HPLC determination of rifampicin and related compounds in pharmaceuticals using monolithic column

A rapid, sensitive and reproducible HPLC method using C18 monolithic column was developed and validated for the analysis of rifampicin (RIF) and its four related compounds, including rifampicin quinone (RQ), rifamycin SV (SV), rifampicin N-oxide (RNO) and 3-formylrifamycin SV (3-FR). Chromatographic separation was achieved by using the mixture of methanol-acetonitrile-monopotassium phosphate (0.075 M)-citric acid (1.0M) (28:30:38:4, v/v) as the mobile phase at a flow rate of 2 mL/min and with UV detection at 254 nm. Calibration curves were obtained in the concentration ranges of 1-40 microg/mL for SV, RNO and 3-FR, 1.5-60 microg/mL for RQ and 5-200 microg/mL for RIF. Limit of quantitation (LOQ) was determined to be 1 microg/mL and the limit of detection (LOD) was 0.2 microg/mL for all studied compounds with a 10 microL injection. The intra-day R.S.Ds. and inter-day R.S.Ds. for the above five compounds were all less than 2.5%. The recoveries of rifampicin from placebo tablets were from 99.7% to 100.5%. The total run time was less than 11 min, as opposed to around 60 min by using C18 particle-packed column. In conclusion, by this developed method, RIF and its related compounds can be determined rapidly with good precision and accuracy in pharmaceuticals.     

Determination of phenolic compounds in Prunella L. by liquid chromatography-diode array detection

Four species of Prunella L. (Prunella vulgaris L., Prunella laciniata L., Prunella grandiflora L. and Prunella orientalis Bornm.) belong to the family of Lamiaceae and representing popular Western and Chinese herbal medicine were examined for the content of phenolic compounds. Phenolic acids (rosmarinic acid, caffeic acid, ferulic acid, chlorogenic acid, protocatechuic acid), flavonoids (rutin, quercetin) in different quantitative proportions depending on extracts were determined by the rapid, selective and accurate method combining solvent/acid hydrolysis extraction and high performance liquid chromatography-diode array detection (HPLC-DAD). Water, methanol, butanol, acetonitrile, ethyl acetate, hexane and their acidic solutions were used to examine the efficiency of different solvent systems for the extraction of phenolic compounds. Acid hydrolysis extraction was established as the most suitable extraction method for phenolic compounds.     

Determination of phenolic compounds in Yucca gloriosa bark and root by LC-MS/MS

On the basis of the biological activities shown by yuccaols and gloriosaols from Yucca schidigera and Yucca gloriosa, the content of yuccaols and gloriosaols in two different parts of Y. gloriosa (roots and bark), was determined for each single compound, and compared with phenolic determination in Y. schidigera bark, concluding that Y. gloriosa bark and roots are rich sources of phenolic derivatives structurally related to resveratrol. LC/ESIMS (liquid chromatography coupled to electrospray mass spectrometry) qualitative and an LC/ESIMS/MS (liquid chromatography coupled to tandem electrospray mass spectrometry) quantitative studies of the phenolic fraction of Y. gloriosa were performed. LC/ESIMS/MS multiple reaction monitoring (MRM) method previously described for yuccaols in Y. schidigera was applied and optimised for separation and determination of gloriosaols and yuccaols in Y. gloriosa. Due to the sensitivity and the repeatability of the assay, we suggest this method as suitable for industrial quality control of raw materials and final products.     

Characterization of phenolic compounds in the Chinese herbal drug Artemisia annua by liquid chromatography coupled to electrospray ionization mass spectrometry

A simple and rapid method has been established for the screening of the main phenolic compounds in Artemisia annua by LC-DAD-ESI-MS(n). A total of 40 phenolic compounds were identified or tentatively characterized in the methanol extract of A. annua, including 8 C-glycosyl flavonoids, 5 O-glycosyl flavonoids, 3 flavonoid aglycones, 21 quinic acid derivatives, 2 benzoicacid glucosides and 1 coumarin. The C-glycosyl flavonoids were reported from A. annua for the first time and they were found to be a new type of main constituents, and might be responsible for its antioxidant and antiviral activity. Quinic acid derivatives were also found to be the major constituents of A. annua.     

Development of a chromatographic fingerprint for the chloroform extracts of Ganoderma lucidum by HPLC and LC-MS

A new high-performance liquid chromatographic (HPLC) fingerprinting method was developed for the quality control of Ganoderma lucidum. Twenty-nine batches obtained from three different origins in China were used to establish the fingerprint. The constituents of these samples were separated with a Kromasil C(18) column (250 mm x 4.6 mm, 5 microm) by linear gradient elution using water-acetic acid (100:0.1, v/v) and acetonitrile as mobile phase components at a flow rate of 0.8 ml/min and detector wavelength at 254 nm. Mean chromatograms and correlation coefficients of samples were calculated by the software "Similarity Evaluation System for Chromatographic Fingerprint of TCM". There were 19 common peaks in this fingerprint. Eleven of these common peaks were tentatively identified with reference to literature data based on their liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS) and UV data. This profile was then successfully used to identify and assess the differences among samples from various origins with the aid of similarity analysis. The diverse similarities among different samples indicated that the quality of G. lucidum was not stable and the products from different areas were inconsistent. All results showed that the developed fingerprint assay was specific and could further serve for quality identification and comprehensive evaluation of G. lucidum.     

反相高效液相色谱法测定人体血浆中表阿霉素的含量

采用反相高效液相色谱法(以柔红霉素为内标)测定血浆中表阿霉素的含量,流动相为5mmol/LH3PO4-甲醇-异丙醇-乙晴(6∶9∶7∶2,pH2.9),YWGC18不锈钢色谱柱,荧光检测波长λex=450nm,λem=530nm.血浆中样品经氯仿-甲醇-异丙醇=5∶2∶3抽提液抽提处理后进样测定.血浆中表阿霉素的浓度在0.03～2.4mg/L范围内峰面积与柔红霉素的峰面积之比呈线性关系(r=0.9998),方法对表阿霉素的平均回收率为89%～90.7%.最低检测浓度为0.01mg/L.本法灵敏度高,线性范围大,符合临床用药的血药浓度监测和药代动力学研究的分析方法要求,并用于40例已服用表阿霉素的癌症病人的血浆药物含量的测定.     

高效液相色谱法测定酪胺含量

目的：建立快速、准确测定酪胺含量的高效液相色谱方法。方法：采用安捷伦HPLC1200系统,色谱柱采用Agilent Eclipse XDB—C18填料（250mm×4．6mm,5um）,流动相：0．02mol／L磷酸氢二钠-乙腈（85：15）用10％磷酸调节pH值为8．3,流速：1．0ml／min;在225nm波长处检测。结果：酪胺在8．0～160ug·ml-1 的浓度范围内呈线性关系,r=0．9999。     

高效液相色谱法测定肌氨肽苷注射液中次黄嘌呤的含量

目的:建立高效液相色谱法测定肌氨肽苷注射液中次黄嘌呤的含量。方法:以十八烷基硅烷键合硅胶为填充剂,磷酸二氢钾(0.01mol/L)-甲醇(100:0.2)为流动相,检测波长260nm。结果:平均回收率为100.56%,RSD=1.57%。结论:该方法简便、准确。     

Quantification of seven phenylpropanoid compounds in Chinese Cinnamomi Cortex and Ramulus by HPLC

In the present study, we developed and validated a high-performance liquid chromatography method for the simultaneousdetermination of seven phenylpropanoid compounds (2-hydroxyl cinnamaldehyde, coumarin, cinnamyl alcohol, cinnamic acid, 2-methoxy cinnamic acid, cinnamaldehyde and 2-methoxy cinnamaldehyde) in Cinnamomi Cortex and Cinnamomi Ramulus. The levels of seven phenylpropanoid compounds in Cinnamomi Cortex and Cinnamomi Ramulus were compared using this method. A total of 48 samples (27 Cinnamomi Cortex and 21 Cinnamomi Ramulus) were purchased in China and analyzed. Quantities of seven phenylpropanoid compounds ranged from 17.5 to 61.6 mg/g in Cinnamomi Cortex and ranged from 9.91 to 23.4 mg/g in Cinnamomi Ramulus. The level of 2-methoxy cinnamic acid in the Cinnamomi Cortex samples was below the LOD, whereas it ranged from 0 to 0.119 mg/g in the Cinnamomi Ramulus samples. The (cinnamyl alcohol+cinnamic acid)/cinnamaldehyde ratios (R₃₄₆) of Cinnamomi Cortex and Cinnamomi Ramulus ranged from 0.0121 to 0.0467 and 0.0598 to 0.182, respectively. This ratio could be used to discriminate Cinnamomi Cortex (<0.05) and Cinnamomi Ramulus (>0.05). The extraction rates (D_n) of seven compounds in boiling water were different, with the lowest dissolution for cinnamaldehyde (<3%) and the highest for cinnamic acid (about 60%).     

高效液相色谱法测定舒筋定痛片中柚皮苷的含量

目的:建立高效液相色谱法测定舒筋定痛中柚皮苷的含量。方法:以十八烷基硅烷键合硅胶为填充剂,甲醇-醋酸-水(35:4:65)为流动相,检测波长283nm。结果:平均回收率为100.06%,RSD为0.97%。结论:方法简便、准确。     

高效液相色谱法测定百花定喘片中橙皮苷的含量

目的:建立高效液相色谱法测定百花定喘片中橙皮苷的含量。方法:以十八烷基硅烷键合硅胶为填充剂,甲醇-醋酸-水(35:4:61)为流动相,检测波长283nm。结果:平均回收率为98.26%,RSD为0.89%。结论:该方法简便、准确。     

高效液相色谱法测定咽炎片中芍药苷的含量

目的:建立高效液相色谱法测定咽炎片中芍药苷的含量。方法:以十八烷基硅烷键合硅胶为填充剂,乙腈-0.1%磷酸溶液(15:85)为流动相,检测波长230nm。结果:平均回收率为98.48%,RSD为1.28%。结论:方法简便、准确。     

高效液相色谱法测定氯硝西泮片含量

目的建立用高效液相色谱法测定氯硝西泮片含量的方法。方法采用C18柱,流动相为甲醇-水(70∶30),柱温30℃,流速:1.0mL/min,检测波长254nm。结果氯硝西泮在进样量0.4~2.0μg的范围内呈良好的线性关系,r=0.9999;平均回收率为100.4%,相对标准偏差RSD为0.69%(n=5)。结论本法专属性强、稳定、准确,适用于氯硝西泮片的含量测定。     

Quantification of estragole in fennel herbal teas: Implications on the assessment of dietary exposure to estragole

Quantification of estragole content in commercial fennel herbal teas was carried out in order to allow for a more accurate estimate of the dietary exposure to estragole. A simple and rapid analytical method, based on Stir Bar Sorptive Extraction and GC-MS, was developed for this purpose. Fennel teas obtained from different types of commercial products were analysed. Concentration levels ranged from 241 to 2058 μg L⁻¹ in teas from teabags, from 9 to 912 μg L⁻¹ in diluted instant teas, from 251 to 1718 μg L⁻¹ in teas from not packaged seeds. Based on these data and considering the daily consumption of three portions of herbal tea, a maximum exposure to estragole for adults of 10 μg/kg bw/day was calculated. The relatively high level observed in diluted instant teas of some brands deserves attention since these products are designed for infant consumption. Estimated exposure in infants was up to 51 μg/kg bw/day for teas from teabags, and up to 23 μg/kg bw/day for instant teas. A generalization of the use of suitable technologies in production processes of instant teas could substantially reduce the exposure to estragole in the vulnerable population groups (infants, young children, pregnant and breastfeeding women) who consume these products.     

Optimization and validation of a dissolution test for selegiline hydrochloride tablets by a novel rapid HPLC assay using a monolithic stationary phase

The present study reports the optimization and validation of a dissolution test for selegiline.HCl tablets using a new high-performance liquid chromatographic (HPLC) method. Rapid separation of the analyte from sample matrix was achieved in less than 60s using a Cromolith RP-18e monolithic column using UV detection at 220 nm. Thorough validation of the assay based on pre-defined criteria included linearity, LOD/LOQ, accuracy, precision, selectivity and ruggedness. The dissolution test was optimized in terms of dissolution medium, basket (type I)/paddle (type II) agitation and rotation speed. Its ruggedness was also validated. The presented analytical and dissolution procedures are currently being applied in the quality and stability control of Cosmopril tablets (5mg/tablet selegiline.HCl, Cosmopharm Ltd., Korinthos, Greece).     

LC-determination of five paraben preservatives in saliva and toothpaste samples using UV detection and a short monolithic column

The present study reports the development and application of an HPLC–UV method for the simultaneous separation and determination of five paraben preservatives (methyl-, ethyl-, propyl-, n-butyl- and iso-butyl-paraben) in real samples. All analytes were separated efficiently in less than 20 min using a simple H₂O:ACN linear gradient and a short monolithic column (50 mm × 4.6 mm i.d.) at a flow rate of 3.0 mL min⁻¹. Phenoxyethanol was used as chromatographic internal standard. The method was validated for linearity, limits of detection and quantification, accuracy and precision. Human saliva and toothpaste samples were analyzed after SPE pretreatment on Licrolut RP-18 cartridges. The detection limits varied between 0.1 and 0.3 mg L⁻¹ in all cases and the percent recoveries between 86 and 113%.     

HPLC测定天山花楸中的苦杏仁苷

目的 采用HPLC法测定天山花楸中的苦杏仁苷.方法 用甲醇超声提取天山花楸,色谱柱为Shim-pack VP C18(150 mm×4.6 mm,5 μm);流动相为甲醇-水(20:80);柱温35℃;流速1.0 ml · min-1;检测波长215 nm.结果 苦杏仁苷进样量0.4～3.2 μg与峰面积的线性关系良好(r＝0.9999),平均回收率96.6%,RSD=2.2%.结论 所建方法可用于测定天山花楸中的苦杏仁苷,精密度好,结果准确可靠.     

Characterization and quantification of phenolic compounds of extra-virgin olive oils with anticancer properties by a rapid and resolutive LC-ESI-TOF MS method

The characterization and quantification of extra-virgin olive oil (EVOO) phenolic compounds by a rapid resolution liquid chromatography (RRLC) method coupled to diode-array and time of flight mass spectrometry (TOF) detection systems was developed. The RRLC method transferred from a conventional HPLC one achieved better performance with shorter analysis times. The phenolic compounds were separated with a C18 column (150 mm × 4.6 mm, 1.8 μm) using water with 0.5% acetic acid and acetonitrile as mobile phases. Good peak resolution was obtained and 19 different phenols were identified in less than 20 min providing a new level of information about the samples in shorter time. The applicability of this analytical approach was confirmed by the successful analysis of three different EVOO varieties (Picual, Hojiblanca, and Arbequina) obtained from different trademarks. Besides identification of the most important phenolic compounds and their quantification in three different ways (RRLC-UV, RRLC-MS and a new approach using the total polyphenol content obtained with Folin Ciocalteau, the relative areas and the response factors), we also described the occurrence of correlations between the phenolic composition of EVOO-derived crude phenolic extracts and their anti-proliferative abilities toward human breast cancer-derived cell lines. When compared with lignans-rich EVOO varieties, secoiridoids-rich EVOO had a significantly strong ability to alter cell viability in four different types of human breast carcinoma cells.     

高效液相色谱法测定己酮可可碱注射液的有关物质

目的：建立一种高效液相色谱法测定己酮可可碱注射液有关物质。方法：以ODS-C_(18)为固定相,甲醇-水(45：55)为流动相,检测波长为274nm。结果：该方法灵敏度高,精密度好、专属性强。结论：该法能有效检测己酮可可碱注射液有关物质。