CN101266233B - Zanthoxylum dissitum Hemsl medicinal materials discrimination method - Google Patents

Zanthoxylum dissitum Hemsl medicinal materials discrimination method Download PDF

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CN101266233B
CN101266233B CN 200710064499 CN200710064499A CN101266233B CN 101266233 B CN101266233 B CN 101266233B CN 200710064499 CN200710064499 CN 200710064499 CN 200710064499 A CN200710064499 A CN 200710064499A CN 101266233 B CN101266233 B CN 101266233B
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dissitum
hemsl
zanthoxylum
medical
material
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CN101266233A (en )
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左之文
朱大安
滕晖
王振勇
祝晨陈
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株洲千金药业股份有限公司
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Abstract

The invention relates to a discrimination method of Zanthoxylum dissitum Hemsl medical material, comprising establishing HPLC standard fingerprint of Zanthoxylum dissitum Hemsl medical material; measuring the fingerprint of the measured sample of the Zanthoxylum dissitum Hemsl medical material using the same method and comparing the fingerprint of the measured sample of the Zanthoxylum dissitum Hemsl medical material and the standard fingerprint of Zanthoxylum dissitum Hemsl medical material to screen the qualified Zanthoxylum dissitum Hemsl medical material in line with the standard fingerprint of Zanthoxylum dissitum Hemsl medical material. The process of establishing the HPLC standard fingerprint of Zanthoxylum dissitum Hemsl medical material comprises: preparing the sample solution and acquiring the HPLC standard fingerprint of Zanthoxylum dissitum Hemsl medical material by determing the common fingerprint features by comparing the fingerprintsof 30 batches of the Zanthoxylum dissitum Hemsl medical material at the conditions that: chromatographic column: Kromasil RP-C18 4.6mm*250mm, 5um; mobile phase: acetonitrile-0.2%triethylamine phosphate solution; using gradient elution: 0min-10min-30min-70min; the ratio by volume of acetonitrile and 0.2%triethylamine phosphate solution is 5:95-10:90-10:90-40:60; detection wave:280nm. The invention provides a new method for quality control and discrimination of the Zanthoxylum dissitum Hemsl medical material.

Description

一种单面针药材的鉴别方法 A method of identifying method sided needle herbs

技术领域 FIELD

[0001] 本发明涉及医药领域,具体的说,涉及一种单面针药材的鉴别方法。 [0001] The present invention relates to the field of medicine, specifically, to a method of identifying medicinal sided needle. 背景技术 Background technique

[0002] 中药现代化是当代中药发展中的一个最热门的研究方向,已成为国内外医药界的一大热点。 [0002] modernization of Chinese medicine is one of the most popular contemporary research in the development of Chinese medicine, has become a focus of world pharmaceutical industry. 其中中药材,中成药的质量标准现代化是中药现代化的一个重要组成部分。 Where the quality standards of modern Chinese herbal medicine, Chinese medicine is an important part of the modernization of traditional Chinese medicine. 由于中药材品种繁多,地区用药习惯各异,古代本草记载简单,误传错用和用药演变等因素,导致中药材同物异名,同名异物以及正品,非正品都入药的混乱现象非常严重。 Due to a wide variety of Chinese herbal medicines, drug habits of different regions, the ancient Materia Medica records simple, with misinformation and wrong medication evolution of other factors, lead to medicines synonym, homonym confusion and genuine, non-genuine medicine are very serious. 更为普遍的是影响中药成分的因素复杂,即使是同一药材,其有效化学成分常因生态环境,采集时间,储存和炮制方法等不同而有差异,这些问题严重影响了中药的质量和用药的安全与有效。 More generally, factors affecting Chinese medicine complex, even for the same medicine, the active chemical ingredient often due to the ecological environment, and processing methods, etc. differ among acquisition time, store, these problems seriously affected the quality of traditional Chinese medicine and medication safe and effective. 如何根据中医药的特点提出一整套具有中医药特点又具有高科技特征的中药材、中成药现代化标准,是一重要课题。 How to propose a set of Chinese herbal medicine has the characteristics of high-tech features but also has the characteristics of Chinese medicine, Chinese medicine modern standards, is an important issue. 建立中药材单味药和复方全组成的指纹图谱是一种具有先进性的切实可行的方法,现在这方法已成为国际上通用的方法,而且指纹图谱已成为国际公认的控制中药或天然药物质量的最有效的手段”。 Establish fingerprint Chinese herbal single herbs and compound the whole composition is a practical method of having advanced, and now this method has become a common international approach and fingerprinting has become an internationally recognized quality control of traditional Chinese medicine or natural medicine the most effective means. "

[0003] 中药指纹图谱是在中医药基础理论指导下针对中药的品种和质量,在现有认知水平基础上,以整体信息和药效指标为依据,采用多种分析方法和技术,建立的客观、整体和多指标的综合评价体系。 [0003] Fingerprinting is under the guidance of the basic theory of traditional Chinese medicine for the variety and quality of traditional Chinese medicine, the existing level of knowledge on the basis of the information and to the overall efficacy based on the indicators, using a variety of analytical methods and techniques, the establishment of objective, overall and comprehensive evaluation system of multi-indices. 中药指纹图谱的建立,将对中药的规范化、标准化、产业化和国际化提供科学的技术平台,对中药现代化的战略行动具有科学的指导意义。 Establish fingerprint of traditional Chinese medicine, Chinese medicine will be standardized, standardization, industrialization and internationalization of scientific technology platforms, scientific guidance for strategic action modernization of Chinese medicine.

[0004] 单面针,Zanthoxylum dissitum Hemsl,芸香科木质藤本,细时为灌木状;茎枝着生略下变的皮刺。 [0004] one side of the needle, Zanthoxylum dissitum Hemsl, Rutaceae woody vine, shrub-like thin one; slightly green shoots forward under varying prickles. 单数羽状复叶;小叶3-9,对生坚纸质至革质,狭矩圆形到卵状矩圆形,长7-16厘米,宽3-6厘米,有时背面中脉上着生有下变的钩状刺,聚伞状圆锥花序,腋生,花4 数;萼片宽卵形;花瓣卵状矩圆形;雄花的雄蕊开花时伸出花瓣外,顶端2-4驻裂;雌花无退化雄蕊。 Single pinnate; leaflets 3-9, Kennedy paper opposite to leathery, oblong narrowly ovate to oblong, 7-16 cm long, 3-6 cm wide, sometimes midvein there bears the - varying the barbed hook, cymose panicles, axillary, flower number 4; sepals broadly ovate; petals ovate oblong; stamens protruding male flower petals time, the crack in the top 2-4; female no staminodes. 瞢美果成熟时淡褐色,外形似蚬。 When ripe fruit hazel Meng Mei, shape like clams. 分布于西南、广东、广西、湖南、湖北、陕西。 Located in the southwest of Guangdong, Guangxi, Hunan, Hubei, Shaanxi. 生山地林中。 Health mountain forest. 根、茎皮及叶均供药用,能祛风活络、散瘀止痛、解毒消肿。 Roots, bark and leaves are for medicinal purposes, can Qufenghuoluo, Sanyu pain, swelling detoxification.

[0005] 单面针不是药典和部颁标准药材,现在的中药成药中以该药材入药的也不多见, 更没有中成药以单面针的主要成分来进行质量控制,因此关于单面针药材的有效成分的研究极少。 [0005] Pharmaceutical sided needle than ministerial standards and herbs, now Chinese medicines to the medicine of medicine also rare, and no medicine at one side of the main component for quality control of the needle, so the needle on one side effective ingredients of herbs rarely. 中药指纹图谱是指某种中药材或中成药中所共有的、具有特征性的某类或数类成分的色谱或光谱的图谱。 Fingerprinting profiles chromatographic or spectroscopic means some Chinese herbal medicine or in common, certain types or categories of components having a characteristic. 在现阶段中药的有效成分绝大多数没有明确的情况下,有效控制中药材或中成药的质量具有重要的意义。 In a clear case of the vast majority of the active ingredients of traditional Chinese medicine this stage there is no effective quality control of herbal medicines or proprietary Chinese medicine has important significance. 日本汉方药主要生产企业在20世纪80年代就已经在企业中采用高效液相指纹图谱控制质量。 Japan Kampo major manufacturers already using HPLC fingerprint quality control in the enterprise in the 1980s. 德国、法国在对银杏叶提取物联合开发的过程中,发现银杏叶提取物的医疗作用是提取物所得物质群的整体作用结果,而对这样一个整体的质量控制,亦采用高效液相指纹图谱方法。 Germany, France, in the process of Ginkgo biloba extract developed jointly found the medical effect of Ginkgo biloba extract is the overall effect of the results of the extract resulting material base, while such an overall quality control, also using HPLC fingerprint method. 美国FDA最近几年制定的植物草药指南中已经明确把指纹图谱作为混合物质群的质量控制方法。 Plant herbs guidelines from the American FDA in recent years has been clearly established by the fingerprint as a quality control method for hybrid substance group. 指纹图谱作为中草药及其提取物质量控制方法,目前已经成为国际共识。 Fingerprints as herbs and their extracts quality control methods, has now become an international consensus. 为有效控制单面针药材的质量,本申请人对不同产地单面针进行了考察比较,为建立单面针药材质量的综合控制方法,特以国家食品药品监督管理局2000年颁发的关于中药材指纹图谱技术要求,对湖南省张家界地区采集的单面针30批药材进行了研究,建立起单面针指纹图谱测定方法。 For the quality control of medicines single needle, the Applicant different origin were studied comparing single needle, single needle for the establishment of a comprehensive quality control method herbs, especially to the State Food and Drug Administration in 2000 issued on Chinese medicine material fingerprinting requirements of the Hunan Zhangjiajie collection of one-sided needle 30 batches of medicines were studied to establish a single needle fingerprints of methods. 发明内容 SUMMARY

[0006] 本发明的目的是提供一种单面针药材的鉴别方法。 [0006] The object of the present invention is to provide a method of identifying medicinal sided needle.

[0007] 为了实现本发明的目的,本发明提供了一种单面针药材的鉴别方法,包括如下步骤: [0007] To achieve the object of the present invention, the present invention provides a method for identification of single-sided needle herbs, comprising the steps of:

[0008] 第一步,建立单面针药材的HPLC标准指纹图谱,包括: [0008] The first step in establishing single needle of HPLC standard fingerprint, including:

[0009] 1)供试品溶液的制备:精密称取单面针药材粉末,经石油醚预处理后,挥干石油醚,残渣中加入甲醇加热回流,合并滤液,水浴蒸干,残渣加甲醇溶解,转移至容量瓶中,定容至刻度,得到供试品溶液; Preparation of [0009] 1) for the test solution: Weigh accurately sided needle medicinal powder, after pre-treatment with petroleum ether, petroleum ether, evaporated to dryness, the residue was added methanol was heated to reflux and the combined filtrate evaporated to dryness in a water bath, the residue in methanol dissolved, transferred to a volumetric flask, dilute to volume, to give the test solution;

[0010] 2)高效液相色谱分析:精密吸取供试品溶液进样,色谱条件:色谱柱为Kromasil RP-C184. 6mmX 250mm, 5 μ m ;HP G1313A自动进样系统;流动相为乙腈_0. 2%磷酸三乙胺溶液;采用梯度洗脱方式:0min-10min-30min-70min,乙腈与0. 2%磷酸三乙胺溶液的体积比为:5:95-10:90-10:90-40:60 ;检测波长:210〜366nm ;得到单面针药材HPLC标准指纹图谱; [0010] 2) HPLC: Precision drawing test solution was injected, chromatographic conditions: column was Kromasil RP-C184 6mmX 250mm, 5 μ m; HP G1313A autosampler system; mobile phase of acetonitrile _ phosphoric acid 0.2% triethylamine; gradient elution mode: 0min-10min-30min-70min, volumes of acetonitrile and 0.2% triethylamine phosphate ratio: 5: 95-10: 90-10: 90-40: 60; detection wavelength: 210~366nm; obtained sided needle on HPLC standard fingerprint;

[0011] 3)测定:精密吸取30批单面针药材的供试品溶液各5-20 μ 1,分别注入液相色谱仪,照高效液相色谱法测定,得到30批单面针药材的HPLC指纹图谱并进行比较,得到由共有特征峰构成的单面针药材HPLC标准指纹图谱。 [0011] 3) Measurement: 30 batches precision drawing the test solution in each medicinal sided needle 5-20 μ 1, were injected into the liquid chromatograph to measure performance liquid chromatography, to give 30-sided needle batch of herbs HPLC fingerprint and compare give sided needle on HPLC standard fingerprint characteristic peaks consisting of a total of. 所述单面针标准指纹图谱中,共有特征峰有8个,计算各个特征峰与总峰面积的积分相对比值(相对峰面积),其中1号峰的平均Rt为3. 364min,相对峰面积为4. 2〜7. 8 % ;2号峰的平均Rt为4. 620min,相对峰面积为0. 8〜1. 6% ;3号峰的平均Rt为10. 772min,相对峰面积为1. 5〜3. 3% ;4号峰的平均Rt为12. 799min,相对峰面积为3. 7〜9. 9% ;5号峰的平均Rt为13. 828min,相对峰面积为4. 5〜16. 3% ;6号峰的平均Rt为34. 015min,相对峰面积为46. 9〜73. ;7号峰的平均Rt为58. 820min,相对峰面积为5. 3〜10. 7% ;8号峰的平均Rt为62. 179min,相对峰面积为2. 7 〜7. 5% ; The standard one-sided needle fingerprints, there are a total of eight characteristic peaks, calculated for each feature and the relative ratio of the integrated peak total peak area (relative area), the average peak 1 wherein Rt is 3. 364min, relative peak area 4. 2~7 to 8%; average peak No. 2 is Rt 4. 620min, relative peak area 8~1 0. 6%; average peak of Rt 3 10. 772min, the relative area of ​​peak 1 . 5~3 3%; average peak of Rt 4 12. 799min, relative peak area 3. 7~9 9%; average peak of Rt 5 13. 828min, the relative peak area of ​​4.5 . ~ 16 3%; average peak of Rt 6 34. 015min, 46. the relative peak areas of 9~73; average peak of Rt 7 58. 820min, relative peak area 7 5. 3~10. %; average peak of Rt 8 62. 179min, the relative peak area of ​​2.7 5% ~ 7.;

[0012] 第二步,以上述相同的方法测定待测单面针药材样品的指纹图谱; [0012] The second step in the same manner as above was measured fingerprint test sample medicinal sided needle;

[0013] 第三步,将第二步所测定的待测单面针药材样品指纹图谱与第一步构建的单面针标准指纹图谱对比,筛选符合单面针药材标准指纹图谱的合格单面针药材。 [0013] The third step, single-sided needle-sided standard fingerprint comparison test needle with medicine fingerprint sample measured First Step Second Step constructed, screened sided meet eligibility criteria sided needle medicine fingerprint needle medicine.

[0014] 所述第一步1)中,单面针药材粉末细度优选为40目。 [0014] The first step 1), the medicinal powder fineness sided needle preferably 40 mesh.

[0015] 所述第一步1)中,加入石油醚加热回流不少于10分钟,其中,Ig单面针药材加入石油醚量不少于10ml,优选Ig单面针药材加入石油醚30mi加热回流30分钟。 [0015] The first step a), the petroleum ether was added was heated at reflux for less than 10 minutes, wherein, Ig-sided needle of not less than medicinal petroleum ether 10ml, preferably Ig-sided needle herbs Petroleum ether was heated 30mi reflux for 30 minutes.

[0016] 所述第一步3)中,优选精密吸取单面针药材的供试品溶液10μ 1。 [0016] The first step 3), preferably precision drawing-sided needle medicine test solution 10μ 1.

[0017] 所述第一步1)中,加入甲醇加热回流1次以上,每次不少于10分钟,其中Ig单面针药材加入甲醇量不少于10ml,优选加入甲醇30ml加热回流2次,每次1小时。 [0017] The first step a) in methanol was added heated to reflux for 1 or more times less than 10 minutes, wherein the Ig-sided needle herbs less than the amount of methanol was added 10ml, preferably from 30ml of methanol was added heated to reflux for 2 1 hour each.

[0018] 所述第一步1)中,残渣优选用甲醇定容至2ml。 [0018] The first step 1), preferably the residue with methanol to 2ml.

[0019] 所述第一步1)中,供试品溶液高效液相分析前优选用0.45μπι微孔滤膜滤过。 Preferably prior to analysis by HPLC 0.45μπι microporous membrane filtration [0019] The first step a), the test solution.

[0020] 所述第一步2)中,检测波长优选为280nm。 [0020] The first step 2), the detection wavelength is preferably 280nm.

[0021] 所述8个共有特征峰的相对保留时间为,1号峰的平均相对保留时间Rt为3. 364min,标准偏差SD为0. 019 ;2号峰的平均相对保留时间Rt为4. 620min,标准偏差SD为0. 008 ;3号峰的平均相对保留时间Rt为10. 772min,标准偏差SD为0. 102 ;4号峰的平均相对保留时间Rt为12. 799min,标准偏差SD为0. 082 ;5号峰的平均相对保留时间Rt为13. 828min,标准偏差SD为0. 101 ;6号峰的平均相对保留时间Rt为34. 015min,标准偏差SD为0. 603 ;7号峰的平均相对保留时间Rt为58. 820min,标准偏差SD为0. 074 ;8号峰的平均相对保留时间Rt为62. 179min,标准偏差SD为0. 256 ; The relative retention time [0021] the eight common peaks, for an average number of peaks of the relative retention times Rt 3. 364min, the standard deviation SD 0.019; number average relative retention time of 2 peaks Rt of 4. 620min, the standard deviation SD 0.008; average peak 3 relative retention time Rt of 10. 772min, the standard deviation SD 0.102; average peak 4 relative retention time Rt of 12. 799min, the standard deviation SD 0.082; number average relative retention time of 5 peaks Rt of 13. 828min, the standard deviation SD 0.101; number average relative retention time of 6 peaks Rt of 34. 015min, the standard deviation SD 0.603; No. 7 the average relative retention time of the peak of Rt 58. 820min, the standard deviation SD 0.074; number average relative retention time of 8 peaks Rt of 62. 179min, the standard deviation SD 0.256;

[0022] 现阶段未发现有对单面针有效成分及其质量控制方法的深入研究,本发明人运用国际通用的对不能明确有效成分的天然药物以指纹图谱来综合控制质量的方法,特以国家食品药品监督管理局2000年颁发的关于中药材指纹图谱技术要求,对湖南省张家界地区采集的单面针30批药材进行了研究,独创性地建立起单面针指纹图谱测定方法。 [0022] at this stage have not found one-sided needle-depth study of the active ingredients and quality control methods, the present invention is the use of internationally accepted methods can not clear the active ingredients of natural drugs to be integrated fingerprint quality control, especially with State food and Drug Administration in 2000 issued about herbal fingerprinting requirements of the Hunan Zhangjiajie collection of one-sided needle 30 batches of medicines were studied to establish a single needle originality fingerprint determination. 该方法通过指纹图谱有效地对不同产地、不同时间、不同批次的单面针药材的有效成分物质群整体进行控制,从而从整体上有效地表征单面针药材质量,达到有效控制单面针药材质量的目的。 The method of the active substance group of different habitats, at different times, different batches of whole herbs sided needle through the fingerprint effectively controlled, thereby effectively characterize the quality of medicine from one side of the needle as a whole, to achieve effective control of single-sided needle the purpose of the quality of medicine.

附图说明 BRIEF DESCRIPTION

[0023] 图1四种检测波长下的HPLC图谱,其中,检测波长的选择(由上至下分别为210nm、254nm、280nm、366nm)。 [0023] HPLC pattern in FIG. 1 four kinds of detection wavelength, wherein the wavelength selective detector (from top to bottom, respectively 210nm, 254nm, 280nm, 366nm).

[0024] 图2单面针液相色谱指纹图谱。 [0024] FIG 2 HPLC Fingerprint sided needle.

[0025] 图330批单面针药材指纹图谱相似性评价结果。 [0025] FIG similarity evaluation results of 330 batches fingerprints of single-sided needle.

[0026] 图4样本9的原始液相色谱图谱。 HPLC profiles of the original [0026] FIG. 9 is 4 samples.

[0027] 图5整体相似度在0. 90以上的样本(17批)指纹共有模式(以平均值方法生成)。 [0027] FIG. 5 overall similarity (mean value generating method) of 0.90 or more in a sample (batch 17) consensus fingerprint pattern.

[0028] 图6相似度大于0. 90的样品中各特征峰峰面积与8个特征峰总峰面积的比例关系。 [0028] FIG 6 wherein each similarity is larger than the ratio between peak area and peak 8 wherein the total peak area of ​​the sample 0.90.

[0029] 图7相似度大于0. 90的样品各特征峰与参照峰峰面积的比例关系。 [0029] FIG. 7 is greater than the ratio between the degree of similarity of each reference characteristic peak peak area of ​​the sample 0.90.

[0030] 图8整体相似度在0. 80以上的样本(21批)指纹共有模式(以平均值方法生成)。 [0030] FIG. 8 global similarity (mean value generating method) of 0.80 or more in a sample (batch 21) consensus fingerprint pattern.

[0031] 图9相似度大于0. 80的样品中各特征峰峰面积与8个特征峰总峰面积的比例关系。 [0031] FIG 9 wherein each similarity is larger than the ratio between peak area and peak 8 wherein the total peak area of ​​the sample 0.80.

[0032] 图10相似度大于0. 80的样品各特征峰与参照峰峰面积的比例关系。 [0032] FIG. 10 is greater than the similarity ratio between the peaks of each sample with the reference peak area is 0.80. 具体实施方式 detailed description

[0033] 以下实施例用于说明本发明,但不用来限制本发明的范围。 [0033] The following examples serve to illustrate the present invention but are not intended to limit the scope of the present invention. 实施例1单面针药材供试样品 Example single-sided embodiment of a test sample needle herbs

[0034] 单面针为芸香科植物单面针Zanthoxylum dissitum Hemsl.的干燥根和茎。 [0034] Rutaceae sided needle-sided needle Zanthoxylum dissitum Hemsl. The dried roots and stems. 药材 Medicinal material

由湖南省株洲千金药业股份有限公司提供,共30批,分别编号为1、2、3........28、29、30。 By Zhuzhou Qianjin Pharmaceutical Co., Ltd., a total of 30 batches, numbered 1, 2 ........ 28, 29.

见表1 : Table 1:

[0035] 表1单面针药材供试样品 [0035] The ingredients in Table 1 in the test sample sided needle

[0036] [0036]

样品号 药材名 产地 采收期 批号1 单面针 商品药材 2003. 11〜12月份 20040308A2 单面针 商品药材 2003. 11〜12月份 20040308B3 单面针 商品药材 2003. 11〜12月份 20040312A Sample No. Name Origin harvest herbs Lot 1-sided needle commodity herbs 2003.11 - December 20040308A2 single needle commodity herbs 2003.11 - December 20040308B3 single needle commodity herbs 2003.11 - December 20040312A

Figure CN101266233BD00071

[0037] 实施例2供试品的制备 Example 2 Preparation of the test [0037] Embodiment

[0038] 药材经粉碎后过40目筛,取各批药材粉末1. 0g,精密称定,置于50ml圆底烧瓶中, 加石油醚30ml加热回流30min,弃去石油醚液,残渣挥干溶剂,再加甲醇30ml加热回流提取2次,每次lh,趁热滤过,合并两次滤液,蒸干,残渣用甲醇定容至2ml,作为供试品溶液, 高效液相分析前用0. 45 μ m微孔滤膜滤过。 [0038] After the medicine was sifted through a 40 mesh sieve, each batch taken medicinal powder 1. 0g, accurately weighed, placed in 50ml round bottom flask 30ml petroleum ether was heated at reflux for 30min, petroleum ether solution was discarded, the residue was evaporated to dryness The solvent plus 30ml methanol was heated at reflux for 2 times, each LH, hot filtration, two filtrates were combined, evaporated to dryness, the residue was diluted with methanol to 2ml, as the test solution, prior to analysis by high performance liquid 0 . 45 μ m microporous membrane filtration.

[0039] 实施例3参照物的选择 Selection of reference Example 3 [0039] Embodiment

[0040] 单面针药材经上述方法提取后,经HPLC分析出现多个色谱峰,由于目前尚没有单面针专属成分的标准品,故经过详细比较多批次单面针药材的指纹图谱,确定一个稳定出现的色谱峰(6号峰,Rt34min)作为参照物(结构尚未鉴定)进行指纹图谱的分析和评价, 经过系统比较分析,证明该方法可行。 [0040] The method of the above-described medicine sided needle after extraction by HPLC analysis plurality of peaks appear, since there is currently no standard one-sided needle-specific component, it passes more detailed fingerprint sided needle batch of herbs, determining peak (peak 6, Rt34min) appears as a stable reference (structure not identified) analysis and evaluation fingerprinting system through comparative analysis, it proves that the method is feasible.

[0041] 实施例4检测方法 Example 4 Detection [0041] Embodiment

[0042] 仪器 [0042] Instruments

[0043] HPllOOSeries 高效液相色谱仪(Agilent Technologies,USA),配置HP G1311A 四元梯度泵,HP G1313A自动进样系统,HP G1315A 二极管阵列检测器,HP G1313真空在线脱气机,HP G1316A柱温箱及HP Chem Station4. 0色谱工作站;Kromasil RP-C18 (4. 6mm X 250mm,5 μ m)色谱分析柱。 [0043] HPllOOSeries high performance liquid chromatography (Agilent Technologies, USA), arranged HP G1311A quaternary pump, HP G1313A autosampler system, HP G1315A diode array detector, HP G1313 vacuum online degasser, HP G1316A Column temperature The box and HP Chem Station4 0 chromatography workstation; Kromasil RP-C18 (4. 6mm X 250mm, 5 μ m) by column chromatography.

[0044] 流动相的选择 [0044] The choice of mobile phase

[0045] 试验过程中共试用了3种流动相系统: [0045] Experimental procedure Three trial CCP mobile phase system:

Figure CN101266233BD00081
Figure CN101266233BD00082
Figure CN101266233BD00083
Figure CN101266233BD00084

[0053] 结果表明,上述梯度洗脱中以梯度③为最佳,谱图上各个色谱峰(280nm)的分离度较好,保留时间适中,因此选用该流动相系统作为单面针药材的HPLC指纹图谱检测的流动相。 [0053] The results show that, in the above-described gradient elution gradient ③ best, each peak (280 nm) of the spectral resolution is better, retention time is moderate, so the choice of the system as the mobile phase of HPLC-sided needle Mobile phase fingerprint detection.

[0054] 检测波长的确定 [0054] Detection wavelength determined

[0055] 试验过程中共选择了210nm、254nm、280nm、366nm四种波长,在这四种检测波长下的HPLC图谱中均有多个色谱峰(见图1,由上至下分别为210nm、254nm、280nm、366nm), 但是检测波长在280nm时色谱峰较多且峰形较好,基线也比较平稳,因此综合考虑,选定280nm为制定指纹图谱的检测波长。 [0055] Experimental procedure CCP selected 210nm, 254nm, 280nm, 366nm four wavelengths in the HPLC profile at four wavelengths are detected a plurality of peaks (see Fig. 1, top to bottom, respectively 210nm, 254nm , 280nm, 366nm), but the detection wavelength at 280nm peak when more and better peak shape, the baseline is relatively stable, and therefore considered, 280nm wavelength detection for the development of fingerprint selected.

[0056] 稳定性试验 [0056] Stability Test

[0057] 以编号为4的药材制备供试品,分别在1、2、4、8、12、24小时后进样,记录各共有色谱峰保留时间和积分面积。 [0057] In Preparation No. 4 test medicine, each sample in the backward 1,2,4,8,12,24 hours, record retention time of each peak and the total integrated area. 考察色谱峰保留时间的一致性,各主要色谱峰保留时间的RSD 分别为0. 06〜0. 15%,同时考察1〜8#特征色谱峰的相似度,各图谱的相似度均达0. 90 以上。 Investigation peak retention time consistent, the RSD major peak retention times were 0.5 06~0. 15%, while # 1~8 investigated characteristic peaks of similarity, similarity of each map reached 0. More than 90.

[0058] 精密度试验 [0058] Precision Test

[0059] 以编号为4的药材制备供试品,1天内连续进样5次,记录各共有色谱峰保留时间和积分面积。 [0059] The number of medicinal 4 Preparation of the test, a sample 5 consecutive days, times, recording each of the total integrated peak area and retention time. 考察色谱峰保留时间的一致性,各主要色谱峰保留时间的RSD分别为0. 04〜 0. 70%。 Examine the consistency of peak retention times, RSD major peak retention time of 0.5 04~ 0.70% respectively. 同时考察1〜8#特征色谱峰的相似度,各图谱的相似度均达0. 90以上。 # 1~8 also investigated characteristic peaks of similarity, similarity of each map reached 0.90 or more.

[0060] 重现性试验 [0060] reproduce the test

[0061] 取编号为4的药材6份,按3项下方法进行供试品制备并检测,考察各色谱峰相对保留时间、峰面积比值的一致性,同时计算各色谱指纹图谱的相似度。 [0061] taken of the number 4 6 parts of herbs, prepared according to the test method and the three detecting investigated relative retention time of each peak, the peak area ratio of consistency, while the degree of similarity calculated for each chromatographic fingerprints. 结果表明,所考察的共有峰的相对保留时间及相对峰面积具有较好的重现性,各主要色谱峰保留时间的RSD分别为0. 08〜0. 16%。 The results show that the examined relative retention time and relative peak area of ​​common peaks with good reproducibility, the RSD major peak retention time 0. 08~0. 16%. 同时考察1〜8#特征色谱峰的相似度,各图谱的相似度均达0. 90以 # 1~8 also investigated characteristic peaks of similarity, similarity of each map reached to 0.90

8上。 8.

[0062] 实施例5单面针HPLC指纹图谱建立 [0062] Example 5 sided needle HPLC fingerprints established

[0063] 指纹特征峰确定 [0063] fingerprint characteristic peaks OK

[0064] 综合分析30批供试品HPLC图谱,确定了8个稳定出现的主要色谱峰作为单面针的特征指纹峰,各批次样品中8个主要色谱峰的绝对保留时间见表2,积分面积见表3,平均保留时间见表4。 [0064] 30 comprehensive analysis of HPLC profiles of the test batches, identified eight major peaks appearing as a stable characteristic fingerprint sided needle peak, the absolute retention time of each batch in Table 2 sample 8 major peaks, integrated area in Table 3, the average retention time shown in Table 4. 代表性指纹图谱见图2 Representative fingerprint shown in Figure 2

[0065] 表230批单面针药材绝对保留时间表 [0065] Table 230 batches of medicines is absolutely one-sided needle retention schedules

[0066] [0066]

Figure CN101266233BD00091

[0067] [0067]

Figure CN101266233BD00092

[0068] 表3 30批单面针药材各特征峰积分面积 [0068] Table 3 sided needle 30 batches of each characteristic peak integration area of ​​medicine

[0069] [0069]

Figure CN101266233BD00101

[0070] [0070]

Figure CN101266233BD00102

[0071] 表4 30批单面针8个特征峰平均保留时间表 [0071] Table 4 8 30 batches sided needle retention schedules average peaks

[0072] [0072]

Figure CN101266233BD00103

[0073] 单面针药材指纹图谱的分析与评价 [0073] Analysis and Evaluation sided needle on fingerprints

[0074] 详细比较30批次单面针的液相色谱图,8个特征指纹的有无及含量差异分别构成各自的特征(见图3),对30批次样品所得的数据进行相似性分析所得结果见表5 : [0074] The detailed comparison of batch-sided needle 30 liquid chromatogram, the presence and the content difference of the 8 characteristic fingerprints constitute respective feature (see FIG. 3), the data of 30 batches of samples were obtained similarity analysis The results obtained are shown in Table 5:

[0075] 表5 [0075] TABLE 5

[0076] [0076]

Figure CN101266233BD00111
Figure CN101266233BD00112

[0078] 从此结果可以看出,所检测30批样本的相似度在0. 378〜0. 946间,各样品差异虽然较大,但也有17批样本的相似度在0.9以上,这17批样品显示了较好的一致性;另有4批样本相似度在0. 8〜0. 9之间,查看这21批相似度在0. 8以上样本的原始液相色谱图, 发现它们均有一共同点:6#峰的表观丰度与峰面积比较高而且差别不明显。 [0078] From this result, it can be seen, the degree of similarity of the sample detected in 30 batches 0. 378~0. 946, for each sample, although the difference is large, but there are 17 sample batches of similarity above 0.9, these 17 batches of samples It shows good agreement; and another four batches of samples 8~0 similarity between 0. 9, 21 batch View this original liquid chromatograph of 0.8 or more degree of similarity of the sample and found that they have a common point: # 6 peak of apparent abundance and peak area is relatively high and the difference is not obvious.

[0079] 另外,样品9的相似度低(小于0. 8)却是由于7#和8#峰的表观丰度均较6#峰的为高,和构建的指纹图谱共有模式中7#、8#与6#峰的比例关系正好相反,因此它和指纹图谱共有模式的相似性较差,见图4。 [0079] Further, the low degree of similarity Sample 9 (less than 0.8) is due to the # 7 and # 8 than in the apparent abundance of peaks peak # 6 is high, and constructed Fingerprint pattern # 7 , # 6 and # 8 peak ratio between the opposite, and it is therefore poor similarity fingerprint pattern, shown in Figure 4.

[0080] 单面针药材指纹图谱共有模式的建立 Establish [0080] single needle Fingerprint common pattern

[0081] 上述分析表明,单面针药材批间质量差异大,以30批样本的指纹图共建的共有模式难以体现样本之间的差异,为此,根据其相似程度,分别建立共有模式并进行评价: [0081] The above analysis shows, among herbs sided needle batch quality differences, to a total of 30 batch mode to build a fingerprint of the sample is difficult to reflect the differences between the samples, for which, according to its degree of similarity, and were established consensus pattern were evaluated:

[0082] 整体相似度在0.90以上的样本(17批)指纹共有模式(以平均值方法生成,见图5): [0082] The overall similarity of 0.90 or more sample (batch 17) consensus fingerprint pattern (average value generating method shown in Figure 5):

[0083] 分别计算满足该相似度要求的17批药材(1、12、14、18、19、20)8个特征峰峰面积的平均数(A)、标准平均偏差(SD),见表6 ;各特征峰峰面积与8个特征峰总峰面积的比值 [0083] calculate the degree of similarity satisfy the requirements of 17 batches of ingredients (1,12,14,18,19,20) 8 wherein the average peak area (A), the mean standard deviation (SD), Table 6 ; wherein each peak area and peak 8 wherein the ratio of the total peak area

表7,图6);各特征峰峰面积与参照峰(6#)峰面积的比值(表8,图7)。 Table 7, FIG. 6); (# 6) the peak area ratio of each peak area to the reference peak characteristics (Table 8, FIG. 7).

[0084] 表6相似度大于0. 90的17批单面针药材峰面 17 Batch ingredients sided needle peak area [0084] Table 6 of similarity is greater than 0.90

[0085] [0085]

Figure CN101266233BD00113

[0086] [0086]

Figure CN101266233BD00114

[0087] 表7相似度大于0. 90的样品中各特征峰峰面积与8个特征峰总峰面积的比值 [0087] Table 7 similarity is larger than the peak area ratio of 8 characteristic peaks of the total peak area of ​​each feature in a sample 0.90

[0088] [0088]

Figure CN101266233BD00121

[0089] *注:峰面积比例计算公式:It = AiAtotal和Ii = AiA6 ;其中Ai表示各特征峰峰面积,Atotal表示某批次药材中8个特征峰的总峰面积,A6表示某批次样品中6#特征峰的峰面积。 [0089] * Note: the ratio of the peak area calculation formula: It = AiAtotal and Ii = AiA6; where Ai represents wherein each peak area, Atotal is the total peak area of ​​a batch of herbs 8 characteristic peaks, A6 represents a batch samples # 6 characteristic peak of the peak area.

[0090] [0090]

[0091] [0091]

[0092] [0092]

Figure CN101266233BD00122
Figure CN101266233BD00131

[0093] 从以上几个图表所显示出的信息可知,这17批样品中各特征峰峰面积与总峰面积及参照峰峰面积的比值较为稳定,即无论这17批样品中各特征峰峰面积的绝对值高低如何变化,它们与总峰面积及参照峰峰面积的比例关系是一致的,这也是这17批样品相似度较高的根本原因。 [0093] From the above exhibited several Chart understood, each of these 17 batches of samples and wherein the ratio of the area of ​​reference peak area peak area to total peak is more stable, i.e., no matter in each batch of samples 17 wherein peak how to change the level of the absolute value of the area, which is consistent with the ratio between the total peak area and peak area of ​​the reference, which is that 17 batches of samples with high similarity of the underlying causes.

[0094] 拟定相似度0. 90以上样品的各指纹峰与总峰面积、参照峰峰面积的比例关系见表9。 [0094] The preparation of the above sample was 0.90 similarity of each fingerprint peak total peak area, peak area ratio with reference to the relationship shown in Table 9.

[0095] 表9 [0095] Table 9

[0096] [0096]

Figure CN101266233BD00132

[0097] 表8相似度0. 90以上样品的各指纹峰与总峰面积、参照峰峰面积的最小比值# 注:划定依据为I-SD,其中I表示多批次样品中同一特征峰与总峰面积或参照峰峰面积比值的平均值;SD表示多批次样品中同一特征峰与总峰面积或参照峰峰面积比值的相对标准平均偏差。 [0097] TABLE each fingerprint peak total peak area was 0.90 or more similarity sample 8, with reference to the minimum peak area ratio # Note: evidence for the designated I-SD, where I represents the same characteristic peak sample batches or the average of the total peak area of ​​reference peak area ratio; the SD represents the same batches of sample peaks with reference to the total peak area or peak area ratio relative standard deviation of the mean.

[0098] 整体相似度在0. 80以上的样本(21批)指纹共有模式(以平均值方法生成,如图8): [0098] In the above overall similarity sample 0.80 (21 batches) consensus fingerprint pattern (average value generating method, as shown in FIG. 8):

[0099] 同前法分析与评价相似度在0. 8以上的21批药材(1、3、4、5、6、7、10、11、12、13、 14、15、16、18、19、20、21、22、23、24、29)的指纹图谱,分别计算这21批药材的8个特征峰峰面积的平均数(A)、标准平均偏差(SD),见表10 ;21批样品中各特征峰峰面积占总峰面积的比例(见表11,图9) ;21批合格样品中各特征峰峰面积与参照峰峰面积的比例关系(见表12,图10) [0099] with the former method of analysis and evaluation of the degree of similarity in 21 batches of 0.8 or more herbs (1,3,4,5,6,7,10,11,12,13, 14,15,16,18,19 , 20,21,22,23,24,29) of the fingerprint, which calculates the average of 21 batches of eight herbs characteristic peak area (a), the mean standard deviation (the SD), see Table 10; 21 batches wherein the ratio of peak area of ​​each sample total peak area (see Table 11, FIG. 9); wherein each peak area to the reference peak area ratio between the sample 21 passing batches (Table 12, FIG. 10)

[0100] 表10相似度大于0. 8的21批单面针药材峰面积 [0100] Table 0.8 of similarity is greater than 10 batches of 21 herbs sided needle peak area

[0101] [0101]

Figure CN101266233BD00141

[0102] 表11相似度大于0. 80的21批样品中各特征峰峰面积占总峰面积的比例 The total peak area ratio of peak area of ​​each batch of samples wherein 21 [0102] Table 11 of similarity is greater than 0.80

[0103] [0104] [0103] [0104]

Figure CN101266233BD00142

[0105] 表12相似度大于0. 80的21批样品各指纹峰与参照峰(6#)峰面积的比值 Ratio [0105] Table 0.80 of similarity is greater than 12 batches of 21 samples of each fingerprint peak reference peak (# 6) peak area

[0106] [0106]

Figure CN101266233BD00151

134567W1112S31516比S20M22S 对29 134567W1112S31516 than 29 S20M22S

0.151 0.034 0.038 0.112 0.075 0.112 0.124 0.053 0.072 0.213 0.117 0.083 0.159 0.088 0.094 0.140 0.046 0.159 0.082 0.218 0.138 0.151 0.034 0.038 0.112 0.075 0.112 0.124 0.053 0.072 0.213 0.117 0.083 0.159 0.088 0.094 0.140 0.046 0.159 0.082 0.218 0.138

0.025 0.007 0.007 0.017 0.013 0.020 0.023 0.010 0.013 0.036 0.017 0.020 0.034 0.023 0.025 0.026 0.010 0.046 0.016 0.039 0.019 0.025 0.007 0.007 0.017 0.013 0.020 0.023 0.010 0.013 0.036 0.017 0.020 0.034 0.023 0.025 0.026 0.010 0.046 0.016 0.039 0.019

0.051 0.019 0.008 0.060 0.025 0.085 0.050 0.014 0.033 0.075 0.037 0.050 0.072 0.049 0.034 0.049 0.016 0.054 0.046 0.068 0.045 0.051 0.019 0.008 0.060 0.025 0.085 0.050 0.014 0.033 0.075 0.037 0.050 0.072 0.049 0.034 0.049 0.016 0.054 0.046 0.068 0.045

0.122 0.048 0.015 0.136 0.050 0.212 0.149 0.028 0.126 0.140 0.083 0.146 0.203 0.111 0.071 0.160 0.036 0.174 0.097 0.390 0.223 0.122 0.048 0.015 0.136 0.050 0.212 0.149 0.028 0.126 0.140 0.083 0.146 0.203 0.111 0.071 0.160 0.036 0.174 0.097 0.390 0.223

0.221 0.056 0.027 0.160 0.053 0.328 0.263 0.016 0.177 0.453 0.131 0.179 0.273 0.152 0.094 0.238 0.038 0.262 0.113 0.604 0.429 0.221 0.056 0.027 0.160 0.053 0.328 0.263 0.016 0.177 0.453 0.131 0.179 0.273 0.152 0.094 0.238 0.038 0.262 0.113 0.604 0.429

0.183 0.059 0.078 0.094 0.083 0.170 0.100 0.060 0.139 0.221 0.171 0.232 0.231 0.173 0.107 0.097 0.084 0.136 0.067 0.355 0.227 0.183 0.059 0.078 0.094 0.083 0.170 0.100 0.060 0.139 0.221 0.171 0.232 0.231 0.173 0.107 0.097 0.084 0.136 0.067 0.355 0.227

0.080 0.033 0.049 0.063 0.038 0.107 0.249 0.060 0.068 0.129 0.154 0.097 0.091 0.137 0.060 0.124 0.062 0.103 0.056 0.103 0.056 0.080 0.033 0.049 0.063 0.038 0.107 0.249 0.060 0.068 0.129 0.154 0.097 0.091 0.137 0.060 0.124 0.062 0.103 0.056 0.103 0.056

平均值 average value

0.110 0.021 0.045 0.130 0.203 1 0.146 0.091 0.110 0.021 0.045 0.130 0.203 0.146 0.091 1

[0107] 以上几个图表所显示出的信息可知,这21批样品中各特征峰峰面积与总峰面积及参照峰峰面积的比值较为稳定,即不管这21批样品中各特征峰峰面积的绝对值高低如何变化,它们与总峰面积及6#内标峰峰面积的比例关系是比较一致的。 [0107] The above chart shows the several information indicates that 21 batches of samples and the reference ratio of the area of ​​each characteristic peak area peak area to total peak is more stable, i.e., no matter which of the 21 batches of samples wherein each peak area the absolute value of the change in the level of how they are proportional to the total peak area # 6 and the internal standard peak area is more consistent.

[0108] 拟定相似度0. 80以上样品的各指纹峰与总峰面积、参照峰面积的比例关系见表13。 [0108] similarity of each fingerprint peak prepare total peak area of ​​sample than 0.80, the ratio of the peak area of ​​the reference relationships shown in Table 13.

[0109] 表13相似度0. 80以上样品的各指纹峰与总峰面积、参照峰峰面积的最小比值 [0109] each fingerprint peak total peak area above the sample 13 0.80 similarity tables, with reference to the ratio of the minimum peak area

[0110] [0110]

Figure CN101266233BD00152

[0111] 小结 [0111] Summary

[0112] 本研究共测试30批单面针药材,研究结果表明由于该药材药用部位、生长期、采收时间等的差异,造成30批药材整体指纹图谱差异极大,为此,考虑到企业的生产实际情况,为扩大单面针药源的选择,采用相似度0. 80以上样品来构建共有模式并作为评价标准。 [0112] This study tested 30 batches of single-sided needle medicine, research results show that due to differences in the parts of medicinal herbs, growing, harvest time, resulting in 30 batches of overall herbs fingerprints great differences, for which, taking into account the actual production situation of enterprises, the expansion of single-sided needle drug source options for using the similarity 0.80 or more to build a total sample mode and as evaluation criteria.

Claims (9)

  1. 一种单面针药材的鉴别方法,包括如下步骤:第一步,建立单面针药材的HPLC标准指纹图谱,包括:1)供试品溶液的制备:精密称取单面针药材粉末,经石油醚预处理后,挥干石油醚,残渣中加入甲醇加热回流,合并滤液,水浴蒸干,残渣加甲醇溶解,转移至容量瓶中,定容至刻度,得到供试品溶液;所述单面针药材是在湖南省张家界地区采集;2)高效液相色谱分析:精密吸取供试品溶液进样,色谱条件:色谱柱为Kromasil RP‑C184.6mm×250mm,5μm;HP G1313A自动进样系统;流动相为乙腈‑0.2%磷酸三乙胺溶液;采用梯度洗脱方式:0min‑10min‑30min‑70min,乙腈与0.2%磷酸三乙胺溶液的体积比为:5∶95‑10∶90‑10∶90‑40∶60;检测波长:210~366nm;得到单面针药材HPLC标准指纹图谱;3)测定:精密吸取30批单面针药材的供试品溶液各5‑20μl,分别注入液相色谱仪, A method of identifying method sided needle herbs, comprising the steps of: a first step the establishment of one-sided needle of HPLC standard fingerprint, comprising: 1) Preparation of test solution for: Precision Weigh sided needle medicinal powder, dried after pretreatment with petroleum ether, petroleum ether, evaporated to dryness, the residue was added methanol was heated to reflux and the combined filtrate evaporated to dryness in a water bath, the residue dissolved in methanol, transferred to a volumetric flask, dilute to volume, to give the test solution; said single face acupuncture medicine is collected in Zhangjiajie Hunan; 2) HPLC analysis: precision drawing test solution was injected, chromatographic conditions: column of Kromasil RP-C184.6mm × 250mm, 5μm; HP G1313A Autosampler system; mobile phase acetonitrile -0.2% phosphoric triethylamine; gradient elution mode: 0min-10min-30min-70min, the volume ratio of acetonitrile and 0.2% triethylamine phosphate solution is: 5:95-10:90 -10:90-40:60; detection wavelength: 210 ~ 366nm; obtained sided needle on HPLC standard fingerprint; 3) Determination: precision drawing test solution of 30 batches sided needle medicinal each 5-20μl, were injected Liquid chromatography, 照高效液相色谱法测定,得到30批单面针药材的HPLC指纹图谱进行比较,得到由共有特征峰构成的单面针药材HPLC标准指纹图谱;所述单面针标准指纹图谱中,共有特征峰有8个,计算各个特征峰与总峰面积的积分相对比值,其中1号峰的平均相对保留时间Rt为3.364min,相对峰面积为4.2~7.8%;2号峰的平均相对保留时间Rt为4.620min,相对峰面积为0.8~1.6%;3号峰的平均相对保留时间Rt为10.772min,相对峰面积为1.5~3.3%;4号峰的平均相对保留时间Rt为12.799min,相对峰面积为3.7~9.9%;5号峰的平均相对保留时间Rt为13.828min,相对峰面积为4.5~16.3%;6号峰的平均相对保留时间Rt为34.015min,相对峰面积为46.9~73.1%;7号峰的平均相对保留时间Rt为58.820min,相对峰面积为5.3~10.7%;8号峰的平均相对保留时间Rt为62.179min,相对峰面积为2.7~7.5%;第二步,以上述相同的 According to HPLC, to give one side of the needle 30 batches of HPLC fingerprints are compared, one side of the needle on HPLC to give standard fingerprint characteristic peaks consisting of total; one surface of said needle in the standard fingerprint, a common feature has eight peaks, each peak to the total integrated peaks area relative ratio calculated, wherein the number average relative retention time of peak 1 was Rt 3.364min, relative peak area from 4.2 to 7.8%; number average relative retention time of 2 peaks Rt is 4.620min, the relative peak area of ​​0.8 to 1.6%; average peak No. 3 is the relative retention times Rt 10.772min, the relative peak area of ​​1.5 to 3.3%; average peak 4 relative retention time of Rt 12.799min, relative peak area of ​​3.7 to 9.9%; average peak number 5 of the relative retention time of Rt 13.828min, relative peak area of ​​4.5 to 16.3%; number average relative retention time of peak 6 was Rt 34.015min, the relative peak area of ​​46.9 - 73.1% ; number average relative retention time of peak 7 was Rt 58.820min, relative peak area from 5.3 to 10.7%; number average relative retention time of peak 8 as Rt 62.179min, relative peak area of ​​2.7 to 7.5%; the second step, to same as above 方法测定待测单面针药材样品的指纹图谱;第三步,将第二步所测定的待测单面针药材样品指纹图谱与第一步构建的单面针标准指纹图谱对比,筛选符合单面针药材标准指纹图谱的合格单面针药材。 The method of measuring the test fingerprint sided needle medicine samples; the third step, the single-sided needle-sided standard fingerprint comparison test needle with medicine fingerprint sample measured First Step Second Step constructed, in line with a single filter Eligible single needle pin medicinal herbs surface standard fingerprint.
  2. 2.如权利要求1所述的方法,其特征在于,所述第一步1)中,单面针药材粉末细度为40目。 2. The method according to claim 1, wherein said first step 1), the medicinal powder sided needle fineness of 40 mesh.
  3. 3.如权利要求1所述的方法,其特征在于,所述第一步1)中,加入石油醚加热回流不少于10分钟,其中,Ig单面针药材加入石油醚量不少于IOml ;加入甲醇加热回流1次以上,每次不少于10分钟,其中Ig单面针药材加入甲醇量不少于10ml。 3. The method according to claim 1, wherein said first step 1), the petroleum ether was added heated to reflux for less than 10 minutes, wherein, an Ig-sided needle of not less than medicinal petroleum ether IOml ; methanol was added heated to reflux for 1 or more times less than 10 minutes, wherein the Ig-sided needle herbs methanol was added an amount of not less than 10ml.
  4. 4.如权利要求3所述的方法,其特征在于,所述第一步1)中,Ig单面针药材加入石油醚30ml加热回流30分钟;加入甲醇30ml加热回流2次,每次1小时。 4. The method according to claim 3, wherein said first step 1), an Ig-sided needle herbs 30ml petroleum ether was heated to reflux for 30 min; 30ml methanol was heated at reflux was added 2 times, each time 1 hour .
  5. 5.如权利要求1所述的方法,其特征在于,所述第一步3)中,精密吸取单面针药材的供试品溶液10 μ 1。 5. The method according to claim 1, wherein said first step 3), the precision drawing-sided needle medicine test solution 10 μ 1.
  6. 6.如权利要求1所述的方法,其特征在于,所述第一步1)中,残渣用甲醇定容至2ml。 6. The method according to claim 1, wherein said first step 1), the residue was diluted with methanol to 2ml.
  7. 7.如权利要求1所述的方法,其特征在于,所述第一步1)中,供试品溶液高效液相分析前用0. 45 μ m微孔滤膜滤过。 7. The method according to claim 1, wherein said first step 1), the test solution prior to analysis by HPLC 0. 45 μ m microporous membrane filtration.
  8. 8.如权利要求1所述的方法,其特征在于,所述第一步2)中,检测波长为280nm。 8. The method according to claim 1, wherein said first step 2), the detection wavelength was 280nm.
  9. 9.如权利要求1所述的单面针药材鉴别方法,其特征在于,所述8个共有特征峰的相对保留时间为,1号峰的平均相对保留时间Rt为3. 364min,标准偏差SD为0. 019 ;2号峰的平均相对保留时间Rt为4. 620min,标准偏差SD为0. 008 ;3号峰的平均相对保留时间Rt为10. 772min,标准偏差SD为0. 102 ;4号峰的平均相对保留时间Rt为12. 799min,标准偏差SD为0. 082 ;5号峰的平均相对保留时间Rt为13. 828min,标准偏差SD为0. 101 ;6号峰的平均相对保留时间Rt为34. 015min,标准偏差SD为0. 603 ;7号峰的平均相对保留时间Rt 为58. 820min,标准偏差SD为0. 074 ;8号峰的平均相对保留时间Rt为62. 179min,标准偏差SD 为0. 256。 9. A blind pin medicine identification method according to claim 1, characterized in that the eight common peaks relative retention time, peak No. 1 average relative retention time of Rt 3. 364min, standard deviation SD to 0.019; number average relative retention time of 2 peaks Rt of 4. 620min, the standard deviation SD 0.008; average peak 3 relative retention time Rt of 10. 772min, the standard deviation SD 0.102; 4 the average number of peaks of the relative retention time Rt of 12. 799min, the standard deviation SD 0.082; number average relative retention time of 5 peaks Rt of 13. 828min, the standard deviation SD 0.101; number average relative retention peaks 6 time Rt of 34. 015min, the standard deviation SD 0.603; number average relative retention time of 7 peaks Rt of 58. 820min, the standard deviation SD 0.074; number average relative retention time of 8 peaks Rt of 62. 179min standard deviation SD of 0.256.
CN 200710064499 2007-03-16 2007-03-16 Zanthoxylum dissitum Hemsl medicinal materials discrimination method CN101266233B (en)

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CN1322951A (en) 2001-06-29 2001-11-21 天津市金士力药物研究开发有限公司 Red sage medicine fingerprint establishing method and standard fingerprint atlas
CN1844912A (en) 2006-01-24 2006-10-11 李振国 Earthworm fingerprint spectrum establishment method and medicinal earthworm identification method
CN1903237A (en) 2006-07-31 2007-01-31 暨南大学 Angelica dehurica water extractive liquid fingerprint, its establishmetn method and application

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CN1322951A (en) 2001-06-29 2001-11-21 天津市金士力药物研究开发有限公司 Red sage medicine fingerprint establishing method and standard fingerprint atlas
CN1844912A (en) 2006-01-24 2006-10-11 李振国 Earthworm fingerprint spectrum establishment method and medicinal earthworm identification method
CN1903237A (en) 2006-07-31 2007-01-31 暨南大学 Angelica dehurica water extractive liquid fingerprint, its establishmetn method and application

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9804136B2 (en) 2014-09-18 2017-10-31 Dionex Corporation Automated method of calibrating a chromatography system and analysis of a sample

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