ANALYTICAL METHOD VALIDATION FOR DETERMINATION OF HEAVY METAL IN CAPSULE SHELL BY USING INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY (ICP-MS)

R K Phadke 1 and V D Gaitonde 2 . 1. Research Scholar, J.J.T University, churu, Jhunjhunu Road, Rajasthan -333001. 2. ProChrome India, A/2 ,Varsha Milan, Sahra road, Andheri (East), Mumbai-400099, India. ................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History


ISSN: 2320-5407
Int. J. Adv. Res. 4 (10), 447-456 448 Q3D have given control stringent limit base on risk assessment. The graphical representations of material used in gelatin production are as follows. Gelatin capsule was first patented by Mr.F.A.B. Mothes, students and Dublanc, a pharmacist, they obtained the patent in 1834, cover a method for producing gelatin capsules consisting of one section, oval-shaped, and covered with a drop of hot concentrated solution of gelatin after charging. The uses of gelatin capsules are spread even produced by many countries in Europe and America restricted they use gelatin capsules patent on a particular company, sparked two new capsule forms. In 1839 in Paris, Garot create a thin layer coated products, gelatin-coated pills. In 1846 another pharmacist, J.C. Lebhubby patented capsule 2 parts which is still used. Many medications enclosed in capsule shell are administered orally. The Pharmacopoeia of the People's Republic of China (2010 version) sets a clear standard for the grade of gelatin that can be used for drug capsule production and requires that pharmaceutical companies only purchase capsules from manufacturers that are licensed. There have been recent reports that some companies in eastern China have been making and selling capsules made from cheaper industrial gelatin prepared from discarded leather. Heavy metal like Chromium, which is a known carcinogen, and can be toxic if ingested in large quantities, is used in the leather tanning process. Consequently 20 to 90 times more Cr was typically found in the leather-derived gelatin than in pharmaceutical/edible grade gelatin.
In current pharmacopeia heavy metals control by heavy metal test by visual observation no any specific instrument technique like flame photometric, atomic abortion spectroscopic, inductively coupled plasma atomic emission spectroscopy and inductively coupled plasma mass spectroscopy was given. In most of Active pharmaceutical ingredient and excipients pharmacopeia mentions that heavy metals to be perform by any of analytical instruments. This instrumentation technique will compulsory applied by pharmacopeia from year 2018. In current scenario most of literatures are given on inductively coupled plasma atomic emission spectroscopy (ICP-AES), however an extensive survey revealed that there were no any quantitative methods for determination of genotoxic heavy metal by inductively coupled plasma mass spectrometry (ICP-MS) in hard gelatin capsule. Hence it was felt necessary to develop an accurate, rapid, sensitive, and specific method for the determination of heavy metals in hard gelatin capsule. We developed simple, fast, linear, accurate, reproducible and robust ICP-MS method. The method was validated by following ICH guideline parameter.   System Suitability Criteria:-1. The correlation coefficient should not be less than 0.99 2. Cumulative %RSD of intensity response of Std-5 and Bracketing standard (Std-5) should not be more than 20.

Calculations:-
Calculate the concentration of element in sample as per following formula: Where, I= Intensity Response of element for Sample. C= Intercept of the linearity curve. m= Slope of the linearity curve.

Specificity:-
Specificity is the ability of a method to measure specifically or selectively the analyte in the presence of components which may be expected to be present in the sample. Specificity was established by analyzing the blank, Test blank, standard and Sample solutions in ICP-MS and observed the interference. The observed interference of blank and Test blank was less than 3.0% hence method is specific refer (Table 3). System suitability (system precision):-Six replicate of Linearity standard solutions 5 was run and find out relative standard deviation and System suitability run the Linearity standard solutions from 1 to 5 and check co-relation coefficient. The %RSD of the six replicate run of Linearity standard solutions 5 was should be below 20% and co-relation coefficient of Linearity standard from 1 to 5 should less than 0.99 refer ( Table 4 and 5).

Limit of detection (LOD) and limit of quantitation (LOQ):-
Five different concentrations of standard where run and find out the slope and STE YX . Base on slope and STE YX determined the LOD and LOQ. The LOD and LOQ for the element found to be 0.006ppm and 0.02ppm for As; 0.005ppm and 0.017 for Cd; 0.002ppm and 0.006 for Hg and 0.019ppm and 0.063ppm for Pb w.r.t test concentration refer (Table 6 ).   Accuracy/ recovery study:-Accuracy of method was determined by doping the respective concentration solution of element in test preparation and find out the content of elements from test preparation. Recovery studies were carried out at concentration LOQ, 50%, 100%, and 200%.The obtained % recovery was well within the limit 70% to150%. For accuracy refer (Table.10

Robustness study:-
The robustness study was carried out by varying the instrument parameter and find out the content of heavy metals. The variation in parameters was change in Dwell time from to 0.1s to 0.11s; Power hold time from 15 min to 16.5 min and 15min to 13.5 min. The obtained results of element were shown in (Table.12, 13 and 14).

Results and Discussion:-
Heavy metals are toxic in nature and have to control in specified limit. In resent ICH Q3D guideline specific limits were given for each element. In current pharmacopeia heavy metals test procedure performed by chemically and observed by visual observation, there was no any specific instrument methods was given in pharmacopeia therefore pharmacopeia team has revised the USP General chapter <232> and <2232>. This change will effective in year 2018. In hard gelatin capsules manufacturing process lots of water and colour dyes were used therefore heavy metals present in sample. To control these heavy metals needs method development. In hard gelatin capsule Arsenic, Mercury, Lead and Cadmium are heavy metals. We tried to develop a heavy metal on atomic absorption spectroscopy (AAS) instrument but due to less sensitivity and stringent limits as well as some limitation of AAS, AAS technique was not feasible. Hence we tried to develop method on inductively coupled plasma mass spectrometer. The sample preparation technique of hard gelatin capsule is very difficult because metals are soluble in waters and gelatin was insoluble in water. Hence sample preparation technique was critical. We use concentrated hydrochloric and nitric acid for sample preparation but sample was not dissolving properly. In sample preparation some issues was observed therefore we digest the sample in microwave digester and run the sample in inductively coupled plasma mass spectrometer (ICP-MS). The results obtained of such sample are getting higher side where as blank inference was more than 3.0%. Therefore again method development was required. We developed new technique using hydrogen peroxide and digest the sample in microwave digester. In this technique the blank interfere was below 3.0%. Base on this technique we further validate the method for specificity, linearity, accuracy, precision and robustness and obtained result are within acceptance criteria. The method was validated by ICH Q2 (R1) guideline parameter. 456

Conclusion:-
The developed analytical method for determination of Arsenic, Mercury, cadmium and Lead as heavy metal in hard gelatin capsule shell by using inductively coupled plasma mass spectroscopy (ICP-MS). The analytical method was specific,Accurate, precision, reproducible, rugged, linear and robust method. The same method has been validated as per ICH guideline Q2 (R1). This method can be use for routine quality control sample analysis or can use for control monitor for heavy metals in the manufacturing process of hard gelatin capsule preparation.