现有技术的不足,本发明的第一个目的在于提供一种林可霉素的衍生化方法,以该衍生化方法对林可霉素进行处理,可以提高在液相中检测林可霉素含量时的灵敏度。
本发明的第二个目的在于提供一种林可霉素衍生物。
本发明的第三个目的在于提供一种林可霉素衍生化方法的应用,用于对林可霉素含量的测定,大大提高测定的灵敏度和效率。
实现本发明的第一个目的可以通过采取如下技术方案达到:一种林可霉素的衍生化方法,衍生化方法为在林可霉素中加入不含活泼氢的反应溶剂和衍生化试剂进行反应。
进一步地,不含活泼氢的反应溶剂为乙腈、四氢呋喃、***和甲苯中的至少一种。
进一步地,衍生化试剂为对甲苯磺酰异氰酸酯。
进一步地,衍生化试剂在使用前以乙腈作为溶剂进行稀释。
进一步地,反应的条件为氮气保护下,温度10-30℃,震荡20-60min。
进一步地,反应后加入终止剂终止反应。
进一步地,终止剂为水、盐酸、甲醇和乙醇中的至少一种。
进一步地,林可霉素和衍生化试剂的摩尔比为1:(1-10)。
实现本发明的第二个目的可以通过采取如下技术方案达到:一种林可霉素衍生物,林可霉素衍生物通过林可霉素的衍生化方法得到;林可霉素衍生物的结构通式为C50H62N6O18S5,化学式如式Ⅰ所示:
林可霉素的衍生化方法为在林可霉素中加入不含活泼氢的反应溶剂和衍生化试剂进行反应。
实现本发明的第三个目的可以通过采取如下技术方案达到:一种林可霉素的衍生化方法的应用,通过林可霉素的衍生化方法得到林可霉素衍生物,然后将林可霉素衍生物用于测定林可霉素的含量;林可霉素的衍生化方法为在林可霉素中加入不含活泼氢的反应溶剂和衍生化试剂进行反应。
进一步地,测定林可霉素的含量的方法为色谱法。
进一步地,色谱法包括高效液相色谱法、高效液相色谱-质谱或薄层色谱法。
相比现有技术,本发明的有益效果在于:
1、本发明林可霉素的衍生化方法对林可霉素进行处理,可以提高在液相中检测林可霉素含量时的灵敏度;
2、本发明林可霉素的衍生化方法可用于对林可霉素含量的测定,具有快速、准确、简便、易于推广、成本低的特点,还具有反应条件温和、副产物少,过量的衍生化试剂不会对样品的检测造成影响等优点。
附图说明
图1为林可霉素衍生物的红外光谱图;
图2为林可霉素标液的高效液相色谱图;
图3为林可霉素经衍化后的高效液相色谱图;
图4为不经衍生化处理直接测定的牛肉含林可霉素高效液相色谱图;
图5为经衍生化处理后测定的牛肉含林可霉素高效液相色谱图;
图6为经衍生化处理后测定的鸡肉含林可霉素高效液相色谱图。
具体实施方式
下面,结合附图以及具体实施方式,对本发明做进一步描述:
一种林可霉素的衍生化方法,衍生化方法为:
对林可霉素进行预处理,如除水或者将甲醇于氮气下吹干;
对甲苯磺酰异氰酸酯以乙腈稀释为对甲苯磺酰异氰酸酯乙腈溶液;
在林可霉素中加入不含活泼氢的反应溶剂和对甲苯磺酰异氰酸酯乙腈溶液,林可霉素和衍生化试剂的摩尔比为1:(1-10),在氮气保护下,温度10-30℃,震荡20-60min条件下进行反应,然后加入终止剂终止,减压旋蒸为油状物,真空干燥。
其中,不含活泼氢的反应溶剂为乙腈、四氢呋喃、***和甲苯中的至少一种。
其中,终止剂为水、盐酸、甲醇和乙醇中的至少一种。
经林可霉素的衍生化方法,得到林可霉素衍生物,结构通式为C50H62N6O18S5,化学式如式Ⅰ所示:
对林可霉素的含量进行测定的方法如下:
在林可霉素中加入不含活泼氢的反应溶剂和衍生化试剂进行反应,得到林可霉素衍生物,然后以高效液相色谱法、高效液相色谱-质谱或薄层色谱法进行测定,得到样品中林可霉素的含量。
林可霉素其本身含有弱紫外吸收,若采用常用的高效液相色谱法进行高灵敏测定,需要引入紫外吸收基团,如果将林可霉素先进行衍生化处理,加入衍生化试剂,特别是对甲苯磺酰异氰酸酯为羟基衍生化试剂,是具有强亲核性的化合物,能与带有羟基的化合物迅速反应,使林可霉素产生具有苯环的强紫外吸收基团,提高了林可霉素在高效液相色谱检测中的灵敏度,具有良好的应用前景。
实施例1:
林可霉素衍生物的制备:
在林可霉素原料药2.35g,加入无水乙腈10mL和对甲苯磺酰异氰酸酯乙腈溶液(4mL对甲苯磺酰异氰酸酯加入10mL乙腈稀释为浓度为2.5mmol/mL),在氮气保护下,温度25℃,震荡50min条件下进行反应,然后加入1mL水终止,涡旋1min,减压旋蒸为油状物,30-35℃下真空干燥箱12h,得到白色或淡黄色固体为林可霉素衍生物1.5g,纯度为96.3%。
1HNMR(600MHz,Chloroform-d)δ9.61(s,1H),8.72(d,1H),7.70-7.76(m,5H),7.59-7.64(m,3H),7.36-7.45(m,6H),7.29(d,J=3.6Hz,2H),5.50(d,J=5.1Hz,1H),5.04(d,J=12.3Hz,1H),4.72-4.81(m,2H),4.51-4.53(m,1H),4.11(d,1H),3.98-4.06(m,2H),3.45-3.59(m,3H),2.81(s,3H),2.40(s,3H),2.37(s,3H),2.35(s,3H),2.32(s,3H),2.08-2.15(m,2H),1.99(s,3H),1.35-1.39(m,2H),1.24-1.29(m,2H),1.16-1.19(t,3H),0.98-1.02(d,J=4.8Hz,3H),0.86-0.89(t,3H);13CNMR(101MHz,DMSO-d)δ171.60(C=O),168.10(C=O),150.60(C=O),150.32(C=O),150.31(C=O),144.78,144.69,136.65,136.29,136.19,129.72,129.33,129.19,127.74,127.70,127.51,127.11,84.65,71.26,69.71,69.00,68.54,67.25,61.32,60.14,49.07,41.07,37.11,35.29,34.09,20.82,20.19,20.17,19.47,13.07,13.03,12.45,12.23。
衍生物的红外光谱图如图1所示。
实施例2:
林可霉素标液中的含量测定(HPLC):
仪器与条件:
美国安捷伦1260型高效液相色谱***,包括四元梯度***、柱温控制箱、紫外可见可变波长检测器、在线真空脱气机以及自动进样***;色谱柱:Agile nt C18(150*4.6mm,3.5μm),ZORBAX;水相:0.1vt%三乙胺水。有机相:乙腈。等度洗脱,水相:有机相=90:10;流速:1.0mL/min;柱温:25℃;检测波长:210nm;进样量:20μL。
实验步骤:
将林可霉素标液用流动相稀释至100μg/mL,过0.22μm有机滤膜,进样20μL供高效液相色谱测定,高效液相色谱(HPLC)图如图2所示,数据如下。信号:VWD1A,Wavelength-210nm
林可霉素标液中加入28.5vt%对甲苯磺酰异氰酸酯乙腈溶液和乙腈各100μL,涡旋30s,室温放置30min,待其充分反应后,加入100μL甲醇终止反应,涡旋混匀,用流动相定容至1mL,过0.22μm有机滤膜,进样20μL供高效液相色谱测定,高效液相色谱(HPLC)图如图3所示,数据如下。
信号:VWD1A,Wavelength-227nm
林可霉素衍生化之后比衍生化之前在高效液相色谱仪上的检测灵敏度高达17倍。
实施例3:
牛肉中林可霉素的含量测定(HPLC):
仪器与条件:
美国安捷伦1260型高效液相色谱***,包括四元梯度***、柱温控制箱、紫外可见可变波长检测器、在线真空脱气机以及自动进样***;色谱柱:Agile nt C18(150*4.6mm,3.5μm),ZORBAX;水相:称取0.385g醋酸铵溶于1L纯水加0.1vt%甲酸。有机相:乙腈。等度洗脱,水相:有机相=40:60(充分混匀后,再加入100mL水,再次进行混匀);流速:1.0mL/min;柱温:25℃;检测波长:227nm;进样量:20μL。
实验步骤:
准确称取5g牛肉置于50mL离心管中,加入林可霉素标液,静置10min后,加入10mL乙腈,涡旋30s,超声10min,中速振荡15min,6500rpm/min离心5min,取上清液置于另一50mL离心管中;组织残留物用10mL乙腈重提一次,合并两次提取后的上清液,氮气挥至约1mL。加入5mL水复溶,之后加入5mL饱和乙腈正己烷除脂,6500rpm/min离心5min,弃去正己烷层,待净化。
净化:
C18固相萃取小柱依次用3mL甲醇,3mL水,3mL十二烷基磺酸钠缓冲液活化,取样品复溶液过柱,流速保持为20滴/min,待样液完全流出后,用3mL水淋洗,弃去全部流出液。减压抽干5min,然后用3mL甲醇洗脱于10mL具塞玻璃试管中。于40℃氮气下挥干,加入28.5vt%对甲苯磺酰异氰酸酯乙腈溶液和乙腈各100μL,涡旋30s,室温放置30min,待其充分反应后,加入100μL甲醇终止反应,涡旋混匀,用流动相定容至1mL,过0.22μm有机滤膜,进样20μL供高效液相色谱测定。
图4为不经衍生化处理直接测定的牛肉含林可霉素高效液相色谱(HPLC)图;图5为经衍生化处理后测定的牛肉含林可霉素高效液相色谱(HPLC)图。
实施例4:
鸡肉中林可霉素的含量测定(HPLC):
仪器与条件:
美国安捷伦1260型高效液相色谱***,包括四元梯度***、柱温控制箱、紫外可见可变波长检测器、在线真空脱气机以及自动进样***;色谱柱:Agile nt C18(150*4.6mm,3.5μm),ZORBAX;水相:称取0.385g醋酸铵溶于1L纯水加0.1vt%甲酸。有机相:乙腈。等度洗脱,水相:有机相=40:60(充分混匀后,再加入100mL水,再次进行混匀);流速:1.0mL/min;柱温:25℃;检测波长:227nm;进样量:20μL。
实验步骤:
准确称取5g鸡肉置于50mL离心管中,加入林可霉素标液,静置10min后,加入7.5mL乙腈,涡旋30s,超声10min,中速振荡15min,6500rpm/min离心5min,取上清液置于另一50mL离心管中;组织残留物用7.5mL乙腈重提一次,8000rpm/min离心10min,合并两次提取后的上清液,氮气挥至约1mL。加入5mL水复溶,之后加入5mL饱和乙腈正己烷除脂,8000rpm/min离心5min,弃去正己烷层,待净化。
净化:
C18固相萃取小柱依次用3mL甲醇,3mL水,3mL十二烷基磺酸钠缓冲液活化,取样品复溶液过柱,流速保持为20滴/min,待样液完全流出后,用3mL水淋洗,弃去全部流出液。减压抽干5min,然后用3mL甲醇洗脱于10mL具塞玻璃试管中。于40℃氮气下挥干,加入28.5vt%对甲苯磺酰异氰酸酯乙腈溶液和乙腈各100μL,涡旋30s,室温放置30min,待其充分反应后,加入100μL甲醇终止反应,涡旋混匀,用流动相定容至1mL,过0.22μm有机滤膜,进样20μL供高效液相色谱测定。
图6为经衍生化处理后测定的鸡肉含林可霉素高效液相色谱(HPLC)图。
对于本领域的技术人员来说,可根据以上描述的技术方案以及构思,做出其它各种相应的改变以及变形,而所有的这些改变以及变形都应该属于本发明权利要求的保护范围之内。
Claims (10)
1.一种林可霉素的衍生化方法,其特征在于,所述衍生化方法为在林可霉素中加入不含活泼氢的反应溶剂和衍生化试剂进行反应。
2.如权利要求1所述的林可霉素的衍生化方法,其特征在于,所述不含活泼氢的反应溶剂为乙腈、四氢呋喃、***和甲苯中的至少一种。
3.如权利要求1所述的林可霉素的衍生化方法,其特征在于,所述衍生化试剂为对甲苯磺酰异氰酸酯。
4.如权利要求1所述的林可霉素的衍生化方法,其特征在于,所述反应的条件为氮气保护下,温度10-30℃,震荡20-60min。
5.如权利要求1所述的林可霉素的衍生化方法,其特征在于,反应后加入终止剂终止反应。
6.如权利要求5所述的林可霉素的衍生化方法,其特征在于,所述终止剂为水、盐酸、甲醇和乙醇中的至少一种。
7.如权利要求1所述的林可霉素的衍生化方法,其特征在于,所述林可霉素和衍生化试剂的摩尔比为1:(1-10)。
9.一种林可霉素的衍生化方法的应用,其特征在于,通过林可霉素的衍生化方法得到林可霉素衍生物,然后将林可霉素衍生物用于测定林可霉素的含量;所述林可霉素的衍生化方法为在林可霉素中加入不含活泼氢的反应溶剂和衍生化试剂进行反应。
10.如权利要求9所述的林可霉素的衍生化方法的应用,其特征在于,所述测定林可霉素的含量的方法为色谱法。
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
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CN1963492A (zh) * | 2006-11-29 | 2007-05-16 | 复旦大学 | 磺酰异氰酸酯柱前衍生化法测定水分含量的方法 |
CN103424481A (zh) * | 2013-05-06 | 2013-12-04 | 华中农业大学 | 林可霉素、克林霉素和大观霉素的残留分析方法 |
CN107957463A (zh) * | 2017-12-29 | 2018-04-24 | 宁夏希望田野生物农业科技有限公司 | 一种土壤中林可霉素残留量的检测方法 |
-
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1963492A (zh) * | 2006-11-29 | 2007-05-16 | 复旦大学 | 磺酰异氰酸酯柱前衍生化法测定水分含量的方法 |
CN103424481A (zh) * | 2013-05-06 | 2013-12-04 | 华中农业大学 | 林可霉素、克林霉素和大观霉素的残留分析方法 |
CN107957463A (zh) * | 2017-12-29 | 2018-04-24 | 宁夏希望田野生物农业科技有限公司 | 一种土壤中林可霉素残留量的检测方法 |
Non-Patent Citations (2)
Title |
---|
周涛: "对甲苯磺酰异氰酸酯柱前衍生化HPLC法测定二甘醇等物质的含量", 《复旦大学硕士学位论文》 * |
王淑梅,等: "《临床输液配伍禁忌》", 30 June 2020 * |
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