Quantification of Total Hydroxypropyl Methylcellulose (HPMC) Levels: Analysis of All Congeners/Isomers as a Single Analytical Peak by HPLC with Corona

CAD.
Darwin Asa, Ian Acworth, Tom Villaseor, David Carreiro, John Christensen, and Eddie Goodall1 ESA Biosciences, Inc., Chelmsford, MA 1ESA Analytical, Aylesbury, England, UK

Abstract
Hydroxypropyl methylcellulose (HPMC) is a non-ionic water soluble ether of methylcellulose. It is commonly used by the pharmaceutical, food, adhesive, paint, printing, and textile industries and has a variety of uses including: an enteric film coating, viscosity control agent, gelling agent, stabilizer, lubricant, binder, emulsifier and suspending agent. It uses throughout these industries is greatly affected by the viscosity of the solution, which typically ranges from 400-15,000 cp. Determination of HPMC is complicated as it is composed of a mixture of isomers and congeners the relative abundance of each affecting solution viscosity. A variety of approaches are available for measuring HPMC forms, but these tend to be complicated, lack selectivity, or suffer from a lack of sensitivity and poor dynamic range (e.g., HPLC-UV; HPLC-ELSD). Recently, a new HPLC detector using Charged Aerosol Detection (Corona CAD) was developed. CAD is an evaporative technique and is based on the charging of aerosolborne analyte particles by nitrogen gas and corona discharge, with subsequent measurement of the charge, derived from the charged analyte particles, by a high sensitivity electrometer. Two isocratic methods were evaluated for the measurement of total HPMC levels (all congeners/isomers are forced to elute as a single analytical peak) using HPMC with viscosity of 4,000cp as the test analyte. Method 1 used a RP approach Asahipak NH2 column with aq acetonitrile as the mobile phase: HPMC eluted at 3.2mins; LOD ~1.5g o.c. Method 2 used a GFC approach Tosoh TSKgel column with aq acetonitrile containing ammonium acetate buffer as the mobile phase: HPMC eluted at 5.5mins; LOD ~800ng o.c. Method 2 proved more robust and showed fewer chromatographic artifacts.

Materials and Methods

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Discussion and Conclusions

Corona Parameters: Gas: 35psi via nitrogen generator Filter: None Range: 100pA

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Method 1
HPLC Parameters: Mobile Phase: Flow Rate: Column: Column Temp: Injection Vol: 90% Aqueous acetonitrile 1.0mL/min Asahipak NH2 40 oC 10L
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As demonstrated in the data presented here, the Corona CAD is readily able to analyze HPMC when it analyzed in total in a single peak analysis using either a RP approach (Method 1) or a GFC approach (Method 2). Using Method 2, the Corona CAD was able to measure less than 1 g on column. The Corona CAD is an excellent detector for the analysis of complex polymer mixtures such as HPMC. It is especially powerful in the case of molecules such as HPMC which lack efficient chromophores for easy analysis. With the Corona CAD, it is possible to measure HPMC with a universal detector even at low levels. To help address the many challenges of universal detection, such as HPMC, ESA has developed the Corona CAD detector. This novel technology offers many benefits to analytical scientists including: High Sensitivity - Low ng limits of detection. Consistent Response Factors - Response magnitude does not significantly depend on analyte properties Broad and Useful Dynamic Range - 4 orders of magnitude (ng - g quantities). Excellent Reproducibility - Typically less than 2% RSD. Broad Applicability - Can be used with a wide variety of HPLC conditions to measure virtually any nonvolatile analyte including proteins, lipids, carbohydrates and small molecules. Ease of Use - Easy setup. Uses minimal bench space and requires only gas input pressure and signal output range to be set.

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Method 2
HPLC Parameters: Mobile Phase: 90% 40mM Ammonium Acetate (pH6.6); 10% acetonitrile Flow Rate: 1.0mL/min Column: Tosoh TSKgel; G3000SWXL; 7.8 x 300mm Column Temp: Ambient Injection Vol: 20L

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Sample preparation The standard, (viscosity of 2% solution ~ 4,000cP) obtained from Sigma (St. Louis, MO), was dissolved in mobile phase. Further dilutions were in mobile phase.

Figure 1. Analysis of HPMC Standard Using Method 1. A = 25g (on column), B = 2.1g (on column).

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Introduction
Measurement of Hydroxylpropylmethyl cellulose (HPMC) is problematic due to the lack of an efficient chromophore on the molecule and the polydispersity and heterogeneity inherent in the preparation of this polymer. Typical samples of this material contain a wide variety of molecules of various sizes and compositions. Measurement techniques used for HPMC involve the use of viscosity or refractive index measurements. Neither of these techniques is particularly useful in assessing the composition of these molecules as they lack the specificity and sensitivity required for useful assessment. Additionally, many of the chromatographic techniques described for analyzing HPMC involve separation of individual components. In some cases, analysis of the overall amount of HPMC present in a sample is the analytical goal and this type of measurement requires a single peak separation scheme for accurate quantitation. To meet the challenges of HPMC analysis, the Corona CAD universal HPLC detector was applied. The Corona CAD possesses superior sensitivity and dynamic range to other universal HPLC detection techniques and would appear to be a good candidate for analyzing HPMC. The Corona CAD was evaluated for its ability to measure HPMC in either seperate or single peak analysis operations.
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The Corona CAD provides universal detection of nonvolatile analytes with response independent of chemical properties, a wide dynamic response range, high sensitivity and good precision. These characteristics, along with reliability and simple operation, make this a superior detector for a wide range of HPLC analyses, especially for polymers like HPMC. For more information about this application, the Corona CAD, or charged aerosol detection visit www.esainc.com.
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Figure 2. Analysis of HPMC Standard Using Method 2. A = 41g (on column), B = 840ng (on column).