Benefits and Pitfalls of HPLC Coupled to Diode-Array, Charged Aerosol, and Coulometric Detections: Effect of Detection on Screening of Bioactive Compounds in Apples
Marcela HolláAneta BílkováPavel JakubecStanislava KoškováHana VlčkováDalibor Šatı́nskýFrantišek ŠvecHana Sklenářová
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The new screening method for rapid evaluation of major phenolic compounds in apples has been developed. Suitability of coupling HPLC/UHPLC separation with the diode-array detection and universal charged aerosol detection with respect to the presence of interfering substances was tested. Characteristics of both detection techniques were compared and method linearity, limits of detection and quantitation, and selectivity of them determined. Student t-test based on slopes of calibration plots was applied for the detailed comparison. The diode-array detection provided the best results regarding sensitivity and selectivity of the developed method in terms of evaluation of phenolics profiles. The response of the charged aerosol detector was negatively affected by co-eluting substances during rapid-screening analyses. Coulometric detection was used for advanced characterization of extracts in terms of antioxidant content and strength to obtain more complex information concerning sample composition. This detection also allowed evaluation of unidentified compounds with antioxidant activity. HPLC/UHPLC separation using a combination of diode-array and coulometric detectors thus represented the best approach enabling quick, yet complex characterization of bioactive compounds in apples.Keywords:
Coulometry
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Objective:To determine the content of Astragoloside IV in Xiaoshuantong Granules by HPLC-ELSD. Methods:HPLC- ELSD separation was achieved. Results:The astragaloside Ⅳ showed good linearity (r=0.9999)in the range of 1.08μg-6.48μg, the average recovery was 100.2% with RSD 0.93%. Conclusions:The method is sensitive,reliable,accurate and repeatability,it can be used for quality control method of Xiaoshuantong granules.
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In the present study, a new analytical method was devised for the simultaneous determination of soluble carbohydrates in soybean seeds using high performance liquid chromatography/evaporative light scattering detection (HPLC/ELSD). The limit of quantification (LOQ) for soybean soluble carbohydrates ranged from 5.6~7.6 mg/kg using the HPLC/ELSD method and from 16.2~33.9 mg/kg using the high performance liquid chromatography/refractive index detection (HPLC/RID) method. Therefore, the HPLC/ELSD method was more sensitive than HPLC/RID. The precision values for retention time and peak area of the HPLC/ELSD method were evaluated by inter-day (n=5) and intra-day (n=10) assays using a standard solution. All precision values (CV<2.5%) for soybean soluble carbohydrates were acceptable and fulfilled international acceptance criteria. All linear calibration curves were obtained with a correlation coefficient of R2>0.999. The contents of soluble carbohydrates for the
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A rapid and simple analysis for the identification/quantification of L-ascorbic acid (AsA) in fruit drinks by high-performance liquid chromatography (HPLC) with a photo-diode array detector is developed. The presence of dehydroascorbic acid is reduced by dithiothreitol before HPLC analysis. Hypersil ODS column (250 × 4.6 mm i.d.) with a mobile phase of 2% (v/v) acetic acid solution is used for the HPLC determination. AsA in sample can be identified by the retention time and absorption spectrum.
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OBJECTIVE To compare the advantages and disadvantages of the HPLC-ELSD and UV method for determination of Polysorbate 80 in Huangqi Injection.METHODS HPLC-ELSD and UV were used for testing;The HPLC separation was carried out on a molecular gel column,using acetonitrile-water(90:10) as mobile phase.The flow rate was 0.6 mL·min-1,temperature for the drift tube was 110 ℃ and the nitrogen flow rate was 2.3 L·min-1.RESULTS There was a relationship between the results of these two methods,that was to say,the result of UV was about 1.54 times that of HPLC-ELSD.Compared with the result of UV,that of HPLC-ELSD was closer to the theoretical value of Polysorbate 80 In Huangqi Injection.CONCLUSION The HPLC-ELSD was simple,accurate and more conducive to control the content of Polysorbate 80 in Huangqi Injection.
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Polyvinyl Alcohol
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复方贞术调脂胶囊（FTZc）是一种用于治疗高脂血症的专利药物,由八味中药组成,含有多活性成分的复方制剂。本文建立了可行的HPLC-DAD-ELSD方法用于FTZc的质量控制,同时测定15个活性成分（红景天苷、特女贞苷、木兰碱、迷迭香酸、丹酚酸B、非洲防己碱、表小檗碱、药根碱、黄连碱、巴马汀、小檗碱、5,7-二甲氧基香豆素、人参皂苷Rg1、人参皂苷Rb1和齐墩果酸）。色谱柱采用Ultimate XB C18,流动相为乙腈–水（0.25%乙酸,0.13%三乙胺）（梯度洗脱）,流速为0.8mL/min,检测方法采用光电二极管阵列检测器的多波长检测模式和蒸发光散射检测器联合应用。方法学验证表明该方法精密度、准确度良好,成功用于不同批次FTZc的测定。结果表明,本文建立的测定多种生物活性成分HPLC-DAD-ELSD方法切实可行、方法可靠,特别适用于复方贞术调脂胶囊的质量评价。
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目的 优化胃康颗粒中人参皂苷Rb1和黄芪甲苷的提取方法，并建立其含量测定方法。 方法 以人参皂苷Rb1和黄芪甲苷的含量作为指标，采用单因素考察法对提取工艺进行优化；采用高效液相色谱-蒸发光散射法（HPLC-ELSD），XBridge®Shield RP18（4.6 mm×250 mm，5 μm）为色谱柱；乙腈-水（32：68）为流动相；柱温为30 ℃；漂移管温度为60 ℃，载气流量为1.7 SLM，建立测定胃康颗粒中人参皂苷Rb1和黄芪甲苷含量的方法。 结果 当采用甲醇回流提取1.5 h，正丁醇提取5次，氨水洗涤2次时，人参皂苷Rb1和黄芪甲苷的提取含量较高；建立的HPLC-ELSD法测定人参皂苷Rb1和黄芪甲苷含量，线性关系良好（r > 0.9997），日内日间精密度均小于1%，加样回收率分别为95.65%和100.57%，稳定性和重复性的RSD均小于3%，含量分别为2.8630 mg/g和0.2576 mg/g，RSD分别为0.62%和1.51%。 结论 优化了胃康颗粒中人参皂苷Rb1和黄芪甲苷的提取方法，建立了可靠、准确、重现性好的测定胃康颗粒中人参皂苷Rb1和黄芪甲苷含量的HPLC-ELSD方法。
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OBJECTIVE To establish a HPLC-DAD-ELSD method for the deter mination of the content of cuscohygrinol,acetylmandelic acid and other related compounds in cuscohygrinolis α-acetylbenzoacetate.METHODS The HPLC method was conducted on a Waters symmetry C18 column(4.6 mm×150 mm,5 μm)and coupled with ELSD and DAD detector at 219 nm.The mobile phase consisted of methanol-0.25% of pentafluoropropionic acid(contains 30 mmol.L-1 of ammonium formate),and the column temperature was at room temperature.RESULTS A good resolution was obtained among cuscohygrinol,acetylmandelic acid,cuscohygrinolis α-acetylbenzoacetate and other related compound.The linear ranges of cuscohygrinol,acetylmandelic acid and cuscohygrinolis α-acetylbenzoacetate were 0.016 32-0.163 2,0.005 65-0.113 and 0.228 2-2.282 g.L-1,respectively.The average recovery of cuscohygrinol,acetylmandelic acid and cuscohygrinolis α-acetylbenzoacetate were 100.1%,101.2% and 99.6% respectively.CONCLUSION A simple,accurate and sensitive HPLC-DAD-ELSD method to determine the content of cuscohygrinol,acetylmandelic acid and other related compounds in cuscohygrinolis α-acetylbenzoacetate was established.
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Objective To establish the HPLC-ELSD of n-butylol froution of Altemanthera philoxeroide.Methods The gradient elution was adopted with a mixture consisting of acetonitrile and water,the flow rate was 1.0 ml·min-1,and the detector was ELSD.Results The constituents of Altemanthera philoxeroide were well separated on HPLC and the HPLC-FPC of 10 batches of samples was established.The results showed that this method had a good repeatability and the 12 common peaks had an exclusive properties.Conclusion The method developed can supply the reference for the quality evaluation of Altemanthera philoxeroide.
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High‐performance liquid chromatography (HPLC) for analysis of phospholipids has traditionally employed ultraviolet detection of the eluted compounds. The evaporative laser light‐scattering detector (ELSD) offers new opportunities for quantitative analysis of lipids. Phospholipids were isolated from crude and degummed oils prepared from soybeans subjected to storage at high moisture content. Analytical and preparative separations of phospholipids by normal‐phase HPLC were accomplished. Major class fractions were analyzed by transmethylation and capillary collumn chromatography for fatty acid composition, and by reverse‐phase C‐18 HPLC (RP‐HPLC) for molecular species composition. The RP‐HPLC‐ELSD system was limited to the analysis of phosphatidylcholine and phosphatidylethanolamine.
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Phosphatidylethanolamine
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