Determination of impurities in heparin by capillary electrophoresis using high molarity phosphate buffers

Wielgos, Todd; Havel, Karalyn; Ivanova, N. T.; Weinberger, Robert

doi:10.1016/j.jpba.2008.11.034

Cited by 47 publications

(37 citation statements)

References 9 publications

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“…CE. Capillary electrophoresis was completed on an Agilent Technologies CE System using a modification of a published procedure (29). The buffer was 1 M Li Phosphate, pH 2.5.…”

mentioning

confidence: 99%

Orthogonal analytical approaches to detect potential contaminants in heparin

Guerrini

Zhang

Shriver

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Heparin is a widely used anticoagulant and antithrombotic agent. Recently, a contaminant, oversulfated chondroitin sulfate (OSCS), was discovered within heparin preparations. The presence of OSCS within heparin likely led to clinical manifestations, most prevalently, hypotension and abdominal pain leading to the deaths of several dozens of patients. Given the biological effects of OSCS, one continuing item of concern is the ability for existing methods to identify other persulfonated polysaccharide compounds that would also have anticoagulant activity and would likely elicit a similar activation of the contact system. To complete a more extensive analysis of the ability for NMR and capillary electrophoresis (CE) to capture a broader array of potential contaminants within heparin, we completed a systematic study of NMR, both mono-and bidimensional, and CE to detect both various components of sidestream heparin and their persulfonated derivatives. We show that given the complexity of heparin samples, and the requirement to ensure their purity and safety, use of orthogonal analytical techniques is effective at detecting an array of potential contaminants that could be present.anticoagulant ͉ capillary electrophoresis ͉ contamination ͉ NMR ͉ oversulfated chondroitin

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“…CE. Capillary electrophoresis was completed on an Agilent Technologies CE System using a modification of a published procedure (29). The buffer was 1 M Li Phosphate, pH 2.5.…”

mentioning

confidence: 99%

Orthogonal analytical approaches to detect potential contaminants in heparin

Guerrini

Zhang

Shriver

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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“…The LOD obtained for OSCS was 0.1% (30 mg/mL) using 30 mg/mL of Hep sample concentration [7]. Moreover, Somsen et al have developed a CE method using 850 mM tris-phosphate at pH 3.0 as background electrolyte using high Hep concentrations (up to 50 mg/mL) and large injection sample volumes.…”

Section: Optimization Of Buffer Type Concentration and Phmentioning

confidence: 99%

“…To solve this problem, different authors worked with smaller diameter capillaries (25 mm). Consequently, higher concentrations of Hep samples (50-60 mg/mL) were necessary for the analysis [7,8] making impossible to test the finished product in which Hep concentration is usually well below 10 mg/mL.…”

Section: Optimization Of Buffer Type Concentration and Phmentioning

confidence: 99%

“…The analytical method reported for the analysis of contaminants like OSCS and the impurities of Hep is NMR [5,6], CE [1,7,8], and HPLC [6,9]. The USP official method uses a polymer-based strong anion exchange column and a 1% limit is set for the galactosamine impurities expressed as dermatan sulfate and other galactosamines along with the absence of OSCS.…”

Section: Introductionmentioning

confidence: 99%

“…[11,12]. Different CE methods have been reported for the analysis of Hep and its impurities and contaminants with the use of different inorganic buffers at concentrations ranging from 50 to 850 mM, acidic pH values and the addition of salts with different cations like sodium, ammonium, lithium, or amines [7,8].…”

Section: Introductionmentioning

confidence: 99%

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Novel and highly sensitive mixed‐polymeric electrokinetic chromatography system for determination of contaminants and impurities of heparin samples

et al. 2010

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A mixed-polymeric electrokinetic chromatography system has been developed for the simultaneous determination of a contaminant like oversulfated condroitin sulfate (OSCS) and impurities expressed as dermatan (Der) in heparin (Hep) samples. The EKC system consisted of 0.5% w/v polymeric β-CD, 0.4% w/v tetronic(®) 1107 and 400 mM tris-phosphate buffer at pH 3.5. The optimized electrophoretic conditions included the use of an uncoated-silica capillary of 50 cm of total length and 75 μm id, an applied voltage of -7 kV, a temperature of 30°C and 200 nm UV-detection. The highly sensitive method developed showed low values of LOD, 0.07% w/w (0.07 μg/mL) (OSCS) and 0.1% w/w (0.1 μg/mL) (Der), and values of LOQ 0.2% w/w (0.2 μg/mL) (OSCS) and 0.3% w/w (0.3 μg/mL) (Der) with a concentration level of Hep sample as low as 0.1 mg/mL. Additional parameters of validation such as specificity, linearity, accuracy, and robustness were evaluated according to international guidelines. Owing to its simplicity, high sensitivity, and reliability, the proposed method can be an advantageous alternative to the traditional methodologies for the analysis of Hep in raw material and specially in finished products because of the low amounts of Hep sample required.

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