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??OBJECTIVE To establish a method for rapid screening and quantification of 24 anti-inflammatory painkillers illegally added in traditional Chinese medicines and health foods by UPLC-quadrupole/electrostaticfield orbitrap high resolution mass spectrometry. METHODS The separation was performed on a Thermo Acquity UPLC system with a Hypersil Gold C₁₈ column with gradient elution of acetonitrile and 20 mmol??L^-1 ammonium acetate solution.Electrospray ionization of positive and negative ions and one and two high precision full scan were used for the MS detection. The qualitative and quantitative analyses were carried out by accurate and precise parent ion mass and product ion mass information. RESULTS The limits of detection(S/N??3) were 10.0-45.0 ??g??kg^-1. The average recoveries were 79.6%-98.3%. Thirty positive samples were detected among a total of 45 samples, resulting a positive rate of 66.7%. The detected components were paracetamol, diclofenac sodium, indometacin, and ibuprofen. CONCLUSION The method is simple, highly accurate, sensitive and selective, and can be used in rapid screening and quantitative analysis of illegally added anti-inflammatory painkillers in traditional Chinese medicines and health foods.     

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??OBJECTIVE To determine two toxic impurities, namely bromoacetic acid and 4-aminobenzonitrile, in the intermediate of dabigatran etexilate by UHPLC-MS. METHODS The separation was performed on a Shimadzu Shim-Pack GIS C₁₈ column (2.1 mm??50 mm,2 ??m) with mobile phase consisting of 0.1% formic acid aqueous solution (A) and 0.1% formic acid methanol (B) by gradient elution at a flow rate of 0.4 mL??min^-1. The detection was achieved by triple quadrupole mass spectrometry with rapid polarity switching using MRM mode. RESULTS The calibration curves were linear in the ranges of 0.2-40 and 0.4-40 ng??mL^-1 for bromoacetic acid and 4-aminobenzonitrile, respectively. The values of LOQ of bromoacetic acid and 4-aminobenzonitrile were 0.1 and 0.4 ng??mL^-1, respectively. The recoveries of bromoacetic acid and 4-aminobenzonitrile were 100.9% and 99.6%, respectively. CONCLUSION The method is accurate, rapid, sensitive, and reliable to determine the two toxic impurities bromoacetic acid and 4-aminobenzonitrile in the intermediate of dabigatran etexilate for quality control.     

LC-MS/MS�ⶨ��Ѫ�����������Ũ�ȼ���ҩ��ѧ�о�

??OBJECTIVE To establish an LC-MS/MS method for the determination of pregabalin, which was used subsequently to investigate the pharmacokinetics of pregabalin in healthy Chinese volunteers. METHODS Gabapentin was used as internal standard. The separation was achieved on a Waters Symmetry C₁₈ column (3.9 mm??150 mm,5 ??m) with a mobile phase consisting of acetonitrile -5 mmoL??L^-1 ammonium acetate and 0.3% formic acid aqueous solution (8/92,V/V) at a flow rate of 1.0 mL??min^-1 within 6.0 min. Pregabalin and the internal standard were measured by a triple-quadrupole mass spectrometer in positive electron atmospheric-pressure chemical ionization (APCI) mode using multiple reaction monitoring (MRM). The extracted ions monitored following MRM transitions were m/z 160.1??142.1 for pregabalin and m/z 172.1??154.1 for the internal standard gabapentin. Plasma samples were pretreated by methanol precipitation. RESULTS The calibration curve of pregabalin in human plasma was linear over the concentration range of (30-5 000) ng??mL^-1. The lower limit of quantitation was 30 ng??mL^-1. The intra- and inter-run precisions at three quality control levels were within 2.0%-6.2%, the relative deviation of the assay was within -10.5%-3.0%. The plasma samples were stable at room temperature (25 ??) for 6 h, at -30 ?? for 27 d and during three freeze-thaw cycles. The determination and the result of incurred sample reanalysis met the requirements. CONCLUSION The method is proved to be convenient, accurate and sensitive. The method is proved to be suitable for the pharmacokinetics of pregabalin in healthy Chinese volunteers after a single oral dose of 75 mg pregabalin capsule.     

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??OBJECTIVE To establish an LC/MSⁿ method for identification of the related substances in etimicn sulfate detected under the chromatographic condition described in Chinese Pharmacopoeia 2010.METHODS The HPLC separation was carried out on a Welch Ultimate LP-C₁₈ column(4.6 mm??300 mm,5 ??m)with mobile phase consisting of 0.2 mol??L^-1trifluoroacetic acid(containing 0.1% propionic acid )-methanol(84??16) at a flow rate of 1.0 mL??min^-1. Thirty percent of the eluent was detected by ion trap mass spectrometry, and the parent ions and the corresponding product spectra of all the related substances in etimicin sulfate were determined and elucidated.RESULTS Addition of 0.1% propionic acid into the mobile phase significantly enhanced the sensitivity of MS detector without altering the chromatographic behavior such as retention time and elution order of the related substances. Twenty-eight related substances were separated and detected by the LC/MSⁿ method in a typical sample. Nine of them were identified with the help of corresponding impurity reference substances and 14 of them were elucidated by MS fragment information, while the other five were not identified due to limitated information. CONCLUSION The established method can be applied to the identification of the related substances in etimicn sulfate detected under the chromatographic condition described in Chinese Pharmacopoeia 2010, which is helpful to the quality improvement and process optimization of etmicin sulfate.     

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??OBJECTIVE To develop an immunoaffinity column clean-up and high performance liquid chromatography coupled with triple quadrupole mass spectrometry(HPLC-MS/MS) method to determine aflatoxins in gelatin drugs. METHODS The analysis was performed by an HPLC-MS/MS system with X-Brigde-C₁₈(3.0 mm??50 mm,3.5 ??m)column. Multiple-reaction monitoring (MRM) was performed to identify and quantify aflatoxin B₁,B₂,G₁ and G₂, which were extracted from Asini Coril Colla, Cervil Cornus Colla, and Testudinis Carapacis Colla with 60% methanol solution. RESULTS Linear calibration curves were obtained with r??0.997 9. The precision of the method was showed by RSDs (n=6) ranging from 1.2% to 4.1%. The recoveries were determined at three concentration levels and ranged from 77.3% to 94.6%. The ranges of LOQs were from 0.5 to 0.8 ??g??L^-1 and the RSDs (n=9) of intra-day precision and inter-day precision were from 1.1% to 2.9% and from 1.5% to 2.8%, respectively. CONCLUSION The method is specific, simple and rapid to detect aflatoxins in gelatin drugs.     

HPLC-ELSD�ⶨ���������ĵ��������˯����Ч������5��������ɷֵĺ���

??OBJECTIVE To establish a method for content determination of ginsenoside Rg₁, ginsenoside Rb₁, saikosaponin a, saikosaponin d, and saikosaponin B₂ in 70% ethanol elution effective fraction of Chaihu plus Longgu Muli decoction by HPLC-ELSD. METHODS A Diamonsil C₁₈ column(4.6 mm??250 mm, 5 ??m) was used as the stationary phase and the mobile phase consisted of acetonitrile and water. Gradient elution was carried out at the flow rate of 1.0 mL??min^-1. The column temperature was maitained at 30 ??. The ELSD detector was operated at 105 ?? with nebulizing gas at the optimum flow rate of 2.5 L??min^-1. RESULTS The average contents of ginsenoside Rg₁, ginsenoside Rb₁, saikosaponin a, saikosaponin d, and saikosaponin B₂ were 0.474%, 1.372%, 1.554%, 0.883%, and 2.073%, respectively. The calibration curves were linear in the ranges of 1.820-9.10, 1.810-9.050, 1.130-10.170, 0.420-2.100, and 3.125-15.625 ??g, respectively. The RSDs of precision, reproducibility and recovery were all less than 3.0%. CONCLUSION The method is rapid, simple, reliable and accurate, and has been successfully used to the quantification of five components in 70% ethanol elution effective fraction of Chaihu plus Longgu Muli decoction, which can provide a basis for the quality evaluation of Chaihu plus Longgu Muli decoction.     

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??OBJECTIVE To develop a high throughput qualitative method to analyze 50 pigments in traditional Chinese medicine by UPLC/QTOF, and provide a strategy for screening of illegally traditional Chinese medicine. METHODS The analysis was carried on an Agilent Poroshell 120 EC-C₁₈ column with 0.1% formic acid solution-acetonitrile as mobile phase for detection of fat-soluble and alkaline pigments or with 10 mmol??L^-1 ammonium formate solution-acetonitrile as mobile phase for detection of acidic pigments respectively, both using gradient elution program at the flow rate of 0.4 mL??min^-1. Q-TOF-MS equipped with ESI ion source was performed in positive ionization mode for detection of fat-soluble and alkaline pigments and in negtive ionization mode for detection of acidic pigments respectively. Qualitative analysis was based on the comparison of the retention time, accurate mass, elemental compositions, and product ions with those of the reference standard. RESULTS The qualitative analysis method was established for detection of 50 pigments, using different medicinal parts of four representative traditional Chinese medicine for method verification. The detection limits for the 50 pigments were 0.008-9.2 mg??kg^-1. One to seven pigments were detected in 12 batches among 83 batches of 26 Traditional Chinese Medicine. CONCLUSION The established method has high throughput and is accurate, and it is suitable for rapid screening of illegally traditional Chinese medicine.     

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??OBJECTIVE To establish a rapid and sensitive method for the determination of vitamin K homologue in human plasma using HPLC-fluorescence detection. METHODS With salting-out assisted liquid/liquid extraction, the fluorescence derivatization reaction of vitamin K homologues was proposed by ethanolic SnCl₂ in acidic medium. An AQ-C₁₈ column was used as the chromatographic column with a mobile phase of a mixture of methanol-water(98??2) at a flow rate of 1 mL??min^-1,the fluorescence detector was set at ??ex/??em=248 nm/418 nm, and the column temperature was maintained at 30 ??. RESULTS The calibration curves of vitamin K1(VK1) and vitamin K2 (VK2) were as followed:Y_a=2.944??_a+0.023(r=0.999 8,1.000-50.036 ng??mL^-1),Y_b=1.195??_b+2.653(r=0.999 7,5.006-100.124 ng??mL^-1), respectively. Their precision RSD, stability RSD, repeatability RSD and freezing-thawing test RSD were all less than 10%.CONCLUSION The method is simple, sensitive and accurate, which can be used for the assay of endogenous vitamin K homologue in human plasma.     

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??OBJECTIVE To develop a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for determination of amikacin, dexamethasone, and ribavirin in human plasma. METHODS With kanamycin as internal standard (IS), samples involve a one-step extraction with methanol of 0.2 mL plasma. The analysis was carried out on the Inertsil HILIC (2.1 mm??150 mm,3.5 ??m) column with gradient elution at a flow rate of 0.4 mL??min^-1. The mobile phase was acetonitrile (containing 0.1% formic acid) and 0.1% formic acid. The determination was performed on a triple quadrupole tandem mass spectrometer by multiple reactions monitoring (MRM) mode via electrospray ionization (ESI) source. RESULTS The standard curves were linear (r?? 0.999) over the concentration range of 1.038-1 038 ng??mL^-1 (amikacin), 1.063-1 063 ng??mL^-1 (dexamethasone), and 1.029-1 029 ng??mL^-1(ribavirin) with the lower limit of quantification(LLOQ) of 1.038,1.063,1.029 ng??mL^-1. The intra- and inter-day precision (RSD) value was below 13.7% and accuracy (relative error) was ranged from -9.2% to 10.9% at all three quality control (QC) level. CONCLUSION The method for determination of amikacin, dexamethasone, and ribavirin in human plasma is rapid sensitive and accurate, as well as suitable to evaluate the and forensic toxicological analysis.     

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??OBJECTIVE To develop and validate an HPLC-MS/MS method for the measurement of 11 antipsychotics in human plasma samples for TDM(therapeutic drug monitoring) or other objectives. METHODS Protein precipitation method with acetonitrile was used to extract the drugs in plasma. Agilent Poroshell 120 EC-C₁₈ column(2.1 mm??50 mm, 2.7 ??m)was used for chromatographic separation. The mobile phase was acetonitrile with 0.1% formic acid-water with 0.1% formic acid gradiently eluted at a flow rate of 0.3 mLmin^-1. The injection volume was 5 ??L. The deprotonated ions of analytes were ionized by electrospray(ESI) and detected in positive ionization by multiple reaction monitoring mode(MRM). RESULTS The total run time of the chromatographic separation and mass detection was 6 min. Excellent linear relationship with correlation coefficient of 0.99 was obtained from 2 to 200 ngmL^-1 for risperidone, paliperidone and olanzapine, from 10 to 1 000 ngmL^-1 for aripiprazole, quetiapine, amisulpride, clozapine, chlorpromazine and sulpride, from 0.4 to 40 ngmL^-1 for haloperidol, and 0.2 to 20 ngmL^-1for perphenazine, respectively. The precision and accuracy varied from 2.7% to 10.8% and from 90.7% to 102.7%, respectively. CONCLUSION This established method is simple, rapid, and economic, fulfilling all criteria for TDM, and could be successfully applied in the routine TDM of antipsychotics in clinic.