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ABSTRACT: An approach for quantitative determination of amino-acid consumption profiling in culture media by high performance liquid chromatography with fluorescence detection (HPLC-FLD) was developed and validated, using o-phthalic dicarboxaldehyde (OPA) as the derivatizing reagent and norvaline as the internal standard. Mobile phase A was 10 mmol/L Na2HPO4Na2B4O7 buffer (pH 7.95), and mobile phase B was acetonitrile-methanol-water (45:45:10, v/v/v). The linear elution program was 5% B at the start and 52% B at the end in 35 min. The 17 free amino-acids (FAAs) were separated satisfactorily in 33 min. Following HeLa cells incubation in conditioned medias of taxol (4 micromol/L) and mitomycin (75 micromol/L), respectively, with control for 24 h, the media 17 amino-acid consumption profilings were determined, and then analyzed by multivariate statistical analysis based on Matlab7.1 software platform. Relation analysis performed by partial least squares-discriminant analysis (PLS-DA) indicated that in comparison with the control group, the media amino-acid consumption profiling can distinguish the two anticancer drugs with different mechanisms, which provides a new perspective for the pre-classification of drug action mechanisms during the screening of new anticancer drugs. Meanwhile, the idea from the outer into the inner has convenient and economic characteristics.
Se pu = Chinese journal of chromatography / Zhongguo hua xue hui 04/2011; 29(4):330-4.
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ABSTRACT: A simple metabolic profiling approach for quantitative analysis of free fatty acids (FFAs) in human plasma by high performance liquid chromatography was described and validated, using alpha-bromoacetophenone as the derivative reagent and heptadecanoic acid (C(17:0)) as the internal standard. The quantitations of 6 predominant FFAs and 6 trace FFAs were achieved. Plasma fatty acid metabolic profiling of 75 diabetic patients was investigated, and then analyzed by multivariate statistical analysis. The linear discriminant analysis (LDA) model was established and validated for the pattern discrimination between Qi-deficiency and Qi and Yin-deficiency, with the hit ratio 94.3%. Stepwise discriminant analysis (SDA) model indicated that arachidonic acid (C(20:4)) and oleic acid (C(18:1)) contained the important information on the two syndromes above, and can be used as potential biomarkers of traditional Chinese medicine (TCM) syndrome. It is of great significance to systematically study the relationship between fatty acid metabolic profiling and TCM syndrome using metabolomics methods, and to improve the credibility and repeatability of clinical diagnosis and treatment system.
Se pu = Chinese journal of chromatography / Zhongguo hua xue hui 06/2010; 28(6):547-50.
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ABSTRACT: A method of hydrophilic interaction chromatography (HILIC) was established for the quantitative determination of trigonelline in Trigonella foenum-graecum L. HILIC analysis was performed on a Waters Atlantis HILIC Silica column (150 mm x 2.1 mm, 3 microm). The mobile phase consisted of acetonitrile-ammonium acetate (pH 4.4) (70:30, v/v), and the flow rate was 0. 4 mL/min. The detection wavelength was set at 265 nm. The method has good linearity in the range of 2.50-100 mg/L for trigonelline (r = 0.999 6). The recoveries were on an average of 102% by adding 29.2 mg/L and 43.8 mg/L with relative standard deviations (RSDs) of 4.17% and 2.28% (n = 3), respectively. The results indicate that the method is simple and rapid for the determination of strong polar trigonelline in Trigonella foenum-graecum L. Furthermore, it significantly reduces the equilibration time compared with ion-pair liquid chromatography (IPLC) recorded in the Pharmacopoeia of China. This new method can be used as a valid method for the quality control of Trigonella foenum-graecum L.
Se pu = Chinese journal of chromatography / Zhongguo hua xue hui 04/2010; 28(4):379-82.
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ABSTRACT: A high performance liquid chromatography-tandem mass spectrometric method has been developed for the simultaneous analysis of three minor curcuminoids in the crude extract of Curcuma longa L. After ultrasonic extraction with ethanol, the sample was separated on a Microsorb C18 column (250 mm x 4.6 mm, 5 microm) with the gradient elution of acetonitrile and 0.1% (v/v) formic acid aqueous solution as mobile phase, then detected by electrospray ionization tandem mass spectrometry in multi-reaction monitoring (MRM) mode. On the basis of analysis of the fragmentation patterns, the identification of the minor ones was achieved in a single experiment using MRM with 8 precursor-product ion transitions for each component. Using this technique, the three minor curcuminoids were observed in C. longa L. for first time and the limit of detection was 0.2 microg/L. The experimental results show that this method is simple, rapid, accurate, sensitive and suitable for the analysis of curcuminoids in the complex samples of traditional Chinese medicine.
Se pu = Chinese journal of chromatography / Zhongguo hua xue hui 05/2009; 27(3):264-9.
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ABSTRACT: Longdan Xiegan Decoction (LXD) is a formulated preparation composed of 10 ingredient herbs, with iridoids as the main bioactive components. In this study, a rapid, simple and reliable method of simultaneous determination of four iridoid glycosides in LXD using high-performance liquid chromatography (HPLC) coupled with electrospray ionization mass spectrometry (MS) was first developed and validated. The four iridoid glycosides references were isolated from LXD extract and purified using a preparative HPLC chromatography. The sample preparation for quantification comprised of a simple ultrasonic extraction and the satisfactory chromatographic separation of the four structurally similar iridoid glycosides was effected in less than three minutes on a CAPCELL PAK C(18) MGII column (3 microm, 100 mm x 2.0 mm), using an elution system of 10% methanol and their concentrations in different batches of LXD and ingredient herbs were simultaneously determined by HPLC-MS/MS using a multiple reaction monitoring (MRM) mode. The method was validated with respect to the overall intra- and inter-day variation (RSD less than 8%) and the limits of quantification for the four iridoid glycosides were 35, 20, 37 and 33 ng/mL, respectively.
Journal of chromatography. A 03/2009; 1216(11):2098-103. · 4.19 Impact Factor
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ABSTRACT: Preparative high performance liquid chromatography (HPLC) are widely used recently. The optimization of its operating conditions is important for improving efficiency and saving operating cost. Now, most works such as linear scale up technology were focused on the optimization of mobile phase. The sample collection region is another important operating parameter in preparative HPLC. In this study, a software was written according to the exponentially modified Gaussian (EMG) model. With this software, the operators of preparative HPLC can easily choose a suitable sample collection region.
Se pu = Chinese journal of chromatography / Zhongguo hua xue hui 08/2008; 26(4):523-5.
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ABSTRACT: Rhizoma Anemarrhenae (Zhimu in Chinese), the dried rhizome of Anemarrhena asphodeloides Bge. (Fam. Liliaceae), is a well-known traditional Chinese medicinal herb and has been used clinically in China for centuries to cure various diseases. However, like other traditional Chinese medicines, the effective constituents of this medicine, especially the assimilation and metabolites in vivo, which are very important to show their effects, have not been systematically studied. In this paper, solid-phase extraction and liquid chromatography-atmospheric pressure chemical ionization mass spectrometry technologies were used to study the constituents absorbed into rat urine and their metabolites after oral administration of Rhizoma Anemarrhenae decoction. A total of 11 compounds, including two xanthones, three of their metabolites and six steroidal saponins, were identified in rat urine sample. They were neomangiferin (1), glucuronide and monomethyl conjugate of mangiferin (2), mangiferin (3), monomethyl conjugate of mangiferin (4), dimethyl conjugate of mangiferin (5), timosaponin N or timosaponin E1 (6), timosaponin BII (7), timosaponin BIII (8), anemarrhenasaponin I or anemarrhenasaponin II (9), timosaponin AII (10) and timosaponin AIII (11). The results would efficaciously narrow the potentially active compounds range in Rhizoma Anemarrhenae decoction, and pave a helpful way for follow-up mechanism of action research.
Biomedical Chromatography 05/2008; 22(10):1066-83. · 1.97 Impact Factor
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ABSTRACT: An analytical method for flavones of Astragalus has been developed and a process of high performance liquid chromatographic (HPLC) condition optimization has been elaborated. The LC retention parameters of 34 components were obtained quickly by using three linear gradient elutions. Overlapping separation range map (OSRM), which is related to the separation quality and the analysis time, was utilized to search the optimised stepwise gradient method for HPLC analysis of flavones of Astragalus. Finally, the optimised stepwise gradient elution conditions were set up with the help of this map. Under the optimised conditions, the sample of flavones of Astragalus was well separated in reasonable time. The method is more convenient than that using a series of isocratic elutions.
Se pu = Chinese journal of chromatography / Zhongguo hua xue hui 12/2005; 23(6):630-2.
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ABSTRACT: Fufang Danshen (FFDS) is a famous typical Chinese complex prescription, which is mainly composed of Radix Salvia miltiorrhiza Bunge (SM) and Radix Panax notoginseng (PN). An HPLC method is developed to analyze SM, PN, and FFDS effectively; the effective analysis is achieved by using a gradient elution procedure with a mobile phase consisting of acetonitrile and 0.025% aqueous phosphoric acid (v/v). Through this method, 33 peaks in FFDS are clearly exhibited, and the components that make up the 33 peaks in FFDS are evaluated. Also, the chemical ingredients are compared between the single herbs (SM and PN) and the complex prescription (FFDS). The result indicate that the chemical ingredients in FFDS are not simply a combination of SM and PN. In addition, the HPLC method is suitable for the routine quality control of SM, PN, and FFDS, which could present a uniform quality control method for single medicines and one of the most commonly used Traditional Chinese Medicine-complex prescriptions.
Journal of chromatographic science 44(10):591-5. · 0.88 Impact Factor
