INTRODUCTION:

Morphine, the main alkaloid of opium, was first obtained from poppy seeds in 1805. It is a potent analgesic, though its use is limited due to tolerance, withdrawal, and the risk of abuse. Morphine is still routinely used today, though there are a number of semi-synthetic opioids of varying strength such as codeine, fentanyl, methadone, hydrocodone, hydromorphone, meperidine, and oxycodone^.1Morphine is used for the management of chronic, moderate to severe pain. Opioids, including morphine, are effective for the short term management of pain. Patients taking opioids long term may need to be monitored for the development of physical dependence, addiction disorder, and drug abuse.²

Morphine is a pain medication of the opiate family that is found naturally in a dark brown resin in poppies (Papaver somniferum).³

Morphine is an opioid agonist used for the relief of moderate to severe acute and chronic pain.

Brand Names:

Arymo, Avinza, Doloral, Duramorph, Embeda, Infumorph, Kadian, M-Ediat, M-Eslon, Msir, Mitigo, Ms Contin, Statex

Generic Name:

Morphine

Volume of distribution:

The volume of distribution of morphine is 5.31L/kg. Morphine-6-glucuronide has a volume of distribution of 3.61L/kg.

Protein binding:

Morphine is 35% protein bound, the metabolite morphine-3-glucuronide is 10% protein bound, and morphine-6-glucuronide is 15% protein bound.

Metabolism:

Morphine is 90% metabolized by glucuronidation by UGT2B7 and sulfation at positions 3 and 6. Morphine can also be metabolized to codeine, normorphine, and morphine ethereal sulfate.

Route of elimination:

70-80% of an administered dose is excreted within 48 hours. Morphine is predominantly eliminated in the urine with 2-10% of a dose recovered as the unchanged parent drug. 7-10% of a dose of morphine is eliminated in the feces.

Morphine is derived from the Opium plant which is known as Papaver somniferum (Poppy). The plant grows upto a height of 1meter and each plant bears 5 to 8capsules. It is an annual plant. The plant is widely cultivated in the states of Rajasthan, Uttar Pradesh and Madhya Pradesh in India.^4-7

Morphine occurs as white crystals. It is the principal alkaloid of opium and of poppy straw. It is an alkaloid manufactured from raw opium or extracted directly from poppy straw.

The unripe capsule of Papaver somniferum is incised to derive opium. Crude opium is a dark brown or grey, irregular mass with characteristic odour and bitter taste and contains several alkaloids namely Morphine (10%), Codeine (0.5%), Thebaine (0.3%), Papaverine (1%), Noscopine (6%), Narcoenia (0.5%) etc.^8-11

MATERIAL AND METHODS:

Sample preparation

Preparation of standard solution 1000-ppm solution of Morphine was prepared in methanol by dissolving 0.1 gm of Morphine in 100ml of methanol. Preparation of Draggendroff spraying reagent (i) 2gm of Bismuthsubnitrate is dissolved in 25ml of glacial acetic acid and to it 100ml of water is added. (ii) 40gm of Potassium iodide is dissolved in 100ml of water. 10ml of (i) is mixed with 10ml of (ii) and 25ml of glacial acetic acid is added. The solution is then diluted with 100ml of water.^12-16

Thin Layer Chromatography Sample-extraction The following extraction technique is suitable for the isolation of the heroin, opium, and crude morphine from seized materials. Dissolve 5mg of sample each 1mg of methanol, and place both a 1 micro liter spot on to the plate.

OBSERVATION:

S. No.	Solvent System	Time to run	Color of	R_fof standard sample	R_fof extracted sample
S. No.	Solvent System	Time to run	spot	R_fof standard sample	R_fof extracted sample
1	Chloroform: Methanol (9:1)	39 min	Orange	0.95	0.95
2	Acetic acid: Ethanol: Water (3:6:1)	56 min	Orange	0.48	0.45
3	Diethyl ether: Acetic acid (6:4)	59 min	Orange	0.19	0.15
4	Ethyl acetate: Methanol: Conc. Ammonia (8.5:1:0.5)	25 min	Orange	0.32	0.3
5	Methanol: Isobutanol (6:4)	40 min	Orange	0.2	0.2
6	Isobutanol: acetic acid: water (4:1:5)	106 min	Orange	0.35	0.32
7	Cyclohexane: Chloroform: Diethyl amine (5:4:1)	41 min	Orange	0.15	0.13
8	Isobutanol: acetic acid: water (8:0.5:1.5)	106 min	Orange	0.35	0.33
9	Chloroform: Methanol: Water (3:5:2)	105 min	Orange	0.25	0.25
10	Ethanol: Water: Ammonia (6:3:1)	77 min	Orange	0.8	0.8
11	Chloroform: Ethanol: Water (2:6:2)	68 min	Orange	0.3	0.25
12	Isopropanol: Chloroform: Ammonia (4.5:4.5:1)	52 min	Orange	0.68	0.65
13	Chloroform: Diethylamine (9:1)	28 min	Orange	0.39	0.39
14	Chloroform: Methanol: Acetic acid (7.2:1.8:1)	54 min	Orange	0.58	0.55
15	Chloroform: Methanol: ammonia (6:3:1)	55 min	Orange	0.76	0.76
16	Chloroform: Methanol: Acetic acid (6:3:1)	44 min	Orange	0.69	0.65
17	Chloroform: Acetone: Diethylamine (7:2:1)	38 min	Orange	0.35	0.34
18	Dioxane: Acetone:Water (7:2:1)	44 min	Orange	0.2	0.2
19	2-propanol: Chloroform: Methanol: Water (5:2:2:1)	64 min	Orange	0.34	0.3
20	Acetone: Isobutanol: Acetic acid: Water (7:1:0.5:1.5)	41 min	Orange	0.41	0.4
21	Chloroform: Dioxane: Ethylacetate: ammonia (2.5:6:1:0.5)	33 min	Orange	0.41	0.41
22	Methanol: ammonium hydroxide (9.5:0.5)	19 min	Orange	0.5	0.5
23	Hexane: Benzene (5:5)	26 min	Nil	Nil	Nil
24	Benzene	39 min	Nil	Nil	Nil
25	Ethylacetate	18 min	Nil	Nil	Nil
26	n-Hexane: Acetone (8:2)	19 min	Nil	Nil	Nil
27	Cyclohexane: Chloroform (7:3)	45 min	Nil	Nil	Nil
28	Ethyl acetate: isopropanol (8.5:1.5)	24 min	Nil	Nil
29	Cyclohexane: Toluene: Diethyl amine (7:5:1.5:1)	32 min	Nil	Nil	Nil
30	Chloroform: acetone (8:2)	26 min	Nil	Nil	Nil
31	Ethylacetate: Chloroform: Acetone (6:2:2)	20 min	Nil	Nil	Nil
32	Acetone: Benzene (5:5)	16 min	Nil	Nil	Ni
33	Cyclohexane: Ethylacetate (5:5)	29 min	Nil	Nil	Nil
34	Ethylacetate: Cyclohexane: Ethanol (5:4:1)	35 min	Nil	Nil	Nil
35	Hexane: Ether: Acetone (8.5:1:0.5)	21 min	Nil	Nil	Nil
36	Hexane: Chloroform (5:5)	20 min	Nil	Nil	Nil
37	Diethylether: Ethylacetate (8:2)	28 min	Nil	Nil	Nil
38	Cyclohexane: Diethylamine (9:1)	26 min	Nil	Nil	Nil
39	Chloroform	29 min	Nil	Nil	Nil
40	Chloroform: Ethylacetate (9:1)	46 min	Nil	Nil	Nil
41	Xylene: Hexane: Diethylamine (6:3:1)	20 min	Nil	Nil	Nil
42	Toluene	17 min	Nil	Nil	Nil
43	Ethyl acetate: iso-propanol (8.5:1.5)	24 min	Nil	Nil	Nil
44	Chloroform: Isopropanol (1:9)	100min	Nil	Nil	Nil
45	Ethanol: Acetone:Water (1:1:8)	45 min	Nil	Nil	Nil
46	Benzene: 2 propanolacetone (6:3:1)	48 min	Nil	Nil	Nil

RESULTS AND DISCUSSION:

The morphine samples were analyzed in UV spectroscopy it showed the maximum absorbance at 287 nm and the result was compared with standard morphine sample.

RESULTS AND DISCUSSION:

The morphine samples were taken from different shops and their different characteristics like its color, texture, R_f value, etc were analyzed.

In the present study, it was found that morphine samples were analyzed.

After the TLC plates were sprayed with the Draggendorff’s reagent, Morphine appeared as orange spot in both samples and standard against a yellow background.

The reaction was instantaneous. Color formation was permanent. R_f value of Morphine extracted from viscera, under experimental conditions was found nearly equal to that of standard used.

All the 46 solvent systems were analyzed and out of which 22 solvent systems were found to be good to run the sample. The 22 as solvent systems which are found as good systems are from 1 to 22 shown in table 1 and solvent system from 23 to 46are not considered good solvent system for detection of Morphine. R_f value of sample in different solvent systems was in between 0.13 to 0.95 and the time taken by solvent system was in between 19 min to 106 min.

Chloroform:

Methanol (9:1) resulted highest R_f value i.e. 0.95 and Methanol: ammonium hydroxide (9.5:0.5) took 19 min. to run the sample which is the least time among the 22 solvent systems. Therefore according to highest R_f value and the least time taken to run the sample, both of them can be considered as good solvent systems. In case any one of 22 solvent systems is not available, other mentioned system can also be used for the analysis of Morphine.^17-18

CONCLUSION:

Alternative methods were sought. TLC proved to be an excellent choice because it is a simple equipment, takes less time for development, simultaneous analyses of large number of samples are possible, early recovery of separated components is possible, separation effects are superior, early visualization of separated compounds, sensitive as extremely sharp delineated spots are obtained in TLC. The main advantage of TLC plate is the low cost. When all the 46 solvent systems were analyzed, it was found that 22 solvent systems are good to run the sample. If any one of the solvent systems is not available, we can choose any one of 22 solvent systems. Beside Draggendorff’s reagent, we can also use Iodoplatinate solution as a spraying reagent.

REFERENCES:

1. I.C.F. Poole, S.K. Poole; Analytical Chemistry. Acta, 216: 1989;109-145.

2. Clarke; 'Isolation and Identification of Drugs'; A.C. Moffat, London; 2nd edn; 160-174 and 790.

3. M.L. Puttermans et al: 'Analytical Chemistry'; 67; 1984.

4. Stewart C.P., Stolman A; 'Toxicology Mechanisms and Analytical methods'; vol.II; 1996; 842.

5. http://www.phytomedical.com/Plant/Morphine.asp.

6. Parikh, 'Textbook of Medical Jurisprudence and Toxicology'. 2001.10.1 and 11.4.

7. Modi, Jaishing P. 'Textbook of Medical Jurisprudence and Toxicology'. M Tripathi Pub; 2001.

8. http://www.narconon.ca/images/opium.jpg.

9. Reddy K.S. Narayan; 'The essentials of forensic Medicine and Toxicology'; 6th edn., 1997; 473-491.

10. http://z.about.com/d/worldnews/1/7/t/3/-/-/2004822. JPG.

11. V. Shankar et al; Forensic Sci. Int.,40; 1989; 45-55

12. Saferstein R; 'Criminalistics-An Introduction to forensic Science', 6th Edn. Prentice Hall, New Jersey; 1998.

13. Dreisbach Robert; 'Handbook of Poisoning': 3rd edn; 225-229.

14. Dreisbach Robet; 'Handbook of Poisoning': 4th edn; 226-228.

15. Oliver John; 'forensic Toxicology'; 119-125.

16. D. McNally William; Toxicology; 1937: 551-573.

17. M. Houck Max. A Siegel Jay; 'fundamental of forensic Medicine and Toxicology'; 335-337.

18. Sharma B.R. 'forensic Science in Criminal Investigation and Trials'; 4th edn; 834-837.

Received on 27.07.2023 Modified on 28.08.2023

Research J. Science and Tech. 2023; 15(4):191-196.

DOI: 10.52711/2349-2988.2023.00031