INTRODUCTION:

Glycopyrronium bromide is a chemically 3-[(2-cyclopentyl-2-hydroxy-2-phenylacetyl) oxy]-1, 1-dimethylpyrrolidin-1-ium (fig.1). Glycopyrronium bromide is a synthetic anticholinergic agent with a quaternary ammonium structure. Glycopyrronium bromide is a synthetic anticholinergic agent and a competitive muscarinic antagonist [1].

Glycopyrronium bromide binds competitively to the muscarinic acetylcholine receptor M1 to M5. Like other anticholinergic (antimuscarinic) agents, it inhibits the action of acetylcholine on structures innervated by postganglionic cholinergic nerves and on smooth muscles that respond to acetylcholine but lack cholinergic innervation.

These peripheral cholinergic receptors are present in the autonomic effector cells of smooth muscle, cardiac muscle, the sinoatrial node, the atrioventricular node, exocrine glands and, to a limited degree, in the autonomic ganglia. Thus, it diminishes the volume and free acidity of gastric secretions and controls excessive pharyngeal, tracheal, and bronchial secretions.

Glycopyrronium bromide is available in form of injection, oral solution, and oral tablets [2]. Glycopyrronium bromide is used in treatment of primary pediatric hyperhidrosis, acute respiratory disease, sialorrhea, peptic ulcer and in practice of anesthesia to reduce salivation [3, 4].

Absorption of CUVPOSA (fasting) was compared to that of a marketed Glycopyrronium bromide oral tablet. The C_max after oral solution administration was 23% lower compared to tablet administration and the AUC₀-inf was 28% lower after oral solution administration. Mean C_maxafter oral solution administration in the fasting state was 0.318 ng/mL, and mean AUC₀-24was 1.74 ng.hr/mL. Mean time to maximum plasma concentration for CUVPOSA was 3.1 hours, and mean plasma half-life was 3.0 hours.

After IV administration, Glycopyrronium bromide has a mean volume of distribution in children aged 1 to 14 years of approximately 1.3 to 1.8 L/kg, with a range from 0.7 to 3.9 L/kg. In adults aged 60-75 years, the volume of distribution was lower (0.42 L/kg +/- 0.22). Metabolism, in adult patients who underwent surgery for cholelithiasis and were given a single IV dose of tritiated Glycopyrronium bromide, approximately 85% of total radioactivity was excreted in urine and < 5% was present in T-tube drainage of bile. In both urine and bile, > 80% of the radioactivity corresponded to unchanged drug. These data suggest a small proportion of IV Glycopyrronium bromide is excreted as one or more metabolites.

Approximately 65-80% of an IV Glycopyrronium bromide dose was eliminated unchanged in urine in adults. In two studies, after IV administration to pediatric patients ages 1-14 years, mean clearance values ranged from 1.01- 1.41 L/kg/hr (range 0.32 – 2.22 L/kg/hr). In adults, IV clearance values were 0.54 ± 0.14 L/kg/hr [5].

Common side effects of Glycopyrronium bromide are stomach/abdominal bloating or pain, persistent constipation, persistent nausea, vomiting, diarrhoea, decreased sweating, dry/hot/flushed skin, fast/irregular heartbeat, mental/mood changes (such as confusion, hallucinations, agitation, nervousness, unusual excitement), difficulty urinating/inability to urinate. Drowsiness, dizziness, blurred vision, dry mouth [6].

Figure 1: Chemical structure of Glycopyrronium Bromide

Analytical methods for Glycopyrronium bromide for estimation in Bulk drug, Pharmaceutical formulation and Biological fluids:

Many different analytical methods have been reported for the estimation of Glycopyrronium bromide in bulk and dosage form as well as in biological fluids.

Spectrophotometric methods:

Jamdar P et al developed and validated a simple, precise and accurate method for the quantitative determination of Neostigmine Methyl Sulphate (NEO) and Glycopyrronium bromide (GLYCO) in bulk and injectable dosage form. The method was based on of Q-absorbance equation method at 241nm isoabsorptive point and at 260nm, which is λmax of Neostigmine Methyl Sulphate. Linearity of response was obtained in the concentration range of 125-425μg/ml for NEO and 25-85μg/ml for Glycopyrronium bromide. This involved method was successfully applied to the marketed formulations and no interference was obtained due to excipients. Validation was performed as per the ICH guidelines to prove the suitability of method for quantitative determination and reproducibility. It was cost effective method because of only distilled water was used as reagent in method development [7].

Abdel Ghany MF et al. developed and validated three specific, sensitive and precise spectrophotometric methods for determination of Indacaterol (IND) and Glycopyrronium (GLY) in their binary mixtures and novel pharmaceutical dosage form for quality control. The proposed methods are considered to be the first spectrophotometric methods for IND and GLY simultaneous determination without prior separation. The developed methods are based on different signal processing techniques of ratio spectra namely; Numerical Differentiation (ND), Savitsky–Golay (SG) and Fourier Transform (FT). The developed methods were linear in concentration range of 1–30 and 10–35 (μg/mL) for IND and GLY, respectively. The accuracy data as percentage recoveries were reported in the range of 99.00%–100.49% with low value of RSD% (< 1.5%) demonstrating an excellent accuracy of the proposed methods [8].

Chromatographic Method:

HPLC method (High Performance Liquid Chromatography):

Venketshwaran TG et al. developed High Performance Liquid Chromatographic method for the assay of Ondansetron-atropine and Ondansetron-Glycopyrronium bromide mixtures in 0.9% sodium chloride injection. The separation and quqntitation of the ondansetron-atropine mix was performed and successfully achieved on an octylsilane column at ambient temperature using a mobile phase of 60:40 v/v 0.01 M phosphate buffer, pH 4-acetonitrile at a flow rate of 1.0 mL/min with detection of the analytes at 254 nm. Linearity was showed for Ondansetron and Atropine in the 266–1332 and 28–138 μg/mL ranges, respectively. Accuracy and precision were reported in the 0.2–5.6% and 0.4–1.8% ranges, respectively, for both drugs. The limits of detection for Ondansetron and Atropine were found to be 2.1 ng/mL and 8.6 μg/mL, respectively, based on a signal to noise ratio of 3 μL and a 20 μL injection. The separation and quantitation of the ondansetron-Glycopyrronium bromide mix was performed and successfully achieved on an octylsilane column at ambient temperature using a mobile phase of 55:45 v/v 0.01 M phosphate buffer (pH 4): acetonitrile at a flow rate of 1.0 mL/min with detection of the analytes at 254 nm. The separation was achieved within 15 min. The method showed linearity for Ondansetron and Glycopyrronium bromide in the 500–2000 μg/mL and 50–200 μg/mL ranges, respectively. Accuracy and precision were in the 2.5–3.7% and 0.1–1.5% ranges, respectively, for both analytes. The limits of detection for Ondansetron and Glycopyrronium bromide were found to be 90 ng/mL and 6.9 μg/mL, respectively, based on a signal to noise ratio of 3 and a 20 μL injection [9].

Parameshwar P et al. developed and well validated a simple, accurate and precise efficient reverse phase high performance liquid chromatography (RP-HPLC) method for estimation of Glycopyrronium bromide in bulk and its tablet dosage forms. Chromatography separation of analytes were carried out on μ Bondapak C-18 column (300 X 3.9mn; 10μm) as stationary phase by using mobile phase consists of Mixture of buffer (sodium sulphate buffer), 1N Sulfuric acid, acetonitrile and methanol in the ratio of 1230:6:470:300 v/v/v/v at a flow rate of 2.3 ml/min and UV detection at 222 nm, column temperature at 30ºC. The retention time for Glycopyrronium bromide was 4.2 minutes. Extraction of the drug from the dosage form carried out by using Mixture of purified water and Acetonitrile (60:40 v/v). The method was linear in the concentration rang of 20μg-80μg/ml. The percentage recovery of Glycopyrronium bromide was reported 99.08%. This condition is applied only for tablet dosage form [10].

Nebiu D et al. developed a simple, rapid and specific ion-pair HPLC method for the determination of (R, R)-Glycopyrronium bromide and its related impurities, and parameters affecting the chromatographic properties of these compounds are discussed. Optimal chromatographic analytes separation was achieved on Nucleosil column as stationary phase at 40ºC, using phosphate buffer pH 2.30 with sodium-1-decanesulfonate (0.01 M): methanol (35:65 v/v) as mobile phase for isocratic elution at a flow rate 1 ml/min. The analytical assay was validated according to ICH guidelines. The method was reported as suitable for in-process control and as stability indicating assay [11].

HPTLC Method (High Performance Thin Layer Chromatography):

Chitlange SS et al developed and validated a selective, accurate, precise and reproducible high performance thin-layer chromatographic (HPTLC) for the analysis of Glycopyrronium bromide in tablet dosage form (Glycopyrronium bromide tablet USP). The chromatographic separation was performed on pre-coated silica gel 60 F254 (Merck) plate as stationary phase and using ethyl acetate: water: glacial acetic acid (7: 2: 1 v/v/v) as mobile phase. Densitometric analysis was performed in reflectance-absorbance mode at 222 nm. R_f value of the Glycopyrronium bromide was found to be 0.34. The linear response data for the calibration plot showed good linear relationship with correlation coefficient 0.99 (1-6 μg/band). The limit of detection and quantitation were reported 220 μg/ml and 0.667 μg/ml respectively. The accuracy of method proven with percentage recovery of Glycopyrronium bromide 99.87% [12].

Bioanalytical methods:

There are few methods available for estimation of Glycopyrronium bromide in biological matrices like plasma, urine.

Rumpler MJ et al. performed A rapid, sensitive, and specific ultra-high-performance liquid chromatography with heated electrospray ionization-tandem mass spectrometry (UHPLC–HESI-MS–MS) method for detection and Quantification of Glycopyrronium bromide in Horse Plasma. This method also involed determination of Glycopyrronium bromide in plasma after oral and intravenous administration of clinically relevant doses to Thoroughbred horses. Calibration was achieved by weighted, linear regression analysis using a deuterated analogue of Glycopyrronium bromide as internal standard (IS). Isolation of Glycopyrronium bromide (GLY) and the internal standard (GLY-d3) from plasma matrices carried out via weak cation exchange using a simple solid-phase extraction technique. Chromatographic analysis was performed and achieved by reversed-phase UHPLC on a C18 Acquity™ column as stationary phase. Analysis of Extracts was carried out in positive electrospray ionization mode and precursor and product ions were detected and quantified by MS–MS using a triple-stage quadrupole (TSQ) instrument. The method was showed a linear in range of 0.125–25 pg/mL (R2 > 0.998). Lower limit of quantification and lower limit of detection were found to be 0.125 pg/mL and 0.025 pg/mL respectively. Recovery of Glycopyrronium bromide obtained in ranged from 78% to 96%, and intra- and inter batch precision were found to be 3.3–14.4% CV and 3.4–14.4% CV respectively. The stability of Glycopyrronium bromide in plasma was up to 170 days at –80°C, through three freeze/thaw cycles, and for up to 48h after extraction under 20°C autosampler conditions [13].

Storme ML et al. developed a quantitative tool for the determination of the quaternary ammonium anti-cholinergic agent Glycopyrronium bromide in human plasma samples. Mepenzolate was used as internal standard. Two step liquid-liquid ion-pair extraction procedure was performed for plasma samples which was simple and relatively fast. The chromatography was performed using the same volatile ion-pair reagent heptafluorobutyric acid (HFBA), takes only 10 minutes. Relative standard deviation of retention times was never above 2.257% (n=36). A quantitative ESI-LC-MS(/MS) (TOF-mass spectrometry) method was used for the absolute quantitation of Glycopyrronium bromide in human plasma in a concentration range from 0.1009 ng/mL up to 100.9 ng/mL, described by the quadratic calibration function (R²0.9995), y = -2.210 * 10^-4(±3.926 * 10-5)*x² + 5.847 * 10^-2(±5.274 * 10^-3)*x + 4.081 *10^-3(±4.816 * 10^-4). The optimized method was fully validated as per US FDA Bio-analytical Method Validation Guidance for Industry. For the three QC concentrations (QC₁0.253, QC₂2.53, and QC₃25.3 ng/mL) and the LLOQ (0.101 ng/mL), precision was reported under 15% (17.95% (n=6) at the LLOQ concentration) and maximum accuracy was achieved 112.4% (88.92% for the LLOQ n=6). Absolute matrix effect (maximum 143.9% ± 11.59, n=3), absolute recovery (better than 41.77% ± 2.218, n=3), relative (inter-subject) matrix effect (maximum 10.89% ± 1.446, n=3) and process efficiency (better than 45.17% ± 5.739, n=3) were performed at the 3 QC concentrations [14].

REFERENCES:

1. Mirakhur RK, Dundee JW. Glycopyrronium bromide: pharmacology and clinical use. Anaesthesia. 1983; 38(12): 1195-1204.

2. The United State Pharmacopoeia USP31-NF 26, United States pharmacopoeial Convention, Rockville, Vol-II; 2008: 2288,2289

3. “Information of Glycopyrronium bromide”, [cited 2016 Oct 5]. Available from:http://www.rxwiki.com/glycopyrronium-bromide

4. CM Slovis, GM Daniels, DR Wharton. Annals of emergency medicine, Journal of Pharmaceutical Research. 1987: 6-11.

5. “Glycopyrronium bromide oral solution”, [cited 2016 Oct 21]. Available from: http://www.accessdata.fda.gov/drugsatfda_docs/label/2010/022571s000lbl.pdf

6. “Drug profile of Glycopyrronium bromide”, [cited 2016 Oct 5]. Available from: http://www.webmd.com/drugs/2/drug-8788-1384/Glycopyrroniumbromide oral/Glycopyrronium bromide-solution--oral/details#side effects

7. Jamadar P, Meshram D. Isoabsorbtive method for the simultaneous estimation of neostigmine methyl sulphate and Glycopyrronium bromide in injection form. FS Journal of Pharmacy Research. 2015; 4: 5-8.

8. Abdel Ghany MF, Hussein LA, Magdy N, Yamani HZ. Simultaneous spectrophotometric determination of Indacaterol and Glycopyrronium in a newly approved pharmaceutical formulation using different signal processing techniques of ratio spectra. Spectrochimica Acta, Part A: Molecular and Biomolecular Spectroscopy. 2016; 157: 251–257.

9. Venketshwaran TG, King DT, Stewart JT. HPLC determination of oldansetron-atropine and oldansetron-Glycopyrronium bromide mixtures in 0.9% sodium chloride injection. Journal of Liquid Chromatography .2006; 18: 2647-2659.

10. Parmeshwar P. RP-HPLC Method Development and Validation for Assay of Glycopyrronium bromide in Tablet. Asian Journal of Pharmaceutical and Clinical Research. 2011; 4(4): 37-40.

11. Nebiu D, Walter M, Lachmann B, Kopelent H, Noe CR. Determination of (R,R)-Glycopyrronium bromide and its related impurities by ion-pair HPLC. Pharmazie. 2007; 62: 406–410.

12. Chitlange SS, Jagadale SS, Chandani SR, Gandhi SP. Development and validation of hptlc method for Glycopyrronium bromide in single dose tablet formulation. Journal of Liquid Chromatography. 2006; 18: 2647-2659.

13. Rumpler MJ, Sams RA, Colahan P. Validation of a Liquid Chromatography–Tandem Mass Spectrometry Method for Quantification of Glycopyrronium bromide in Horse Plasma. Journal of Analytical Toxicology. 2011; 35: 656-664.

14. Strome ML, TKnidt R, Goeteyn W, Bocxlaer JV, Reynjens KM. Quantitative determination of Glycopyrronium bromide in human plasma by liquid chromatography-electrospray ionization mass spectrometry: The use of ion pairing agent during both.. Journal of Chromatography B. 2008; 876: 24-30.

Received on 08.08.2017 Accepted on 25.10.2017

Asian J. Pharm. Ana. 2017; 7(4): 239-242.

DOI: 10.5958/2231-5675.2017.00039.4