提高液相色谱法检测食品中食品添加剂准确性的对策

针对日常工作中高液相色谱法检测食品中添加剂时容易出现的问题，从实验前的准备工作、样品处理、分析过程、实验后处理四个方面分析了可能出现的问题，总结了一些应对措施，对日常监督工作有一定的指导意义。

快速高分离度液相色谱法测定贝类中软骨藻酸残留量

介绍了用快速高分离度液相色谱（RRLC）测定贝类中记忆丧失性贝类毒素软骨藻酸的方法。样品以V（甲醇）：V（水）=1：1的溶液提取，经LC—SAX强阴离子柱固相萃取净化，以ZORBAX SB-C18柱（1．8μm，2．1×50mm）分离，乙腈＋0．1％三氟乙酸水溶液等度洗脱，1min内完成检测。用RRLC外标法定量分析。在10，15，20，40mg／kg4个添加水平下进行了验证试验，方法在0～18mg／L范围内有良好的线性关系，相关系数大于0．999，回收率为79．3％～106％。

反相高效液相色谱法测定栀子药材及其制剂中栀子甙的含量

目的:反相高效液相色谱法测定振子药材及其制剂中栀子甙的含量。方法:采用C_(18)ODS柱,甲醇-水(30:70)为流动相,238nm为检测波长。结果:栀子甙浓度在 0.317～2.54μg.ml^(-1)范围内,呈良好的线性关系(γ=0.9999),平均回收率为98.27％,RSD为1.96％(n=5)。结论:本 法简便,快速,专属性强。

高分离度快速液相色谱法测定黄芪药材中毛蕊异黄酮含量

目的:采用高分离度快速液相色谱法测定黄芪药材中毛蕊异黄酮的含量。方法:色谱柱:安捷伦Zorbax SB-C_(18)柱(50mm×4.6mm,1.7μm);流动相:0.1%甲酸水溶液(A)-乙腈(B),梯度洗脱,流速:0.6 ml·min^(-1);柱温:25℃;检测波长:280nm。结果:毛蕊异黄酮在6.05～121.00μg·ml^(-1)浓度范围内与峰面积呈良好线性(r=0.9999),平均回收率为96.17%(RSD=2.84%,n=6)。结论:该方法简便、快速、准确,重复性好,可用于黄芪药材的质量评价。

高效液相色谱法测定琥珀酰明胶注射液中明胶的含量

目的 建立高效液相色谱法测定琥珀酰明胶注射液中明胶的含量。方法 采用高效液相色谱法，3根凝胶色谱柱串联，流动相为0．6％磷酸盐，流速0．8ml／min；进样量100μl；柱温：30℃；采用示差折光检测器。结果 琥珀酰明胶在3．0-12mg．ml^-1浓度范围内呈良好的线性关系，r=0．9999（n=5），平均回收率为99．58％，RSD；0．16％。结论 本方法较紫外分光光度法更方便、快捷，且可与琥珀酰明胶注射液相对分子量及相对分子量分布测定同时进行。可作为中间产品的质量控制方法。

反相高效液相色谱法测定达格列净的有关物质

目的建立反相高效液相色谱法(RP-HPLC)测定达格列净的有关物质。方法采用Kromasi L 100-5-C18色谱柱(4.6 mm×250 mm,5μm);以水为流动相A,乙腈为流动相B进行线性梯度洗脱;流速为1.0 ml/min;检测波长为224 nm;柱温为30℃。结果达格列净与相关杂质均分离度良好;达格列净与特定杂质Dapa-7e、Dapa-22的线性范围分别为0.1747~4.3686μg/ml、0.2320~5.7997μg/ml和0.1970~4.9260μg/ml,r均>0.999,方法准确度均>99.0%。结论本方法简便、准确、迅速,可用于达格列净中有关物质的测定。

液相色谱-质谱联用法测定牛奶中4种四环素类药物残留量

目的建立牛奶样品中四环素、土霉素、金霉素、美他环素药物残留量的测定方法 ,进一步探讨四环素类药物的电喷雾质谱规律。方法生物样品经固相萃取后 ,采用HypersilBDS柱分离 ,以乙腈 水 甲酸 (体积比 37 5∶6 2 5∶1 2 )作为流动相 ,采用电喷雾离子源 ,以正离子检测方式进行一级、二级质谱分析。结果牛奶中四环素、土霉素及美他环素的检测限可达 0 0 5 μg/mL,金霉素的检测限可达 0 1 μg/mL。四环素、土霉素及美他环素的浓度在 0 1～ 5 0 μg/mL内 ,金霉素浓度在0 2～ 1 0 0 μg/mL内均呈线性。结论该方法专属性强 ,灵敏度高 。

反相高效液相色谱法测定乙酰麦迪霉素血药浓度

应用反相高效液相色谱法测定乙酰麦迪霉素血清浓度。色谱柱为 ODS O_(18)柱,流动相为甲醇一水(80:20,v/v),流速1.0 ml/min,检测波长254 nm,柱温25℃。线性范围在10～300 ng/ml,检测下限为10 ng/ml 高、低浓度平均回收率分别为101.02%和105.44%,日内,日间测定 RSD 分别为5.12%(n=10)和7.38%(n=7)。应用本法测定健康人单剂量日服乙酰麦迪霉素干糖浆后的血药浓度,结果满意。

高分离度快速液相色谱法测定不同产地枳实中主要黄酮类成分的含量

目的:建立高分离度快速液相色谱法(RRLC)测定枳实中主要黄酮类成分含量的方法。方法:RRLC色谱条件为Zobax Extend-C18(4.6mm×50mm,1.8μm)色谱柱,流动相为甲醇-2%乙酸(33∶77);检测波长283nm;流速1mL·min-1;柱温30℃。结果:27个批次样品中有19个批次中的主要黄酮类成分为橙皮苷、柚皮苷、新橙皮苷和柚皮芸香苷,按各成分含量比例它们又可以分为两组,一组以柚皮苷和新橙皮苷含量较高,另一组则以柚皮芸香苷和橙皮苷含量较高;其余8个批次的主要黄酮类成分为橙皮苷和柚皮芸香苷,不含有柚皮苷和新橙皮苷,并都以橙皮苷含量较高。各样品主要黄酮成分含量之和为12.4%～38.07%。结论:RRLC方法稳定可靠,分析时间是HPLC的三分之一。枳实中黄酮类成分的种类和各成分的含量比例是不同的。

高分离度快速液相色谱法测定小儿清热利肺口服液中绿原酸和牛蒡苷的含量

目的:建立同时测定小儿清热利肺口服液中绿原酸和牛蒡苷含量的方法。方法:采用高分离度快速液相色谱法。色谱柱为ZorbaxSB-C18柱,流动相为0.1%冰醋酸溶液-乙腈(梯度洗脱),检测波长0～5.5min为327nm,5.5～7.0min为280nm,流速为1.0mL.min-1,柱温为20℃,进样量为1.0μL,采集频率为1s。结果:绿原酸、牛蒡苷进样量分别在0.014602～0.29204、0.051396～1.02792μg范围内与峰面积积分值有良好的线性关系(均r=0.9999);平均加样回收率分别为99.56%、99.96%,RSD分别为1.22%、1.09%(n=6)。结论:本方法操作简便易行,可在保持分离度的前提下缩短分离时间,减少溶剂消耗,节约分析成本,提高日常检验工作的效率,可用于控制该药的生产工艺及质量。

高分离度快速液相色谱法测定黄芪药材中毛蕊异黄酮苷含量

目的:采用高分离度快速液相色谱(RRLC)法快速测定黄芪药材中毛蕊异黄酮苷的含量。方法:采用RRLC法,色谱柱:安捷伦Zorbax SB–C18柱(4.6 mm×50.0 mm,1.8μm);流动相:0.3%甲酸水溶液(A)-乙腈(B),梯度洗脱程序:(0～6.0min,15%～20%B;6.0～14.0 min,20%～45%B;14.0～18.0 min,45%～65%B;18.0～20.0 min,65%～100%B)流速:0.6mL·min-1;柱温:25℃;检测波长:280 nm。结果:毛蕊异黄酮苷在5.6～112.0 mg·L-1浓度范围内与峰面积呈良好线性(r=0.999 8),平均回收率为97.78%(n=6,RSD=2.87%)。结论:RRLC法简便、快速、准确,重复性好,可用于黄芪药材中毛蕊异黄酮苷的质量评价。

HPLC法测定血瘀证患者口服川芎丹参煎剂后阿魏酸血药浓度

目的 对以往建立的人血清中阿魏酸浓度测定的 HPL C法加以改进 ,并测定了血瘀证患者口服川芎丹参煎剂后阿魏酸的血药浓度。方法 以香豆素为内标 ,甲醇 -水 -醋酸 (40∶ 5 9.7∶ 0 .3)为流动相 ,Diamonsil(TM)色谱柱 (15 0 m m× 4 .6 m m,5μm ) ,流速 1.0 m L / min,以二极管阵列检测器测定 32 2 nm波长下的数据 ,以沸水浴法加甲醇处理血清样品。结果 阿魏酸和内标分离完全 ,阿魏酸在 32 .18～ 96 5 .38ng/ m L 范围内线性关系良好 (r=0 .9981) ,日内和日间 RSD<10 % ,平均回收率为 (99.3± 5 .4 ) % ,最低检测限为 1.6 ng/ m L,最低检测浓度为 15 .7ng/ m L。结论 此法简单、快速、灵敏而准确 。

抗癌药CA4P与不同种属血浆蛋白结合的规律

目的研究抗癌药Combretastatin A4 phosphate（CA4P）与不同种属血浆蛋白结合的规律。方法采用平衡透析法分离结合药物和游离药物，采用HPLC法测定Combretastatin A4（CA4）的浓度。结果经24h的透析，CA4在3种浓度下与大鼠血浆的蛋白结合率分别是77．86％、74．70％、64．46％；与Beagles犬血浆的蛋白结合率分别是81．70％、81．32％、79．68％；与人血浆的蛋白结合率分别是84．55％、77．04％、71．70％。结论CA40．918—36．720μg·ml-1，是一种中等程度蛋白结合的药物，虽随CA4浓度的增加结合率略下降，但不同浓度的CA4与大鼠、犬和人血浆蛋白的结合率无统计学差异（P〉0．05）。

HPLC同时检测酸性橙Ⅱ和碱性嫩黄O色谱条件优化

该研究建立高效液相色谱法同时检测食品中酸性橙Ⅱ和碱性嫩黄O的方法,对高效液相色谱中色谱柱、柱温、流动相种类及比例、流速、波长等影响因素进行了研究及优化,得到色谱检测条件为:色谱柱为Kromasil C_(18)(4.6 mm×150 mm,5μm);流动相为甲醇-50 mmol/L乙酸铵溶液=50∶50(V/V);流速:1.0 m L/min;检测波长450 nm;柱温35℃。在此最佳条件下,碱性嫩黄O及酸性橙Ⅱ的分离度好,且峰型正常。结果表明,该方法适合食品中碱性嫩黄O及酸性橙Ⅱ的检测。

Stability of Baicalin Aqueous Solution by Validated RP-HPLC

Aim In the present study a RP-HPLC method was developed and validated toinvestigate the stability of baicalin aqueous solution. Methods The influences of temperature and pHon the stability of baicalin aqueous solution were investigated by classic homoiothermicacceleration test, and the pH for the most stable solution was determined. Results The time whenbaicalin suffered 10% loss was found to be 18.1 h, and the degradation activation energy of baicalinwas 79.1 kJ·moL^(-1) . The pH at which baicalin is most stable is 4.28. Conclusion The temperatureshould be kept at a lower level and the pH should be adjusted to near that for the most stablesolution in the production of baicalin preparations.

Enantioseparation of Zolmitriptan by HPLC

Aim To establish a HPLC method for the separation of the enantiomers of zolmitriptan. Methods The separations were performed on Chiralcel OJ column with hexane-ethanol-diethylamine（85：15：0.2） as mobile phase at a flow rate of 0.8 mL·min^-1 and detecttion wavelength of 227 nm at 35 ℃. Several related parameters for separation were studied. Results Baseline separation （Rs 〉 1.5） was easily obtained in the case, and the R-isomer impurity in zolmitriptan was determined. Conclusion The method developed in this study has been successfully applied for quality-control purposes.

HPLC Determination of Harpagoside and Cinnamic Acid in Radix Scrophulariae

A gradient HPLC method was established for the determination of harpagoside and cinnamic acid in Radix Scrophulariae (Xuanshen) and a proposition was put forward for the lowest content of the characteristic and active constituent (harpagoside) for qualified Radix Scrophulariae. The experimental conditions were as follows: Ultrasphere ODS column (250 mm 4.6 mm, 5 mm), mobile phase: acetonitrile-water (containing 1.0% acetic acid) (20:8050:50) (20 min), flow-rate 1 mLmin-1, room temperature, detection wavelength 278 nm. Twenty-eight samples of Xuan-shen (Radix Scrophulariae) from different districts of China were analyzed and the contents of harpagoside and cinnamic acid in Xuan-shen were 0.041~0.244% and 0.012~0.068% respectively. The recoveries (RSD)% were 97.13(0.80)% for harpagoside and 99.38(0.51)% for cinnamic acid. The method is simple and accurate. It can be used for the quality control of Radix Scrophulariae. We propose that the content of harpagoside in qualified Radix Scrophularia should be no less than 0.05%.

Evaluation on Uncertainty of Determining Aspartame in Beverage by HPLC

[Objective] The aim was to evaluate the uncertainty of determining aspartame in beverage by high performance liquid chromatography （ HPLC）. [Method] The content of aspartame in beverage was determined by HPLC, then the source of uncertainty in the whole determination process was analyzed, and each component of uncertainty was evaluated and combined. [ Result] Through 6 repeated determinations by the method in GB/T 22254-2008 ＂Determination of Aspartame in Food＂, the average content of aspartame in beverage was （0.806 ±0.038） g/kg, and k =2. The main sources of uncertainty to affect the process were the sample weighting process, the preparation process of standard solution introduced by sample constant volume and the uncertainty introduced by fitting standard curve. ①The uncertainty of standard work-solution. The combined uncertainty of standard work-solution was 0.013 9, among them the uncertainty introduced by standard sample purity was 0.005 8, the standard uncertainty introduced by standard material weighting was 1.49 ×10^4, the relative uncertainty introduced by glass apparatus calibration in the preparation process of aspartame standard reserving solution was 0. 007 88, and the uncertainty introduced by glass apparatus calibration in the preparation process of standard work-solution was 0. 009 9. ②The uncertainty introduced by the preparation process of sample specimen. Among them the relative standard uncertainty introduced by sample weighting process was 0. 009, and the uncertainty introduced by sample constant volume was 0.000 78. ③The uncertainty introduced by the fitting process of standard curve. Among them the relative uncertainty of curve fitting was 0.002 46, the uncertainty introduced by the determined results of aspartame was 0.017 0, the total combined standard uncertainty was 0.023 9, and the expanded standard uncertainty was 0.019. [ Conclusion] The uncertainty components of standard solution, standard curve and repeatability are the main sources of uncertainty, while those of sample weighting and sample constant volume account for little proportion.

Pharmacokinetics of Scutellarin in Dogs

Aim To establish an RP-HPLC method for the determination of scutellarin in plasma and study its pharmacokinetics in dogs. Methods Scutellarin was given to dogs by intravenous injection and determined by RP-HPLC, the mean plasma concentration-time curve was plotted and pharmacokinetic parameters were calculated by program 3p87. Resu;ts The concentration-time curve of scutellarin could be fitted to three-compartment model with T1/2 pi, T1/2 α and T1/2 β being 1.05 ± 0.80 min, 6.99 + 2.76 min and 51.61 + 28.78 min, respectively, Vc being 880.1 + 508.3 mL, CL being 189.6 + 53.8 mL@ min- 1, and AUC0-90 and AUC0-∞ being 574.43 + 133.95 μg@ min@ mL - 1 and 599.34 ± 132.00μg@ min@mL- 1, respectively. Conclusion The fact that the concentrations of scutellarin in plasma declined rapidly after the medication suggested that the T1/2 of scutellarin should be taken into account in drug administration and preparation development.

Qualitative and Quantitative Analysis of Rhizoma of Polygonum cuspidatum

Aim To establish reliable methods for evaluating the quality of rhizoma of Polygonum cuspidatum( Huzhang in Chinese). Methods TLC and HPLC were employed for the chemical identification and content determination,respectively. Results A qualitative TLC method and a quantitative HPLC method with piceid as the reference substance were established, respectively. With piceid as the reference substance and ethyl acetate-methanol-formic acid-water ( 19:3:0.5:1) as the mobile phase, a TLC method for the identification of Huzhang from the commonly used crude drugs of the same family was also set up. Conclusion The established TLC method can reasonably appraise the quality of the drug and easily distinguish Huzhang from the other commonly used crude drugs of the same family. The HPLC method for determining piceid is simple, reproducible, accurate, and feasible.