अरुण भोई द्वारा गुन्हे कथा मालिका

अरुण भोई द्वारा गुन्हे कथा मालिका

नमस्कार वाचक रसिकहो. हि दिपावली आपणा सर्वांना सुख, समाधान, आरोग्यदायी व समृद्धीची जावो. मित्रहो दरवर्षी चोखंदळ वाचक दिवाळी अंकांची वाट पहात असतात आणि वाचतात. दिवाळीच्या फराळाप्रमाणे विविध प्रकारच्या कथा, कादंबऱ्या, कविता, व्यंगचित्रे इ.चा आनंद घेतात.

अरुण भोई आता वाचकांच्या जिज्ञासा पूर्तीसाठी सत्य घटनांवर आधारित गुन्हेगारी विश्वातील, जगभरातील गाजलेले मती शून्य करणारे गुंतागुंतीचे गुन्हे, त्यांचा तपास व न्यायदान याचा भंडारा आपणा समोर उघडत आहे. साध्या, सोप्या मराठी भाषेत आपल्या वाचनासाठी मांडत आहे. वाचकांसाठी एकानंतर एक अशा अनेक गुन्हे कथा, गुन्हे करण्याची गुन्हेगाराची पद्धती, गुन्हे तपास, त्यास न्यायसहायक विज्ञानाची जोड व न्यायदान प्रक्रिया याविषयीचे ज्ञान समृद्ध करतील.

यास वाचकांचा भरभरुन प्रतिसाद मिळेल यात शंका घेण्याचे कारण नाही.

अरुण भोई: फॉरेन्सिक एक्स्पर्ट

SEMI – QUANTITATIVE DETERMINATION OF ANTHRACENE USING TLC.

SEMI – QUANTITATIVE DETERMINATION OF ANTHRACENE USING TLC.

ARUN BHOI, Forensic Expert

(Former Head of Division, Directorate of Forensic science Laboratories, State of Maharashtra (INDIA) and Former Senior Scientific Executive and OSD, Goa State Forensic Science Laboratory, Goa (INDIA)

e- J. Foren. Crime Inv. 2021, 23-35, 1, Art. 1.

ABSTRACT

In India, Anthracene powder has a great concern with the bribery cases. Therefore, TLC method has been described to analyse anthracene semi – quantitatively. Thin-layer chromatography with sulphanilic acid chromogenic reagent was used for this purpose. The Beer’s law obeyed good for the quantities in the range of ’05 μg to 50 μg’ of anthracene.

KEY WORDS

Anthracene, TLC, thin layer chromatography, Anthracene determination, Tetracene

INTRODUCTION

 A chemical substance ‘Anthracene’ powder is a good tool in the hands of the police personnel attached to the ‘Anti-corruption Bureau’ to catch hold of the culprit, a bribe receiver. The Anthracene powder has been applied to the currency notes and/or relevant articles concerned with the bribery cases. In the process of planned bribe transaction trap, as soon as the money or so has been accepted by the receiver the applied powder gets transferred to his hands. If the money has been accepted and put in a pocket or handkerchief or table drawer, etc. the Anthracene powder falls on or adheres to the surfaces came in contact. Then the relevant articles and powder particles have been collected and seized by the anti-corruption bureau officials for the forensic examination and to ascertain the presence of the powder on the hands and clothes of the culprit and at the places where the currency notes have been put. It necessitates the forensic expert to detect the presence of the anthracene and to determine its quantity found.

The anthracene powder can be seen as a blue-violet fluorescence under ultra-violet light (1). The impurity of tetracene associated with anthracene is observed as yellow particulates with greenish fluorescence (2, 3). Ultra-violet spectroscopy has been used to obtain data for pure anthracene (4, 5). Thin – layer Chromatographic methods (6, 7) explain the separation of constituting impurities present in commercial anthracene. Bhoi, et. al. have carried out the forensic examination of anthracene traces on seizers in anti – corruption offence by TLC, UV and GLC techniques (8). Herein, the TLC technique described by Bhoi, et. al. (8) has been adopted to determine the anthracene semi- quantitatively.

EXPERIMENTAL

Reagents

All solvents and reagents used were of AR Grade. Glass distilled water, ethyl alcohol (95%) were used wherever required.

Sulphanilic acid diazo – salt chromogenic reagent (0.2%)

This reagent was prepared by adding equal quantities of 0.2 %  sulphanilic acid solution in 1N HCl and 0.3 % aqueous solution of sodium nitrite and allowing the mixture to set for half an hour (9).

Standard anthracene solution

The standard anthracene solution was prepared by dissolving 5 mg of anthracene in 5 ml of ethyl alcohol (95%) in a stoppered 5 ml capacity measuring cylinder.

PROCEDURE

Thin Layer Chromatography

A clean TLC plate of the size of 10 cm x 20 cm was taken. It was coated with the slurry of Silica Gel G (E. Merck) (1:2 w/w, SGG:Water) to the thickness of 0.25 mm and activated to a temperature 110⁰c for an hour. The quantities 5, 10, 20, 30, 40 and 50 μg of standard anthracene equidistantly spotted on the activated TLC plate, 1.5cm above the bottom edge. The TLC chamber containing 100 ml of chloroform as a developing solvent kept for saturation in a dark room for half an hour. The anthracene spotted TLC plate was then carefully placed in a TLC chamber and was allowed to get developed for 10 cm from the spot line. Then, the plate was immediately removed and allowed to get dried at room temperature.

Dried plate was sprayed with the chromogenic reagent, diazo – salt of sulphanilic acid formed by intermediate reaction of sodium nitrite solution (aqu.) and acidic solution of sulphanilic acid. Thereafter the sprayed plate was heated for 10 – 15 minutes in an oven pre-adjusted at 110⁰C temperature. The prominent bluish-violet spot/s (Fig. -1) were observed for the standard anthracene at R_f-value 0.76 ± 0.03. The shape of the spot was roundish – oval.

Purity check

The separate TLC experiment was carried for knowing the purity of anthracene samples available at the hand. The greenish spot observed at R_f-value 0.88 ± 0.03 was accounted for tetracene which could have come as an impurity with anthracene (R_f-value 0.76 ± 0.03) in commercial sample. The Figure -1 shows developed spots for (A) Commercial anthracene, (B) Random anthracene sample and (C) Standard anthracene sample used for semi – quantitative determination.

Figure – 1: The coloured developed spots observed on TLC for (A) Commercial anthracene,

 (B) Random anthracene sample, (C) Standard anthracene.

Measurement of areas of respective spots

For the semi – quantitative determination of anthracene the TLC experiment was carried out as described above. The respective spot developed for 05, 10, 20, 30, 40 and 50 microgram quantities of standard anthracene are shown in Figure – 2.

Figure – 2: (Black and White Image): Spots developed for 05, 10, 20, 30, 40 and 50 microgram quantities of standard anthracene.

The experiment was carried out in four sets. The areas of every spot were carried out using the mathematical equation ‘’. The diameters ‘D1 and D2’ of every spot assigned for 05, 10, 20, 30, 40 and 50 microgram were measured and from that,  respective radii ‘r1’ and ‘r2’ were found out. By using above equation for every spot from all the four sets, spot areas were calculated. The findings have been depicted set-wise as below:

SET – I

Quantity Spotted (μg)	Diameter – 1   (mm)	Diameter – 2  (mm)	Radius – 1  (mm)	Radius – 2  (mm)	Area of Spot  (mm2)
05	5.0	6.0	2.5	3	23.6
10	6.0	7.5	3.0	3.25	30.6
20	8.0	11.0	4.0	5.5	69.1
30	8.0	13.0	4.0	6.5	82.7
40	9.0	15.0	4.5	7.5	106
50	9.0	18.5	4.5	9.25	130.8

SET – I : Microgram quantities of Anthracene spotted on TLC and its corresponding areas

SET – II

Quantity Spotted (μg)	Diameter – 1  (mm)	Diameter – 2  (mm)	Radius – 1 (mm)	Radius – 2  (mm)	Area of Spot  (mm2)
05	5.0	6.0	2.5	3	23.6
10	5.5	8.0	2.75	4.0	34.6
20	8.0	11.0	4.0	5.5	69.1
30	8.5	12.0	4.25	6.0	80.1
40	8.5	15.0	4.25	7.5	100.2
50	9.0	18.5	4.5	9.25	130.8

SET – II : Microgram quantities of Anthracene spotted on TLC and its corresponding areas

SET – III

Quantity Spotted (μg)	Diameter – 1  (mm)	Diameter – 2  (mm)	Radius – 1 (mm)	Radius – 2  (mm)	Area of Spot  (mm2)
05	5.0	6.5	2.5	3.5	25.5
10	5.5	8.0	2.75	4.0	34.6
20	7.0	10.5	3.5	5.25	55.32
30	8.0	13.0	4.0	6.5	81.7
40	9.0	15.0	4.5	7.5	106
50	9.0	18.0	4.5	9.0	127.3

SET – III : Microgram quantities of Anthracene spotted on TLC and its corresponding areas

SET – IV

Quantity Spotted (μg)	Diameter – 1  (mm)	Diameter – 2  (mm)	Radius – 1 (mm)	Radius – 2  (mm)	Area of Spot  (mm2)
05	4.5	7.0	2.25	3.5	24.7
10	5.5	8.5	2.75	4.25	36.7
20	6.5	10.0	3.25	5.0	51.1
30	8.0	12.5	4.0	6.25	78.6
40	8.0	14.0	4.0	7.0	88
50	10.0	17.0	5.0	8.5	133.5

SET – IV : Microgram quantities of Anthracene spotted on TLC and its corresponding areas

From the areas of the respective spots obtained and mentioned in the last column of SET – I, SET – II, SET – III and SET – V, the average area for every spot viz. 05, 10, 20, 30, 40 and 50 microgram quantities of standard anthracene are tabulated as below in Table -1.

Quantity of Anthracene Spotted (μg)	Average Area of Spot (mm2) from  SET- I, II, III, IV
05	24.4
10	34.1
20	61.2
30	80.8
40	100.1
50	130.6

Table – 1: Showing microgram quantities of standard anthracene spotted and eluted on TLC and its respective average areas of four sets.

Calibration Graph: Calibration Equation:  Correlation Coefficient

The calibration graph for determining the presence of anthracene semi – quantitatively, the values of quantity of anthracene applied on TLC and its respective average area calculated and mentioned at Table – 1 were taken into consideration for obtaining the calibration curve. The computer program was used for the same and to get the calibration equation, as also, the correlation coefficient. The details of these are as shown below in Figure – 3. The X-axis represents the quantities of anthracene and Y-axis represents corresponding average areas of the spot.

Figure – 3: X-axis and Y-axis showing microgram quantities of anthracene applied on TLC and corresponding average areas of spot (mm²), forming a straight line calibration curve.

RESULTS AND DISCUSSION

The TLC technique described here helps to detect and identify anthracene obtained from variety of sources. It does helps to detect impurities like tetracene coming with anthracene. Diazo-salt of sulphanilic acid plays better role for it, which imparts prominent bluish-violet colour to anthracene located at R_f-value 0.76 ± 0.03. Calibration graph obtained is linear and  Beer’s law has been followed nicely, in the range of 05 μg to 50 μg of the anthracene used for spotting. The calibration equation found to be, ‘y= 2.305x + 12.31’ and the correlation coefficient is ‘0.995’. The data used here is a pretty old. However, the technique described could detect  1 μg quantity of anthracene. The method may also find its role to detect anthracene semi-quantitatively, in/on desired samples.

ACKNOWLEDGEMENT

Authors thanks are due to, Director, Directorate of Forensic Science Laboratories, Mumbai, M. S., INDIA.  

REFERENCES

1. The Condensed Chemical Dictionary, Arthur and Elizabeth Rose, 1966.
2. Allen’s Commercial Organic analysis, Vol. III, Fourth Edition, J and A Churchill, London, 1910, P-273, 277.
3. Merck Index, Eighth edition, Merck and Co., Inc. U. S. A., 1976.
4. Ultra-violet and Visible Spectroscopy, Chemical Applications, Second Edition, C. N. R. Rao, 1967, p- 71.
5. Organic Electronic Spectral Data, Vol. I, Mortier J. Kamlet, Inter Science Publishers Ltd., New York, London, 1946-1952, p- 548.
6. Thin –layer Chromatography, A Laboratory Hand Book, Edited by Egon Stahl, Second Edition, Springer International Student Edition,, New York, 1969, p- 668-669.
7. Kurtt Randerath, Thin Layer Chromatography, second Revised and Enlarged Edition, Translated by D. D. Libman, 1966, p- 262 -264.
8. A. G. Bhoi and K. A. Ambade, Journal of Indian Academy of Forensic Sciences, 1991, 30 (1), p-19-26.
9.  Allen’s Commercial Organic analysis, Vol. III, Fourth Edition, J and A Churchill, London, 1910, P-137 – 140.

THERMAL STUDIES OF ORGANOCHLORINE INSECTICIDE BENZENE HEXACHLORIDE (BHC)

THERMAL STUDIES OF ORGANOCHLORINE INSECTICIDE BENZENE HEXACHLORIDE (BHC)

Arun G. Bhoi (Senior Forensic Expert)

Dy. Director (Former, I/C), Directorate of Forensic Science Labs., State of Maharashtra, (INDIA).

OSD, Consultant (Former), Goa State Forensic Science Laboratory, Verna, Goa (INDIA).

Founder and Head, Arun’s Institute of Forensic Sciences, Research and Education, Pune, Maharashtra (INDIA).

E – mail: arun.bhoi@gmail.com

(e – J. Foren. Crime Inv. 2017, 14 – 22, 1, Art – 1)

Received on: January 2017.

Accepted on: 24^th March 2017.

Published on: 27^th March 2017.

 

ABSTRACT

The thermal decomposition of recrystallized BHC and dusting powder (DP) containing 50 % was studied using thermogravimetry (TG) and differential thermal analysis (DTA) techniques. The behavior of isomeric constituents has also been studied. From thermogravimetric curves, the order (n) and activation energy (Ea) have been elucidated. Additives present in DP have accelerated the rate of the decomposition reaction. However, it did not affect the melting endothermic DTA peaks.

KEYWORDS

Benzene hexachloride, gamma-BHC, Thermogravimetry, DTA, Order, Activation Energy, isomers, Coats and Redfern equation.

INTRODUCTION

BHC (Hexachlorocyclohexane, C₆H₆Cl₆ ) is a synthetic organic chlorine insecticide. It is a stomach poison and contact insecticide with some fumigant action. It has high toxicity towards insects and fleas. Especially, BHC is very much effective against the fleas “Xenopsyllo” which acts as the carrier for plague organisms “Pastewalla postis” from rodents (rats) to human beings. Hence it is extensively used for eradicating diseases like plague and proved to be of great public health importance. It is marketed in the form of emulsion concentrates (EC), wettable dispersible powders (WDP), dust and smoke generators. Some of the countries like UK, Germany, Japan, Poland, Portugal, Spain have restricted its use [1]. The undesirable musty odor of BHC is one of the reasons to restrict its use as agriculture insecticide. However, it is selectively used for protecting seeds and controlling plant pests. Knowingly or unknowingly, many times this organochlorinated insecticide is misused in homicidal and suicidal cases.

BHC consist of eight isomers such as a, b, g, d, e and so on. a- Isomer constitutes major percentage in BHC. However, g- isomer is the prime active ingredient [2]. A number of methods have been reported for the detection and estimation of BHC. For example thin layer chromatography [3-5], spectrometry [6], gas chromatography [7-10] and classical methods [11]. However, least attention has been paid to thermo-analytical methods and kinetics involved therein. Hence, BHC has been chosen for the TG-DTG and DTA studies. The effect of additives (anticaking and deflocculating agents) on the decomposition of BHC has also been studied. Activation energy (Ea) and the order (n) have been deduced by Coats and Redfern equation [12].

EXPERIMENTAL

Thermal properties of BHC are studies using TGA and DTA units installed in our laboratory.

Recrystallised BHC and dusting powder containing 50% BHC passing through 75-micron sieve were used for thermal analysis.

The following specifications were used for TG/DTA studies:

Sample size                                          60-80 mg

Rate of heating                                    5°C/min

Temperature range of study            ambient to 950°C

Atmosphere                                         Static air or ambient

Reference material for DTA               MgO

RESULTS AND DISCUSSION

The thermogravimetric curve for BHC indicates that the weight-loss begins at 90°C and it proceeds through a number of decomposition/weight-loss steps. The first weight – loss step is observed in the temperature range 90°C to 120°C and accounts for 7% weight-loss. The endothermic DTA peak at 80°C is because of melting of g-isomer (Fig.- 1). Hence the loss could be attributed to the volatilization of g-BHC isomer. The presence of other BHC isomers with g-isomer precisely supports the shift in its melting point to the lower temperature.

The observed 8.5% weight –loss in the temperature range 170°C to 230°C corresponds to the simultaneous volatilization of remaining part of gamma-isomer, delta-isomer, and the fraction of the alpha-isomer. The DTA endotherm at 135°C corresponds to the melting of d-BHC and the inflection at 155°C corresponds to the melting of a-BHC. Heating of BHC (isomeric mixture) to 170°C – 180°C isomerizes it to individual isomer. This isomeric change has also been observed at 180°C as DTA endotherm. An endothermic DTA peak at 220°C corresponds to the melting of e-isomer and it could also be contributing partially to the loss occurring in the range described herein (Fig.-1).

All the weight-losses discussed above are clearly depicted on DTG curves (Fig. -1).

The 13% weight –loss is seen in the range 240°-280°C. In this range dehydrochlorination of BHC occurs.

-HCl                                        -HCl                                        -HCl

C₆H₆Cl_{6 —————->}C₆H5Cl5—————–> C₆H₄Cl_{4  ——————>-} C₆H3Cl₃

BHC         Penta – Chlorocyclohexne (CCH)    Tetra- CCH     Trichlorocyclohexane

Hence the observed loss could be attributed partially to the release of HCl. The weight-loss resulted by dehydrochlorination has also been supported by DTG curve. A characteristic odor of pentachloro and tetrachloride products was noticed above 240°C during experimentation. The chloride was tested with the AgNO₃ solution.

The TG curve shows a major weight-loss (73% approx.) in the range 280°-340°C (Fig. 1) leaving sample holder empty. The prominent DTA endotherm at 312°C corresponds to the melting of b-isomer. The b-isomer melts at 312°C and it sublimes after melting [2]. Furthermore, when heated to decomposition BHC emits highly toxic fumes of phosgene.COCl₂ [13]. In the thermoanalytical process above 310°C, the released phosgene gas was detected with p-dimethylaminobenzaldehyde and diphenylamine test paper [2]. Hence the major weight-loss resulted suddenly in the range of 280°-340°C is a composite result of sublimation of b-isomer, rapid release of phosgene and volatilization of residual BHC and products formed therein. This has also been supported by a prominent and sharp peak at 315°C on DTG curve.

From thermogravimetric data, the kinetic parameters activation energy (Ea) and order (n) for the respective decomposition/weight-loss steps have been elucidated using Coats and Redfern equation [12]. The results are presented in Table 1.

The dusting powder contained 50% of BHC and remaining portion consisted anticaking and deflocculating agents like diatomaceous earth, silica, silicates, powdered limestone and sodium tripolyphosphate. The dusting powder was also studied thermogravimetrically. In thermal studies of dusting powder, losses of BHC occurred comparatively at lower temperature. The major loss was observed up to 260°C for dusting powder (Fig. 2). However, the total loss occurred at 340°C in case of recrystallized BHC. This is because in the case of dusting powders the additives could be accelerating the decomposition and thus enhancing the reaction processes. The presence of additives has also increased surface area for the reactions. Therefore, the speedy losses would have occurred by decomposition, volatilization, and sublimation of BHC leaving additives. These steps are also clearly observed on DTG curve. However, this process has not made any remarkable changes on endothermic DTA peaks (Fig. 2). In this range, no interference was observed on TG, DTG and DTA curves by the presence of additives. However, above 340°C, TG-DTG showed many of the decomposition/weight-loss steps corresponding to additives. The exothermic DTA peaks above 340°C could also be attributed to the decomposition and phase changes of additives present therein.

CONCLUSIONS

The present study was undertaken to know the thermal behavior of polychlorinated material isomeric BHC. The TG, DTG and DTA studies throw light on the presence of different isomers in BHC and on the inter-isomerisation and transformations of constituting BHC isomers. It also helped to study decomposition changes and kinetics involving therein. These thermo-analytical methods would also prove helpful to know the presence of different isomers quantitatively. It can also be concluded that additives from dusting powders with 50% BHC only accelerates the changes and remain unchanged till all the BHC is decomposed. Therefore, TG-DTG and DTA methods would prove to be a promising and precise tool for the analysts working in different fields for studying polychlorinated- isomeric compounds.

ACKNOWLEDGEMENTS

Author’s thanks are due to Head, Department of Chemistry, University of Pune, Pune, Maharashtra (INDIA).

REFERENCE

1. K. Gupta and D.K. Salunkhe. Modern Toxicology, Vol. II, The Adverse Effects of Xenobiotics. First Ed., Metropolitan Book Co. Pvt. Ltd., New Delhi, 1985, p.15-17.
2. Martha Windholz. The Merck Index, An Encyclopedia of Chemicals and Drugs, Merck and Co., Inc., U.S.A., 1976, p.719, 955.
3. Alan Curry. Poison Detection in Human Organs, Third Ed., Charles C. Thomas, Publisher, U.S.A., 1976, p. 225-226.
4. Egon Stahl. Thin Layer Chromatography, A Laboratory Handbook, 2^nd, Springer International Student Edition, Springer-Verlag, New York, 1969, p. 643.
5. A. Khan and J. Paul. Microchem. J., 1979, 24, 333-340.
6. Wood. Analyst, 1960, 85, 21.
7. Pesticides, GC Analysis and Standards, Bulletin 758A, Supelco, Inc., Bellefonte, Pennsylvinia, 1976 p. 1-3.
8. British Standards Institution, BS-5202 Part 10. 1982.
9. J. de F. Maunder, H. Egan and J. Roburn. Analyst, 1964, 89, 157.
10. J. de F. Maunder, H. Egan, E.W. Godly, E.W. Hammond, J. Roburn and J. Thomson. Analyst, 1964, 89, 168.
11. S. Sree Ramulu. Methods of Pesticide Analysis, Oxford, and IBH Publishing Co., New Delhi, 1979, p. 52-55.
12. W. Coats and J. P. Redfern. Nature, 1964, 201, 68.
13. Irving Sax. Dangerous Properties of Industrial Materials, Fifth Ed., Van Nostrand Reinhold Company, London, 1979, p. 716.

Table 1: Thermal Data of BHC

Temperature range                       Order                    Activation energy (Ea)

(°C)                                                     (n)                                        (KJ/mole)

_______________________________________________________________________

90-120                                                  0.00                                        102.00

170-230                                                0.00                                          24.19

240-280                                                0.00                                        150.50

290-340                                                0.00                                        130.30

_______________________________________________________________________

Figures 1 and 2 show the thermal curves for recrystallized BHC and Dusting Powder (50% BHC) as below:

AFIS: STUDIES FOR THE FREQUENCY OF APPEARANCES OF FINGER PRINT PATTERNS IN HUMAN FINGERS.

AFIS: STUDIES FOR THE FREQUENCY OF APPEARANCES OF FINGER PRINT PATTERNS IN HUMAN FINGERS.  

B. Lakshamanan

Inspector of Police (FP) (Spl. Grade) (Ex.)

Tamilnadu Finger Print Bureau, Chennai (India).

First Expert, FPB, FSL, Verna, Goa (India).

Arun G. Bhoi (Forensic Expert)

Assistant Director (Ex.), Directorate of Forensic Science Labs., Mumbai, M. S. (India).

Sr. Scientific Executive, Forensic Science Laboratory, Verna, Goa (India).

Arun’s Institute of Forensic Sciences, Research and Education, Pune (India).

E-mail: arun.bhoi@gmail.com

( e-J. Foren. Crime Inv. 2014, 10-13, 1, Art. 1)

Received On: November, 2013.

Accepted On (Modified): January, 2014.

ABSTRACT

The frequency of patterns of fingerprint impressions was studied using AFIS (Automated Finger Print Identification System). Further, the data obtained was comparatively evaluated with the results of Scotland Yard  Committee (1905) and that of Fingerprint Bureau, Calcutta, West Bengal (1953). The differences in the percentage of frequencies of appearances of different fingerprint patterns like arc, loop, whorl etc. are explored, herein, using database of about twenty lakhs of fingerprint impressions.

KEYWORDS

Finger print, impression, pattern, AFIS, arch, loop, whorl, composite.

INTRODUCTION

A  team  of  finger  print experts from Scotland yard (1905 ) had  conducted  a  research study  to know the  percentage  of  frequencies  of  finger  print  patterns  of 50,000 impressions  from  5000 individuals. Subsequently, during the year 1953, S. K. Chatterjee, Central Finger  Print Bureau, Calcutta, West Bengal had also conducted a  research experiments to study the percentage of  frequencies of finger print patterns among  50,000 impressions  from  5000 individuals (Finger, Palm and Sole Prints, 1953,  1^st Edition). Here, the frequency of distribution of different finger print patterns formed by ridges, in about twenty lakhs of impressions is explored using database from AFIS – Automated Fingerprint Identification System and was compared with both of the earlier studies.  

PROCEDURE Database made available from AFIS was used for the comparative studies of percentage distribution – frequency of finger print patterns in lakhs of impressions.

RESULTS AND DISCUSSION

Scotland Yard Committee set up in 1905 exhaustively studied the 50,000 impressions from 5000 individuals to find the percentage distribution of frequencies of finger print patterns, such as, arch, loops, whorls (Fig. 1) and composites etc.

Figure – 1: Showing finger print patterns  (a) Arch, (b) Loop and (c) Whorl.

( a ) Arch

( b ) Loop

( c ) Whorl

The findings of Scotland Yard Committee (1905) are as reported below  in Table– 1.  

Table-1: The percentage distribution of frequencies of finger print patterns (1905):

ARCH	4.866%
LOOPS	69.696%
WHORLS	20.066%
COMPOSITES	5.372%
WHORL + COMPOSITES	25.4385%
AMPUTATED OR INDECIPHERABLE	0.498%

The observations of S. K. Chatterjee , Director,  Central Finger  Print Bureau, Calcutta , West Bengal (1953, Finger, Palm and Sole Prints – 1^st Edition) regarding percentage of  frequencies of finger print patterns derived from 50,000 impressions  of 5000 individuals are cited below in Table – 2:

Table-2: The percentage distribution of frequencies of finger print patterns (1953):

ARCH	4.838%
LOOPS	69.414%
WHORLS (Including Twinned loop Pattern)	19.83%
COMPOSITES (Excluding Twinned loop Pattern)	5.486%
WHORL + COMPOSITES	25.316%
DAMAGED	0.42%

Both  these studies  revealed  almost the same  results  with the percentage  of  frequencies  of  pattern  in human fingers. Finger Print Bureaus, all over the world, have accepted results of  these  studies of  the frequency  of  patterns  of  human fingers, as reliable. As a matter fact, these studies were based on a very small number of samples i. e. 5000 individual  samples, only. However, in this advanced computer era, where it is possible to study any number of finger prints in few seconds, it seems that, no attempts have been made to study frequency of patterns in human fingers, available in lakhs. Hence, efforts have been made to study lakhs of fingerprint patterns to find its frequency in human fingers.  More than 2  lakhs of individual  samples  with  a total   of  20,01,080 impressions  were taken in to consideration  through Semi Automated  Finger  Print  Identification  System  Database for percentage  of  frequency  of  finger  print  patterns  in  human  fingers.  The observations are reported in Table – 3.

Table-3: Percentage  of  frequency  of  Finger  Print  patterns (AFIS)

ARCH	2.235%
LOOPS	55.816%
WHORLS	30.245%
COMPOSITES	10.285%
WHORL + COMPOSITES	40.530%
SCAR	1.259%
AMPUTATED	0.177%

Apart  from analysis  of  20,01,080 finger impressions  through Semi Automated  Finger  Print  Identification  System  Database from one of the States and 2099290, 3795317, 1187433 finger impressions  from other States, wherein AFIS/FACTS installed were also, considered for the studies. The distribution of percentage of frequency of fingerprint patterns in human fingers in total number of  20,99,290 impressions was observed as in Table – 4.

Table-4: Percentage distribution of fingerprint pattern in 20,99,290 impressions.

ARCH	3.358 %
LOOPS	52.543%
WHORL + COMPOSITE	38.253%
SCAR	6.350%
NONE	0.846%

Table – 5 shows the results  of  percentage  of  frequency of  finger  print  pattern  in human  finger  in 37,95,317  impressions.

Table-5: Percentage distribution of fingerprint pattern in 37,95,317 impressions.

ARCH	3.607%
LOOPS	58.277%
WHORL + COMPOSITE	38.068%
SCAR	0.046%

Results  of  percentage  of   frequency of  finger  print  pattern  in human  finger  in  a total sample impressions 11,87,433 are detailed in Table- 6.

Table-6: Percentage distribution of fingerprint pattern in 11,87,433 impressions.

ARCH	3.376%
LOOPS	56.984%
WHORL + COMPOSITE	39.563%
SCAR	0.074%

The average distribution of total of above 70,82,040 finger print patterns in human fingers (2006) is tabulated in the following Table – 7 :

Table-7: Percentage distribution of frequencies of fingerprint patterns (2006)

ARCH	3.319%
LOOPS	56.35%
WHORL + COMPOSITE	39.232%
SCAR	1.919%
None	0.02%

In AFIS no separate icon  is  considered  for  composite pattern except that of accidental. Hence, the above  tables  does  not  disclose  the  percentage  of  frequencies  for  composite  pattern.  Hence, the distribution of 20,01,000 fingerprint impressions explored above (Table – 3),  the  percentage  of  occurrence  for composite  pattern comes  around  10.285%. If this percentage (10.285%) has been taken into consideration and applied to a total of the fingerprint impressions studied by deducting the same from the calculated ‘Whorl’ percentage the final   tally of frequency of finger print pattern should be as in Table – 8.

Table – 8: The   final tally of frequency of finger print pattern (Modified)

ARCH	3.319%
LOOPS	56.35%
WHORL	28.947%
Composite	10.285%
SCAR	1.919%
None	0.02%

The findings of Scotland Yard team (1905), 1953 studies and that of AFIS (2006) are comparatively, depicted in Table – 9.

Table – 9: Inter comparison of the three distribution calculations:

Sr. No.	Finger Print  Pattern	Distribution (%) based  on findings of  Scotland yard                       Expert committee (1905)  (50,000 impressions)	Distribution based  on studies of  Sh. Chatterjee, West Bengal (1953)      (50,000 impressions)	Distribution based  on AFIS(2006)              ( 70,82,040 impressions).
	(1)	(2)	(3)	(4)
1	Arch	4.866%	4.838%	3.319%
2	Loop	69.696%	69.414%	56.35 %
3	Whorl	20.066%	19.83  %	28.947%
4	Composite	5.372%	5.486%	10.285%
5	Scar	0.438%	0.42  %	1.919%
6	None	—		0.02  %

An  analysis  of  the  above  three  studies  reveals  that,  the  percentage  of  Arch pattern  was  nearly  4.8% in both the studies of 1905 and 1953. Whereas,  it  is  only  3.319% based  on  the  AFIS studies i.e. 1.5%  less  than  the  former  two  studies.  Since,  the  AFIS  study  was  made  among  70,82,040 impressions, it is put forth that, the  AFIS may be relied  on  and  hence  the  chances  of occurrence  of  Arch  Pattern including  Tented arch pattern  in  human finger impressions should  be  considered  as  3.319% .

 As  far  as,  loop is concerned  both  the  studies  predicted  a  frequency  of  about 69%, whereas, the  AFIS  study  reveals  the  frequency  of  loop pattern 56%, only. This shows a reduction of about 13 % in percentages of appearances of loop pattern as observed in 1905 and 1953 studies.

In case of whorl, the  earlier  two studies  estimated  its presence as 20%, whereas,  the  AFIS  studies  revealed an  increase  of 8%.

 Existence of composite pattern is about 5% as reflected in 1905 and 1953 findings. However, the study through semi-automated finger print identification system (NCRB)   revealed it as 10.285% resulting in increase of about 5% to that of earlier studies.

CONCLUSION

The results discussed above are based on studies of 70,82,040  sample finger print impressions using computerized AFIS, therefore, it claims to be accepted for the frequency of appearances / distribution of finger print patterns in human fingers. These studies have been extended to find match for the chance print in a minimal time with suggestive changes in AFIS, specifically, concentrating on studies of loop, whorl and ridge counts.

ACKNOWLEDGEMENTS

 Author’s thanks are due to the Heads of Institutions, FPBs.

NEWS : TOXOCON – 7

TOXOCON -7
TOXOCON – 7 conference with the theme ‘Marine Envenomation and seafood Poisoning’ was recently organized by the Department of Forensic Medicine, Goa Medical College, Bambolin, Goa (India) under the Indian Toxicology Society on 2nd and 3rd of March 2013. The keynote address ‘ Marine Envonomation – A Global View’ was delivered by Dr. Ken Winkel, Director, Australian Venom Research Unit, University of Melbourne, Australia.
As a matter of fact, even though, India has thousands of kilometers of coastal area, people, fishermen and most of the medical professionals are not aware of marine envenomation occurring through stings and bites, as also, poisoning and its symptoms caused by deadly venomous jellyfish, shellfish, cone fish, blue ringed octopus, sting ray fish, sea snakes etc. Realizing the facts and in line of the theme various topics such as, Marine Venomous Creatures, Marine Envenomation and its Clinical Features, Seafood Poisoning, its Diagnostic methods and Medical Management, Industrial Toxic Contaminants, Medico-legal Aspects and Precautions to be taken in the preparation and consumption of Seafood were discussed in detailed.
Mr. Arun Bhoi (Forensic Expert) with Dr. Ken Winkel at venue.
Mr. Arun Bhoi (Forensic Expert) with Dr. Madhu Ghodkirekar, Member of Organizing Committee and Treasurer TOXOCON – 7.

TLC DETECTION AND RECOVERY OF ORGANOPHOSPHORUS INSECTICIDE PHOSALONE FROM VISCERA USING COPPER (II) ACETYL ACETONATE.

TLC DETECTION AND RECOVERY OF ORGANOPHOSPHORUS INSECTICIDE PHOSALONE FROM VISCERA USING COPPER (II) ACETYL ACETONATE.

Arun G. Bhoi

Arun’s Institute of  Forensic Sciences, Research and Education, Pune -411 028 (India).

Email : arun.bhoi@gmail.com

(e-J. Foren. Crime Inv. 2013, 9, 2, Art. 1)

Received on : 10^th Jan. 2013.

Accepted on : 16^th Jan. 2013.

 

ABSTRACT

Phosalone (Zolone) has replaced many of the chlorine containing insecticides used for controlling pests on agricultural crops. A number of reagents are reported for the detection and determination of organophosphorus insecticides but these are not specific for the phosalone.  The use of copper(II)   acetylacetonate as a specific spray reagent is reported here for the detection of phosalone on TLC. Phosalone after elution on TLC plates is hydrolysed using sodium hydroxide solution and then sprayed with copper acetylacetonate reagent. Phosalone appears as a brown spot. Other insecticides do not interfere in the detection. The detection limit for phosalone is ca. 2μg. The recovery of phosalone from viscera was found to be 85%.

KEY WORDS : Phosalone, Zolone, insecticide, thin-layer chromatography, TLC, hydrolysis, copper acetylacetonate, recovery, viscera, forensic.

INTRODUCTION

Phosalone (Zolone; phosphorodithioic acid, S-ester with 6-choloro-3-(mercaptomethyl)-2-benzoxazolinone) is a chlorine containing dithiophosphoric acid derivative insecticide. LD₅₀ for phosalone is 120-175 mg/kg orally in rats [1].  It has higher chemical stability and less toxicity for animals as compared to many of the chlorine and phosphorous containing insecticides. Therefore, phosalone has been preferentially used for the protection of seeds, seedlings and agriculture crops. Unfortunately, this has also been misused in many of the homicidal and suicidal cases.

Number of gas chromatographic [2,3], spectrophotometric [4-6] and thin-layer chromatographic [7,8] methods are described in the literature for the detection of phosalone. However, these methods are tedious and need a well equipped instrumental laboratory. The use of copper (II) acetylacetonate as a spray reagent is described here for specific detection of phosalone on silica gel G coated TLC plates.

EXPERIMENTAL

Reagents:

All reagents used were of analytical reagent grade. Glass distilled water  and 95% ethyl alcohol were used throughout.

Sodium hydroxide solution (10%):    10g of NaOH was dissolved in small quantity of distilled water and made  up to 100 ml.

Copper (II) acetylacetone solution (0.25%): 250 mg of copper (II) acetylacetonate was dissolved in chloroform and made up to 100 ml.

Phosalone Standard Solution (1mg/ml) : 10 mg of commercial grade phosalone (99%) was dissolved in 9.9 ml of 95% ethyl alcohol.

PROCEDURE

A standard glass TLC plate of the size of 10cm x 20cm was coated with a slurry of silica gel G in distilled water (1:2 ) to a thickness of 0.25mm. Coated plates were dried in air  and activated at 110 degree centigrade for minimum 1 h. Activated plates were cooled. A 2 μl quantity of a standard solution of phosalone (1mg/1ml) was spotted on TLC plate. The plate was then developed in a pre-saturated flat bottom TLC chamber using n-hexane and acetone (8:2, v/v) as a developing solvent mixture. After the solvent had travelled 10 cm distance on the TLC plate it was taken out, air dried and sprayed with 10% of sodium hydroxide solution and subsequently with 0.25% copper (II) acetylacetonate reagent solution.  A prominent brown spot was visualized at R_f – value 0.60 on the TLC plate for phosalone.

RECOVERY EXPERIMENT OF PHOSALONE FROM VISCERAL MATERIAL

An standard alcoholic solution (1 ml) equivalent to 1 mg of phosalone was added to 50 g  of minced visceral material (stomach – intestine with contents and liver, spleen, kidneys, lungs), homogenized and was kept in situ for about 72 hours. For the extraction of phosalone, the contents were soaked in 100 ml of diethyl ether at least for 15 minutes with intermittent stirring. The ether layer was separated out using separating funnel. The visceral material was re-extracted using 50 ml of diethyl ether, twice. All the ether extracts were combined in a steel capsule and the solvent was allowed to get evaporated at room temperature. The residue (phosalone) left was re-dissolved in 1 ml of an ethanol. A 10 micro liter quantity of it was spotted on activated TLC plate along with 10 micro liter quantity of each of the standard phosalone solutions containing known concentrations of 7.5, 8.0, 8.5, 9.0, 9.5 and 10 mg per 10 ml in ethanol. Te plate was then developed as described in the procedure section and sprayed with 10% sodium hydroxide solution and copper (II) acetylacetonate reagent. The intensity of the brown spot of the residue obtained from visceral material visually agreed with the spot resulting from a standard solution containing 8.5 mg of phosalone per 10 ml. Thus the recovery for phosalone was ca. 85%.

RESULTS AND DISCUSSION

Alkaline sodium hydroxide solution when sprayed on TLC plate it rapidly hydrolyses Phosalone producing sodium salt of 6-chloro-benzenexaazolone, diethyl dithiophosphoric  acid and formaldehyde. It is postulated that the sodium salt formed, in turn, reacts with copper acetylacetonate and produces the brown coloured complex. The reagent used is selective for phosalone. The detection limit is calculated as 2 microgram. Other organophosphorus insecticides such as thimet, phosphamidon, fenitrothion, nuvan, monocrotophos, ekalux, solvirex and organochlorine insecticides such as, BHC, DDT, endosulfan, aldrin, endrin, dieldrin did not give any coloured spots. Dimethoate, malathion, methyl parathion and pyrethroids insecticides did not interfere by way of colour and R_f – values. Carbamate insecticides have been studied, separately, with the same reagents.

Since many of the reagents described for the detection of the insecticides did not respond to phosalone, the reagent studied here can be used for the detection and semi-quantitative determination of phosalone in the extracts of biological material (viscera) submitted in forensic laboratories and commercial samples.

AKNOWLEDGEMENTS

Author’s thanks are due to Prof. (Dr.) S. B. Kulkarni, Head (Ex.), Dept.  Of Chemistry, University of Pune, Maharashtra (India) and the Director, Directorate of Forensic Science Labs. M. S., Mumbai (India).

REFERENCES

[1] Martha W., The Merck Index, An Encyclopedia of Chemicals and Drugs, Merck and Co., Inc., USA, 1976, p-955.

[2] Spenser W. F., Iwata Y., Kilgore W. W. and Knaak J. B.; Bull. Environ. Contam. Toxicol., 1977, 18, 656.

[3] Their H. P., Stijve T. and Discrense H.; Lebenmittelchem. Gerichtl. Chem., 1989, 43, 121 (Ger.).

[4] Syoyama M., Miyachi Y. and Sakakibara J.; Bunseki Kagaku, 1976, 25, 179 (Japanese).

[5] Novikova K. F. and Mel’fser F. R.; Zh. Analit. Khim., 1971, 26, 1651 (Russ.).

[6] Makarova S. V., Khenelnitsakaya V. N. and Eliseeva M. A.; Zar. Lab., 1974, 40, 792 (Russ.).

[7] Rodinova T. V., Belskaya G. G., Ivanov V. M. and Makarova S. V.; Zh. Anal. Khim.; 1987, 42, 1125 (Russ.).

[8] Sevelkar M. T., Patil V. B. and Katkar H. N.; J. Assoc. Off. Ana. Chem., 1991, 74, 545.

 

 

ACCIDENTAL LIGATURE STRANGULATION (ALS) : SURVIVAL OF A MINOR

ACCIDENTAL LIGATURE STRANGULATION : SURVIVAL OF A MINOR

Vilas P. Wadile (M. D.), Sarika V. Wadile (MBBS, DGO)

Jeevak Hospital, Dhule, M. S. (India)

Deepali D. More (MBBS, DCH)

More Hospital, Dapoli -Ratnagiri, M. S. (India)

Arun G. Bhoi

Arun’s Institute of  Forensic Sciences, Research and Education, Pune -411 028 (India).

Email : arun.bhoi@gmail.com

(e-J. Foren. Crime Inv. 2013, 9, 1, Art. 1)

Received on : 12^th Dec. 2012.

Accepted on : 16^th Dec. 2012.

ABSTRACT

Accidental ligature strangulation (ALS) is uncommon, as also, survival of a victim in such cases is rare. The ALS is reported here which occurred to a minor girl while travelling on a M/cycle, however, she survived through it. The prominent ligature mark was observed on the backside of her neck. Observations revealed the presence of the petechiae, too.

KEY WORDS

Accident, ligature, strangulation, petechiae, forensic.

INTRODUCTION

Few cases of accidental ligature strangulation (ALS) are reported in the literature. Kohli et. al. [1] and Aggarwal et. al. [2] have discussed accidental strangulation resulted by chunni in a rickshaw. ALS occurred by massage device has been reported by Deidikar R. D. [3]. Gupta et. al. [4] have reported the case of ALS by sari in M/cycle. ALS by crop thrasher [5], electric grinder [6] and ironing machine [7] are also explored in the literature. Strangulation resulted in domestic violence is reported by Richard [8]. Suicidal ALS is explained by Li D. R. et. al. [9]. Human hair has also, stood the cause of strangulation [10]. The case of incomplete strangulation [11] is explained by Amit et. al. Saurabh and others [12] have reported the survival in an accidental strangulation.

We report here the survival of a minor girl went through a  strangle in incomplete accidental ligature strangulation.

HISTORY

A father was carrying his daughter aged about 10 years on the M/cycle to a nearby place. Being winter days the girl had tied headscarf around her head. She was sitting backside of her father on the M/cycle. On the way of destination the M/cycle got a jerk and bumped heavily by which the girl fell down as she was sitting comfortably behind her father on the M/cycle. While falling down the girl’s headscarf got entangled in a rear wheel of M/cycle and she got strangled. By the motion of the wheel the headscarf got pulled heavily resulting in strangling of  girl’s neck. Father immediately stopped the M/cycle and tried to remove the headscarf from the wheel forcibly to make his daughter free from the trap.

EXPERIMENTAL

PROCEDURE : OBSERVATIONS

Strangulation leads to the constriction of a body organ or duct which leads to ligature and the stoppage of the body fluids like blood. Usually ligature marks (bruising of skin) are caused by materials like ropes, wires, cords, garrote, clothes like sari, chunni and scarf involved in the act of strangulation. In severe cases bleeding, stoppage of respiration and blood flow is noticed. Injuries to head, damage to cartilage may also, be seen.

In the subject matter presented here, the patient’s father briefed that she showed slight signs of suffocation when the incidence took place. Examination of a patient showed presence of a prominent ligature mark to the backside of the neck with peeling of skin (epithelial layer) exposing inner muscular tissues, there from (Fig.1).

Figure 1: Prominent ligature mark to the neck back with peeled skin and exposed tissues.

Redness was also, seen around the neck periphery, on the cheek and face. Petechiae (redness due to blood) was seen in patient’s eyes. It was very much prominent in the left eye as in Fig.2.

Figure 2:  Petechiae seen in patient’s eyes with prominence in the left eye.

The ligature was found slightly extending to the frontal throat portion and curved, too. (Fig.3).

Figure 3:   Ligature mark curved and extending towards throat.

The patient examined, herein, did not show swelling nearby ligature, head injuries, bleeding through ears. ENT and surgical findings were normal. No dislocation/damage was noticed to the cartilage bone.

TREATMENT  

Patient was treated with anti-inflammatory and antibiotic drugs, only and  showed the speedy recovery and found very normal in a week’s time.

RESULTS, DISCUSSION AND CONCLUSION

The slight suffocation noticed by patient’s father must have been resulted by the partial constriction of a wind pipe. The mark produced on neck back is seen prominently as ligature strangulation had got enhanced by abrasion – friction resulted by headscarf while the father tried to remove it forcibly from around the neck of the patient girl. Further, the pressure generated at ligature strangulation site had suppressed the blood flow around neck periphery resulting in redness on the face, cheek and neck. Petechiae (redness due to blood) seen in patient’s eyes was also, the result of the same.

It is ascertained that the falling of a girl in head down – face up position created deep abrasion – ligature mark encircling the back portion of the neck as in Fig.1. The ligature found on the neck back slightly extending to the frontal throat portion and curved (Fig.3) is definitely rare and is suggestive of accidental ligature strangulation.

The patient’s immediate survival is  because of immediate steps taken by her conscious father to stop the speedy M/cycle and to make the girl free from the entangled scarf in the M/cycle by removing it out from the rear wheel which restricted the stopping of respiration and blockage of blood flow.

The proper treatment of anti-inflammatory and antibiotic drugs given to the minor brought her to the normalcy in a short span of a time. Thus, the risk of accidental strangulation which could have lead to fatal and complete strangulation was turned in the saving of the life of a minor girl.

REFERENCES

[1]. Kohli A., Verma S. K. and Agarwal B. B., Forensic Sci. Int. 1996 March 5, 78, 1, 7-11.

[2]. Aggarwal N. K. and Agarwal B. B.,  Med. Sci. Law 1988 July, 38, 3, 263-5.

[3]. Deidikar R. D., Am. J. Forensic Med. Pathol. 1999 Dec., 20, 4, 354-6.

[4]. Gupta B. D., Jain C. B. and Datta R. B., Med. Sci. Law 2004 Oct., 44, 4, 359-64.

[5]. Dixit P. G. and Kukde H. G., Journal of Forensic and Legal Medicine, 2008 May, 15, 4, 263-5.

[6]. Shetty Mahabalesh and Shetty Suresh B., J. Clinical Forensic Medicine 2006 April, 13, 3, 748-750.

[7]. Dogan K. H., Demirci S., Gunaydin G. and bunken B., J. Forensic Sci. 2010 Jan, 55, 1, 251 -3.

[8]. Investigating Domestic Violence Strangultion, Richard, Bluesheepdog.com, 2007 Nov. 9.

[9]. Zhao D., Ishikawa T., Quan L., Li D. R., Michine T and Maeda H., Leg. Med (Tokyo) Nov. 2008, 10, 6, 310-15.

[10]. Milkovich S. M., Owens J, Stool D., Chen X. and Beran M., Int. J. Pediatr. Otorhinolaryngol. 2005Dec., 69, 12, 1621-8.

[11]. Agarwal Amit, Ninave Sudhir, Shrivastav Tripti, Sunkara Anil and Agarwal Sachin, Journal of Forensic and Legal Medicine, 2009 Aug., 16, 6, 350-1.

[12]. Chattopadhyay Saurabh and Pal Indranil, Journal of Forensic and Legal Medicine, 2008 Jan., 15, 1, 53-55.

Communications. Getting Published Shortly. ‘CRIME FORENSICS’ (In Marathi). By Arun Bhoi.

Image

‘CRIME FORENSICS’
(In Marathi).
By Arun Bhoi.

HPTLC DETECTION AND DIFFERENTIATION OF CARDIOVASCULAR DRUG ATORVASTATIN WITH CLOPIDOGREL AND ASPIRIN.

HPTLC  DETECTION  AND DIFFERENTIATION OF  CARDIOVASCULAR  DRUG  ATORVASTATIN  WITH  CLOPIDOGREL  AND  ASPIRIN.

Arun G. Bhoi

Arun’s Institute of  Forensic Sciences, Research and Education, Pune -411 028 (India).

Email : arun.bhoi@gmail.com

(e-J. Foren. Crime Inv. 2012, 6-8, 1, Art. 1, 26^th Aug. 2012)

 

ABSTRACT

Atorvastatin drug helps in decreasing cholesterol and thus reduces the risk of heart attack. HPTLC, high performance thin-layer chromatography is described here  for the detection of atorvistatin.  HPTLC glass plates of the size of 10 cm x 10 cm,  pre-coated with silica gel 60F₂₅₄   as a stationary phase and solvent mixture Bexane : Acetone : Ethyl alcohol (8:1:1, v/v/v) as a mobile phase were preferred for it.  UV detection was carried at 254 nm. Atorvastatin was detected at R_f-values 0.25 ± 0.02. The LOD for these drugs was found to be 50 ng per spot. The solvent system used here nicely differentiates Atorvistatin from Aspirin and Clopidogrel having R_f-values  0.38 and 0.93, respectively.

KEY WORDS : Atorvastatin, clopidogrel, aspirin, cardiovascular, drug, HPTLC, UV, detection.

INTRODUCTION

Atorvastatin is a  drug of choice prescribed for cardiovascular disease. It minimizes chances of heart attack as it lowers amount of cholesterol in the blood. Panchal et. al. [1] has described use of HPLC and HPTLC methods for the determination of atorvastatin with ramipril in combined capsule dosage form. Atorvastatin was determined at 210 nm by HPTLC – densitometry. Easy and speedy HPTLC method  is described here for the detection of  Atorvistatin. Susceptibility of clopidogrel bisulphate to acids, alkali, oxidation and heat using HPTLC was explored by Himani et. al. [2]. Mitacos et. al. [3]  have validated clopidogrel using liquid chromatography in pharmaceutical preparations. Estimation of aspirin   has been explained in the literature using RP-HPLC [4,5,6], Rapid HPLC [7], HPTLC [8] and spectrophotometry [9]. Bexane : Acetone : Ethyl alcohol (8:1:1, v/v/v) as a mobile phase is used herein for the differentiation and detection of Atorvastatin, Clopidogrel and Aspirin.

EXPERIMENTAL

Equipments:

The following material / equipment was used for the experiment.

1. Merck, HPTLC glass plates pre-coated with silica gel 60 F 254 ( 10 cm x 10 cm).
2. Camag Development Chamber ( small size, twin type ).
3. Camag UV Viewing Chamber equipped with 254 nm and 366 nm wavelength exposure facility.

 

High Performance Thin-layer Chromatography (HPTLC):

Reagents: All reagents and solvents used were of analytical reagent grade.

95% ethyl alcohol and glass distilled water was used wherever required.

Standard Atorvistatin Solution : Atorvistatin Tablets IP (X’tor^®-10), each  containing Atorvistatin Calcium  powder equivalent to 10mg of  Atorvistatin  marketed by lpca Laboratories Ltd., Bharikhola, Sikkim, INDIA, were used for studies.

Standard Atorvistatin Solution (1mg/1ml) : Atorvistatin Calcium  powder equivalent to 10mg of  Atorvistatin  from Atorvistatin Tablets IP (X’tor^®-10) dissolved in small quantity of 95% of ethyl alcohol and made up to 10 ml with the 95% ethyl alcohol.

Standard Clopidogrel and Aspirin Solutions:

Clopitab A 150 capsules, each  containing clopidogrel bisulphate powder equivalent to 75 mg of clopidogrel and aspirin in tablet form, two tablets of 75 mg each (i.e.150 mg of aspirin), marketed by LUPIN LTD., MUMBAI, INDIA, were used for studies.

Clopidogrel Standard Solution ( 1mg/ml) : Clopidogrel bisulphate powder equivalent to 10 mg of clopidogrel was weighed from powder contents of capsule (Clopitab A 150), dissolved in small quantity of 95% of ethyl alcohol and made up to 10 ml with the 95% ethyl alcohol.

Aspirin Standard Solution ( 1mg/ml) : Two tablets of aspirin were separated out carefully, from the contents of  Clopitab A 150 capsule, finely powdered and quantity equivalent to 10 mg of aspirin was weighed, dissolved in small quantity of 95% ethyl alcohol which was further made up to 10 ml using the same.

PROCEDURE

Camag, HPTLC glass plate pre-coated   with silica gel 60 F 254 of the size of 10 cm x 10 cm was taken for the experiment. 0.05, 0.1, 0.2 and 0.5 μg quantities of Atorvistatin standard solution (1mg/1ml) were spotted on the HPTLC plate using micro capillaries. Further, 0.05, 0.1, 0.2 and 0.5 μg quantities of each of the clopidogrel and aspirin standard solutions were, also, spotted on the HPTLC plate. The HPTLC plate was eluted in a previously saturated small sized, twin trough Camag HPTLC chamber using 5 ml quantity of  solvent mixture Bexane : Acetone : Ethyl alcohol ( 8:1:1, v/v/v ) . The solvent was allowed to travel to a height of  5 cm. Then the HPTLC plate was taken out, air dried and viewed in a UV chamber at 254 nm wavelength.

RESULTS AND DISCUSSION

The developed and air dried HPTLC plate was then observed at wavelength 254nm using Camag UV viewing chamber. Atorvistatin showed its presence at at R^f-value 0.25±0.02 by fluorescence quenching phenomenon.  Aspirin showed its presence at Rf-value 0.38±0.02 and Clopidogrel  was found present at Rf-value 0.93 ±0.02. The chosen solvent system  showed very nice separation on HPTLC plate for cardiovascular drugs Atorvastatin, Aspirin and Clopidogrel thus differentiating these from each other. The LOD for these drugs is cal. 50 ng. However, the spot visualized for Clopidogrel has progressed towards the solvent front. Further, this system fails differentiate Carvedilol ( R_f =0.26 ) from Atorvastatin ( R_f =0.25±0.02  ) and hence the former drug has been ignored to get mentioned in detailed, herein.  Thus, HPTLC method is easier, simple, specific and speedy one to detect and differentiate cardiovascular drugs Atorvistatin, Aspirin and Clopidogrel, in combined formulations and biological material.

ACKNOWLEDGEMENT

Author’s thanks are due to Director, Directorate of Forensic Science Laboratories, Mumbai, State of Maharashtra, India.

REFERENCES

[1] Panchal S. J., Suhagia B. N.; JAOAC Int. Sept.-Oct 2010, 93(5), 1450-1457.

[2] Himani Agarwal, Niraj Kaul, Paradkar A. R. and Mahadik K. R.;  Talanta, 2003, 61(5), 581-589.

[3] Mitakos, A. and Panderi, I.; J. Pharm. Biomed. Anal., 2002, 28, 431-438.

[4] Gandhimathi, M., Ravi, T. K., Abraham, A and Thomas, R.;  J. Pharm. Biomed. Anal., 2003, 32, 1145-1148.

[5] Shah, D. A., Bhatt, K. K., Mehta, R. S., Shankar, M. B., Baldania, S. L. and Gandhi, T. R.; Ind. J. Pharm. Sci., 2007, 69(4), 571-574.

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THERMAL STUDIES OF CHLORAL HYDRATE

THERMAL  STUDIES  OF  CHLORAL  HYDRATE  

Arun G. Bhoi

Arun’s Institute of  Forensic Sciences, Research and Education, Pune  – 411028, State of Maharashtra, (India).

Email : arun.bhoi@gmail.com

(e-J. Foren. Crime Inv. 2012, 5, 1. Art. 1, 1^st Jan. 2012)

ABSTRACT

Cases of liquor tragedies occur worldwide. This is because the consumption of illicit liquors which are either prepared and sold by adding chloral hydrate or it gets contaminated with the poisonous material like methyl alcohol.  It is covered and  penalized in Maharashtra (India) under Bombay Prohibition Act – 1949. The chloral hydrate induces the hypnotic effect and the latter causes blindness and/or death. Therefore, the thermal analysis  of the chloral hydrate is carried out and its TGA¸ DTG and DTA properties are explored in this work.  The kinetic parameters reaction order and activation energy are calculated using ‘Coats and Redfern’ equation.

KEY WORDS

Illicit liquor, chloral hydrate, hypnotic, thermal analysis, TGA, DTG, DTA, reaction order, activation energy, Coats – Redfern equation.

INTRODUCTION

             Chloral hydrate 2,2,2 – trichloroethane 1,1 – diol is a sedative and hypnotic material. It is illegally added to the alcoholic beverages and its abuse leads to addiction (1). It is famous as “Knockout Drug”. In gastrointestinal tract chloral hydrate gets oxidized to tri-chloroacetic acid and partly reduced to trichloroethanol.  The latter is responsible for hypnotic action (2). Being a sedative  it is widely used in medicinal preparations and sold in the form of capsules, liquid formulations. It is, also, used as supposatory (3), rubefacient (2) and on treatment of asthma. Forensic scientist has to examine variety of samples from medico-legal cases comprising chloral hydrate when submitted for analysis by police / custom officers. Different methods are reported in the literature for analyzing chloral hydrate and its preparations such as ‘Fujiwara Test’ (4), spectrophotometry (5), IR spectrophotometry (6,7) and gas chromatography (8, 9). Least attention has been paid to its thermal studies and hence chosen to study TGA, DTG and DTA properties.

EXPERIMENTAL

Equipments  

Thermal properties of chloral hydrate are studied by using TGA and DTA units developed and installed in the Department of Chemistry, University of Pune, Maharashtra, India.

Reagents

Chloral hydrate used for the experiment was of the Analytical Reagent Grade.

Procedure

            The following specifications were used for the TGA (Thermogravimetric Analysis), DTG (Differential Thermogravimetry) and DTA (Differential Thermal Analysis) studies:

1. Sample Size                                               :  100 mg
2. Sample Holder                                          :  Quartz Cups
3. Temperature Range for Study                   :  Room Temp. to 800°C
4. Rate of Heating                                         :  3-4 °C / min.
5. Thermocouple Used                                  : Chromel – alumel.
6. Atmosphere                                               :  Static Air.

RESULTS AND DISCUSSION

            TGA, DTG and DTA studies of chloral hydrate, a monoclinic, perfect 001 crystal explains the thermal behavior of poly-halogen  compounds. On TG curve slow rate of loss is seen in the range of 60° – 90°C. It can be attributed to the crystalline transition and vaporization of chloral hydrate. Maximum loss on TG curve is seen in the range of 90° – 110°C (Fig. 1). It could be attributed to the decomposition of chloral hydrate to tri-chloro-acetaldehyde and water and its simultaneous vaporization.

Cl₃C.HC(OH)_{2 ———–>} 90° – 110°C _{———–>} Cl₃CHO + H₂O

The further loss in the range of 110°C to 150°C could be attributed to the subsequent decomposition of choral resulting in the release of chloro compounds / gases and CO / CO₂

The corresponding endothermic peaks for decomposition are also, noticed on DTG curve. The steep portion on ‘TG’ curve and ‘DTG’ peak corresponds to maximum slope.

The kinetic parameters, reaction order ( n ) and activation energy (Ea ) are deduced using ‘Coats and Redfern’ equation. These are n = 0.750 and Ea = 133.1 kj/mole.

The DTA studies are equally informative. The earlier endothermic peak is due to liquefaction, vaporization and decomposition of chloral hydrate and partly due to vapourization of products and helps to understand the decomposition reaction. The prominent endothermic peak could depict the decomposition of chloral and evolution of gases formed. The residual carbon gets converted into CO/CO₂ in the temperature range 730° – 750°C and the loss is 100%.

CONCLUSION

The results are reproducible, however, negligible differences are noticed from sample to sample as the quality of material does play the role, here. Thus, thermal analytical aid proves to be a good tool for the analysts and forensic experts in determining and estimating the chloral hydrate in variety of samples.

ACKNOLEDGEMENT

            Author’s thanks are due to Dr. G. N. Natu, Prof. S. B. Kulkarni of Department of Chemistry, University of Pune, Maharashtra, India and Director, Directorate of Forensic Science Laboratory, State of Maharashtra, India. The author’s thanks are also, due to ITAS, Mumbai, Maharashtra (India).

REFERENCES

1. The Merck Index : An Encyclopedia of Chemicals and Drugs. Ninth Edition, Merck and Co., Inc. Rahway, N. J., U.S.A., 1976, p- 260.
2. Satoskar R. S. and Bhandarkar S. D., Pharmacology and Pharmacotherapeutics, Eleventh Edition, Popular Prakashan Bombay, 1988, p- 87.
3. Alfred Goodman, Gilman L. S. and Alfred Gilman, The Pharmacological Basis of Therapeutics, Eighth Edition, Pergamon Press, New York, 1990, p- 365.
4. Fujiwara K., Sber. Abh. Naturf. Ges. Rostock, 1916, 6, p- 33.
5. Mahal H. S., Barve V. P. and Kamat S. S., Analyst, 1972, 97, p- 877-879.
6. Clrke E.G.C., Isolation and Identification of Drugs, The Pharmaceutical Press, London, 1974, p- 245.
7. DMS Working Atlas of Infra-red Spectroscopy, Butterworths and Co. (Pub.) Ltd., London, 1972.
8. Amin D., Al-Asley M. S., Talanta, 1981, 28(12), p- 955-956.
9. Archer A. W. and Haugas E. A., J. Pharma. Pharmae., 1960, 12, p- 754.
10. Coats A. W. and Redfern J. P., Nature, 1964, 201, p- 68.