CN109342580A - 一种测定板山岗颗粒中（r，s）－告依春含量的方法 - Google Patents
一种测定板山岗颗粒中（r，s）－告依春含量的方法 Download PDFInfo
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- CN109342580A CN109342580A CN201811091446.4A CN201811091446A CN109342580A CN 109342580 A CN109342580 A CN 109342580A CN 201811091446 A CN201811091446 A CN 201811091446A CN 109342580 A CN109342580 A CN 109342580A
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- 238000004166 bioassay Methods 0.000 title abstract description 8
- 238000002360 preparation method Methods 0.000 claims abstract description 20
- 239000000126 substance Substances 0.000 claims abstract description 15
- 239000000243 solution Substances 0.000 claims abstract description 14
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 9
- 239000007788 liquid Substances 0.000 claims abstract description 9
- 239000012085 test solution Substances 0.000 claims abstract description 9
- 238000001514 detection method Methods 0.000 claims abstract description 5
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O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims 2
- 239000000945 filler Substances 0.000 claims 1
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound 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CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims 1
- 239000000377 silicon dioxide Substances 0.000 claims 1
- 239000010231 banlangen Substances 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract description 3
- 241000628997 Flos Species 0.000 abstract description 2
- 241000933211 Helicteres jamaicensis Species 0.000 abstract description 2
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Abstract
板山岗颗粒由板蓝根、山芝麻、岗梅等三味药材经科学方法提取配制而成。本发明公开了一种采用二元或二元以上泵体高效液相色谱仪测定板山岗颗粒中(R，S)‑告依春含量的检测方法。属医药技术领域，其包括对照品溶液的制备、供试品溶液的制备、色谱条件等步骤。本发明的方法是：采用本文建立的HPLC高效液相色谱法，对板山岗颗粒中的(R，S)‑告依春的含量进行测定，专属性强，稳定性好，色谱柱为C18色谱柱，流动相为甲醇‑水，流速为1.0ml/min，等度洗脱。该方法快速灵敏，专属性强，操作简便，对设备的要求较低，稳定且重现性好，可以用于板山岗颗粒产品的质量控制。
Description
技术领域
本发明属一种板山岗颗粒有效成分的检测方法，更具体地涉及一种能对板山岗颗粒的有效成分- (R，S)-告依春进行定量分析的方法。
背景技术
感染性疾病是当前严重威胁人类健康的主要病种之一，全球气候的变暖，生态环境的变化，人类现代 行为的改变，不断助长新的病原微生物的滋生，其中以病毒性感染和细菌性感染占绝大多数。各种流感病 毒所致的呼吸道感染俗称“感冒”，是侵害人类的最常见疾病。感冒虽然一般病情不会十分严重，但还是会 对人们的工作生活带来非常不利的影响，有时会引起严重的并发症，甚至危及生命。而由于各类微生物结 构和入侵部位的特殊性以及致病物质特别是感冒病毒的易变性，导致新药研发的速度远远赶不上病原体变 异的发展速度，因此试图用一药一方来解决所有感染性疾病非常困难的。当西医临床治疗出现瓶颈限制时， 很多医药工作者将目光投到中医药、民族医药等传统医药学领域。现代医学的“病”是建立在解剖、生理、 病理等基础上的微观具体化的认识。而传统医学则是把人体的内脏和体表各部分组织、器官看成一个有机 的整体，在阴阳、五行、脏腑等理论的指导下，对疾病的某一阶段进行本质的宏观抽象化的认识，着重强 调于人体内部的协调完整性。因此中医治疗各类感染性疾病，主要根据祛邪扶正的理论指导，而贯彻中医 整体观和辨证论治思想，这是临床能取得疗效的关键。正是基于这种中医整体观和辨证论治的思想，在上 世纪80年代，南宁第一人民医院的医药工作者，应用中医药理论对该院大量临床处方开展筛选，结合广 西特有的各类中药材，反复应用实践，自主研发创制出了“板山岗颗粒”。
板山岗颗粒由板蓝根、山芝麻、白点称(岗梅)3味药材组方，具有清热解毒、凉血等多种功效，临 床上主要用于治疗外感、咽喉炎、扁桃体炎、腮腺炎等感染性疾病。该制剂一经投入临床，就深受患者的 欢迎，它们以其独特的临床疗效、显著的预防保健作用和相对较低的治疗费用，在维护和增进人们健康方 面带来了巨大的优势。有资料显示，这种医院制剂在近三十年的销售期间，均位列南宁市第一人民医院呼 吸道系统中成药品种销售数量的前五位。同时，由于制剂剂型服用方便，中药成分毒性较低，副作用较少， 特别适合应用于儿童、老年人等特殊群体。但目前板山岗颗粒未设定含量测定的质量控制指标，因此通过 本发明，我们建立了对板山岗颗粒中(R，S)-告依春的含量测定方法。该测定方法简便快捷、准确度高、 专属性强，采用本方法可以准确的把握有关药品质量，降低用药风险，提高产品质量。
发明内容
本发明要解决的技术问题是提供一种对板山岗颗粒中的(R，S)-告依春进行准确测定的质量控制方法。
本发明以如下技术方案解决上述技术问题。
具体方法为：
色谱柱：Kromasil 100-5 column C18色谱柱(250mm×4.6mm，5μm)，理论塔板数按(R，S)-告依春峰计 算应不低于5000psi；流动相：甲醇∶水(12∶88)；检测波长：244nm。流速：1.0mL/min；柱温：30℃。
样品测定：照《中国药典》2015年版第四部通则0512高效液相色谱法测定。
对照品溶液的制备：精密称取(R，S)-告依春对照品适量，加水制成每1mL含0.040mg的溶液，即得。
供试品溶液的制备：取板山岗颗粒约5g，精密称定，置具塞三角锥形瓶中，精密加入水25ml，称定重 量，超声处理30min，放冷至室温，再次称定重量，用水补足减失的重量，摇匀，过滤，再微孔滤膜滤过， 取续滤液，即得。
测定法：分别精密吸取对照品溶液与供试品溶液10μL，注入液相色谱仪，测定，按外标法计算(R，S) -告依春含量即得。
本发明的测定板山岗颗粒中(R，S)-告依春含量的方法，可以精确测定板山岗颗粒中的(R，S)-告依 春的含量，有效控制板山岗颗粒的质量。
附图说明
图1是中检院(R，S)-告依春对照品高效液相(HPLC)测定图谱。
图2是180801批板山岗颗粒(R，S)-告依春高效液相(HPLC)测定图谱。
图3是180802批板山岗颗粒(R，S)-告依春高效液相(HPLC)测定图谱。
图4是180803批板山岗颗粒(R，S)-告依春高效液相(HPLC)测定图谱。
具体实施方式
下面将结合实施例，进一步阐明本发明，所举实例只用于解释本发明，但不限定本发明的范围。
利用HPLC法测定板山岗颗粒中(R，S)-告依春含量按照以下步骤进行：
1.仪器与材料
1.1仪器与试药
仪器：VARIAN 210高效液相色谱仪，包括二元泵，自动进样器，紫外检测器，Galaxie工作站；万分 之一电子分析天平(型号为BSA124S，赛多利斯仪器有限公司)。
试剂：高效液相层析用试剂为色谱纯；水为超纯水。
(R，S)-告依春对照品：批号：11753-201505，由中国食品药品检定研究院提供；
1.2供试样品
板山岗颗粒，广西南宁市第一人民医院提供，批号：180801，180802，180803；
2.方法与结果
2.1色谱条件
液相色谱仪：VARIAN-LC210，检测器：紫外检测器，色谱：Kromasil 100-5 columnC18(250mm×4.6mm， 5μm)；流动相：甲醇-水(12∶88)；检测波长244nm；流速：1.0mL/min；柱温：30℃；进样量10μL。
2.2对照品溶液的配制
精密称取(R，S)-告依春以水为溶剂配制成40μg/mL的标准品储备液。测定时用标准品储备液配制 成16μg/mL的对照品溶液。
2.3供试品溶液的配制
供试品溶液的配制：取板山岗颗粒约5g，精密称定，置具塞三角锥形瓶中，精密加入水25ml，称定 重量，超声处理30min，放冷至室温，再次称定重量，用水补足减失的重量，摇匀，过滤，取续滤液适量， 进样前微孔滤膜过滤，即得。
2.4阴性样品溶液的配制
阴性样品溶液的配制：取不含板蓝根的阴性样品5g，精密称定，置具塞三角锥形瓶中，精密加入水 25ml，称定重量，超声处理30min，放冷至室温，再称定重量，用水补足减失的重量，摇匀，过滤，取续 滤液适量，进样前微孔滤膜过滤，即得。
2.5线性关系的考察
不同浓度对照品溶液的配制及测定：分别精密吸取标准品储备液(40μg/mL)1、2、3、4、5、6、7、 8mL，置于10mL容量瓶中，用超纯水稀释至刻度，摇匀，过微孔滤膜过滤，依法测定。以峰面积(Y)为 纵坐标，进样量(X，μg)为横坐标绘制成标准曲线，计算得出线性方程：Y＝120.047X+0.159(R2＝0.9996)。 结果表明：(R，S)-告依春进样量在0.04μg～0.32μg之间时，其进样量与其峰面积之间有着良好的线性关系。
2.6精密度试验、稳定性试验和重现性试验
取(R，S)-告依春对照品溶液(16μg/mL)连续重复进样6次，记录色谱图，考察样品峰面积的变化 情况。其RSD％为0.79％，表明该含量测定方法的精密度良好。
取同一供试品溶液(批号：180801，下同)5.0201g，依“2.3”方法处理后按“2.1”条件，分别于配制 后0、1、2、4、8、12、24h注入液相色谱仪进行测定，记录色谱峰面积，考察溶液放置后峰面积的变化 情况。其RSD％为1.57％，表明该供试品溶液在24h内稳定。
精密称取同一批次板山岗颗粒5g，共6份，按“2.3.”配制方法制备。依法测定，测得(R，S)-告依春 平均含量为0.0651mg/g，RSD％为1.10％，表明重现性良好。
2.7加样回收率试验
加样回收液的配制：精密量取已知(R，S)-告依春含量的板山岗颗粒2.5g(含(R，S)-告依春0.0651 mg/mL)，分别精密加入对照储备液(40μg/mL)2ml、4ml、6ml，低、中、高浓度各3份，按“2.3”配制方法 制备，得加样回收供试液。依法测定，计算回收率，计算其结果如表1。
表1 (R，S)-告依春的加样回收结果表
由上述结果显示：(R，S)-告依春平均回收率为97.97％，RSD为1.51％，表明此测定方法的加样回收 率符合要求。
2.8样品测定
样品溶液的配制：取板山岗颗粒样品3批，分别按上述方法制备对照品溶液和供试品溶液，分别以本 文相对应的色谱条件，以外标法计算(R，S)-告依春的含量，计算其结果如表2。
表2 板山岗颗粒三批样品含量测定结果表
3.结论
用上述高效液相色谱条件测定本品中的(R，S)-告依春含量，结果专属性强、测定结果准确、在规定 时间内测定方法稳定、可用于本品(R，S)-告依春的含量控制。
Claims (4)
1.色谱条件：
色谱柱：十八烷基硅烷键合硅胶为填充剂的色谱柱，如Kromasil 100-5 column C18色谱柱(250mm×4.6mm，5μm)，理论塔板数按(R，S)-告依春峰计算应不低于5000psi；
流动相：甲醇∶水(5～25∶60～100)；检测波长244nm；流速：0.6～1.20mL/min；柱温：25～45℃。
2.对照品溶液的制备：取(R，S)-告依春对照品适量，加水制成每1mL含0.030～0.055mg的溶液，即得。
3.供试品溶液的制备：取本品约3～8g，精密称定，置具塞三角锥形瓶中，精密加入水25ml，称定重量，超声处理30min，放冷至室温，再次称定重量，用水补足减失的重量，摇匀，过滤，再微孔滤膜滤过，取续滤液，即得。
4.测定法：照《中国药典》高效液相色谱法试验，精密吸取对照品溶液与供试品溶液5～10μL，注入液相色谱仪，测定，按外标法计算(R，S)-告依春含量即得。
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