Determination of Benzo[a]pyrene in Edible Oil by High Performance Liquid Chromatography-Fluorescence Detector (HPLC-FL)

In this study, an optimized high performance liquid chromatography-fluorescence detector (HPLC-FL) method for the determination of benzo[a]pyrene in edible oil was established. HPLC was performed with Thermo Fisher Scientific C18 column (250 mm×4.6 mm, 5 μm) as the chromatographic column and acetonitrile and water as the mobile phase, and the excitation wavelength and emission wavelength of fluorescence detector were 286 and 430 nm, respectively. The response was high, and the linear range was 0.5-10.0 ng/ml. The lowest limit of detection was 0.11 ng/ml, and the average recovery was 92.5%. This method is suitable for quantitative analysis of benzo[a]pyrene content in edible oil.

Development of Quantitative Analysis Method of Carbendazim in Tomatoes by High-Performance Liquid Chromatography with Fluorescence Detection(HPLC-FLD)

Carbendazim belongs to the benzimidazole fungicides,which can be used for control lots of fungi pathogens.High-performance liquid chromatography is frequently used for the analysis of carbendazim in all kinds of samples.In most occasions,the developed methods were applied for the simultaneous detection of a huge number of pesticides.Thus,an analytical method via UPLC-FLD was developed,and the sample preparation process was optimized by studying the effect of extraction solvent,approach,time and purification absorbent on the recovery rate of carbendazim.The results showed the optimized method for analysis was ultrasonication-assisted extraction with acetonitrile for 1 min,and subsequent purification by C18.In this occasion,the established analytical method of carbendazim in tomato samples displayed good linearity,accuracy and precision.

Determination of 5-Fluorouracil in Human Plasma by High-Performance Liquid Chromatography (HPLC)

5-Fluorouracil (5-FU) has a broad spectrum of anti-tumor activity, widely applied to the treatment of cancers. However, it is necessary to determine the plasma concentration of 5-FU in clinical practice due to its narrow therapeutic index. Therefore, a simple, economic and sensitive high-performance liquid chromatography (HPLC) method was developed and validated for the determination of 5-FU in human plasma. Ethyl acetate was chosen as extraction reagent. Chromatographic separation was performed on a Diamonsil C18 column (250 mm × 4.6 mm i.d., 5 μm) with the mobile phase consisting of methanol and 20 mmol/L ammonium formate using a linear gradient elution at a flow rate of 0.8 mL/min. 5-FU and 5-bromouracil (5-BU) were detected by UV detector at 265 nm. The calibration curve was linear over the concentration range of 5—500 ng/mL and the correlation coefficient was not less than 0.992 6 for all calibration curves. The intra- and inter-day precisions were less than 10.5% and 4.3%, respectively, and the accuracy was within ±3.7%. The recovery at all concentration levels was 80.1±8.6%. 5-FU was stable under possible conditions of storing and handling. This method is proved applicable to therapeutic drug monitoring and pharmacokinetic studies of 5-FU in human.

Determination of Organic Acids in Root Exudates by High Performance Liquid Chromatography: I. Development and Assessment of Chromatographic Conditions *1

Methods for determining nine low molecular weight organic acids in root exudates were developed by using reversed phase high performance liquid chromatography with UV (ultraviolet) detection at 214 nm. The mobile phase was 18 mmol L -1 kH 2PO 4 adjusted to pH 2.25 with phosphoric acid and the flow rate was 0.3 mL min -1 . The analytical column was a reversed phase silica based C 18 column (Shim pack CLC ODS). The root exudates were collected through submerging the whole root system into aerated deionized water for 2 hours. The filtered exudate solutions were concentrated to dryness by rotary evaporation at 40 °C, dissolved in 10 mL mobile phase. The chromatographic conditions of organic acid determination were analyzed. The results showed that there was a high selectivity and sensitivity in the organic acid determination by reversed phase high performance liquid chromatography. Coefficients of variation for organic acid determination were lower than 10% except lactic acid. The recoveries were consistently between 80.1% to 108.3%. Detection limits were approximately 0.05 to 4.5 mg L -1 for organic acids except succinic acid with the detection limit of 7.0 mg L -1 . Phosphorus deficiency may contribute to the release of organic acids in soybean root exudates especially malic, lactic and citric acids.

Separation of Ephedrines Using 1-Butyl-3-methylimidazolium-tetrafluoroborate Ionic Liquids as Eluent in High-performance Liquid Chromatography (HPLC)

A simple, effective HPLC method for separation of ephedrines was achieved by using 1-butyl-3-methylimidazolium-tetrafluoroborate ionic liquid solution (0.1% v/v) as eluent at pH 3.0. The involved mechanism may be due to that the imidazolium cations can effectively shield the silanol groups of alkylsilica surface, thereby decreasing band tailing and increasing the separation efficiency.

Determination of AF and AFG in Red Ginseng by High Performance Liquid Chromatography with Evaporative Light Scattering Detector (HPLC-ELSD)

[Objective]The paper was to establish a method for determining AF and AFG in red ginseng.[Method]A new simple,rapid and sensitive method for simultaneous determination of two amadori compounds,arginyl-fructose(AF)and arginyl-fructosyl-glucose(AFG),in extracts of three kinds of ginseng preparations was developed and validated using high performance liquid chromatography with evaporative light scattering detector(HPLC-ELSD).Two target analytes were efficiently separated by Prevail CTM18 column(4.6 mm×250 mm,5μm)at the flow rate of 0.8 mL/min within 15 min of single chromatographic run.[Result]Under optimized conditions,the detection limits were 0.015 and 0.02 mg/mL for AF and AFG,respectively.Calibration curves of peak area for two analytes were linear over three orders of magnitude with the correlation coefficients greater than 0.999.The average recoveries,precision,reproducibility and stability for two analytes(AF and AFG)were 99.5% and 100.9%,0.43% and 0.47%,0.46% and 0.43%,0.41% and 0.49%,respectively.[Conclusion]This method was successfully applied for quantifying AF and AFG in red ginseng and the method was efficient,sensitive and accurate.

Sensitive and Rapid Quantification of Felodipine by High-performance Liquid Chromatography-tandem Mass Spectrometry(HPLC-MS/MS) and Its Pharmacokinetics in Healthy Chinese Volunteers

To investigate the pharmacokinetics of felodipine in the plasma of healthy Chinese volunteers, 30 healthy volunteers received a single oral dose of 5 mg of extended release felodipine tablets. The felodipine was extracted from the matrix with a liquid-liquid extract procedure and analyzed by high-performance liquid chromatography-tandem mass spectrometry in the multiple reaction monitoring（MRM） mode using an electrospray ion source with positive ion detection. The method was validated over a felodipine concentration range of 0. 05-10.00 ng/mL in human plasma. Its main pharmacokinetic parameters values were： ρmax = （ 1.67 ± 0. 84 ） ng/mL, occurring at （ 3.93 3± 2. 49 ） h; the plasma elimination half-life： （23. 08 3± 9. 48） h and the area under the plasma concentration versus time curve： （29. 94 ± 14. 39） ng · h/mL. The validation results demonstrated that this method showed a satisfactory precision and accuracy across the calibration range. The procedure involved minimal drug administration, sample preparation, and a 2. 5-min chromatographic run time. It was well suited to clinical studies of the drug involving large numbers of samples.

QuEChERS结合HPLC检测辣椒及其制品中天然辣椒素、二氢辣椒素和合成辣椒素

建立一种QuEChERS结合高效液相色谱快速检测辣椒及其制品中的天然辣椒素、二氢辣椒素和合成辣椒素的方法。样品用乙腈提取,无水硫酸镁、十八烷基硅烷(C18)、乙二胺-N-丙基硅烷化硅胶、石墨化炭黑复合吸附剂净化。采用Agilent Eclipse Plus C18(150 mm×4.6 mm,5μm)分离,以0.4%磷酸水溶液和90%乙腈溶液为流动相进行梯度洗脱,荧光检测器进行检测。结果表明:天然辣椒素、二氢辣椒素和合成辣椒素在0.20~100 mg/L范围内线性关系良好,决定系数(R2)均大于0.999。天然辣椒素的方法检出限为0.014~0.069 mg/kg,定量限为0.046~0.23 mg/kg;二氢辣椒素的方法检出限为0.023~0.12 mg/kg,定量限为0.077~0.39 mg/kg;合成辣椒素的方法检出限为0.018~0.088 mg/kg,定量限为0.059~0.30 mg/kg。在干辣椒、鲜辣椒、香菇酱、辣椒油4种基质中,3个不同添加水平下的加标回收率为91.6%~117.2%,相对标准偏差为0.2%~6.1%。该方法简便、准确,不仅适用于干、鲜辣椒,而且适用于高油、高蛋白等辣椒制品中天然辣椒素、二氢辣椒素和合成辣椒素含量的检测。

Modernization of Chinese herbal compound and the high performance liquid chromatography tandem mass spectrometry (HPLC-MS)

Chinese herbal compound is playing an important role on curing human diseases.And it has been a trend that Chinese herbal compound is being used all over the world in 21 century.However,our Chinese herbal compound is facing serious challenge for the lack of canonical system of quality criterion for Chinese herbal compound so it has been a urgent problem to set up the quality control standards and reveal therapeutic basis of Chinese herbal compound.In order to give full play to the advantages of Chinese herbal compound,modern scientific and technological is used to research of Chinese herbal compound,especially the high performance liquid chromatography tandem mass spectrometry(HPLC-MS),because it is high sensitive,rapid,and obtain more information.It is very necessary that HPLC-MS is uesed to elucidate the effective components of basic substances of Chinese Herbal Compound,and endow traditional Chinese medicine with modern scientific connotation.

Analysis of Amino Acids in Houttuynia Cordata Thunb. by High Performance Liquid Chromatography-Diode Array Detector (HPLC-DAD)

[Objectives] The biologically active compounds of Houttuynia cordata Thunb. as a medicinal and edible plant are well documented. However, amino acid as an important indicator of the value of H. cordata is little reported. [Methods] Amino acids of the H. cordata were analysed by pre-column derivatization with 2,4-dinitrochlorobenzne(CDNB) coupled to HPLC/DAD. The optimized conditions of the amino acids derivatized by CDNB were followed: 110 ℃ of hydrolysis temperature, 1∶12 of the equivalent proportion between amino acid and CDNB, 80 min of derivatization reaction time and 80 ℃ of derivatization reaction temperature. The method was used to determine the amino acids of H. cordata samples from 11 different geographical sites in Guizhou. [Results] The results suggested that the contents of 17 amino acids were quite variable at different H. cordata samples. [Conclusions] This study will provide a basis for the quality evaluation of H. cordata nutrition.

The New Type of Liquid Chromatography—Colloidal High-Performance Liquid Chromatography (CHPLC): Further Applications

The recent paper by I. Brondz (2018), “One-Step Procedure for Direct Purification of Pediocin-Like Bacteriocins and Cationic Antimicrobial Peptides from Complex Culture Medium on an Analytical, Semipreparative, and Preparative Scale. A New Type of Liquid Chromatography—Colloidal High-Performance Liquid Chromatography (CHPLC)” (International Journal of Analytical Mass Spectrometry and Chromatography, 6, 41-49, https://doi.org/10.4236/ijamsc.2018.63004), described a new type of CHPLC. This technique allows colloidal liquids and suspensions to be chromatographed directly despite the presence in the liquids of material such as bacteria, fungi, and other soft and hard particles. The significance of this development lies in enabling the single-step cleanup and concentration of the target substance from a complex mixture of soluble molecules in the presence of insoluble particles by high performance liquid chromatography (HPLC). The technique also allows the use of viscose liquids (as described in Brondz (2018) that are not suitable for analysis by conventional HPLC. In the previous paper, emphasis was placed on describing the applications of the techniques for the preparation of target substances such as small peptides, bacteriocins, bacitracin, and lysosome. Normally, the industrial preparation of these substances requires multistep procedures, which are time- and labor-consuming, and typically results in significant loss of target material and specific activity. In the present paper, the application of CHPLC for the isolation of alkaloids from crude raw material such as opium cake is demonstrated. In the opium cake, large amounts of hard vegetable particles and even sand corns are present together with the target alkaloids. Despite this, isolation by CHPLC of the desired compound was achieved in a single step by using a water/ethanol-based liquid. Isolation of alkaloids from such raw material normally requires a multistep procedure that includes the preparation of insoluble tartrate or picrate complex and this process includes dissolving the substance in flammable organic liquid. The isolation described here was performed in a single step by using the water/ethanol-based liquid.

HPLC法分析不同提取方法对桦褐孔菌中3种成分含量的影响

建立高效液相色谱检测桦褐孔菌粗提物中原儿茶酸、原儿茶醛和紫萁酮3种化合物含量的方法,对比评价超声辅助提取法、回流提取法、微波辅助提取法、酶解提取法4种不同提取方法对桦褐孔菌粗提物中活性成分含量的影响。结果表明,该检测方法精密度较高,3种化合物的精密度的相对标准偏差(relative standard deviation,RSD)分别为0.14%、0.06%、0.20%;重复性RSD分别为1.67%、1.98%、2.22%;在24 h内稳定性较好,其RSD分别为0.20%、0.61%、0.25%。对比不同提取方法对3种化合物的影响,发现采用超声辅助提取法得到的目标化合物产量较高,适用于桦褐孔菌中主要活性成分的提取。该研究建立的HPLC法检测桦褐孔菌粗提物中原儿茶酸、原儿茶醛和紫萁酮含量的结果准确。

HPLC法测定不同产地山慈菇中秋水仙碱含量及提取方法的探讨

以不同产地的山慈菇为样品,运用高效液相色谱法(HPLC)来测定样品中秋水仙碱的含量,并比较了不同的提取方法对结果的影响。采用高效液相色谱法测定山慈菇中秋水仙碱的含量,色谱柱:ODS C18(250 mm×4.6 mm,5μm),流动相:甲醇-水(54∶46),检测波长:243 nm,流速:1.0 mL/min;柱温:35℃;进样体积:5μL。秋水仙碱在0.16281~10.420μg/mL范围内线性关系良好(R2=0.9997),平均回收率为102.09%,RSD为3.73%。该研究发现山慈菇样品的产地和处理方法也会影响秋水仙碱的含量差异。因此,需要在实际提取中进行针对性的方法选择和优化。通过比较甲醇、50%甲醇、70%乙醇、三氯甲烷等不同有机溶剂在提取过程中的效果,得出了以50%甲醇为有机溶剂,超声30 min的提取工艺能够得到更高的提取率。这些研究结果有助于制定科学合理的山慈菇提取工艺和质量控制标准,为其广泛的开发利用提供理论依据和技术支持,也有助于为山慈菇生物活性成分的提取和分析提供参考。

通肠清胰合剂HPLC指纹图谱建立及多指标成分的含量测定

目的建立通肠清胰合剂的HPLC指纹图谱,并对5种成分进行含量测定。方法采用Agilent ZORBAX SB?C_(18)色谱柱(250 mm×4.6 mm,5μm),以乙腈?p H 5.0磷酸盐缓冲液为流动相,梯度洗脱,检测波长为230 nm和250 nm,流速为1.0 m L/min,柱温为30℃,进样量为10μL。通过中药色谱指纹图谱相似度评价系统(2012版)建立10个批次的通肠清胰合剂HPLC指纹图谱,同时进行相似度评价。结合软件SPSS 27.0、Origin Pro 2024b进行聚类分析和主成分分析。结果黄芩苷、芍药苷、大黄酸、厚朴酚、和厚朴酚分别在一定浓度范围内线性关系良好(r≥0.9998),平均回收率和RSD分别在96.39%~97.96%、1.54%~2.37%(n=6)之间。10批通肠清胰合剂指纹图谱显示存在12个共有峰,相似度为0.984~1.000。10批样品可聚为3类,其中S1~S7为第一类,S10为第二类,S8和S9为第三类。指认了5个成分,分别为大黄酸、芍药苷、黄芩苷、厚朴酚、和厚朴酚。结论本研究所建立的通肠清胰合剂HPLC指纹图谱及成分含量测定方法准确可靠、重复性强,为通肠清胰合剂的质量控制提供了科学依据。

影响HPLC在食品检测中准确性的因素及控制对策

为了提高高效液相色谱法(High Performance Liquid Chromatography,HPLC)在食品检测中的准确性,本文分析了样品前处理、仪器设备、检测人员和实验室环境等因素对检测结果的影响。分析认为,样品前处理不当、仪器设备老化、人员操作不规范以及环境条件不稳定是导致检测不准确的关键因素。针对这些问题,提出应根据食品成分特点优化样品前处理、完善设备维护制度、加强检测人员的专业培训和严格控制实验室环境条件的对策,以供参考。

基于HPLC指纹图谱与化学计量学研究常山柚橙幼果与枳实的相似性

目的探索常山柚橙幼果与药典基源枳实的相似性情况。方法采用高效液相色谱法建立常山柚橙幼果与枳实的指纹图谱,采用中药色谱指纹图谱相似度评价系统软件进行相似度分析,利用化学计量学对常山柚橙幼果与枳实全组分比较,并对14种化合物进行定量分析。结果①枳实间相似度与聚类分析结果受基源、产地的影响较大,甜橙与其他基源枳实的相似度仅有0.2;不同产地的酸橙相似度在0.802~0.986,相对离散;②常山柚橙幼果与部分酸橙、代代、臭橙和朱栾聚为一类,相似度>0.9;③变量重要性投影值(VIP)法筛选出21个对质量有重要作用的共有峰,主要有新橙皮苷、柚皮苷、伞形花内酯、川陈皮素、水合橙皮内酯和橙皮苷等。定量分析结果显示:常山柚橙幼果和枳实中富含柚皮苷和新橙皮苷,橙皮苷与芸香柚皮苷次之;不同基源枳实间主成分含量差异很大,特别是甜橙,常山柚橙幼果则相对稳定。结论常山柚橙幼果与大部分基源枳实具有较好的相似度,作为枳实药用具有一定的可行性。

Analysis of Pesticide Raid^(█) in Feed of Wistar Rat by High-Pressure Liquid Chromatography (HPLC)

The distribution of pesticide by-product in tissues of wistar rats were analyzed using high pressure liquid chromatography. The limit of detection of the HPLC was 0.1 μg. Results show bioaccumulation factor of pesticide “Raid?” in lipid, up to three times that of the feed at the first concentration and gradually decreased as the concentration increased in the muscle > (0.7), brain > (0.5) and liver > (0.3) as indicated in the text. At higher concentration of 961 μg/g, bioaccumulation factor decreased in the lipid to 1.2 and 0.6 in the muscle, 0.03 in the brain and 0.08 in the liver respectively. High Pressure Liquid Chromatography (HPLC) analysis of raid extract suggests the presence of micprothrin and palethrin. The implications are numerous, but simply put that accidental ingestion of chlorinated hydrocarbon as in “Raid?” may involve convulsions, collapse and coma after only brief excitation and ataxia at the onset.

Simultaneous determination of five nucleosides and nucleobases in Panax notoginseng using high-performance liquid chromatography

Aim To quantitatively determine five nucleosides and nucleobases, including cytidine, uridine, guanosine, adenosine and uracil in different parts of Panax notoginseng. Methods Separation was performed on a Zorbax SB-Aq column using a gradient elution with mobile phase of 8 mmol^L-1 ammonium acetate aqueous solution （A） and methanol （B）. The assay was carried out at a flow rate of 1 mL·min^-1 at 25 ℃ with the diode-array detection at 260 nm. Results Cytidine, uridine, guanosine, adenosine and uracil had good linearity in the ranges of 1.79 - 57.40 μg·mL^-1 （r^2 = 1.0000）, 3.30 - 105.60 μg·mL^-1 （r^2 = 1.0000）, 3.09 - 98.80 μg·mL^ -1（r^2 = 0.9999）, 2.77 - 88.60 μg·mL^-1 （r^2 = 1.0000） and 0.38 - 12.30 μg·mL ^-1 （r^2 = 1.0000） with average recoveries of 93.9%, 96.5%, 92.7%, 93.2% and 98.8%, respectively. The content of cytidine, uridine, guanosine, adenosine and uracil in different parts of P. notogingeng were significantly different. Conclusion This is the first report on quantitative determination of nucleosides and nucleobases in P notoginseng.

HPLC法测定复方氨酚烷胺颗粒中的对氨基酚杂质研究

目的建立复方氨酚烷胺颗粒中对氨基酚(PAP)杂质的高效液相色谱(HPLC)检测方法及质控要求。方法采用HPLC法,C18色谱柱(250 mm×4.6 mm,5μm),流动相为0.5%乙酸铵溶液水-甲醇(80∶20),DAD检测器,波长为245 nm,柱温为30℃,对复方氨酚烷胺颗粒中的对氨基酚杂质进行检测。结果采用本文建立的HPLC法可有效检测出复方氨酚烷胺颗粒所含的杂质对氨基酚,分离度可达4.3,检测限达0.2μg/ml,定量限为8μg/ml下回收率为98.96%。结论运用本文建立的HPLC法可以对含有对乙酰氨基酚成分的感冒药中杂质进行质量控制。

Simultaneous Determination of Tetramethylpyrazine and Aspirin in a New Compound Formulation by Liquid Chromatography

Aim To establish a reversed-phase liquid chromatographic (LC) method forsimultaneous determination of tetramethylpyrazine (TMP) and aspirin in a new compound formulation.Methods Chromatographic separation of the two drugs was achieved on a Diamonsil C_(18) column, usinga binary mixture of methanol-1.5% acetic acid (35:65, V/V, pH = 3.1) as mobile phase at a flow rateof 1.0 mL·min^(-1). Results Separation was completed in less than 12 min. Benzoic acid was used asthe internal standard. Recoveries at levels corresponding to 80 % to 120 % of the label claim ofthe formulation ranged from 99.6 to 100.3 % for aspirin and from 99.9 to 101.3% for TMP. The linearrange was 12.6 - 150.9 μg·mL^(-1)(r= 0.9997, n = 5) for aspirin and 25.0- 300.0 μg·mL^(-1) (r =0.9999, n = 5) for TMP. Conclusion The method developed can be used for the simultaneousdetermination of TMP and aspirin in pharmaceutical preparations.