2D-LC as an on-line desalting tool allowing peptide identification directly from MS unfriendly HPLC methods.


:The increasing interest in peptides and proteins in pharmaceutical research and development has led to many challenges for the researchers tasked with characterizing and analyzing these larger molecules. Due to the more complicated impurity profile of peptides and proteins, multiple liquid chromatography techniques are often needed to achieve comprehensive analysis. However, many of these separation conditions require buffers, salts or additives that render them incompatible with mass spectrometry (MS) detection. Previous researchers have demonstrated proof of concept for the use of two dimensional liquid chromatography (2D-LC) to provide convenient second dimension online desalting of components purified in the first chromatographic dimension. In this paper, we evaluated the Agilent heart-cutting 2D-LC system connected with an Agilent Q-TOF mass spectrometer to address this frequently encountered analytical challenge. On this 2D-LC/MS system, fractions containing the compounds of interest are separated by the first dimension using an MS incompatible mobile phase, then sent to a second dimension HPLC method where fast desalting using an MS compatible mobile phase is performed prior to MS analysis. The system allows for fast and direct collection of MS information for chromatographic peaks eluted in MS incompatible mobile phases, without requiring difficult, time consuming and error-prone translation of chromatographic methods from MS incompatible to MS compatible eluents, or off-line fraction collection and preparation. Several examples showing the application of the approach to complex mixtures containing peptides with impurities and positional isomers are presented.


J Pharm Biomed Anal


Luo H,Zhong W,Yang J,Zhuang P,Meng F,Caldwell J,Mao B,Welch CJ




Has Abstract


2017-04-15 00:00:00












  • Validation of a UPLC-PDA method to study the content and stability of 5-chloro 8-hydroxyquinoline and 5,7-dichloro 8-hydroxyquinoline in medicated feed used in swine farming.

    abstract::A new, rapid, simple and specific method to determine 5-chloro 8-hydroxyquinoline (5-HQ) and 5,7-dichloro 8-hydroxyquinoline (5,7-HQ) stability in swine feed was optimized and validated. A system consisting of an ACQUITY UPLC BEH C18 column (1.7 μm, 2.1 mm × 100 mm), a mobile phase of acetonitrile-0.1% o-phosphoric ac...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Castillo Rodríguez MA,Rodríguez Patiño G,Revilla Vázquez AL,Rodríguez de San Miguel E,López-Arellano R

    更新日期:2019-03-20 00:00:00

  • Separation and structural elucidation of a new tadalafil analogue diethylaminopretadalafil included as an adulterant in a dietary supplement.

    abstract::A new tadalafil analogue was detected and isolated from a dietary supplement, the structure was elucidated by means of nuclear magnetic resonance (NMR) and mass spectroscopy. The compound was determined to be diethylaminopretadalafil, which might be derived from a precursor in the synthesis of tadalafil. ...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Zhang G,Yu Y,Wu X,Li J

    更新日期:2014-06-01 00:00:00

  • Release of proteolytic activity following reduction in therapeutic human serum albumin containing products: detection with a new neoepitope endopeptidase immunoassay.

    abstract::Botulinum type A toxin (BoNT/A) is defined by its specific endopeptidase cleavage of SNAP25 between Gln(197) and Arg(198) under reducing conditions. The neurotoxin is widely used for therapeutic or cosmetic purposes, but should not contain other toxin serotypes or unwanted protease activities. Using a neoepitope endop...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Jones RG,Liu Y,Halls C,Thorpe SJ,Longstaff C,Matejtschuk P,Sesardic D

    更新日期:2011-01-05 00:00:00

  • Investigation of fatty acid esters to replace isopropyl myristate in the sterility test for ophthalmic ointments.

    abstract::Several pharmacopoeias recommend the membrane filtration method for the sterility test of ophthalmic ointments. Isopropyl myristate, a fatty acid ester that exhibits high toxicity mainly against Gram-negative microorganisms, is indicated as a solvent for ointments. In this study, six fatty acid esters (diethyl adipate...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Cardoso VM,Solano AG,Prado MA,Nunan Ede A

    更新日期:2006-11-16 00:00:00

  • Investigation of tryptophan-related yellowing in parenteral amino acid solution: Development of a stability-indicating method and assessment of degradation products in pharmaceutical formulations.

    abstract::Parenteral amino acid solutions containing tryptophan tend to develop a yellow colouration upon storage. Hence, the aim of the present study was to find out whether tryptophan degradation products are the reason for the yellowing. The degree of discolouration and tryptophan degradation was examined by visual examinati...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Unger N,Ferraro A,Holzgrabe U

    更新日期:2020-01-05 00:00:00

  • Quantitative determination of zidovudine diaryl phosphate triester pro-drugs in rat plasma by high-performance liquid chromatography-electrospray ionization tandem mass spectrometry.

    abstract::A rapid, simple, and sensitive high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS) method was developed and validated for quantitative analysis of 3'-azido-3'-deoxythymidine (zidovudine, AZT) diaryl phosphate triester pro-drugs, in rat plasma using 2',3'-dideoxy-2'3...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Zhu M,Xie B,Tang G,Hu A,Fang M,Wu Z,Zhao Y

    更新日期:2008-12-15 00:00:00

  • Determination of possible impurities in piracetam using FTIR spectroscopy.

    abstract::Piracetam (2-oxo-1-pyrrolidine acetamide (PAm)) may contain some structurally related impurities deriving from synthesis or degradation, mainly 2-oxo-1-pyrrolidine acetic acid (PAc). A rapid and sensitive Fourier transform infrared (FTIR) spectrophotometric method was developed for determination of possible impurities...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Karamancheva I,Staneva T

    更新日期:2000-01-01 00:00:00

  • The use of mixed-mode ion-exchange solid phase extraction to characterize pharmaceutical drug degradation.

    abstract::Solid phase extraction (SPE) has been utilized extensively in the pharmaceutical industry for the isolation of pharmaceuticals from interfering biological matrices and the purification and concentration of impurities and degradation products present in analytical samples. The work described herein involves the novel u...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Landis MS

    更新日期:2007-09-03 00:00:00

  • Isotope-coded derivatization based LC/ESI-MS/MS methods using a pair of novel reagents for quantification of hydroxycinnamic acids and hydroxybenzoic acids in fermented brown rice product.

    abstract::Hydroxycinnamic acids (HCAs) and hydroxybenzoic acids (HBAs) are antioxidant phytochemicals found in rice and effective for the prevention of human diseases including cancer. FBRA, which is a functional food manufactured by fermenting brown rice and rice bran with Aspergillus oryzae, has been demonstrated to have chem...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Ogawa S,Takafuji K,Tsubuku S,Horie Y,Ikegawa S,Higashi T

    更新日期:2017-08-05 00:00:00

  • Determination of tramadol and metabolites by HPLC-FL and HPLC-MS/MS in urine of dogs.

    abstract::Tramadol is a centrally acting analgesic drug used in veterinary and human clinical practice. Its metabolism has been largely characterized in human being but is still long to be comprehended in several animal species, especially in the dog. The aim of the present study was to develop and validate a new analytical pro...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Saccomanni G,Del Carlo S,Giorgi M,Manera C,Saba A,Macchia M

    更新日期:2010-10-10 00:00:00

  • An LC-MS/MS analytical method for the determination of uremic toxins in patients with end-stage renal disease.

    abstract::End-stage renal disease (ESRD) is the last stage of chronic kidney disease, characterized by the progressive accumulation of uremic toxins (UTs). Hemodialysis is the standard approach to remove UTs from the body. Creatinine and urea levels are important indices of hemodialysis effectiveness, but the utility of those m...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Ma YR,Xin MY,Li K,Wang H,Rao Z,Liu TX,Wu XA

    更新日期:2020-11-30 00:00:00

  • A new validation approach applied to the GC determination of impurities in organic solvents.

    abstract::Organic solvents such as methanol, acetone, dichloromethane or toluene are frequently used in the pharmaceutical industry. The manufacturing of new active pharmaceutical ingredients (APIs) under GMP conditions commands to control adequately the quality of the different ingredients happening in the synthesis. Organic s...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Jacobs P,Dewé W,Flament A,Gibella M,Ceccato A

    更新日期:2006-02-13 00:00:00

  • Optimization and validation of a CE-LIF method for amino acid determination in biological samples.

    abstract::There is increasing evidence that amino acids and related compounds are among the most important metabolites for living systems and their quantitative analysis is a continuous challenge for clinical and biochemical laboratories. The lack of chromophore group and their high polarity make the analytical process more dif...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Lorenzo MP,Navarrete A,Balderas C,Garcia A

    更新日期:2013-01-25 00:00:00

  • Disposition of human drug preparations in the horse. II. Orally administered fencamfamine.

    abstract::A gas chromatographic method to measure urinary levels of the central nervous system stimulant fencamfamine and some of its metabolites is described. When 100 mg fencamfamine was given orally to four horses the parent drug could not be detected in the urine. After enzymatic hydrolysis of the urine the major human meta...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Delbeke FT,Debackere M

    更新日期:1992-09-01 00:00:00

  • Cell-based assays in combination with ultra-high performance liquid chromatography-quadrupole time of flight tandem mass spectrometry for screening bioactive capilliposide C metabolites generated by rat intestinal microflora.

    abstract::Many plant-derived glycosides are used as medications. It is common that these glycosides show poor intestinal absorption but their metabolites generated by intestinal microflora demonstrate strong bioactivity. Hence, it is crucial to develop a method for the identification and characterization of the metabolites, and...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Cheng Z,Huang M,Chen G,Yang G,Zhou X,Chen C,Zhang Y,Xu Y,Feng Y,Zhang L,Jiang H

    更新日期:2016-02-05 00:00:00

  • Evaluation of the European Pharmacopoeia method for control of residual solvents in some antibiotics.

    abstract::Residual solvents (RS) are volatile organic chemicals that are used or produced during the manufacturing process of drug substances or excipients. The European Pharmacopoeia (Ph. Eur.) limits the amount of RS in pharmaceuticals, considering the International Conference on Harmonization (ICH) of Technical Requirements ...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Sitaramaraju Y,Riadi A,D'Autry W,Wolfs K,Hoogmartens J,Van Schepdael A,Adams E

    更新日期:2008-09-10 00:00:00

  • HPLC method for simultaneous determination of retinol, alpha-tocopherol and coenzyme Q10 in human plasma.

    abstract::A simple HPLC method with UV detection is proposed for the simultaneous determination of three lipophilic vitamins: all-trans-retinol, alpha-tocopherol and coenzyme Q(10) (ubiquinone) in human plasma. The following chromatographic conditions were used: RP-18 column, a mobile phase consisted of methanol -n-hexane 72:28...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章,随机对照试验


    authors: Karpińska J,Mikołuć B,Motkowski R,Piotrowska-Jastrzebska J

    更新日期:2006-09-18 00:00:00

  • Determination of the chiral isomers of CGS 26214, a synthetic thyromimetic agent, in human plasma using microbore chiral chromatography-tandem mass spectrometry.

    abstract::CGS 26214 is a racemic compound having cholesterol-lowering activity in rats, dogs, and monkeys. This compound has two equipotent chiral components CGS 28934(-) and CGS 28935(+). An analytical challenge was to develop a sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the analysis of the ...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Majumdar TK,Martin LL,Melamed D,Tse FL

    更新日期:2000-09-01 00:00:00

  • A novel hydrophilic interaction liquid chromatography method for the determination of underivatized amino acids in alimentary supplements.

    abstract::Amino acids playing important roles in metabolic processes are often included in dietary supplements whose use has largely expanded over the last 20 years not only in patients with particular deficiencies, but also in athletes and even common people that want to enrich their regular daily diet. In the present study, a...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Themelis T,Gotti R,Gatti R

    更新日期:2017-10-25 00:00:00

  • Forced degradation study of racecadotril: Effect of co-solvent, characterization of degradation products by UHPLC-Q-TOF-MS/MS, NMR and cytotoxicity assay.

    abstract::Racecadotril, an enkephalinase inhibitor, was subjected to hydrolysis (acidic and alkaline), oxidation, photolysis and thermal stress, as per ICH specified conditions. The drug showed extensive degradation under acidic, basic hydrolysis and oxidative stress conditions whereas, it was stable under other stress conditio...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Chiguru V,Lingesh A,R S,N S

    更新日期:2016-09-05 00:00:00

  • Method development for dihydralazine with HPLC-MS/MS--an old but tricky substance in human plasma.

    abstract::An HPLC-MS/MS method was developed and validated for the determination of dihydralazine in human plasma. HPLC-MS/MS has not been used before in a published paper and provides better sensitivity and selectivity. Therefore a much easier sample preparation than published before is feasible (protein precipitation). As thi...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 临床试验,杂志文章


    authors: Mascher DG,Tscherwenka W,Mascher HJ

    更新日期:2007-01-17 00:00:00

  • Introduction of a carbon paste electrode based on nickel carbide for investigation of interaction between warfarin and vitamin K1.

    abstract::In this paper a novel electrochemical sensor based on nickel carbide (Ni3C) nanoparticles as a new modifier was constructed. Ni3C nanoparticle was synthesized and characterized by scanning electron microscopy, X-ray diffraction and first-principles study. Electrochemical impedance spectroscopy (EIS) and cyclic voltamm...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Torkashvand M,Gholivand MB,Taherpour AA,Boochani A,Akhtar A

    更新日期:2017-05-30 00:00:00

  • Identification of in vitro metabolites of ethylphenidate by liquid chromatography coupled to quadrupole time-of-flight mass spectrometry.

    abstract::Ethylphenidate is a new potent synthetic psychoactive drug, structurally related to methylphenidate. Using human liver microsomes and cytosol, we have investigated for the first time the Phase-I and Phase-II in vitro metabolism of ethylphenidate. The structure of the metabolites was elucidated by hybrid quadrupole tim...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Negreira N,Erratico C,van Nuijs AL,Covaci A

    更新日期:2016-01-05 00:00:00

  • Use of reversed-phase liquid chromatography for determining the lipophilicity of alpha-aryl-N-cyclopropylnitrones.

    abstract::The relationship between a reversed-phase high-performance liquid chromatography (RP-HPLC) retention parameter and various calculated log P-values of our previously synthesized alpha-aryl-N-cyclopropyl-nitrone derivatives was investigated. The RP-HPLC experiments were carried out with acetonitrile-water and methanol-w...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Balogh GT,Szántó Z,Forrai E,Gyorffy W,Lopata A

    更新日期:2005-10-04 00:00:00

  • Evaluation of the content and stability of the constituents of mother tinctures and tinctures: the case of Crataegus oxyacantha L. and Hieracium pilosella L.

    abstract::As a part of our investigations on the content and stability of herbal drug preparations, we evaluated the content and stability of tinctures and mother tinctures of Hawthorn leaves and flowers and Hawkweed. Hawthorn preparations are mainly used by patients with cardiac diseases; Hawkweed is employed for the treatment...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Bilia AR,Eterno F,Bergonzi MC,Mazzi G,Vincieri FF

    更新日期:2007-05-09 00:00:00

  • Live cell extraction and HPLC-MS analysis for predicting bioactive components of traditional Chinese medicines.

    abstract::A novel strategy for predicting bioactive components in traditional Chinese medicines (TCM) using live cell extraction and high performance liquid chromatography-diode array detection-mass spectrometry (HPLC-DAD-MS) analysis was proposed. The hypothesis is that when cells are incubated together with the extract of TCM...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Li SL,Li P,Sheng LH,Li RY,Qi LW,Zhang LY

    更新日期:2006-05-03 00:00:00

  • In vitro metabolism of cyadox in rat, chicken and swine using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry.

    abstract::In this work, metabolites of cyadox (CYX) produced by liver microsomes, primary hepatocytes and intestinal microflora systems of rat, chicken and swine were identified and elucidated using ultra-performance liquid chromatography/quadrupole time-of-flight tandem mass spectrometry (UPLC/Q-TOF-MS). Expected and unexpecte...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Wu H,Li L,Shen J,Wang Y,Liu K,Zhang S

    更新日期:2012-08-01 00:00:00

  • Characterization of three main degradation products from novel oral anticoagulant rivaroxaban under stress conditions by UPLC-Q-TOF-MS/MS.

    abstract::Drugs of long-term use may cause the accumulation of chemical compounds in human body. Therefore, the evaluation and structure characterization of synthesis and degradation impurities is substantial to guarantee drug safety and successful pharmaceutical therapy. The present work evaluated the anticoagulant rivaroxaban...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Wingert NR,dos Santos NO,Nunes MA,Gomes P,Müller EI,Flores ÉM,Steppe M

    更新日期:2016-05-10 00:00:00

  • On-chip assay for determining the inhibitory effects and modes of action of drugs against xanthine oxidase.

    abstract::Xanthine oxidase (XO) is a key enzyme that can catalyze the conversion of xanthine to uric acid, causing various diseases in humans. We have developed a high-throughput chip-based assay that uses a photodiode array (PDA) microchip system to explore the inhibitory effects of drug analogs on XO. Inhibitory activities of...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Naoghare PK,Kwon HT,Song JM

    更新日期:2010-01-05 00:00:00

  • Elucidation of pharmacokinetics of novel DNA ligase I inhibitor, S012-1332 in rats: Integration of in vitro and in vivo findings.

    abstract::S012-1332 is the first DNA ligase I inhibitor that demonstrated in vivo anti-breast cancer activity. The present study aimed to assess the in vivo pharmacokinetics of S012-1332 in rats and interpret them with in vitro findings. A sensitive and selective liquid chromatography-tandem mass spectrometry bioanalytical meth...

    journal_title:Journal of pharmaceutical and biomedical analysis

    pub_type: 杂志文章


    authors: Riyazuddin M,Valicherla GR,Husain A,Hussain MK,Shukla M,Katekar R,Gupta AP,Singh P,Banerjee D,Hajela K,Gayen JR

    更新日期:2019-01-05 00:00:00