Determination of Oleanolic Acid in Achyranthes aspera L. of Different Production Areas by HPLC

[Objectives] To determine the content of oleanolic acid in 10 batches of Achyranthes aspera L. from different production areas,and to establish a high performance liquid chromatography( HPLC) method for the determination of oleanolic acid in A. aspera. [Methods]Agilent C18 liquid chromatography column( 250 mm × 4. 6 mm,5 μm) was used for gradient elution with acetonitrile∶ water = 61∶ 39 as the mobile phase. The flow rate was 1 mL/min,the detection wavelength was 210 nm,and the column temperature was 35℃. [Results] The oleanolic acid injection volume showed a good linear relationship with the peak area in the range of 0. 382-7. 640 μg. The linear equation of oleanolic acid was: A = 530. 76 C,R = 1. 000 0; the range of sample recovery rate was 96. 03%-102. 73%,and the RSD value was 2. 16%( n =9). The content of the oleanolic acid was the highest in A. aspera produced in Guilin City of Guangxi( 0. 92%),and the lowest oleanolic acid content was in sample of Sitang in Nanning City of Guangxi( 0. 27%). It is recommended that the oleanolic acid content of A. aspera should not be lower than 0. 20%. [Conclusions]The HPLC method is accurate,reliable,easy to operate,and has good resolution. It is suitable for the determination of the content of oleic acid in A. aspera. It can provide reference for the quality control and standard drafting of A. aspera.

SIMULTANEOUS DETERMINATION OF VITAMIN B COMPLEX (THIAMINE, RIBOFLAVIN, NIACIN, PYRIDOXINE AND FOLIC ACID) IN FOODS BY HPLC

A high performance liquid chromatographic assay has been developed for the simultaneous determination of five water soluble vitamins:thimine, riboflavin, niacin, pyridoxine and folic acid in cereal products and fresh vegetables. Food samples were hydrolyzed in 0.4 mol/L HCl, autoclaved at 120℃ 15 psi for 20 minutes, using a μ-Bondapak column (3.9×300mm, Waters Co.), a mobile phase of methanol-water (30:70) (0.005 mol/L heptanesulfonic acid) and a flow rate of 1.0 ml/min gave the most satisfactory separation of the five water soluble vitamins. A double channel deteetion was used: four vitamins (B_5, folic acid, B_2, B_1) were detected hy UV spectrophotometry (254 nm) first, pyridoxine (B_6) was detected by fluoromelry (EX 290nm, EM 395 nm) afterwards. Detection limits were 2, 5, 5, 2and 5ng, linear ranges were 5-10ng, 5-50ng, 5-40ng, 5-50ng and 10-50ng for B_1, B_2, B_5, B_6 and folic acid respectively. Recoveries were 92-100%(B_5), 51-52% (folio acid), 103—105% (B_2), 99—100% (B_6) and 91.3—102% (B_1) respectively. In comparison with a reference method and checking with food composition tables, very satisfactory results were obtained by this method.

DETERMINATION OF ORGANIC ACIDS IN SOY SAUCE BY HPLC

Organic acids in 3 kinds of soy sauces were analyzed by HRLC.Column:C18 ()Bondapak(10μm,3.9mm i.d.* 30cm;mobile phase:0.5%(NH_4)_2HPO_4(pH=2.9);flow rate:1.0 ml/m.PT-C18 cartridge was used for the sample preparation.Recoveries of organic acids are between 96-111%.

HPLC法同时测定杠板归不同部位中5种成分的含量

目的建立高效液相色谱法同时测定杠板归不同部位(茎、叶、根)中5种成分的含量。方法采用Luna?-C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈-0.1%磷酸水为流动相梯度洗脱,流速为1.0 mL/min,检测波长为320 nm,柱温为30℃,测定杠板归不同部位中绿原酸、咖啡酸、芦丁、阿魏酸、槲皮素的含量。结果绿原酸、咖啡酸、芦丁、阿魏酸、槲皮素分别在0.046~4.6μg/mL,0.212~21.2μg/mL,0.055~5.5μg/mL,0.084~8.4μg/mL,0.114~11.4μg/mL范围内质量浓度与峰面积呈线性关系良好(r>0.9990),平均加样回收率为97.65%~102.09%,RSD为1.76%~2.44%。不同部位中5种成分含量差异较大,绿原酸和咖啡酸2个成分在叶中含量高,且远远高于根和茎中的含量;芦丁和阿魏酸主要存在于茎和根中,叶中未检测到;槲皮素在茎中含量最高,叶中次之,根中最低。结论建立的HPLC方法准确可靠、重复性好、专属性强,可用于杠板归不同部位中5个成分含量的同时测定。

Determination of Chlorogenic Acid in Stems of Mussaenda pubescens Ait.f.by High Performance Liquid Chromatography

[Objectives] This study was conducted to establish a method for determination of chlorogenic acid in Mussaenda pubescens Ait.f.[Methods]HPLC used SHISEIDO C_(18) MG Ⅱ column( 5 μm,4.6 mml.D.× 250 mm) as chromatographic column and acetonitrile-0.4% phosphoric acid solution( 9∶91) as mobile phase.The separation was performed at a flow rate of 1.0 ml/min and a column temperature of 30 ℃,and the detection wavelength was set at 327 nm.[Results]Chlorogenic acid had a good linear relation in the range of 0.254 7-2.547 5 μg( R2= 0.999 9).The recovery rate was 97.8%,RSD = 1.82%( n = 9).[Conclusions]The content of chlorogenic acid in M.pubescens was determined by ultrasonic extraction and HPLC.The method was simple,stable and reliable and could be used for the quality control of M.pubescens.

Extraction Process and Content Determination of Caffeic Acid in Laggera alata from Different Production Areas of Guangxi

[Objectives] To establish a method for determining the content of Laggera alata( D. Don) Sch. Bip. Ex Oliv. using caffeic acid the target component,and to compare the content of caffeic acid in the medicinal materials of L. alata in different production areas of Guangxi.[Methods]The content was determined by Inertsil~ODS-3 chromatographic column C_(18)( 4. 60 mm × 250 mm,5 μm,mobile phase: acetonitrile-0. 1% phosphoric acid( 22∶ 78),detection wavelength: 320 nm,flow rate: 1. 0 m L/min,column temperature: 30℃,and injection volume: 10 μL. [Results] The caffeic acid showed a good linear relationship in the range of injection volume of 0. 025 92-0. 259 2 μg( R =0. 999 5). The average recovery rate was 98. 33%( RSD = 1. 85%). L. alata in different production areas of Guangxi contained the caffeic acid,and there was a great difference in the caffeic acid. L. alata in Baise had the highest content of caffeic acid,while that in Guilin had the lowest content of caffeic acid. [Conclusions]This method can accurately determine the content of caffeic acid and is expected provide a scientific basis for the development and utilization of herbal medicine L. alata.

HPLC-MS/MS法同时测定清开灵口服液中7种成分的含量

目的 建立高效液相色谱-质谱联用(HPLC-MS/MS)法同时测定清开灵口服液中7种成分含量。方法 采用Waters ACQUITY UPLC BEH C_(18)色谱柱(2.1 mm×10 mm,1.7μm);以含10 mmol·L^(-1)乙酸铵和0.1%甲酸的水溶液(A)及甲醇(B)为流动相,梯度洗脱;电喷雾离子源,正、负离子多反应监测(MRM)模式进行定量分析。结果 腺苷、绿原酸、咖啡酸、栀子苷、黄芩苷、猪去氧胆酸和胆酸质量浓度分别在0.100 4~3.213、0.784 5~8.982、0.998~3.194、0.622 5~19.92、25.05~300.6、2.513~30.15和7.775~93.30μg·m L^(-1)范围内与峰面积呈良好线性关系(r≥0.999 0);平均回收率(n=6)分别为100.9%、98.74%、101.2%、100.2%、100.8%、99.97%和98.94%,RSD分别为1.58%、0.59%、1.78%、1.25%、0.65%、1.69%和1.07%。15批样品中腺苷、绿原酸、咖啡酸、栀子苷、黄芩苷、猪去氧胆酸和胆酸的含量范围分别为0.12~0.18、0.19~0.24、0.06~0.09、0.34~0.37、4.54~4.85、0.49~0.67和1.82~2.19 mg·m L^(-1)。15批样品测定结果较为接近。结论 该方法具有良好的灵敏度、准确性和重复性,可为清开灵口服液的质量控制和后续研究提供参考及数据支持。

Extraction Process and Content Determination of Caffeic Acid in Zhuang Medicine Cryptolepis buchananii

[Objectives]To establish a method for determining the content of caffeic acid in Zhuang Medicine Cryptolepis buchananii.[Methods]The content of caffeic acid in C.buchananii leaves was determined by high performance liquid chromatography(HPLC).[Results]The results of HPLC determination showed that the RSD values were less than 3%,and the content of caffeic acid in 10 batches of C.buchananii leaves was in the range of 0.0965%-0.4772%.[Conclusions]This method is accurate and reliable,has good linear relationship and high precision,and can be used for quality evaluation of C.buchananii.

HPLC法测定蒙药材金露梅中没食子酸含量

目的:建立高效液相法测定金露梅中没食子酸含量。方法:采用Kromasil 100-5-C18(250 mm×4.6 mm,5μm)色谱柱,以甲醇-0.1%磷酸(10∶90)为流动相,流速为1.0 mL/min,检测波长273 nm,柱温30℃,进样量为10μL。结果:没食子酸在3.44~137.52μg/mL范围内线性关系良好,相关系数r=1.000;平均回收率为99.4%(RSD%=1.588%);6批不同产地金露梅中没食子酸含量在3.156~11.537 mg/g之间。结论:所建立的高效液相法稳定可靠,可用于金露梅中没食子酸的含量测定。

在线衍生HPLC快速测定聚酯纤维中TPA与IPA的含量

聚酯纤维在氢氧化钾-乙醇溶液中解聚,解聚液用水定容后采用在线衍生高效液相色谱法(HPLC)分析苯二甲酸,建立了快速测定聚酯纤维中对苯二甲酸(TPA)与间苯二甲酸(IPA)含量的方法。结果表明:聚酯纤维在65℃氢氧化钾-乙醇溶液中振荡处理35min后完全解聚;在190~360nm检测波长范围内,TPA和IPA的最大吸收波长分别为196、212nm,次吸收波长分别为241、220nm,流动相为乙腈-水(0.1%甲酸,V_(乙腈)∶V_(水)=15∶85)时的保留时间分别为6.038、7.776min;TPA和IPA的检出限分别为0.50、1.58mg/L,加标回收率高,相对标准偏差小。该方法样品前处理简单,分析快捷,结果可靠。

HPLC法测定乳苣不同部位菊苣酸含量

采用超声法提取乳苣不同部位总多酚,建立了测定乳苣不同部位菊苣酸含量的HPLC法,色谱条件为:岛津LC-16型高效液相色谱仪,SPD检测器,C18色谱柱(4.6 mm×150 mm,5μm),柱温为40℃,流动相为甲醇-0.1%磷酸,梯度洗脱,流速为1.0 mL·min^(-1),检测波长为280 nm,进样量为10μL。结果表明,菊苣酸浓度在11.6~58.0μg·mL^(-1)范围内与峰面积线性关系良好(R^(2)=0.9994),平均加标回收率为95.30%(n=6);乳苣根、茎、叶、全草中菊苣酸含量分别为23.70 mg·g^(-1)、37.20 mg·g^(-1)、114.24 mg·g^(-1)、102.50 mg·g^(-1),乳苣不同部位菊苣酸含量大小顺序为:叶>全草>茎>根。该方法简便、快速、准确,适用于乳苣中菊苣酸含量的测定。

藏药五味清热汤散的HPLC指纹图谱及5种成分含量测定方法的研究

目的 建立藏药五味清热汤散的HPLC指纹图谱并测定指标成分没食子酸、没食子酸甲酯、诃子联苯酸、诃子酸、鞣花酸的含量。方法 采用Ultimate XB-C18液相色谱柱(4.6 mm×250 mm,5 μm),以0.1%磷酸水溶液(A)-甲醇(B)为流动相,梯度洗脱,流速1 mL·min^(-1),检测波长270 nm,柱温30℃,进样量10 μL。结果 五味清热汤散指纹图谱中标识出18个共有峰,指纹图谱相似度>0.99;指标成分没食子酸、没食子酸甲酯、诃子联苯酸、诃子酸和鞣花酸线性范围分别为2.76~55.17 μg·mL^(-1)(r=0.9998)、1.56~31.13 μg·mL^(-1)(r=0.9997)、5.51~110.17 μg·mL^(-1)(r=0.9999)、7.27~145.31 μg·mL^(-1)(r=0.9998)、0.90~18.08 μg·mL^(-1)(r=0.9997);精密度、重复性、稳定性试验的RSD均小于2.0%(n=6);平均加样回收率分别为98.27%(RSD=0.72%)、98.31%(RSD=0.94%)、96.48%(RSD=0.73%)、96.86%(RSD=0.80%)、96.07%(RSD=0.81%)。8批样品中没食子酸、没食子酸甲酯、诃子联苯酸、诃子酸和鞣花酸的含量分别为7.85~8.25、4.02~4.23、16.85~17.77、23.55~24.51、3.09~3.18 mg·g^(-1)。结论 建立的指纹图谱和指标成分含量测定方法具有简便、准确、稳定、快速等优点,可为五味清热汤散的质量评价提供参考。

HPLC法测定雪地茶中鳞片衣酸和羊角衣酸的含量

目的:建立高效液相色谱法(HPLC)同时测定雪地茶中2种缩酚酸(鳞片衣酸、羊角衣酸)含量的方法。方法:采用Eclipse XDB-C_(18)(4.6 mm×250 mm,5μm)色谱柱,乙腈(A)-0.075%磷酸水溶液(B)为流动相,梯度洗脱(0~40 min,35%A→55%A;40~41 min,55%A→100%A;41~49 min,100%A;49~60 min,100%A→35%A),流速为1.0 mL·min^(-1),检测波长213 nm,柱温40℃,进样量10μL。结果:鳞片衣酸在0.15~0.55 mg·mL^(-1)范围内线性关系良好(R^(2)=1),方法的平均回收率为94.15%,RSD为0.21%(n=6)。羊角衣酸在0.05~0.55 mg·mL^(-1)范围内线性关系良好(R^(2)=0.9997),方法的平均回收率为97.83%,RSD为0.16%(n=6)。结论:建立的该含量测定方法结果准确,重复性和分离度好,可用于同时测定雪地茶中两种缩酚酸的含量,为雪地茶药材的质量控制提供了科学依据,对其质量标准的建立具有一定的参考意义。

HPLC法检测复方金银花颗粒剂型中绿原酸的含量

采用高效液相色谱法(HPLC)检测复方金银花颗粒中绿原酸的含量。采用C18反相色谱柱,以水∶乙腈溶液为流动相,等度洗脱,检测波长327 nm。以绿原酸浓度(μg/mL)为横坐标,峰面积为纵坐标,建立绿原酸标准曲线y=14 862x-254 314,相关系数r^(2)=0.999。通过峰面积计算复方金银花颗粒剂型中绿原酸的含量为60.78μg/mL,RSD值为1.72%。本方法操作简单、快速、灵敏,重现性良好,线性关系良好,可用于绿原酸的质量控制。

HPLC法测定小儿肺热咳喘口服液中3种防腐剂的含量

目的:建立小儿肺热咳喘口服液中3种防腐剂苯甲酸、山梨酸、糖精钠的含量测定方法。方法:采用高效液相色谱法(HPLC),色谱柱为Thermo syncronis C_(18)(250 mm×4.6 mm, 5μm),流动相为乙腈-0.2%磷酸溶液(19∶81),流速1 mL/min,柱温30℃,检测波长320 nm,进样量10μL。结果:苯甲酸、山梨酸、糖精钠在5~60μg/mL范围内与其峰面积均呈良好的线性关系(r>0.999),平均加样回收率分别为99.72%(RSD=1.57%,n=6),97.30%(RSD=1.70%,n=6),101.37%(RSD=0.99%,n=6);苯甲酸、山梨酸、糖精钠精密度试验的RSD分别为0.07%、0.21%、0.18%。结论:实验采用的HPLC法简便、准确、专属性强、重复性好,可用于液体口服制剂中苯甲酸、山梨酸、糖精钠的含量测定。

HPLC法同时测定野木瓜片中木通苯乙醇苷B和绿原酸的含量

目的:建立同时测定野木瓜片中木通苯乙醇苷B和绿原酸含量的高效液相色谱法。方法:采用Thermo BDS HYPERSIL C_(18)色谱柱(4.6 mm×250 mm,5μm),柱温为35℃;以甲醇-0.1%磷酸水溶液为流动相,梯度洗脱;流速为1.0 mL•min^(-1);检测波长为327 nm;进样体积为10μL。结果:木通苯乙醇苷B和绿原酸分别在0.51~20.60μg•mL^(-1)(r=1.000)和0.52~20.63μg•mL^(-1)(r=1.000)内呈良好线性关系;平均回收率分别为100.3%(RSD为1.1%)和105.9%(RSD为1.4%)。测定5批次样品,木通苯乙醇苷B和绿原酸含量分别为0.083~1.115 mg•g^(-1)和0.161~1.204 mg•g^(-1)。结论:本法可为野木瓜片的质量评价标准提高提供参考依据。

HPLC-DAD波长切换法测定阿司匹林双嘧达莫片的含量及游离水杨酸

目的:建立HPLC-DAD波长切换法测定阿司匹林双嘧达莫片的含量及游离水杨酸的方法。方法:采用Agilent ZORBAX SB C 18色谱柱(250 mm×4.6 mm,5μm),以乙腈-四氢呋喃-冰醋酸-水(20∶5∶5∶70)为流动相,流速1.0 mL·min^(-1),柱温30℃,检测波长276 nm(阿司匹林)、284 nm(双嘧达莫)和303 nm(水杨酸)。结果:阿司匹林、双嘧达莫和游离水杨酸分别在38.37~383.71、9.60~96.05和1.02~10.21μg·mL^(-1)范围内线性关系良好。平均回收率分别为99.88%(RSD=0.23%)、99.01%(RSD=0.46%)和100.99%(RSD=0.75%)。本法与标准方法含量测定结果无明显差异。含量均匀度A+2.2S在4.7~6.4。结论:建立的含量测定方法,操作简单,更适用于阿司匹林双嘧达莫片的质量控制。

HPLC法测定金丹附延颗粒中肉桂酸和桂皮醛的含量

目的:建立金丹附延颗粒中肉桂酸和桂皮醛的HPLC含量测定方法。方法:采用C 18色谱柱,流动相为乙腈和0.1%磷酸溶液,检测器检测波长设置为290 nm,输液泵流速1.0 mL·min^(-1),色谱柱柱温设置为25℃。结果:肉桂酸、桂皮醛分别在0.2079~10.3938μg·mL^(-1)、0.2025~10.1270μg·mL^(-1)范围内线性关系良好(r=1.0000),回收率分别为100.3%、102.6%,回收率RSD分别为1.57%、1.82%。结论:该方法简单、稳定,可用于金丹附延颗粒中肉桂酸和桂皮醛的含量测定。

HPLC法测定内蒙古不同产地蒙药材接骨木中熊果酸和齐墩果酸的含量

目的:采用高效液相色谱(HPLC)法测定内蒙古不同产地蒙药材接骨木所含熊果酸和齐墩果酸的量。方法:采用Promosil C18色谱柱(250mm×4.6mm,5 um),以乙腈-甲醇-0.5%醋酸铵溶液(59∶14∶27)作检测流动相,等度洗脱,柱流速为1.0mL·min^(-1),检温40℃,波长210nm。结果:1.齐墩果酸的平均含量为0.302 mg/g、熊果酸的平均含量为0.585 mg/g;呈良好线性关系,熊果酸在1.048~3.143ug/ul范围内r值可达0.9995,齐墩果酸在0.778~2.333 ug/ul范围内r值可达0.9996;熊果酸、齐墩果酸平均加样回收率为102.83%和99.02%,RSD为1.08和1.88。结论:1.HPLC法是检测蒙药材接骨木中主要成分(熊果酸和齐墩果酸)的有效科学手段。检测结果准确度高,精密度好,可用于蒙药材接骨木的质量评价。2.在内蒙古不同产地的蒙药材接骨木中霍林郭勒市达来胡硕所采接骨木中熊果酸和齐墩果酸的含量最高。

HPLC直接测定血清阿魏酸──方剂血样预处理新方法(Ⅰ)

建立了沸水浴去血清蛋白质后用ＨＰＬＣ直接测定阿魏酸（ＦＡ）方法：方剂的血样预处理新方法分别用二维、三维ＨＰＬＣ和分光光度计定性测定ＦＡ后，用内标（香豆精）法定量。甲醇－乙酸－水（３８：０．３：６１．７）为流动相，Ｃ_(８)（ＯＤＳ_(２)）柱（１５０ｍｍ×４．６ｍｍ，５μｍ）为固定相时，ＦＡ的最低检测限 ６ ｎｇ；沸水浴 １０ ｍｉｎ的最低检测血清浓度是 ２０．２ ｎｇ／ｍＬ，线性范围 ３３． ７～２ １５６．８ ｎｇ／ｍＬ，ｒ＝０．９９９３；方法平均回收率（９３．５９±１．９３）％，日间及日内精密度ＲＳＤ值均为８．４４％。上法已用于人服川芎汤后经时变化的血清ＦＡ浓度测定。