WO2021128665A1 - 一种生物素中间体双氨基物的hplc检测方法 - Google Patents

一种生物素中间体双氨基物的hplc检测方法 Download PDF

Info

Publication number
WO2021128665A1
WO2021128665A1 PCT/CN2020/085714 CN2020085714W WO2021128665A1 WO 2021128665 A1 WO2021128665 A1 WO 2021128665A1 CN 2020085714 W CN2020085714 W CN 2020085714W WO 2021128665 A1 WO2021128665 A1 WO 2021128665A1
Authority
WO
WIPO (PCT)
Prior art keywords
biotin
hplc detection
detection method
mobile phase
phosphate buffer
Prior art date
Application number
PCT/CN2020/085714
Other languages
English (en)
French (fr)
Inventor
吕国锋
王昌泽
单国红
杭姣
王振振
李小军
段文杰
韦念想
Original Assignee
上虞新和成生物化工有限公司
浙江新和成药业有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 上虞新和成生物化工有限公司, 浙江新和成药业有限公司 filed Critical 上虞新和成生物化工有限公司
Publication of WO2021128665A1 publication Critical patent/WO2021128665A1/zh

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/86—Signal analysis
    • G01N30/8624—Detection of slopes or peaks; baseline correction
    • G01N30/8631—Peaks

Definitions

  • the invention belongs to the technical field of biotin intermediate content detection, and specifically relates to an HPLC detection method for a diamino compound of a process intermediate in biotin synthesis.
  • Biotin (the following formula a) will produce a diamino compound (the following formula b) during the synthesis process, which is an intermediate process of urea acid to biotin, and d-biotin can be obtained by carbonylation and post-treatment.
  • the detection methods in the prior art all use biotin as the target product for detection, which leads to the neglect of diamino compounds that can be converted into biotin. Because the diamino compound appears near the dead time, it is not easy to determine its residue, which affects the overall yield of biotin.
  • the detection methods of biotin mainly include microbiological method, liquid chromatography, enzyme-linked immunoassay, spectroscopy and electrochemical methods.
  • the operation of the microbiological method is time-consuming, laborious, and costly; although enzyme-linked immunoassay and spectroscopy are simple to operate, fast in analysis, and highly accurate, their market cost is high and it is difficult to promote and apply in production.
  • the above methods are all based on the detection of biotin, and the detection of impurities lacks accuracy and sensitivity.
  • Chinese patent document CN200810162138.6 discloses a method for quantitatively detecting the content of d-biotin and its impurities.
  • the chromatographic conditions when the invention adopts high-performance liquid chromatography detection are: a reversed-phase C18 column, a mobile phase of trifluoroacetic acid aqueous solution-acetonitrile with a mass concentration of 0.05%, a flow rate of 0.5-1.5 mL/min, a flow rate of 210-220 nm Detection wavelength, the volume ratio of trifluoroacetic acid aqueous solution and acetonitrile in the mobile phase is 60-80:40-20; dissolve and dilute the impurity-containing d-biotin with the mobile phase, and measure by sample injection.
  • the structures of these impurities A-H are as follows:
  • the detection method has high sensitivity (the detection limit is as low as 0.5ng), which meets the needs of biotin quantitative detection and helps to improve the quality control of biotin.
  • the test product is prepared by dissolving biotin and various impurities, and it directly uses the mobile phase as the solvent. It can be seen from the peak figure that the impurity diamino compound (structure C) peaks earlier (about 1.5-2 min).
  • the detection method of the invention is suitable for the quality detection of the final product, and in the actual production process, the reaction solvent for synthesizing biotin is usually methanol, toluene, hydrobromic acid, etc. When this method is used to directly detect the reaction solution, there will be a problem that the diamino compound C overlaps with the solvent peak, and its content cannot be accurately detected. Therefore, the above method is not directly applicable to the direct detection of the reaction solution in the biotin preparation process.
  • the purpose of the present invention is to provide an HPLC detection method for the biotin intermediate bisamino compound.
  • the detection result of this method is accurate, reliable and easy to operate.
  • the detection method can better grasp the content of the diamino compound, so as to effectively control the conversion rate, thereby increasing the overall yield of biotin.
  • An HPLC detection method for biotin intermediate bisamino compound which comprises the following steps: dissolving a sample containing biotin and bisamino compound in a mobile phase and then performing detection under the following chromatographic conditions: chromatographic column C18-WCX mixed column;
  • the detection wavelength is 195-220nm
  • the test time is 16-20min
  • the mobile phase is acetonitrile-phosphate buffer.
  • the phosphate buffer is potassium dihydrogen phosphate buffer.
  • the phosphate buffer is 0.01-0.03 mol/L. More preferably, it is 0.015 mol/L.
  • the volume ratio of acetonitrile to phosphate buffer in the mobile phase acetonitrile-phosphate is: 60%-80%: 40%-20%, preferably the volume ratio of acetonitrile-phosphate is 70%: 30%.
  • equal concentration elution is used during the HPLC detection process.
  • the detection wavelength is 200 nm.
  • the structural formula of the biotin is as follows (a), and the bisamino compound is as follows (b):
  • the ratio of the sample containing biotin and diamino compound to the mobile phase is: 0.05 g: 10 mL.
  • the injection volume 10 ⁇ L
  • the column temperature is 30° C.
  • the flow rate 1.0-1.5 mL/min.
  • the formula for calculating the content of the diamino compound is:
  • a diamino the peak area corresponding to the diamino compound in the sample solution chromatogram
  • ⁇ Ai the sum of the peak areas corresponding to each component in the sample solution chromatogram.
  • the detection method provided by the present invention can stabilize the peak time of the bisamino compound to be tested at 10-12 min, solves the problem of overlapping peaks of the bisamino compound and the solvent, and makes the monitoring more accurate and reliable.
  • the detection method provided by the present invention can more accurately grasp the content of the impurity diamino compound in biotin, can better control the reaction end point, effectively convert the diamino compound into biotin, and increase the yield of biotin on average Around 10%.
  • the detection method provided by the present invention has high precision, and the relative standard deviation ranges of the chromatographic peak area and retention time of the diamino compound are 0.31%-1.20% and 0.14%-1.94%, respectively, as verified by repeatability experiments.
  • the detection limit of this method is low, and the minimum detection limit of diamino compounds is 1.2ng.
  • Figure 1 is a chromatogram of the HPLC detection result of the standard bisamino compound of the present invention at 195 nm;
  • Fig. 2 is a chromatogram of the HPLC detection result of the bisamino compound standard substance of the present invention at 200 nm;
  • Fig. 3 is a chromatogram of the HPLC detection result of the bisamino compound standard substance of the present invention at 210 nm;
  • Fig. 4 is a chromatogram of the HPLC detection result of the bisamino compound standard substance of the present invention at 220 nm;
  • Figure 5 is a chromatogram of the HPLC detection result of the diamino compound in the reaction solution at 195 nm;
  • Figure 6 is a chromatogram of the HPLC detection result of the diamino compound in the reaction solution at 200 nm;
  • Figure 7 is a chromatogram of the HPLC detection result of the diamino compound in the reaction solution at 210 nm;
  • Figure 8 is a chromatogram of the HPLC detection result of the diamino compound in the reaction solution at 220 nm.
  • Acetonitrile is a chromatographic reagent, and other reagents are analytical reagents
  • Buffer preparation Weigh 0.015moL of dipotassium hydrogen phosphate, dissolve it in 1L of deionized water, filter it with a 0.22 ⁇ m microporous membrane, and use it now.
  • the preparation process of biotin in this patent is as follows: the bromine on the uric acid generates monobenzyl and is quickly converted into biotin, but at the same time, the ring opens due to excessive reaction to generate diamino compound, and the diamino compound regenerates biotin through photochemical cyclization.
  • the reaction solution after biotin synthesis refers to the reaction solution (photochemical reaction solution) at the end of the photochemical cyclization reaction.
  • the specific preparation examples of biotin are given as follows:
  • Injection volume 20 ⁇ L
  • Test results The chromatograms of the HPLC test results of the above reaction solution at different wavelengths are shown in Figure 5-8.
  • a mobile phase is used to gradually dissolve the diamino standard solution at room temperature, and the sample is injected for detection and analysis.
  • the calculated minimum detection limit of the diamino compound is 1.2 ng.

Abstract

一种生物素中间体双氨基物的HPLC检测方法，该检测方法包括如下步骤：用流动相溶解生物素合成过程中的环合反应液后，在以下色谱条件下进行检测：色谱柱为C18-WCX混合柱，检测波长为195-220nm，测试时间为16-20min，流动相为乙腈-磷酸盐缓冲液。该检测方法可以使待测物质双氨基物的出峰时间稳定在10-12min，解决了双氨基物与溶剂峰出峰重合的问题，使得监测更加准确、可靠。

Description

一种生物素中间体双氨基物的HPLC检测方法 技术领域
本发明属于生物素中间体含量检测技术领域，具体涉及一种生物素合成中的过程中间体双氨基物的HPLC检测方法。
背景技术
生物素(如下式a)合成过程中会产生一种双氨基物(如下式b)，该物质是脲酸生成生物素的中间过程物，经羰基化及后处理可得d-生物素。
Figure PCTCN2020085714-appb-000001
现有技术中的检测方法，都以生物素为检测的目标产物，导致能转化为生物素的双氨基物被忽视。因为双氨基物出现在死时间附近，不易确定其残留，从而影响生物素的整体收率。目前，生物素的检测方法主要有微生物法、液相色谱法、酶联免疫分析法、光谱法和电化学法等。微生物法操作费时、费力，成本高；酶联免疫法和光谱法虽然操作简单、分析速度快且准确度高，但市场造价较高，难以在生产上进行推广应用。且以上方法都是基于生物素的检测，而对于杂质的检测欠缺准确性和灵敏性。
经检索，中国专利文献CN200810162138.6公开了一种定量检测d-生物素及其杂质含量的方法。该发明采用高效液相色谱法检测时的色谱条件为：采用反相C18柱，质量浓度为0.05％的三氟乙酸水溶液-乙腈的流动相，0.5-1.5mL/min的流速，210-220nm的检测波长，流动相中三氟乙酸水溶液与乙腈的体积比为60-80∶40-20；用流动相溶解含杂质的d-生物素并稀释，进样测定，通过高效液相色谱仪检测d-生物素及其各个杂质的含量。这些杂质A-H结构如下所示：
Figure PCTCN2020085714-appb-000002
该检测方法具有较高的灵敏度(检测限低至0.5ng)，满足了生物素定量检测的需要，有助于提高生物素的质量控制。
但是该发明提供的实施例，供试品是采用生物素及各杂质溶解后配制而成，其直接采用了流动相作为溶剂。由其出峰图可以看出，杂质双氨基物(结构C)出峰较早(大约1.5-2min左右)。该发明的检测方法适用于最终产品质量检测，而在实际生产过程中，合成生物素的反应溶剂通常是甲醇、甲苯、氢溴酸等。当采用该方法直接检测反应液时，则会出现双氨基物C与溶剂峰重合，无法准确检测其含量的问题。所以，上述方法并无法直接适用于生物素制备过程中反应液的直接检测。
在生物素制备过程中，需要随时监测中间体及产物的含量，以提高产物收率。为此，亟需提出一种具有针对性的用于双氨基物的定量检测的方法。
发明内容
为了解决现有技术中存在的上述问题，本发明的目的在于提供一种生物素中间体双氨基物的HPLC检测方法。该方法检测结果准确、可靠、操作简便。而且利用该检测方法可以更好的掌握双氨基物的含量，以便于有效控制其转化率，进而提高生物素的整体收率。
为了实现上述发明目的，本发明提供了如下技术方案：
一种生物素中间体双氨基物的HPLC检测方法，其包括如下步骤：采用流动相将含有生物素和双氨基物的样品溶解后在以下色谱条件下进行检测：色谱柱C18－WCX混合柱；
检测波长为195-220nm，测试时间为16-20min，流动相为乙腈-磷酸盐缓冲液。
所述磷酸盐缓冲液为磷酸二氢钾缓冲液。
优选地，所述磷酸盐缓冲液为0.01-0.03mol/L。进一步优选为0.015mol/L。
优选地，流动相乙腈-磷酸盐中乙腈与磷酸盐缓冲液的体积配比为：60％-80％：40％-20％，优选乙腈-磷酸盐体积配比为70％：30％。
优选地，HPLC检测过程中采用等浓度洗脱。
优选地，所述检测波长为200nm。
优选地，所述生物素的结构式如下(a)，双氨基物结构式如下(b)：
Figure PCTCN2020085714-appb-000003
优选地，所述含有生物素和双氨基物的样品与流动相的比例为：0.05g：10mL。
优选地，进样体积：10μL、柱温为30℃；流速：1.0-1.5mL/min。
优选地，双氨基物的含量计算公式为：
Figure PCTCN2020085714-appb-000004
式中：A 双氨基—样品溶液色谱图中双氨基物对应的峰面积；ΣAi—样品溶液色谱图中各组分对应的峰面积之和。
与现有技术相比，本发明所取得的有益效果是：
(1)本发明提供的检测方法可以在使待测物质双氨基物的出峰时间稳定在10-12min，解决了双氨基物与溶剂峰出峰重合的问题，使得监测更加准确、可靠。
(2)本发明提供的检测方法能够更准确的掌握生物素中杂质双氨基物的含量，可更好的控制反应终点，将双氨基物有效转化为生物素，使生物素的收率平均提高10％左右。
(3)本发明提供的检测方法精密度高，经重复性试验验证，双氨基物的色谱峰面积和保留时间的相对标准偏差范围分别为0.31％-1.20％和0.14％-1.94％。
(4)该方法检出限低，双氨基物的最小检测限度为1.2ng。
附图说明
图1为本发明双氨基物标准物在195nm处的HPLC检测结果色谱图；
图2为本发明双氨基物标准物在200nm处的HPLC检测结果色谱图；
图3为本发明双氨基物标准物在210nm处的HPLC检测结果色谱图；
图4为本发明双氨基物标准物在220nm处的HPLC检测结果色谱图；
图5为反应液中双氨基物在195nm处的HPLC检测结果色谱图；
图6为反应液中双氨基物在200nm处的HPLC检测结果色谱图；
图7为反应液中双氨基物在210nm处的HPLC检测结果色谱图；
图8为反应液中双氨基物在220nm处的HPLC检测结果色谱图。
具体实施方式
下面通过具体实施例对本发明的方法进行说明，以使本发明技术方案更易于理解、掌握，但本发明并不局限于此。下述实施例中所述实验方法，如无特殊说明，均为常规方法；所述试剂和材料，如无特殊说明，均可从商业途径获得。
实施例1
1、仪器与试剂
仪器型号：戴安液相色谱仪，U3000
试剂：乙腈为色谱纯试剂，其他试剂为分析纯试剂
2、标准品溶液的配制
双氨基物标准物溶液配制：取适量的氨基物标准物溶于流动相中，必要时超声处理使完全溶解。
缓冲液的配制：称取0.015moL的磷酸氢二钾溶于1L去离子水中，并用0.22μm微孔滤膜过滤后备用，且现配现用。
3、待测样品的配制
用毛细管取0.05g的生物素合成后的反应液至15mL取样瓶内，加10mL流动相溶解，超声后备用。
本专利中生物素的制备过程为：脲酸上溴生成单苄并快速转化为生物素，但同时因过度反应而开环生成双氨基物，双氨基物通过光化环合重新生成生物素。所述生物素合成后的反应液是指光化环合反应结束时的反应液(光化反应液)。如下给出生物素的具体制备实施例：
在带有精馏塔的500mL四口烧瓶中，投入双苄基生物素(30g，70mmol)和300mL一定浓度的氢溴酸，于130℃下搅拌反应，反应过程中及时从精馏塔顶分离溴化苄。用HPLC监测反应完全后，冷却至80℃，加入100mL甲苯萃取分层，得到的水层中含有生物素和过度反应生成的双氨基物，减压回收其中的氢溴酸。回收毕，加入氢氧化钠溶液调节pH至9-10，于-5℃冰浴下滴入固体光气甲苯溶液(50mL，16mmol)，滴加固体光气过程中持续加入氢氧化钠溶液维持反应体系pH为9-10。滴加毕继续保温搅拌30min，取样监测反应结果。该样品即为光化反应液，需要静置5-10min，取水层进行检测。
4、检测波长的选择
称取双氨基标准物，用流动相配制成合适浓度的溶液，在紫外190-400nm波长范围内扫描，结果显示，在200nm处，双氨基物的响应值最高，因此检测波长优选200nm。不同波长下双氨基物的高效液相检测结果色谱图如图1-4所示。
5、色谱柱条件
色谱柱：AcclaimTM Mixed-Mode WCX-1；
流速：1.0mL/min；
柱温：30℃；
测试时间：20min；
进样体积：20μL；
流动相：乙腈-磷酸二氢钾缓冲液(70％：30％)。
6、洗脱方式：等浓度洗脱。
7、检测结果：如上反应液在不同波长的高效液相检测结果色谱图如图5-8所示。
对比图5-8可以明显看出，波长在195-220nm条件下，双氨基物出峰时间稳定在10-12min，均与其它杂质较好的分离开。而且反应液中的所有杂质均在16min左右完全出峰。检测时间短。
如上反应液中各杂质的含量测定结果如下表1所示。
Figure PCTCN2020085714-appb-000005
195nm虽然响应更好，但基线开始变差；220nm，各种峰响应开始改变，综合来讲200nm测试结果更佳。
方法学考察
1、精密度考察
取100倍稀释的双氨基物标准品，在如上检测条件下进行7次重复性试验，得到的双氨基物的色谱峰面积和保留时间的相对标准偏差范围分别为0.31％-1.20％和0.14％-1.94％，表明本方法精密度良好。
2、检测限
在本发明中的色谱条件下，用流动相在室温条件下逐步溶解双氨基标准品溶液，进样检测，进行分析，计算的双氨基物的最小检测限度为1.2ng。
最后应当说明的是，以上内容仅用以说明本发明的技术方案，而非对本发明保护范围的限制，本领域的普通技术人员对本发明的技术方案进行的简单修改或者等同替换，均不脱离本发明技术方案的实质和范围。

Claims (10)

  1. 一种生物素中间体双氨基物的HPLC检测方法，其包括如下步骤：
    用流动相溶解生物素合成过程中的环合反应液后在以下色谱条件下进行检测：色谱柱C18-WCX混合柱；
    检测波长为195-220nm，测试时间为16-20min，流动相为乙腈-磷酸盐缓冲液。
  2. 根据权利要求1所述的HPLC检测方法，所述磷酸盐缓冲液为磷酸二氢钾缓冲液。
  3. 根据权利要求1所述的HPLC检测方法，所述磷酸盐缓冲液为0.01-0.03mol/L。
  4. 根据权利要求1所述的HPLC检测方法，流动相乙腈-磷酸盐中乙腈与磷酸盐缓冲液的体积比为：60％-80％：40％-20％。
  5. 根据权利要求4所述的HPLC检测方法，流动相乙腈-磷酸盐中乙腈与磷酸盐缓冲液的体积比为70％：30％。
  6. 根据权利要求1所述的HPLC检测方法，HPLC检测过程中采用等浓度洗脱。
  7. 根据权利要求1所述的HPLC检测方法，所述生物素的结构式如下(a)，双氨基物结构式如下(b)：
    Figure PCTCN2020085714-appb-100001
  8. 根据权利要求1所述的HPLC检测方法，所述含有生物素和双氨基物的样品与流动相的比例为：0.05g：10mL。
  9. 根据权利要求1所述的HPLC检测方法，进样体积：20μL、柱温为30℃；流速：1.0-1.5mL/min。
  10. 根据权利要求1所述的HPLC检测方法，双氨基物的含量计算公式为：
    Figure PCTCN2020085714-appb-100002
    式中：A 双氨基—样品溶液色谱图中双氨基物对应的峰面积；ΣA i—样品溶液色谱图中各组分对应的峰面积之和。
PCT/CN2020/085714 2019-12-27 2020-04-20 一种生物素中间体双氨基物的hplc检测方法 WO2021128665A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201911373716.5 2019-12-27
CN201911373716.5A CN110887913B (zh) 2019-12-27 2019-12-27 一种生物素中间体双氨基物的hplc检测方法

Publications (1)

Publication Number Publication Date
WO2021128665A1 true WO2021128665A1 (zh) 2021-07-01

Family

ID=69753330

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2020/085714 WO2021128665A1 (zh) 2019-12-27 2020-04-20 一种生物素中间体双氨基物的hplc检测方法

Country Status (2)

Country Link
CN (1) CN110887913B (zh)
WO (1) WO2021128665A1 (zh)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110887913B (zh) * 2019-12-27 2020-10-20 上虞新和成生物化工有限公司 一种生物素中间体双氨基物的hplc检测方法
CN114371243A (zh) * 2021-12-28 2022-04-19 湖南中晟全肽生化有限公司 一种d-生物素的HPLC检测方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101504393A (zh) * 2008-11-14 2009-08-12 浙江医药股份有限公司新昌制药厂 d-生物素及其杂质含量的HPLC测定方法
JP2015118029A (ja) * 2013-12-19 2015-06-25 国立大学法人東北大学 生体試料中のビオチンまたはその関連物質の測定方法
CN105418633A (zh) * 2015-12-04 2016-03-23 蚌埠丰原医药科技发展有限公司 一种从d-生物素中分离杂质的方法
CN110887913A (zh) * 2019-12-27 2020-03-17 上虞新和成生物化工有限公司 一种生物素中间体双氨基物的hplc检测方法

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030077616A1 (en) * 2001-04-19 2003-04-24 Ciphergen Biosystems, Inc. Biomolecule characterization using mass spectrometry and affinity tags
CN103772410B (zh) * 2013-12-31 2016-08-31 浙江新和成股份有限公司 一种d-生物素的合成方法
CN109503619A (zh) * 2018-12-25 2019-03-22 浙江新和成股份有限公司 一种d-生物素的合成方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101504393A (zh) * 2008-11-14 2009-08-12 浙江医药股份有限公司新昌制药厂 d-生物素及其杂质含量的HPLC测定方法
JP2015118029A (ja) * 2013-12-19 2015-06-25 国立大学法人東北大学 生体試料中のビオチンまたはその関連物質の測定方法
CN105418633A (zh) * 2015-12-04 2016-03-23 蚌埠丰原医药科技发展有限公司 一种从d-生物素中分离杂质的方法
CN110887913A (zh) * 2019-12-27 2020-03-17 上虞新和成生物化工有限公司 一种生物素中间体双氨基物的hplc检测方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CHASTAIN, J.L. ; BOWERS-KOMRO, D.M. ; MCCORMICK, D.B.: "High-performance liquid chromatography of biotin and analogues", JOURNAL OF CHROMATOGRAPHY A, ELSEVIER, AMSTERDAM, NL, vol. 330, 1 January 1985 (1985-01-01), AMSTERDAM, NL, pages 153 - 158, XP026557023, ISSN: 0021-9673, DOI: 10.1016/S0021-9673(01)81971-3 *
XUEFAN CHEN, CHUCHU HU, PANPAN ZHAO, GUOGANG ZHENG: "Determination of Related Substances in Biotin by HPLC", CHINESE JOURNAL OF PHARMACEUTICALS, SHANGHAI YIYAO GONGYE YANJIUYUAN,SHANGHAI INSTITUTE OF PHARMACEUTICAL INDUSTRY, CN, vol. 45, no. 11, 1 January 2014 (2014-01-01), CN, pages 1066 - 1069, XP055827309, ISSN: 1001-8255, DOI: 10.16522/j.cnki.cjph.2014.11.004 *

Also Published As

Publication number Publication date
CN110887913B (zh) 2020-10-20
CN110887913A (zh) 2020-03-17

Similar Documents

Publication Publication Date Title
WO2021128665A1 (zh) 一种生物素中间体双氨基物的hplc检测方法
Riggin et al. Liquid chromatographic method for monitoring therapeutic concentrations of L-dopa and dopamine in serum.
CN105572241B (zh) 测定血液和尿液中苯丙胺类、氯胺酮、哌替啶和美沙酮的方法
CN106442771A (zh) 一种测定十四烷基硫酸钠及其有关物质的检测方法
CN104502477B (zh) 一种三氯乙醛废硫酸中有机物的分析方法
CN101393196B (zh) 一种同时测定人血浆中多种局部麻醉药物浓度的方法
CN109307716B (zh) 一种依匹哌唑有关物质的检测方法
CN113295805A (zh) 一种药物中水合肼的检测方法
CN102565208B (zh) 一种硫酸依替米星新的检测方法
Gong et al. Purity assessment of luteolin reference material candidates by coulometric titration method
CN113533548A (zh) 一种化工产品中1-乙烯基咪唑的检测方法
CN110954629A (zh) 一种呋塞米中糠胺含量测定的控制方法
CN114778743B (zh) L-脯氨酸中痕量手性异构体d-脯氨酸的检测方法
Zhao et al. A fast and sensitive HPLC–MS/MS analysis and preliminary pharmacokinetic characterization of ergone in rats
CN112557520B (zh) Tgr-1中tgr-1-对应异构体的检测方法
CN105301118A (zh) 一种枸橼酸莫沙必利有关物质的检测方法
Ma et al. Development of highly sensitive analysis method for histamine and metabolites in pregnant women’s fingernail by UPLC-ESI-MS
CN108558964B (zh) 一种α-熊果苷的纯化方法
CN112578037A (zh) 一种检测醋酸乌利司他中间体ⅰ中乌双缩酮含量的方法
CN105572063A (zh) 一种基于氯化血红素可控聚集的水胺硫磷便捷检测方法
CN111044636A (zh) 米卡芬净含量的分析方法
CN112213411B (zh) 一种双醋瑞因中基因毒性杂质的检测方法
CN115097022B (zh) 一种甘油法制二氯丙醇过程中的杂质含量的测定方法
CN111122742B (zh) 一种待测样品中二巯基聚乙二醇残留量的检测方法
CN108760906A (zh) 一种新精神活性物质pFPP的检测方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20904359

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20904359

Country of ref document: EP

Kind code of ref document: A1