参考QB/T 5715-2022高效液相色谱法测定发酵液中衣康酸含量-医疗器械网

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参考QB/T 5715-2022高效液相色谱法测定发酵液中衣康酸含量

来源：东曹（上海）生物科技有限公司分类：行业标准 2024-04-28 10:00:08 16阅读次数

衣康酸（itaconicacid）又称甲叉丁二酸、亚甲基丁二酸，是世界上5大有机酸之一。由于其含不饱和双键，具有活泼的化学性质，能进行各种加成反应、聚合反应和酯化反应，是化学合成和化工生产的重要工业原料，在化纤、塑料、橡胶、医药、食品、合成树脂及表面活性剂领域具有广泛的应用。

本应用参照QB/T 5715-2022《发酵液中衣康酸的测定高效液相色谱法》，采用C18色谱柱TSKgel ODS-100V（4.6 mm I.D.×25 cm，5μm）对衣康酸进行定量分析，本方法可满足上述行标中的定量要求。

01

溶液配制

① 衣康酸标准溶液（2 mg/mL）：准确称取20 mg衣康酸标准，用流动相溶解并定容至10 mL，此溶液衣康酸浓度为2 mg/mL，于4℃保存，有效期6个月。

② 衣康酸标准工作曲线溶液：使用上述2 mg/mL标准溶液，使用流动相逐级稀释至5 mg/L、10 mg/L、20 mg/L、50 mg/L、100 mg/L、200 mg/L。

③ 流动相：0.1%磷酸水溶液:甲醇=87:13 (体积比)

02

分析条件

仪器：Thermo Ultimate 3000

色谱柱：TSKgel ODS-100V（4.6 mmI.D.×25 cm, 5 μm）

流速：0.8 mL/min

柱温：45℃

进样量：5 μL

检测器：UV@210nm

03

分析结果

图1 约0.1mg/mL衣康酸溶液的色谱图

图2 衣康酸浓度-峰面积校正曲线

使用阴离子交换色谱法分离完整AAV衣壳和空AAV衣壳

采用SEC色谱柱测定人促红素中二聚体及高分子量物质

采用亲和色谱柱TSKgel Boronate-5PW测定单抗糖化比例

参照USP129修订案建立单抗的SEC-UHPLC分析方法

TSKgel色谱柱使用中的10大常见典型问题及解答

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[8]未定义数组索引: page
[2]Declaration of app\common\model\TechNewsCategory::getList($filter = Array) should be compatible with app\common\model\BaseModel::getList($filter = Array, $sort = Array)
[8]未定义数组下标: 0
[2]Declaration of app\common\model\TechNews::getAllList($filter = Array, $limit = 10, $order = Array, $with = Array) should be compatible with app\common\model\BaseModel::getAllList($filter = Array, $with = Array, $limit = 0, $sort = Array, $field = 'a.*', $join = Array)
[2]Declaration of app\common\model\Company::getAllList($filter = Array, $limit = 20) should be compatible with app\common\model\BaseModel::getAllList($filter = Array, $with = Array, $limit = 0, $sort = Array, $field = 'a.*', $join = Array)
[8]未定义数组索引: center
[8]Only variables should be passed by reference
[2]Declaration of app\common\model\TechNewsComment::getAllList($filter = Array, $limit = 16) should be compatible with app\common\model\BaseModel::getAllList($filter = Array, $with = Array, $limit = 0, $sort = Array, $field = 'a.*', $join = Array)
[8192]Function create_function() is deprecated
[8192]Function create_function() is deprecated
[8]未定义数组索引: area
[8]未定义数组索引: st
[2]Declaration of app\common\model\Topic::getList($filter = Array) should be compatible with app\common\model\BaseModel::getList($filter = Array, $sort = Array)
[2]Declaration of app\common\model\Topic::getAllList($filter = Array, $field = 'id,md5,tit...', $limit = 0) should be compatible with app\common\model\BaseModel::getAllList($filter = Array, $with = Array, $limit = 0, $sort = Array, $field = 'a.*', $join = Array)
[2]Declaration of app\common\model\TopicInfoList::getAllList($filter = Array, $limit = 8) should be compatible with app\common\model\BaseModel::getAllList($filter = Array, $with = Array, $limit = 0, $sort = Array, $field = 'a.*', $join = Array)
[2]Declaration of app\common\model\News::getAllList($filter = Array, $limit = 16, $order = Array, $with = Array) should be compatible with app\common\model\BaseModel::getAllList($filter = Array, $with = Array, $limit = 0, $sort = Array, $field = 'a.*', $join = Array)
[8]未定义数组索引: center
[8]未定义数组索引: cid
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[8]未定义数组索引: center
[8]未定义数组索引: center
[8]未定义数组索引: wtt_list

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[ SQL ] SELECT * FROM `user_info` WHERE `user_id` IN (273,197,252,316,473,714,250) [ RunTime:0.000888s ]
[ SQL ] SHOW COLUMNS FROM `topic` [ RunTime:0.000859s ]
[ SQL ] SELECT `id`,`md5`,`title`,`created_time` FROM `topic` WHERE `review_status` = 1 ORDER BY `id` DESC LIMIT 4 [ RunTime:0.000200s ]
[ SQL ] SHOW COLUMNS FROM `topic_info_list` [ RunTime:0.000926s ]
[ SQL ] SELECT * FROM `topic_info_list` WHERE `topic_id` = 1 ORDER BY `id` DESC LIMIT 2 [ RunTime:0.000206s ]
[ SQL ] SHOW COLUMNS FROM `news` [ RunTime:0.000933s ]
[ SQL ] SELECT * FROM `news` WHERE `id` IN (864,849) [ RunTime:0.000241s ]
[ SQL ] SELECT `id`,`md5`,`title`,`created_time` FROM `tech_news` WHERE `review_status` = 1 AND `is_show` = 1 ORDER BY `id` DESC LIMIT 10 [ RunTime:0.000427s ]
[ SQL ] SHOW COLUMNS FROM `user` [ RunTime:0.001044s ]
[ SQL ] SELECT `a`.* FROM `user` `a` INNER JOIN `tech_news` `b` ON `a`.`id`=`b`.`user_id` GROUP BY `a`.`id` ORDER BY `b`.`id` DESC LIMIT 10 [ RunTime:0.004579s ]
[ SQL ] SELECT * FROM `user_info` WHERE `user_id` IN (592,413,444,224,13,239,598,697,394,285) [ RunTime:0.000908s ]
[ SQL ] SHOW COLUMNS FROM `seo` [ RunTime:0.000906s ]
[ SQL ] SELECT * FROM `seo` WHERE `column_type` = '16' AND `page_type` = 'tech_news' AND `info_id` = 3694 LIMIT 1 [ RunTime:0.000197s ]
[ SQL ] SELECT * FROM `tech_news_belong_category` WHERE `tech_news_id` = 3694 [ RunTime:0.000904s ]
[ SQL ] SELECT * FROM `tech_news_belong_category` WHERE `tech_news_id` = 3668 [ RunTime:0.001178s ]
[ SQL ] SELECT * FROM `tech_news_belong_category` WHERE `tech_news_id` = 3452 [ RunTime:0.000999s ]
[ SQL ] SELECT * FROM `tech_news_belong_category` WHERE `tech_news_id` = 3314 [ RunTime:0.000990s ]
[ SQL ] SELECT * FROM `tech_news_belong_category` WHERE `tech_news_id` = 2129 [ RunTime:0.000976s ]
[ SQL ] SELECT * FROM `tech_news_belong_category` WHERE `tech_news_id` = 1815 [ RunTime:0.000995s ]
[ SQL ] SELECT * FROM `tech_news_belong_category` WHERE `tech_news_id` = 3821 [ RunTime:0.001024s ]
[ SQL ] SELECT * FROM `tech_news_belong_category` WHERE `tech_news_id` = 1821 [ RunTime:0.001000s ]
[ SQL ] SELECT * FROM `tech_news_belong_category` WHERE `tech_news_id` = 1600 [ RunTime:0.001006s ]
[ SQL ] SELECT * FROM `tech_news_belong_category` WHERE `tech_news_id` = 2207 [ RunTime:0.001043s ]
[ SQL ] SELECT * FROM `user` WHERE `id` = 273 LIMIT 1 [ RunTime:0.000174s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `follow_user_id` = 592 LIMIT 1 [ RunTime:0.000188s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `user_id` = 592 LIMIT 1 [ RunTime:0.000141s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `follow_user_id` = 413 LIMIT 1 [ RunTime:0.000157s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `user_id` = 413 LIMIT 1 [ RunTime:0.000131s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `follow_user_id` = 444 LIMIT 1 [ RunTime:0.000114s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `user_id` = 444 LIMIT 1 [ RunTime:0.000155s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `follow_user_id` = 224 LIMIT 1 [ RunTime:0.000125s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `user_id` = 224 LIMIT 1 [ RunTime:0.000115s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `follow_user_id` = 13 LIMIT 1 [ RunTime:0.000151s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `user_id` = 13 LIMIT 1 [ RunTime:0.000142s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `follow_user_id` = 239 LIMIT 1 [ RunTime:0.000128s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `user_id` = 239 LIMIT 1 [ RunTime:0.000157s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `follow_user_id` = 598 LIMIT 1 [ RunTime:0.000169s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `user_id` = 598 LIMIT 1 [ RunTime:0.000545s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `follow_user_id` = 697 LIMIT 1 [ RunTime:0.000167s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `user_id` = 697 LIMIT 1 [ RunTime:0.000190s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `follow_user_id` = 394 LIMIT 1 [ RunTime:0.000151s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `user_id` = 394 LIMIT 1 [ RunTime:0.000163s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `follow_user_id` = 285 LIMIT 1 [ RunTime:0.000222s ]
[ SQL ] SELECT COUNT(*) AS tp_count FROM `user_follow` WHERE `user_id` = 285 LIMIT 1 [ RunTime:0.000121s ]

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