A Sensitive Reversed-Phase High-Performance Liquid Chromatography Method for the Quantitative Determination of Milk Xanthine Oxidase Activity

A new reversed-phase high performance liquid chromatography method was developed to quantitate the activity of xanthine oxidase involved in milk fat globule membrane with xanthine as the substrate and the separation of product (uric acid). The increment of uric acid in the reaction system was used to calculate the total activity of XO. The optimized assay conditions, linearity of detection, recovery of uric acid and chromatogram were developed in text, indicating this method is simple, rapid and efficient. It is an alternative potential method for the determination of the activity of XO in milk.

Simultaneous Determination of Amlodipine with H₁-Receptor Antagonists by Reversed Phase High Performance Liquid Chromatography and Application to Interaction Studies

A rapid, fast and precise method has been developed and validated for the simultaneous determination of amlodipine with H1-receptor antagonists (cetirizine, fexofenadine, and buclizine) from dosage forms. The chromatography was performed on a Purospher? Star, C18 (5 mm, 250 × 4.6 mm) column using acetonitrile: buffer (0.01 mM) (40:60, v/v, pH adjusted to 3.0), as a mobile phase. The mobile phase was pumped at a flow rate of 1.0 mL·min-1 and UV detection was performed at 240 nm. The method was validated for linearity, accuracy, precision and specificity. The method was applied to study the interaction between amlodipine and H1-receptor antagonists. These interactions were carried out in simulated gastric juice (pH 1), simulated full stomach (pH 4), blood pH (pH 7.4) and simulating GI (pH 9). The interacting drugs were heated at 37℃ with intermit-tent shaking and the samples were withdrawn every thirty minutes for three hours and drug contents were analyzed by RP-HPLC techniques. In most cases the in vitro availability of amlodipine was decreased. It was observed that the change in in vitro availability was pH dependent.

Quantitative analysis by reversed-phase high-performance liquid chromatography and retinal neuroprotection after topical administration of moxonidine

AIM:To determine moxonidine in aqueous humor and iris-ciliary body by reversed-phase high performance liquid chromatography(RP-HPLC),and to evaluate the retinal neuroprotective effect after topical administration with moxonidine in a high intraocular pressure(IOP)model.METHODS:The eyes of albino rabbits were administered topically and ipsilaterally with 0.2%moxonidine.A RPHPLC method was employed for the identification and quantification of moxonidine between 2 and 480 min,which presented in the aqueous humor and iris-ciliary body.Flash electroretinography(F-ERG)amplitude and superoxide dismutase(SOD)level were measured between day 1 and day 15 after topical administration with moxonidine in a rabbit model of high IOP.Histological and ultrastructural observation underwent to analyze the changes of retinal morphology,the inner retinal layers(IRL)thickness,and retinal ganglion cell(RGC)counting.RESULTS:Moxonidine was detectable between 2 and 480 min after administration,and the peak concentration developed both in the two tissues at 30 min,0.51μg/m Lin aqueous humor and 1.03μg/g in iris-ciliary body.In comparison to control,F-ERG b-wave amplitude in moxonidine eyes were significantly differences between day 3 and day 15(P<0.01)in the high IOP model;SOD levels were significantly higher at all time-points(P<0.01)with a maximum level of 20.29 U/mgprot at day 15;and RGCs were significantly higher(P<0.05).CONCLUSION:Moxonidine is a viable neuroprotective agent with application to high IOP model.All layers of retina,including RGC layer,retinal nerve fiber layer and INL,are more preserved after moxonidine administration.SOD plays a neuroprotective role in ocular hypertension-mediated RGC death.

RP-HPLC法同时测定紫苏叶提取物没食子酸、迷迭香酸、木犀草素含量研究

目的建立安徽地区紫苏叶中没食子酸、迷迭香酸和木犀草素成分的含量同时检测方法。方法采用反相高效液相色谱法,Symmetry C18(250 mm×4.6 mm,5μm)色谱柱,以甲醇(A)-0.1%磷酸溶液(B)为流动相梯度洗脱,流速1.0 mL/min,柱温30℃,检测波长272 nm同时检测没食子酸、迷迭香酸和木犀草素含量。采用SPSS 17.0对测定结果进行分析,比较本地主产区紫苏叶中各成分的含量差异。结果没食子酸、迷迭香酸和木犀草素分别在33.80~101.2μg/mL,33.60~100.8μg/mL,34.00~102.0μg/mL呈线性关系,平均加样回收率分别为98.04%(RSD=1.50%,n=6),96.40%(RSD=0.53%,n=6)和96.49%(RSD=1.89%,n=6)。结论此方法简单方便,稳定可重复,可作为安徽地区主产地紫苏叶没食子酸、迷迭香酸和木犀草素成分含量的检测依据。

RP-HPLC法同时检测石榴皮3种多酚类提取物含量研究

采用RP-HPLC法,Symmetry C18(250 mm×4.6 mm,5μm)色谱柱,以甲醇(A)-0.2%磷酸溶液(B)为流动相梯度洗脱,流速1.0 mL/min,柱温30℃,检测波长272 nm同时检测安石榴苷、鞣花酸和没食子酸含量。采用SPSS 17.0对测定结果进行分析,比较不同品种石榴皮中各成分的含量差异。结果表明,安石榴苷、鞣花酸、没食子酸分别在13.120~209.92μg/mL,13.325~213.20μg/mL和13.275~212.40μg/mL与峰面积呈线性关系,平均加样回收率为97.58%(RSD:1.02%,n=6),99.31%(RSD:0.94%,n=6)和98.63%(RSD:0.70%,n=6)。白玉石榴皮中安石榴苷、鞣花酸的含量分别为7.13%和1.00%,比红酸石榴皮中的含量高。安徽地区石榴皮中安石榴苷、鞣花酸、没食子酸3种成分含量同时检测方法的灵敏度高、准确度好、重现性一致,可用于安徽本地石榴皮部位中安石榴苷、鞣花酸、没食子酸成分含量同时检测。

PEAK IDENTIFICATION FROM INTERACTION INDEX IN REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

The interaction index c which was derived from the fundamental retention equationlogk’ =a + cC_B in reversed--phase high--performancc liquid chromatography (RP-HPLC) quan-titatively describes the difference in the interaction between solute--strong. solvent andsolute--weak solvent; it has shown to be a constant for a specific solute even when columnsystems with different C18 packings are used. The theoretical basis for peak identificationby using interaction index has been proposed, which was based on the a,c values on stan-dard C18 column by utilizing linear a-a plots on column pairs and the linear relationship be-tween parameters a and c for the structural related compounds. Through the establishmentof parameters a,c data based on the standard C18 column for a certain type of compounds,the retention of thesc compounds on various C18 columns can be predicted. Typical exam-ples have been given to verify the correctness of this method.

基于RP-HPLC双波长法的蓝芩口服液中主要成分的检测分析

目的基于反相高效液相色谱(RP-HPLC)双波长法测定蓝岑口服液中主要成分[包括栀子苷、(R,S)-告依春、盐酸小檗碱和黄岑苷]的检测价值。方法采用色谱柱Shimadzu C18柱,检测波长0~12 min为245 nm,12~20 min为275 nm;流速为1.0 ml/min;柱温为30℃;进样量为20μl;栀子苷、(R,S)-告依春、盐酸小檗碱检测波长240 nm,黄芩苷检测波长为275 nm。结果4种成分浓度与峰面积的线性回归方程分别是Y=1011X-31.41、Y=6408X-1.798、Y=4401X+28.61、Y=3755X-82.02,且在各自浓度线性范围内均呈现良好线性关系;平均加样回收率分别为98.31%、97.77%、98.33%和97.84%,相对标准差分别为1.12%、1.33%、0.74%和1.21%。结论基于RP-HPLC双波长法可以同时测定蓝岑口服液中栀子苷、(R,S)-告依春、盐酸小檗碱和黄岑苷的含量,且具有良好的稳定性、重复性和精密性,可以作为一种可靠检测方式应用于蓝岑口服液的质量控制中。

RP-HPLC法测定消炎退热颗粒中菊苣酸的含量

目的:建立以RP-HPLC法测定消炎退热颗粒中菊苣酸的含量测定。方法:岛津C18(250×4.6mm,5μm),流动相:乙腈-0.2%磷酸溶液(16:84),检测波长326nm,柱温30℃。结果:菊苣酸线性范围在2.345μg/mL~23.44μg/mL呈良好的线性关系,r=0.9999,平均回收率102.3%,RSD为2.8%。结论:该方法简便、准确、重现性好,为消炎退热颗粒质量评价提供了可靠的依据。

RP-HPLC法对乳制品中主要牛奶蛋白的分离及定量测定

建立一种基于反相高效液相色谱(RP-HPLC)的分析方法,对牛奶及其乳制品进行定量分析。此方法可一次分离牛奶及乳制品中的7种主要蛋白成分(κ-CN、αs2-CN、αs1-CN、β-CN、α-La、β-LgB、β-LgA),解决了加工后乳制品中变性蛋白定量不准确的问题和κ-CN、αs2-CN以及乳清蛋白中α-La、β-LgB和β-LgA难以分离的困难。各组分分离度均大于1,达到基线分离,回收率达88.9%～97.1%,相对标准偏差为0.8%～5.1%。该方法不需对样品进行离心,在测定清蛋白组分时,也不需提前去除优势蛋白酪蛋白,与传统的检测方法相比,具有准确、灵敏和快速等特点,可用于牛奶及其乳制品的质量控制和定量检测。

RP-HPLC法同时测定延胡索药材中延胡索乙素和原阿片碱

建立了反相高效液相色谱(RP HPLC)法同时测定延胡索药材中延胡索乙素与原阿片碱含量的方法,并对不同产地延胡索药材中延胡索乙素与原阿片碱含量进行了考察.结果表明,使用PhenomenexLunaC18色谱柱,甲醇 磷酸盐缓冲溶液(pH=7 68,V∶V=65∶35)流动相,在检测波长281nm下,测定延胡索乙素与原阿片碱的线性范围分别为0 13～4 68μg和0 09～3 24μg,平均回收率分别为101 86%和98 13%,RSD分别为1 42%和1 36%.结果表明,此法简便、准确、快速,可用于药材中延胡索乙素与原阿片碱含量的测定,也可用于含这两种生物碱的药材的质量评价;两种生物碱含量在不同产地及来源延胡索药材中有较大差别.

RP-HPLC法测定东北地区6种红树莓果实中有机酸组成

建立一种反相高效液相色谱法分离和测定6种红树莓果实中草酸、酒石酸、柠檬酸、DL-苹果酸和乳酸5种有机酸的方法。色谱条件为：采用Sino Chrom DS-BP C18色谱柱（150 mm×4.6 mm,5μm）,流动相为甲醇-0.01 mol/L KH2PO4溶液（p H 2.60,97∶3,V/V）,流速0.6 m L/min,柱温30℃,检测波长210 nm。结果表明：在此条件下5种有机酸都被有效地分离,各种有机酸的质量浓度与峰面积在测定范围内呈良好的线性关系,标准曲线相关系数在0.999 3-0.999 9之间;精密度实验相对标准偏差在0.20%-1.53%（n=5）范围内;回收率为98.83%-105.42%,相对标准偏差为0.06%-1.00%。测得6种红树莓果实中有机酸以柠檬酸为主,含量为1 058.41-1 825.45 mg/100 g,草酸、乳酸、DL-苹果酸含量较低,未检测到酒石酸。该方法简单、高效、准确、重复性好,可用于红树莓果实中有机酸的分离测定。

RP-HPLC法测定蔬菜、水果及食用菌中9种农药残留的研究

建立了反相高效液相色谱同时测定蔬菜、水果和食用菌中9种农药（吡虫啉、啶虫脒、多菌灵、甲基硫菌灵、嘧霉胺、除虫脲、灭幼脲、辛硫磷、阿维菌素）残留的可变波长检测器检测的分析方法。样品经乙腈提取,PSA粉固相分散萃取净化,取净化液氮吹至近干后,以1 mL甲醇定容,采用C18（250 mm×4.6mm,5μm）色谱柱分离,以甲醇-乙腈（体积比3∶1）和水二元流动相梯度洗脱,采用紫外可变波长检测器（270,258,275,254,280,245 nm）检测,流速1.0 mL/min,柱温40℃。结果表明9种农药在0.05～10.0mg/L浓度范围内线性关系良好;相关系数均大于0.999;检出限为0.006～0.07 mg/kg;平均加标回收率为81.0%～115.2%,相对标准偏差（RSD）为0.5%～10.2%。该方法具有快速、灵敏、准确、重现性好以及操作简单等特点,适用于蔬菜、水果以及食用菌中上述9种农药残留的分析。

RP-HPLC法测定苹果树枝 叶中根皮苷的含量

建立了一种快速测定苹果树枝、叶中根皮苷含量的RP-HPLC检测方法。色谱条件为Waters Symmetry C18柱(150mm×4·6mm,Φ5μm),流动相为甲醇和pH为2·6的磷酸水溶液(50∶50),等度洗脱,柱温为30℃,287nm紫外检测,流速为0·6mL·min-1,在10min内可实现苹果树枝、叶中根皮苷的快速分离。分析结果表明,根皮苷在0·30～4·50μg范围内呈良好的线性关系,相关系数R2=0·9993,加标回收率98·95%,RSD为1·22%,说明该方法准确可靠。

反相高效液相色谱法(RP-HPLC)测定冷却猪肉中生物胺

应用反相高效液相色谱法（RP-HPLC）建立同时测定冷却猪肉中多种生物胺的方法。样品经0.4 mol/L高氯酸提取后用丹磺酰氯柱前衍生，流动相为乙腈和水，采用梯度洗脱，流速为1.0 mL/min，荧光检测器激发波长（Ex）340 nm，发射波长（Em）515 nm。结果表明：采用该方法色胺、苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺8种生物胺能在25 min内得到很好分离。线性范围为0.5-20μg/mL（R^2〉0.99），回收率在82.43%-97.14%之间，相对标准偏差RSD小于10%。

RP-HPLC法测定血浆中阿魏酸哌嗪浓度

目的:建立血浆中阿魏酸哌嗪浓度的反相高效液相色谱法。方法:血浆标本以西沙比利为内标,经酸化和二氯甲烷提取后,以0．02 mol·L^(-1)磷酸二氢钠(pH2．7)-甲醇(45:55)为流动相,经碳十八烷基键合硅胶柱分离,在紫外310nm处检测。结果:阿魏酸哌嗪标准曲线范围0．016～16 mg·L～(-1),(r=0．9999),最低定量限为0．016 mg·L～(-1),高、中、低4种浓度的日内精密度均＜4%,日间精密度均＜6%,方法回收率93．8%～102．3%。结论:本法简单可靠,可用于临床阿魏酸哌嗪的药动学和药效学研究。

RP-HPLC法分析测定人参、西洋参、三七中的皂苷类化合物

利用反相高效液相色谱(reversed-phase high performance liquid chromatography,RP-HPLC)技术对人参、西洋参和三七不同方法提取物中的8种皂苷进行定量分析。采用乙醇回流、温浸法和超声辅助提取法提取3种药材中的有效成分,并结合高效液相色谱技术进行分析。结果表明:不同提取方法比较,超声辅助法所提取的3种药材中皂苷含量较高;3种五加科人参属的药材中,三七提取物中主要皂苷含量较高,人参提取物中的皂苷种类较多。

DNFB柱前衍生化RP-HPLC测定大黄种子氨基酸的含量

采用柱前衍生RP-HPLC法测定大黄种子中氨基酸的含量。6mol／L盐酸水解得到氨基酸，以2，4-二硝基氟苯（DNFB）为柱前衍生化试剂，梯度洗脱。色谱柱为Gemlmi—NX C18（4．6mm×250mm，5μm）；流动相A相为0．1M乙酸钠水溶液，B相为乙腈-水（1：1），柱温37℃，检测波长360nm。结果表明，三种正品大黄种子中均含有17种氨基酸，总量在11．30％-19．26％。该方法灵敏、准确，有良好的重复性和稳定性。

RP-HPLC同时测定人血清中维生素A和维生素E

目的：测定不同病理生理状态下人血清中维生素Ａ、维生素Ｅ的浓度，寻求弄清与某些疾病的关系，为临床治疗学、营养学等研究提供参考。方法：取血清２００μｌ加入含内标的无水乙醇液２００μｌ，振荡，加入正己烷５００μｌ，离心，取正己烷层１００μｌ进样；色谱条件为：色谱柱ＷａｔｅｒｓμＢｏｎｄａｐａｋＣ１８柱（３．９ｍｍ×３０ｃｍ），流动相甲醇水（９６∶４），内标维生素Ａ醋酸酯，检测波长３３０ｎｍ，２９２ｎｍ。结果：维生素Ａ在８６．８０４８～９７２．２１３０μｇ·Ｌ－１浓度范围内呈线性，ｒ＝０．９９９５，维生素Ｅ在１．２６６５～２５．３３１２ｍｇ·Ｌ－１浓度范围内呈线性关系，ｒ＝０．９９９６；维生素Ａ和维生素Ｅ的平均回收率分别为９６．４０％（ＲＳＤ＝３．９８％）和１０３．６７％（ＲＳＤ＝４．２６％），测定了２１８例人血清中维生素Ａ、维生素Ｅ的浓度。结论：方法快速、灵敏、准确，样品处理简便易行。

运用RP-HPLC同时测定火龙果中7种有机酸和Vc含量的方法研究

以紫红肉火龙果为试验材料,对影响有机酸分离的主要因素进行研究,最终建立了运用反相高效液相色谱法(RP-HPLC)同时快速测定火龙果果实中草酸、酒石酸、苹果酸、乙酸、乳酸、柠檬酸、琥珀酸7种有机酸及Vc含量的方法。采用色谱条件为:柱温30℃,流速1 m L/min,色谱柱为Kromasil柱(250 mm×4.6 mm),波长215 nm,缓冲液0.05 mol/L H3PO4-Na2HPO4;测定结果显示,供试火龙果含有7种有机酸和Vc,苹果酸是其最主要的有机酸、其次是柠檬酸。此方法可以在10 min内完成1次测定,具有分析速度快、灵敏度高、重复性好等优点,为果实中有机酸及Vc的测定及相关代谢研究提供参考。

RP-HPLC法测定细叶杜香叶中4种活性成分的质量分数

为了建立测定细叶杜香叶中秦皮乙素、秦皮素、金丝桃苷和槲皮素质量分数的方法,采用反相高效液相色谱法(色谱条件:色谱柱为Inertsil ODS-SP(5μm4,.6mm×250mm,柱号:9KI92609)、流动相为V(甲醇)∶V(0.2%醋酸)=43∶57、流速1.0 mL.min-1、检测波长350 nm),测得秦皮乙素、秦皮素、金丝桃苷和槲皮素4种成分的平均加样回收率分别为99.71%9、8.62%1、00.13%9、8.33%,其相对标准偏差RSD分别为1.45%1、.53%、1.04%1、.69%。结果表明:反相高效液相色谱法操作简便、准确,具有良好的重复性,可用于细叶杜香中此4种成分的质量分数的测定。