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CHAPTER 4

O m S ^ ( R y jl I0 9 fS ( R ^ V L ^ S

The pharmacognostical and phytochemical studies on the stem .bark of Alstonia scholaris (Linn.) R.Br. (Chatiun), Balanites aegyptiaca (Linn.) Delile (Hingot), Bombax ceiba Linn. (Semul) and Morus alba Linn. (Shehtoot) have been conducted, Pharmacognostical studies include detailed description of macroscopical and microscopical characters of stem bark, histochemical tests, ash and extractive values determination, fluorescence analysis, reactions of the powdered bark with different chemical reagents, thin layer chromatographic studies, inorganic elemental analysis and quantitative organic analysis. In phytochemical studies, the preliminary tests for the detection of different phytoconstituents, extraction, isolation, purification of compounds and their spectral and chemical analysis have been carried out among the investigated species and the results are given below:

1 Alstonia scholaris fLinn.) R. Br„ fChatSyii)A: Pharmacognostical studies Macroscopical characters

Fresh bark samples of A. shcolahs were collected from Ghaziabad, Uttar Pradesh. The bark was in pieces of 4-6 cm. long, 1.5-4 cm broad and up to 1-8 mm thick, these were curved, often quilled. The outer surface was rugged with distinct white-coloured lenticles; colour iron grey to mouse grey before scraping with scalpel and mouse grey after scraping, cracks and fissures with clean cut edges, wrinkles irregular. Inner surface was rosy buff to cinnamon coloured, corrugations absent. Transverse surface shows much lignification in both the surfaces. The taste' is bitter and acrid and odour is not characteristic (Plate-11, Fig.-A&B).

Microscopical charactersThe transectional view of the bark shows that the cork consists of 10-20-30

layers of tangentially elongated, brick-shaped and thin walled parenchymatdus cells

measuring 22.8-31,4-38.0 jam in length and 15.2-20.0-22.8 im in width (Plate-Ill, Fig.-A). The cork cambium or phellogen is notited to be of single layer; the cells are tangentially elongated and thin wailed parenchymatous. Phelloderm in outer cortex is

5-8 layered. Measurements of the different cells of phelloderrn were taken, these were found 26.6-38.0-45.4 fim in length and 11.4-22.8-30.4 f.im in width (Plate-Ill, Fig.-B). Cortex is divisible into outer and inner zones. The outer zone is distinguished by having large sclereids of 32-35-46 layered; dark cell contents, abundant crystals and starch grains. Starch grains have been found uniform in distribution, simple, oval in shape, lamellae distinct. Crystals have been found mostly rhomboidal type, measuring from 15.3-23.3-28.5 |am in length and from 12.0-15.0-21.5|im in width. Scelerenchymatous tissue is of two types: phloem fibres and sclereids. Sclereids are not thick walled; length and width of sclereids are from 42.6-60.5-80.5 |.im and 22.8-

26.4-32.4 p,m respectively. Latex cells frequently show their presence in the periderm region; pericycle is fibrous, 6-8-10 fibres have been noticed in each patch. Secondary phloem is from 560-644-728 (im deep, Phloem fibres are with narrow apex, 165-229-

286 i-im in length and 30.4-35.6-42.8 {.im in width. Phloem rays are multiseriate, 280- 430-540 p.m long; 56-70-98 fj,m wide; upright cells vary from 40-45-58 jam in length

and 32-40-52 p,m width; rays have been found in abundance near the cambium 2.5- 3.0-3.5 /mm (Table-5, Plate-Ill and IV).

Microscopy of powdered barkPowder of the crude drug is brown in colour, coarse and free-floating. Small

amount qf powdered material (sieved through 40 mesh) when placed on microscopic slide; after mixing with few drops of 40% w/v aqueous chloral hydrate after, heating gently under Bunsen Burner with few drops of 1% alcoholic phloroglucinol and by mixing one drop of concentrated hydrochloric acid, shows the presence of abundant cork cells, a few sclereids; frequent rhomboidal crystals; few phloem fibres (Plate-lV, Fig.-L); sclereids often in chains,

Histochemical tests

Transverse sections of the stem bark have been treated with commonly used chemicals and reagents for the tests of starch, saponins, tannins, lignins, alkaloids, flavonoids, calcium oxalate and proteins and the results have been presented in Table-7.

C hapter ~ Four O bserva tion s & R esults

( 1 , ....... — — ------------ ---------- Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

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Ash valuesPercentage of total ash, water-soluble ash. acid-insoluble ash and sulphated

ash were determined by conventional methods and were noted to be 20.72%, 14.62%, 1.46% and 14.2% respectively (Table-8 and Graph-1).

Total extractive valuesAlcohol soluble matters, water soluble matters and crude fibres were

determined and found to be 17.32%, 23,61% and 25,66% respectively (Table-6).

Successive extractive values f

The percentage of extracts in different solvents, viz. petroleum 80°C), diethyl ether, benzene chloroform, ethanol and water were determined. The maximum extract was found in water (21,61%) and the minimum in chloroform (2.41%). In petroleum ether, diethyl ether, benzene and ethanol, the percentage of extracts, were recorded 3.25%, 2,60%, 3.60% and 5,60% respectively (Table-9 and Graph'2).

Moisture contentMoisture content (loss in weight on drying at 105*^0) was determined by the

imethod given in "The Unani pharmacopoeia of India", and found to be 63.20% (Table-6),

Fluorescence analysisThe fluorescence analyses of the entire bark, powder of the bark, ethanolic

extract and aqueous extract of the bark were carried out and the results haVe been presented in Table-10.

Behaviour of the powdered bark towards some chemical reagents.The yellowish brown powder was treated with routinely used acids/ reagents

and the characteristic changes were observed and the results are summarized in Table-11.

C h a p ter- - F o u r ___________ __________________________________O bserva tion \ & Residl.s

'■'.•mv- Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

Quantitative estimation' of organic constituentsIn quantitative estimation of organic constituents, the total alkaloids,'steroids

and resins are found to be 0.20%, 0.078% and 0.044% respectively (Table-6).

Thin layer chromatographic studiesThin layer chromatographic studies of the different extracts were studied. The

benzene extract showed maximum (six) spots on T.L.C. plate coated with silica gel, followed by five spots in methanolic extracts, four spots in hexane and ethanolic extracts and three spots in petroleum ether and water extracts. The minimum (two) spots were found in chloroform and acetone extracts. The solvent systems used and R.F. values are given in Table-12 (Plate-XV, Fig-A au aa, Fig~B as, 34, as, Fig-C ae, a?, as).

inorganic elemental analysisNitrogen, phosphorus, potassium, sulphur, calcium and iron contents were

estimated in the young bark of A scholaris during January 2001 to December 2001 at the intervals of one month. The maximum concentrations of nitrogen (2.86%±0,020), phosphorus (G.88%+0.012), potassium (3.92%±0.041), sulphur

(0.Q48%±0.008), calcium (3.42%±0.038) and iron (0.72%±0.08Q) were found in the months of May, August, August, May, September and August respectively, then the concentrations were found less in the following months and the minimum concentrations of nitrogen (2.58%±0,061), phosphorus (0.80%±0.008), potassium

(3.80%±0,080), sulphur (0.042%±0.002), Calcium (3.34%±0.G40) and Iron (0.61 %±0.002), were estimated in the months of September, December, January, October, February and April respectively (Table-13).

B; Phytochemicai stydies Prelsminary phytochemicai screening

Tests for the detection of different organic constituents were carried out. Our results showed the presence of reducing sugar, steroids, alkaloids, proteins, flavonoides, resins and glycosides Tannins and saponins did not show their presence in any extract (Table-14).

C hapter - Four____________________________________________________________ ______________________________________________________________________________________ O bserva tions & R esu lts

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Pharmaco-anatomy Laboratory, DepartmentofBotany, Faculty o f Science, Jamia Hamdard

Extraction and isolation of compoundsStem bark of A. scholaris, was dried in air and finally in an oven at a

temperature below 45°C and then coarsely powdered. The dried and powdered stem bark (1.760 kg) was extracted with ethanol in soxhlet apparatus. The extract was concentrated under reduced pressure to get a dark brown viscous semi-solid mass (300 gm). It was analyzed chemically for the determination of the presence of different chemical constituents. The concentrated mass was dissolved in minimum amount of Me-OH and absorbed on silica gel to form slurry. The slurry was air dried and subjected to silica gel column chromatography prepared in petroleum ether. The column was eluted with petroleum ether chloroform and methanol in order to increasing polarity to isolate the following compounds.

1.1 Aistrinine [AS-1]Elution of the column with petroleum ether (fraction 1-9) afforded colorless

crystals of AS-1 re crystallized from chloroform: methanol (1:1), 4.90 gm (0,28% yield). Rf. : 0,82 (n-hexane: ethylacetate, 9:1)m.p. : 139-14fCIRvmaxIKBr) : 2959, 2840, 1730, 1600. 1462, 1380, 1274, 1123,10.72, 742 cm’^^HNMR (CDCia) : Table-45

NMR (GDCb) : Table-45+ve FAB MS miz {m l in i) : 480 [M]^ (C32H48O3) (6,9), 438 (90,8), 423(18.3), 408 (22.7), 393 (24.1), 232 (5.8), 214 (5.3), 191 (11.6), 188 (18.1).

Chapter ~ Four O bserva tions & R esulis

1.2 Alstotrinine [AS-2]Further elution of the column with petroleum ether (fraction 10-26) furnished

colurless crystals of AS-2 recrystallized from chloroform; methanol (1 :1 ), 4.70gm (0.26% yield).

0.54 (n-hexane: ethylacetate, 9:1)129-131°C

3440, 2960, 2873, 1640, 1463, 1360, 1274, 1124,

Rf.m.p,IR vmax (KBr)

1072, 1040. 742 cm'^

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Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Xainia Hamdard

Chapter - ■ Four Ohservationn & Resuh.s

■'H NiiRICDCIsl : Table-46NMR ICDCia) : Table-46

+veFABiiS mlz (rei. in t| : 438 [M f (C30H4602) (22.5), 423 (31,8),408 (55.6), 392 (11.2), 378 (11.1), 232 (18.6), 217 (22.8) 214 (13.5), 206 (21.7), 202(41.6), 191 (21.6), 188 (32.5), 173 (29.1), 158 (31.7).

1 .:Elution of the column with petroleum ether-CHCls (3:1) (fraction 5-15)

furnished colurless crystals of AS-3 recrystallized from chloroform: methanol (1:1), 1 .2 0 gm (0.068% yield).

0.36 (n-hexane: ethylacetate, 9:1)108-110°C 292 nm (loge= 2,95)

3452, 2960, 2820, 1640, 1462, 1380, 1273, 1123,1072, 742 cm"''

^HNMR'CCDCig) ; Table-47NMR (CDCial : Table-47

+veFABMS m/z (rei, in t) : 412 [M f (C29H48O) (27.6), 397 (41,2), 394(33.6), 382 (41.6), 379 (16.2), 367 (11,5), 351 (9.6), 339 (10.2), 288 (6.1), 273 (5,2), 271 (14,9), 255 (16.8), 240 (11.9), 213 (17.8), 198 (16.8), 192(10.1), 178 (18,6), 174(22.3), 164 (24,8), 160 (31.1), 159 (16.9), 149 (18,3)146 (42.8), 145 (67.6), 138 (7.8), 131 (62.7), 124 (26.1), 120 (68.2), 106 (70.8), 105 (100).

1.4p-

Elution of columr) with petroleum ether; chloroform (4:1) (fraction 16-23) furnished a colourless amorphous powder of As-4 recrystallized from chloroform: methanol (1:1), 1.1 Ogm (0.040% yield).

Rf.m.p.UV Xmax (MeOH)

IR vmax (KBr)

810, 785 cm'^

0.40 (n-hexane-ethylacetate, 9; 1)132-134°C205 nm (loge= 4.15)

3430, 2930, 2855, 1620, 1460, 1375, 1055, 955,

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pharraacQ-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

i i lR fCDCial 5 5.34 (1H, d. J = 4,64 Hz, H-6 ), 3.52 (1H, brs,

w1/2, 18.5 Hz, H-3a), 1.25 (3H, brs, Me-19), 0.93 (3H, d. J - 6.50 Hz, Me-21), 0.84 (3H, t, J = 6,03 Hz. Me-29), 0.82 (3H, d, J = 6.00 Hz, Me-26), 0.80 (3H, d, J = 6.0Hz, Me-27), 0.68 (3H, brs. Me-18).

NMRfCDCy : 537,25 (C~1), 31.66 (C-2), 71,82 (C-3), 42.3 (C^4),140.71 (C-5). 121.73 (C-6), 29.69 (C-7), 31.9 (C-8), 51.23 (C-9), 36.14 {G-10), 21.07 (C-11)., 39.77 (C-12), 40.48 (C-13), 56.76 (C-U), 24.29 (C-15), 28.23 (C_16), 56.05 (C-17), 11.97 (C-18), 19.38 (C-19), 36.51 (C-20), 18.77 (C-21), 33.94 (C-2^), 26.08 (C-23), 45.83 (C-24). 19.15(0-25), 19.81 (C-26), 19.02 (C-27), 23..06 (C-28), 11.85 (C-29).+ve FAB MS m/z frel. int.) ; 414 [m]^ (CsgHsoO) (50.5), 400 (5.4), 396(22.2), 381 (13.6), 329 (18.3), 303 (21.8), 273 (12,1), 256 (14.2), 231 (11.1), 213(17.3), 198 (6.9), 173 (8.9), 163 (15.4), 161 (15,8), 159 (16.8), 145 (22.7), 133 (17.8), 123 (9.8), 69 (32.5), 67 (12.3), 57 (50.1), 55 (55.8).

1.5 {5-Sitosterol glycoside |AS~5]

Elution of column with chloroform: methanol (9:1) (fraction 29-38) furnished a colourless amorphous powder of AS-5 (2 ' crop) recrystallized from Chloroform: methanol (1:1), O.SOgm (0.034% yield).Rf. : 0.76 (Toluene: ethylformate: formic acid, 50:40:10)m.p. : 257-259°CU V W : 328 nm (logf;= 4.15)

IRvmax(CCl4) : 3590, 3540, 2965, 2875, 2362, 1810, 1700, 1652,1460, 1375, 1072, 785 cm'^'H NMR (CDCIs) : table-48

NMR (CDCI3) ; Table-48+ve FAB MS m/z frel, in t) : 575 [M]’ (CssHeoOe) (N.O.), 414 [M-sugarf.4.8, 397 (32.1), 381 (3.4) 279 (5.3), 255 (6.2), 242 (7.6), 213 (2.6), 198 (4.9), 191(13.6), 186 (12.8), 178 (6.5), 174 (5.6), 164 (7.8), 154 (36.3), 149 (10.2), 146 (4.5), 188 (9.8), 135 (23.2), 131 (6 .6), 124 (10.2), 120 (6 .8), 115 (8.3), 107 (16.2), 106(19.3), 97 (18.2), 91 (22.1), 83 (22.0), 81(35.11), 71 (13.2), 69 (21.6), 55 (43.0), 43 (100 .0).

C hapter ~ Four _______________________________O bservations & Results

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Pharmaco-anatomy Laboratory, Departinent o f Botany, Faculty o f Science, Jamia Haindard

Acid hydrolysis of compoynd AS-5The compound AS-4 was refluxed with 2N HCl in 80% MeOH 15 mi for 4 hrs.

After cooling the reaction mixture was poured in crushed ice and the hydrolysate was then extracted with EtOAc to give the aglycon, m.p. 130-140°C. CO-TLC comparable. The neutralized and concentrated aqueous hydrolysate showed the presence of glucose on comparison with authentic sugar on silica gel.

C hapter - F our O bserva tions & Resu/fs

Pharraaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hainciard

Chapter-Four O bserva tion s R esu lts

Clieiiriical compounds Isolated from the stem l3 arlc of Alstonia schoiaris

AcO

'O H

‘ 32 46* 3 * 30 46 2

Alstrinine Alstotrisiiffle

Isostigmastero! P“Sitosterol

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Pliamiaco-anatonay Laboratory, Department of Botany, Faculty o f Science, Jarnia Hamdard

2. Balanites aegyptiaca (Linn.) Deliie (Hingot)A: Pharmacognostical stydies Macroscopical characters

The shape of the bark of the trunk region of B. aegyptiaca is curved. The outer surface is grey and rough showing lenticles. In older barks, sometimes small circular patches called corky worts are found. The inner surface is yellowish and smooth showing reticulate structures of fibres. The taste is bitter and the smell is pungent (Plate-VI, Fig.-A&B).

t

Microscopical charactersThe periderm consists of three parts, the phellem, which is the cork, consists

of 10-30 layers of rectangular and thin-walled parenchymatous cells measuring 15,2- 26.6-34.2 /.im in length and 15.2-19.0-22.8 |.im in width, the phellogen or cork cambium is single layered and the phelloderm is 1 0 -2 2 layered, thin-walled parenchymatous cells measuring 22.8-26.4-32.5 ).im in length and 12.0-15.0-21.5 /.im in width. Following, the secondary cortex is usually composed of a wide zone of compact, thin-walied, medium-sized, and polygonal to oval parenchymatous cells. There are some stone cells interspersed in this region. Stone cells are rectangular to somewhat oval with highly thickened and iignified walls bearing numerous sirnple pits are measured from 76.0-95.0-110.2 |j.m in length and 63-72.2-79,8 jim in width. The starch grains are found in few cells of secondary cortex which are small, simple and oval to rounded in shape, pericycle is fibrous, 8-12-16 fibres / patches. The medullary rays are multiseriate in the innermost region of secondary phloem and Bi- or tri- seriate medullary rays are rarely observed. The ray cells are thin walled, radially

4

elongated, parenchymatous and individual cell is measured from 57.0-76.0-95.0 jim

in length and 26.6-28.8-34.2 fxm in width but gradually increasing in dimension towards outer ends. The secondary phloem is generally represented by sieve tubes, companion cells, phloem parenchyma and phloem fibres. The phloem parenchyma is

thin-walled, polygonal and is 19.0-41.8-53.2 jarp long and 15.2-22.5-32.0 p.m wide in dimension (Table-15, Plate-VII).

C hapter - Four_______________________________________________ O bsetyation^i & Resiihs

Pharmaco-anatomy Laboratoiy, Department o f Botany, Faculty o f Science, Jarnia Harndard

Microscopy of povi/dered barkPowder of the crude drug is light brown coloured, coarsed and free flowing.

The taste is acrid and odour is pungent. Small amount of the powdered material (sieved through 40 mesh) is placed on microscopic slide; mixed with few drops of 40% w/v aqueous chloral hydrate and heated gently under Bunsen burner. Few drops of 1 % alcoholic phloroglucinol are added to this and warmed by mixing one drop of concentrated hydrochloric acid, the slides are then mounted in glycerin and are observed under microscope which shows the presence of fragments of cork, secondary cortex made up of parenchymatous cells and also associated with the stone cells. The stone cells are medium-sized, lignified horizontally striated with small lumen; ray cells usually found attached with phloem parenchyma. Many phloem fibres and starch grains were also observed during the present investigation (Plate- VII).Histochemical tests

Transverse sections of the stem bark were treated with different chemicals and reagents and found positive tests for starch, saponins, lignins, tannins, alkaloids, flavonoids, proteins and calcium oxalate crystals. Details of the tests are given in Table-17.

Ash valuesAsh analysis of the bark was carried out by conventional methods. Percentage

of the ash, water soluble ash, acid insoluble ash and sulphated ash was determined and the results are presented in Table-18 and Graph-3.

Total extractive valuesAlcohol soluble extractives, water soluble extractives and crude fibres content

were determined and the results are shown in Table-16.

«

Successive extractive valuesThe successive extractive values of the bark in different solvents, viz.;

petroleum ether (60-80°G), diethyl ether, chloroform, benzene, ethanol and water were determined and the results summarized in Table-19 and Graph-4.

C hapter - Four_______________________________________________________ ___________________ O h serva tion s & R esidls

A..,.,..,-, pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Scicncc, Jamia Haindard

78

Moisture contentMoisture content (loss on drying in weight at 105°C) was determined by tine

method given in the Unani Pharmacopoeia of India, Part 11" and results are depictedin the Table-16.

Fluorescence analysis ,the fluorescence analysis of the entire bark, powdered bark, ethanolic and

aqueous extracts of the bark was conducted. The colours of the entire bark, powder as such, powder after different treatments, ethanolic extract and aqueous extrats at different,pH, and were observed in daylight and under U.V, light (Table-20).

<

Behaviour o f the powdered bark towards some chemical reagentsThe light brown powder of B. aegyptiaca bark was treated with different

chemical reagents. The mixture of the powdered drug and chemicals were allowed to warm and cool down for two hours, The actual colour of the powdered drug and the changes in colour of powdered drug were noted and presented in Table-21.

Quantitative estimation of organic constituentsQuantitative estimation of organic constituents e.g. total phenolics, tannins and

saponins percentage in the bark of B. aegyptiaca were calculated and the results are presented in Table-16.

Thin layer ctiromatograpiiic studiesThin layer chromatography of various extracts of the bark of B. aegyptiaca was

examined and Rf values are calculated and presented in Table-22 ((Plate-XV, Fig-A bi, b2, Fig-B ba, b4, bs, Flg-C be, by, be).

4

Inorganic elemental analysisInorganic elemental analysis for the estimation of nitrogen, phosphorus,

potassium. Sulphur, Calcium and Iron were carried out in the young bark of B. aegyptiaca from January 2001-December 2001 at the interval of one month. The high

concentration of nitrogen (5.83%±0.051). phosphorus (0.89%±0.010), Potassium

(2.66%±0.041). Sulphur (0.028%±0.002), Calcium (1.93%±0.019) and Iron

/ r f lV . : 79- pharmaco-anatomy Laboratoiy, Department o f Botany, Faculty o f Science, Jamia Mamdard

Chapter - - F our __________________________________________ O bserva lio m c & Ra.sitlis

(0.79%±0.032) were calculated in the months of November, October, September, May, May and June respectively. The concentrations were found in the decreasing trend in the following months and the minimum concentrations of nitrogen (5.46%±0.113), Phosphorus (0.85%±0.0155), Potassium (2.57%±0,047), Sulphur (.021%±0.001) Calcium (1.85%±0,011) and iron (0.71%±0.019) were calculated in the months of September, May, June, August, March and February respectively Table-23.

B: Phytocliemical studies«

Preliminary pliytoctiemica! screeningQualitative tests were conducted to detect the presence of different

phytoconstituents in various extracts of the drug, showed positive results for reduqing sugar, tannins alkaloids, saponins, steroids, proteins, flavonoids and glycosides and negative result for resin (Table-24).

«

Extraction and Isolation of compoundsStem bark of B. aegyptiaca, was dried in air and finally in an oven at a

temperature below 45°C and then coarsely powdered. The dried and powdered stem bark (2.5 kg) was extracted with ethanol in soxhlet apparatus. The extract was concentrated under reduced pressure to get a dark brown viscous semi-solid mass (200 gm). It was analyzed chemically for the determination of the presence of different chemical constituents. The concentrated mass was dissolved in minimum amount of Me-OH and absorbed on silica gel to form slurry. The slurry was air dried and subjected to silica gel column chromatography prepared in petroleum ether. The column was eluted with petroleum ether chloroform and methanol in prder to increasing polarity to isolate the following compounds

2.1 Compound BA-1Elution of the column with petroleum ether (fraction 1-30) furnished colourless

amorphous powder of BA-1 recrystallized from CHCI3-MeOH (1:1), 300 mg (0.021% yield).

Rf. ; 0.60 (n-Hexane-EtOAc, 9:1)m.p. : 78-80°C

C hapter - F our O bserva tion s & Rexiillx

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S',;,,.,.,, phamiaco-anatoray Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

IR vmax (KBr| : 2919, 2850, 1463, 1379, 1371, 805, 721 cm'^

■'H NMR fCDCIg) : S 2.32 == 17.65 Hz. H~10 a). 1.58 (2H.rn, H2-9). 1.43 (2H. brs, H2- I I) . 1.25 (30H, brs, 15 x CH2), 0.88 (3H, d, J = 7.20 Hz, Me-28), 0.85 (3H, t, J " 8,7 Hz, Me-1), 0.82 (3H. t, J = 8.7 Hz, Me-27).

NMR (CDCial : 8 32.81 .(C-10), 29.71 (24 x CH2), 22.32 (C-27),21.13 (C-28), 14.31 (C-1).+ve FAB MS m/2 (rel. in t) : 394 [M f (CasHss) 8.7, 155 (68.9), 127 (27.3).

C hapter - F our _______________________ O bserva lion s & Results

Elution of the column with nnethanol (fraction 1-30) furnished colourless crystals of BA-2, recrystaiiized from EtOH, 530 mg (0.021% yield).Rf. : 0,83 (n-Butanol-AcOH-water, 5:4:1)

m .p . : 160-162°CIRvmaxfKBr) 3405, 3324, 2950, 2930, 2912, 1512, 1452, 1380,1383, 1133, 1050, 700 cm'^1u MBBO ^ . , g g q j d, J = 6.0 Hz, H-1), 4.65 (1H, d, J =

15.0 Hz, H-1'), 4.65 (1H, m, H-5), 4.50 (1H, m, H-5'), 4.45 (1H, m, H-4), 4.43 (1H, m, H-4'), 4.36 (1H, m, 1+3), 4.32 (1H. m. H-3), 4.32 (1H, dd, J = 6.0, 12.0 Hz, H-2), 2.24 (1H, m, H2-2 '), 3.62 (2H, brs, H2-6), 3.24 (2H, brs, H2-6 ').

c

NMR fCDCIa) : 6 101.23 (C-1),' 9553 (C-1'), 83.83 (C-5, C-6’),76.50 (C-5), 76,33 (C-5'), 72.65 (C-2), 72.46 (0-2'), 72.02 (C-3), 70.97 (C-3’), 70.14 (C-4, C-4'), 62.49 (C-6), 60.01 (C-6 ').

Elution of the column with MeOH (fraction 31-52) furnished colourless crystals of BA-3, recrystallized from ethanol, 420 mg (0.016% yield).Rf. : 0.73 (n-Butanol-AcOH-water, 5:4:1)

m.p. : 182-185°CIR vmax (KBr) : 3399, 2937, 1631, 1417, 1379, 1051 cm’’’

NMR (DMSO-dg) : 5 5.20 (1H, d, J = 8.4 Hz, H-1), 5.17 (1H, d', J = 3.3Hz, H-1'), 5.04 (1H, d, J - 9.0 Hz, H-1"), 4.66 (1H, d, J = 6.6 Hz, H-1'"), 4.45 (2H, m, H-5. H-5'), 4.24 (2H, m, H-5", H*5'"), 3.96 (1H, m, H-'2), 3.78 (1H, m, H-2'), 3.75 (1H, m, H-2"), 3.62 (1H. m. H-2'"), 3.54 (2H. m. H-3, H-3'), 3.42 (2H, m, H-3". H-3”’), 3.18

. . 8 1

Phanmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

(2H. m. H-4, H-4’), 3.14 (2H, m. H-4”, H-4'"), 3.11 (2H, m, H2-6 ). 3.03 (2H, m. H2-6 '), 1.06 (3H. d, J = 5.7 Hz, Me-6"), 0.87 (3H, d, J == 4.2 Hz, Me-6'").■'"C NMR fCDCia) : Table~49.-s-ve FAB liS mfe frei, int.) : 634 [M f (C24H42 O19) (5.6).

Chapter - Four Observations & Results

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Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

Chapler - Four Ohxervalion.s A Resnhs

Chemicai compounds isolated from the stem bark of Balanites aegyptiaca

CH,

CH3'(CHj)g-CH (CH2),|a CH3

10-Methyl-n-licptacossine

HO CH^OH

OHHO

O

OH

O O OH

HO OH

2 22 11Sucrose

HO

OHHO I

O

OH

HO

OOH

O

OH

O

CHyCHg

OOH

-O

OH

CH2CH3I O OH

OH

OH

24 42 19

DigliifcotlirliainHoside

r-'' 83

Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia tlamdard

Chapter - Four Ohservalions & Rexiil^«

3. Bombax ceiba Liriri. (Sernui)A: Pharmacognostical studies llacroscopicai characters

The sample of Semul bark was cut into pieces of 2-8 cm long, 1.5-5 cm broadI

and 0.5-2 cm thick; these were curved longitudinally. The outer surface is grey,?

covered, with hard sharp conical prickles and bears horizontal lenticels. The inner surface is rough longitudinally striated, brown to black in colour. The bark is acrid in taste and has no characteristic smell (Plate-lX, Fig.-A&B).

Microscopical ciiaractersThe stem bark of the B. ceiba is composed of 12,5 percent periderm and 85.5

percent secondary phloem (37.5 percent non-conducting phloem and 50 percentconducting phloem). In periderm, the cork zone is composed of many layers oflongitudinally elongated somewhat rectangular cells, arranged in radial rows withslightly wavy and thick walls. Each cell of the cork is measured from 38.0-40.4-60.8

fim X 19.8-22.8-30.4 |am (Plate-X, Fig,-A). The cork cambium or phellogen is 3-8

layered, its cells are tangentially elongated and thin walled parenchymatous (Plate-X,Fig.-A), Following, the phelloderm cells are rectangular and somewhat oval andparenchymatous and are mostly filled with tannins, measuring 26.6-38.0-41,8 jam x22,8-26.6-30.4 fim (Plate-X, Fig.-A), The secondary cortex is usually composed ofwide zone of compact, thin walled medium sized, rectangular, polygonal to ovalparenchymatous cells which are measured from 58,0-65,0-78,0 j,tm x 15.0-19.0-22.8

jim. There are some stone cells interspersed in this region (Plate-X, Fig.-B). Stonecells are oval with thickened and lignified walls bearing numerous pits and havenarrow lumen. Their size vary from 49.4-57.0-64.6 jim x 38.0- 41.8-49.4 fim. Fewstone cells are noticed to possess prismatic crystals of calcium oxalate (Plate-X, Fig.-BCD&E). Besides, the starch grains are also found in few cells of secondary cortex,which are small simple and rounded in shape (Plate-X, Fig.-C&D). The medullaryrays are bi, tri, or multiseriates in the innermost region of secondary phloem (Plate-X,Fig.-F, Piate-XI, Fig.-G&H) but uniseriate medullary rays are also observed (plate-XI,Fig.-H&l). The ray cells are thin walled, radially elongated, rectangular,

parenchymatous and individual ceil is measured from 45.0-69.5-75.2 jim x 28.5-30.2-

33.2 jim but gradually increasing in dimensions towards outer ends. The sieve tube84

PhaiTOaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

members and companion cells are found in groups in this species. The sieve tube cells have simple plates on their slightly or oblique end walls and several sieve areas of varying size on lateral walls, It is also noticed that some ray cells •become scelerenchymatous and attached with stone cells so that it become continuous and extended to 3-4 rays in tangential direction. Many phloem fibre patches are interspersed in secondary cortex and each patch contains 10-30 fibres (Plate-Xl, Fig.-E). A thin walled parenchymatous, 7-9 layered, cambium cleady can be seen the junction of wood and bark (Table-25, Plate-Xl, Fig.-l&J),

Microscopy of powdered barkPowder of the bark is reddish brown, coarsed and free floating, without any

characteristic smell and taste, The small amount of the powdered material (sieved through 40 mesh) is placed on microscopic slide; mixed with a few drop of 40%w/v aqueous chloral hydrate and heated gently on Bunsen Burner, Few drops of 1% alcoholic phlorogucino! are added to this and warmed by mixing one drop of concentrated hydrochloric acid. The slides are then mounted in glycerin and observed under microscope, which shows the presence of fragments of cork, secondary cortex having parenchymatous cells and sometimes associated with stone

4

cells. Many free stone cells are also observed. Stone cells are medium-sized, lignified, horizontally striated with broad lumen. Cork cambium pieces are mostly attached with the cork cells but occasionaliy with phelloderm. Prismatic crystals of tannins and starch grains were also observed during thepowder analysis. Few phloem fibres with narrow lumen and tapering ends were also observed during the present investigation (Table-25, Plate-Xl, Fig.-K).

Histochemical testsTarnsections of the stem bark of B. ceiba were treated with routinely used

chemicals and reagents which showed the presence of starch, tannins, lignins, alkaloids, flavonoides and proteins, and the absence of saponins, lipids and.calcium oxalate (Table-27).Ash values

The percentage of total ash, water-soluble ash,. acid-insoluble ash, and sulphated ash were calculated and the results are summarized in Table-28 and Grapg-5.

. 85 ■

Chapter - Four Ohservatiom & Results

Pharmado-anatomy Laboratoiy, Department o f Botany, Faculty o f Science, Jamia Hamdard

Total extractive valueTotal water-soluble extractive, alcohol soluble extractives and crude fibres

w/ere determined and the results are depicted in Table-26.

Successive extractive valuesThe percentage of extracts of the stem bark of the B. ceiba was determined in

the petroleum ether, solvent ether, chloroform benzene, ethanol and water. The results showed that the maximum percentage of extract was in water (8.80%) and the minimum was in petroleum ether (6O-8O0C) (1.26%). The percentage of extract in ether chloroform, benzene and ethanol were found to be 2.61%, 1.60%, 1.40% and 4.00% respectively (Table-29 and Graph-6).

I

Moisture content

Moisture content (loss in weight on drying at 1 Q5°C) was determined by the conventional method and results are represented in Table-26.

Fluorescence analysisThe fluorescence characters of the entire bark, powdered bark, ethanolic

extract of the bark and aqueous extracts of the bark at pH-5, pH-7 and pH-9 were observed in day light as well as under U.V. light, The results are depicted in Table- 30.

Behaviour of the powdered bark towards some chemical reagentsThe reddish brown powder of Bombax ceiba bark was treated with different

acids/chemical reagents. The mixture of powdered drug and chemicals were allowed to warm and cool for two hours. The actual colour of the powder drug and changes in the colour of powdered drug was noted and presented in Table-31. .Quantitative estimation of organic constituents

In quantitative estimation of organic constituents, the total alkaloids, tannins and resins were found to be 3.13%, 2.92% and 0.08% respectively (Table-26).

Chapter - Four Obsen’aiions & RexiiIts

86

'fesf- Pharmaco-anatomy Laboratoiy, Department o f Botany, Faculty o f Science, Jamia Hamdard

Thin layer chromatographic studiesThe thin layer chromatographic study of stem bark of B. Ceiba was conducted.

The results showed that the extract in petroleum ether (60-80°C) has maximum four spots on a T.L.G. plate coated with silica gel. followed by three spots in the extracts of hexane, methanol and water. The extracts' in benzene, chloroform, acetone and ethanol showed two spots in each. The solvent system used and R.f. values are given in Table-32 (Plate-XV, Fig-A c i, C2. Fig-B C3, C4, C5, Fig-C ce, c/. ca).

Inorganic elemental analysisQuantitative estimation of nitrogen, phosphorus, potassium, sulphur, calcium

and iron were carried out in the young stemi bark of the B. ceiba during 'January 2001- December 2001 at the interval of one month. The maximum concentrations of nitrogen (1.87%±0.003), phosphorus (G.88%+0.010), potassium (4.65%±0.027),

sulphur (0.037%±0.002), calcium (1-93%±0.019) and iron (0.88%±0,032) were observed in the months of May, February, May, September, May and October respectively, then the concentrations were found to be decreased in the following months and the minimum concentrations of nitrogen (1,53%±0.048), phosphorus

(0.83%±0.027), potassium (4.55%±0.030), sulphur (0.031 %±0.002), calcium

(1.85%±0,011) and iron {0.80%±0.042) were estimated in the months of July, May, July, May, March and March respectively (Table-33).

B: Phytochemical studies Preliminary phytochemical screening

The different extract of the powdered bark of B. ceiba was subjected to standard chemical tests for detection of various components. It showed the presence of reducing sugar, tannins, alkaloids, steroids, proteins, flavonoids, resins and glycosides and the absence of saponins (Table-34).

Extraction and isolation of chemical constituentsThe collected stem bark of B. ceiba was dried in air and finally in an oven at a

temperature below 45°C and then coarsely powdered. The dried and powdered stem bark (1.250 kg) was extracted with ethanol in soxhlet apparatus. The extract was concentrated under reduced pressure to get a dark brown viscous semi-solid mass (200 gm). It was analyzed chemically for the determination of the presence of

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Chapter - Four Observation!! & ResuHs

Pharmaco-anatorny Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

different chemical constituents. The concentrated mass was dissolved in minimum amount of IVle-OH and absorbed on silica gel to form slurry. The slurry was air dried and subjected to silica gel column chromatography prepared in Petroleum ether. The column ,was eluted with petroleum ether chloroform and methanol in order to increasing polarity to isolate the following compounds,

3.1 Compound [BC-1]Elution of the column with petroleum ether-ch loro form (1:3) (fraction 61-75)

afforded colorless crystals of BC-1 recrystallized from chloroform-methanol (1;1), 1250 mg (0.10% yield). '

0.72 (Toluene-ethylacetate-formic acid, 5:4:1)

Chapter - Four Oh.servation^' & Rexult.'^

Rtm.p.UV A,max

158-160°C274 nm (log e 4,9)

3374, 2950, 2923, 2845,1698, 1635. 1465, 1382,!R vmax (KBr)

1274, 1124, 1073, 289, 763 cm■‘H NfiR (CDCisI : 5 4.62 (1H, d, J == 3.33 Hz, H2~29a), 4.49 (1H, d, J =3.18 Hz, H2-29b), 3.15 (IN, dd, J==6.04, 8.85 Hz. H-3a), 1,69 (3H, brs, Me-30), 0.96 (3H, brs. Me-23), 0.89 (3H, brs, Me-25), 0.87 (3H, brs, Me-24), 0.78 (3H, brs, Me-26), 0.76 (3H, brs, Me-27), 0.72 (3H. brs, Me-28).

NMR (CDCis) : Table-504-veFABPS mlz (rel. In t| : 426 [M f (C30H50O) (19.6), 272 (11.3), 25,8(12.6), 231 (11.7), 218 (13.6), 217 (27.6), 208 (20.7), 190 (32.5), 189 (39.1), 177(22.5), 163 (26.3), 154 (37,5), 149 (38.9), 136 (78.1), 122 (95.6), 107 (100).

3.2 Ceibanaphthaquinone [BC-2]Elution of the column with CHCb-MeOH (1:1) (fraction 1-10) furnished reddish

crystals of BC-2, recrystallized from chloroform-methanol (1:1), 720 mg (0.057% yield).

Rf- ■ 0.68 (Toluene-ethylacetate-formic acid, 5:4:1)m.p. : 258-260°CUV Xmax ; 297 nm (log s 5.2)

IRvmax(KBr) : 2960, 2931, 1701. 1620, 1464, 1381, 1274, 1123,1072, 742 cm

/ r t i \ , ■ ' ' 88

Phannaco-anatomyLaboratoiy, Department o f Botany, Faculty ofScience, Jamia Hamdard

■•h NMR (CDGia) : 6 7.39 (1H, s, H~3), 7.22 (1H, s, H-6), 7.05 (1H, s,H-9), 2,71 (2H, brs. H2- II) , 2.26 (6 H, brs, Me~17, Me-18), 2.16 (2H, m, H2- I2 ), 1.37 (2H, m, H2-13), 1.26 (2H, m, H2- I 4 ), 1.06 (2H, m, Hs-IS), 0.83 (3H. t, J = 6.6 Hz, Me- 16).+ve FAB IWS m/z (rei. in t| : 270 [M f (C18H22O2) (18.6). 242 (4.2), 214(11.3), 168 (100).

Chapter - Four Ohservations & Re.nilts

8 9

Pharraaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

chapter ~ Four ObservaHom (If Resnhs

Chemical compounds isolated from the stem bark of Bombax ceiba

H 0 ‘ /

C30H50O

Conipoiiiid BC-1

oCH,

CH3

(CH2)gCH3

O

Ceibanaphthaquinone

I y y ' 90

PhamaccKinatotny Laboratory, Department of Botany, Faculty of Science, Jarrtia Hamdard

4. Moms alba Linn. (Shehtoot)A: Pharmacognosticai studies ftflacroscoplcai characters

The samples of the bark of M. alba were collected and cut into pieces of 3-5 cm. long, 1-3 cm broad, and up to 1-5 mm thick; these were recurved longitudinally. The outer surface is iron gray in colour and deep fissured. The fissures were V- shaped. The successive layers of periderm are parallel to the cambium. The outer crust of compact rhytidome roughs off in the form of very minute scale or even small particles peels off in the form of thin strips. The inner surface is smooth longitudinally straited and light yellow colured, The taste ;s bitter and acrid and the odour is pungent (Plate-XIII, FIg.-A&B).

Microscopical charactersTtie bark is primarily divisible into (I) the outer dead crust madp up of

rhytidome (about 30 percent) and (ii) the inner bark consisting of secondary phloem. The later is further divisible into conducting and non-conducting phloem zones. The conducting phloem occupy 25 percent of the total bark thickness. On the whole about 46 percent of the total bark is occupied by the non-conducting phloem. Rhytidome portion forms 29% of the bark (Plate-XIV, Fig.-A). In the periderm, the cork consists of many layered rectangular and thick-walled parenchymatous cells. Each cell is measured from 26.6-30.4- 49.4 ^m x 12.5-22.8-30.4. |Lim (Plate-XIV, Fig.-B). The cork cambium or phellogen is not seen in transverse section. Phelloderm cells are somewhat rectangular and comparatively thin walled and vary in size from 19.8-30.4- 41.8. |.im 11.4-15.2-26.6 jam, Plenty stone cells are also interspersed in this region.

«Following, the non-conducting phloem region is composed of rectangular to somewhat oval and parenchymatous cells mostly filled with starch grains; some stone cells are also present in this region. Stone cells are oval in shape and are measured from 26.6-30.4-38.0 jam x 19.2-22.8-26.6 jim. Out of 25 percent total conducting phloem about 10 percent is made up of thin walled parenchymatous cells and in remaining portion of conducting phloem, patches of fibres are interspersed (Plate-XIV, Fig.-E). The fascicles of fibre are tangentially arranged in the nonconducting phloem whereas in conducting region the fibres occur as isolated elements or in small groups (Ptate-XiV, Fig.-E). The fibres constitute about 80

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Chapter - Four Observation^ & Rexidts

Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jaraia Hamdard

percent of the conducting and about 15 percent of the non-conducting phloem. Few to many fibres are present in a fascicle. Sieve tube elements tend to form.groups, which are flanked radially with axial parenchyma and laterally with ray parenchyma. The presence of nacreous wall thickenings demarcated them from phloem parenchyma, which contains some coloured content or solitary and clustered crystals (Plate-XIV, Fig.-C). The medullary rays are multiseriate in the innermost region of conducting phloem (Plate-XiV, Fig.-E). Ray cells are thin walled, radially ebngated, parenchymatous and individual cell is measured 62.0-68.4-73.0 (.im x 29.3-41.0-45.2 /.im (Plate-XIV, Fig.-D). A thin walled parenchymatous 6-7 layered, cambium is visible at the junction of wood and bark (Plate-XIV, Fig.-E).

Microscopy of powdered barkPowder of the crude drug is grey coloured, coarsed and free flowing. The taste

Is acrid and odour is pungent. Small amount of the powdered material (sieved through 40 mesh) is placed on microscopic slide; mixed with few drops of 40% W/v aqueous chloral hydrate and heated gently under Bunsen Burner. Few drops of 1% alcoholic phluroglucinol are added to this and warmed by mixing one drop of concentrated hydrochloric acid. The slides are then mounted in glycerin and observed under microscope, which shows the fragments of cork cells, phelloderm cells and phloem parenchyma filled with starch grains were also observed (Plate-XIV, Fig.-F). Cork cambium is not visible during powder study. Pieces of phloem fibres with narrow lumen and pointed ends were also found. Sclerieds and stone cells are also observed in the powder. Stone cells were smaller than sclerieds. Groups of rounded starch grains were also observed during investigation (Table-35).

Histochemfcal tests

Transections of stem bark of M. alba were treated with routinely used chemicals and reagents, gave positive tests for starch tannins, lignins, alkaloids, flavonoids and proteins while saponins gave negative test. (Table-37).

Ash values

The percentage of total ash, acid-insoluble ash, water-soluble ash and sulphated ash were estimated and results are presented in Table-38 and Graph-7.

Chapter - Four Observations & Remits

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Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hanidard

Total extractive valuesTotal water-soluble extractives, alcohol soluble extractives and crude fibres

were calculated and results are presented in Table-36.

Successive extractive valuesThe percentages of extractive values were found to be 4.04% in petroleum

ether (60-80°G), 3.40% in diethyl ether, 2.59% in chloroform, 4,20% with benzene, 12.04% with ethanol and 9.60% in water (Table 39 and Graph-8).

Moisture contentMoisture content (loss in weight on drying at 105°C) has been determined by

the method given in the "The Unani Pharmacopoeia of India Part II" and the result is shown in Table-36.

Fluorescence analysisFluorescence analysis of the entire bark, powdered bark, ethanolic and

aqueous extracts of the bark have been carried out. The entire bark, powdered bark, ethanolic and aqueous extracts of the bark was observed in daylight and under U.V. light. Further the bark powder was treated to different organic solvents and solutions were again observed in normal daylight and under U,V, light and the observations are pooled in Table-40.

Behaviour of Powdered bark towards some chemical reagentsThe behaviour of the powdered bark on treatment with conc. sulphuric acid,

conc. hydrochloric acid, glacial acetic acid, picric acid, ferric chloride solution, iodine solution sodium hydroxide solution and potassium hydroxide solution were observed and the results are presented in Table-41.

T h i n l a y e r c h r o m a t o g r a p h i c studiesThin layer chromatographic studies were conducted. The extract in hexane

showed maximum (six) spots on a T.L.C. plate coated with silica gel. followed by fivespots in , acetone extract, three spots in each pWoleum ether (60-80°C), chjoroform and methanol extracts and minimum two spots in each ethanol and water extracts.

Chapter - Four ___________ ___________________Observations & Results

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Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science. Jamia Hamdard

The solvent systems used and Rf. values recorded are given in Table-42 (Plate-XV,

Fig-A di, da, Fig-B da, d4, dg, Fig-C de, dr, ds).

Inorganic elemental analysisNitrogen, phosphorus, potassium, sulphur, calcium and iron contents were

estimated in the young stem bark of M. alba during January 2001-December 2001 at the interval of one month. The maximum concentration of nitrogen (1.87%±0.039), phosphorus (0.97%±0.016), potassium (5.32%±0.039), sulphur (G.058%+0,008), calcium (2.94%±0.005) and iron (0.65%±0.062) were found in the mdnths of December. April, August, December, October and September respectively, then the concentration were found to be decreased in the following months and the minimum concentration of nitrogen (1.69%±0.016), phosphorus (0.94%±0.010), potassium

(5.26%±0.007), sulphur (.052%±0,001), calcium (2.80%±0.010) and iron

(0.53%±0.031) were estimated in the months of May. November, Jui^e July, December and February respectively (Table-43),

' IB: phytochemicai studies Preliminai’y phytochemicai screening

Hexane, petroleum ether {60-80°C), benzene, chloroform, acetone, methanol and aqu'eous extracts of the powdered bark were subjected to standard chemical tests for the detection of different phytoconstituents. Results showed the presence of carbohydrates (reducing sugars), tannins, alkaloids, steroids, proteins, flavonoids, resins and glycosides and the absence of saponins (Table~44).

Extraction and isolation of .chemical constituentsStem bark of M. alba was procured, dried in air and finally in an oven at a

temperature below 45°C and then coarsely powdered. The dried and powdered stem bark (2 kg) was extracted with methanol in soxhlet apparatus. The extract was concentrated under reduced pressure to get a dark brown viscous semi-solid mass

»(300 gm). It was analyzed chemically for the determining the presence of different chemical constituents. The concentrated mass was dissolved in minimum amount of Me~OH and absorbed on silica gel to form slurry. The slurry was air dried and subjected to silica ge! column chromatography prepared in petroleum ether. The


Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, JamiaHanidard

Chapter - Four Obxervations & Results

column was eluted with petroleum ether chloroform and methanol in order to increasing polarity to isolate the following compounds.

4.1 Albanylacetate [MA-i]Elution of the column with petroleum ether-chloroform (1:1) (fraction 61-75)

yielded colorless crystals of MA-1 recrystallized from chloroform; methanol (1:1), 1.32gm (0.066% yield).

0.82 (n-hexane: ethylacetate, 9:1)178-180°C 285 nm (log g 5.2)2960, 2930, 2862, 1729, 1610, 1563, 1380, 1274,

Rf.m.p.UV Xmax

IR fKBr)1123, 1072, 742 cm^

NMR (CDCIsl 6 5,12 (1H, dd, J = 5.30, 5,39 Hz. H-12), 450 (1H,

dd, J = 3.71, 9.82 Hz. H-3a), 2.04 (3H, brs, COCH3), 1.06 (3H. brs, Me-25), 1.01 (3H, brs, Me-23), 0.98 (3H, brs, Me-27), 0.91 (3H,brs, Me~24), 0.B7(3H, brs, Me-26), 0.86 (3h, brs, Me-28), 0.81 (3H, d, J = 6.11 Hz, Me-29), 0.78 (3H, d, J = 5.07 Hz, Me-30)■'Y. NMR (CDCis) ' : Table-51+ve FAB MS m(z (rel. int.) : 468 [M]" (C32H52O2) (32.1), 408 (5.41), 250(4.7), 218 (100), 203 (32.5), 190 (17.8), 188 (30.9), 175 (22.6), 173 (20.5), 160(19,6), 145 (33.2), 135 (46.5), 119 (53.2), 105 (52.8).

4.1 Albalupenoic acid |MA-2|Elution of the column with petroleum ether-chloroform (1:3) (fraction 26-35) yielded colorless crystals of MA-2, recrystallized from chloroform: methanol (1:1), 500 mg (0 .020% yield).

0.69 (toluene-EtOAc-HCOOH, 5:4:1)220-222°C 276 nm (log s 5.3)3373, 2960, 2931, 2840, 1701, 1610, 1463, 1381,

Rf.m.p.UV Xmax IR vmax (KBr)

1274, 1123, 1072,742 cm

NMR (CDCIa) 5 4.73 (1 H, brs, H2-29a), 4.60 (1H,brs,H2-29b),

3.18 (IN, dd, J = 5.1, 5.4 Hz. H-SP), 1.68 (3H, brs, Me-30), 1.25 (3H, brs, Me-23),

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Pharmaco-anatomy Laboratory, Department of Botany, Faculty o f Science, Jamia Hamdard

1.13 (3H, brs, Me-25), 0.98 (3H, brs, Me-26), 0.85 (SH.brs, Me-24), 0.77 (3H, brs, Me-27).

NMR (CDCisI ' : Table-51+ve FAB IIS m/z frel. int.) ; 456 [M]^ (C30H48O3) (6.9). 441 (22.3), 438(6.1), 412 (6 .8),394 (5.6), 374 (5.2), 329 (13.3), 290 (3.6), 288 (15.6), 276 (4.6), 248 (21.9), 236 (3.5), 207 (8.9), 203 (19.5), 190 (7.8), 189 (17.5), 180 (3.5), 176 (25.3) 174 (8 .6), 168 (7.1), 166 (13.5), 153 (100), 152 (14.9), 150 (18.3), 140 (17.5), 139(27.1), 138 (48.1), 136 (73.6), 135 (17.8), 125 (9.8), 122 (13.7), 108 (21.9).

Chapter - Four ______ Observaiions & Resulix

9 6

piiannaco-anatomy Laboratoiy, Department o f Botany, Faculty o f Science, Jainia Hamdard


Chemical compounds isolated from the stem baric of Moms alba

AcO

32* 32 2

Albanylacetate

COOH

30<»8®3

AlbalHpeiioic acid

a <,

9 7

•>' Pliarmacn-anatomy Laboratory, Department of Botany, Faculty o f Science, Jainia Hamdard

Bark 1 Bark 2 Bark 3 Bark 4Plant 1 20.4 27.4 90 20.4Plant 2 30.6 38.6 34.6 31.6Plant 3 45.9 46.9 45 43.9Plant 4 41.2 42.5 98.7 44.2

■ Plant 1■ P lan t! □ Plants

_ □ Plant 4

Bark 1 Bark 2 Bark 3 Bark 4

A !M ) g(R A(EKS

Tabie-S: Quantitative microscopy of the stem bark of ASstonia scholaris

S. N. iridiviciuai cells Length fin |j,m) ±SD (Range)

Width (in |im) ±SD (Rangel

1. Cork cells 31.4±3,14

(22.8-38,0)

20.2±3.21

(15.2-22.8)2. Phelloderrn cells 38.0±2.16

(26.6-45.4)

22.8+2.33.

(11.4-30.4)3. Ray cells 430±7.94

(280-540)

70±3.86

(56-98)4. Phloem fibres 229+11.59

(165-286)

35,6±1,72. .

(30.4-42,8)5. Sclerelds 60.5±5.38

(42,6-80.5)

26.4±3.36

(22,8-32.4)S. Crystals 23.3±1.78

(15.3-28.5)

15.0±2.93.

(12.0-21.5)

Measurements represent the average values of observations

Tah!e.s unci Graphs

Table-6: Preliminary e¥aluation of the stem bark of Aistonia schoiaris _

Organoleptic cliaracters

Colour Iron grey to mouse grey

Odour Not characteristic

T aste Bitter and acrid ,

Texture Rough

Physico-chemical constants (% w/w)

Loss in weight on drying at 105°C 63.20

Solid content 58.57

Extractive values (% w/w)

Water soluble matters 23.61

Alcohol soluble matters 17.32

Crude fibers ' 25.66

pH values

pH of 1% aqueous solution 6,07

pH of 10% aqueous solution 5.68

Quantitative estimation of total alkaloid, steroids and resins (% w/w) <

Total alkaloids 0.200

Steroids 0.078

Resins 0.044

Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Haindard

Tables and Graphs

Tai3 le-7 ; Histochemicai *tests of the stem bark of Alstonsa scholaris

Reactions-j----------------------- p------- -------

Colour Oisservations Inferences

Section placed in weak iodine solution

Dark bluish patches appeared

Starch present

Section'placed in a drop ofH2SO4

Bluish black to dark violet; no yellow colour or red colour appeared

Saponins or lipids absent

Section placed in 10% FeCl3+ a little Naz CO3

No blue or black colour appeared

Tannins absent .

Section placed in a drop' of phluroglucinol then a dropof HCI

Reddish-brown to rose-red colour appeared

Lignins present

Section placed in Dragendorff’s reagent

A Orange colour appeared Alkaloids present'

Section placed in a drop of 5% aqueous KOH solution

A strong yellow colourappeared

Flavonoids present

Section placed in castic alkali+ HCI

Yellowish patches appeared

Calcium oxalate present

Section placed in 5% tartaric acid in 95% ethanol for 2-3 days washed and then treated with iodine

Yelloish brown colour appeared

Proteins present

^Colour reactions mentioned are on the basis of average observation recorded

Pharmaco-anatomy Laboratory, Department o f Botany. Faculty o f Science, Jamia Hamdard

Tables and Graphs

Table-8 ; Ash values of the stem bark of Aistonia schoSaris

S. Wo. Deteniiinants Average values of ash r/owlw)

1 . Total ash 20,72

2 . Water soluble ash 14.62

3. Acid insoluble ash 1.46

4. Sulphated ash 14.20

Values are average of five determinants

Tab!e-9: Successive extractive values of the stem bark of Aistonia scholar'is

S. No. Solvents Colour Average values of extractives (%w/w)

1 . Petroleum ether (60- 80°C)

Pale cream 3,25

2 . Ether Butter scotch 2.60

3. Chloroform Light yellow 2.41

4. Benzene Off white........ .... ... . ... '

3,12

5. Ethanol 95% Light yellow 5.60

6 . Chloroform water Yellow 21.60


Pharmaco-anatomy Laboratoiy, Department o f Botany, Faculty o f Science, Jamia Hamdard

Tables and Graphs

TabIe-10; Fluorescence ainalysis o f the stem isark of Alstonsa scholaris

Materials / Treatments Colour in day light

ObservatiModifyingcolour

on underColourQuality

UV lightDegree of Radiance

Entire dry barkA) Upper surfaceB) Lower surface

Mouse grey Cinnamon

Dark purple Purple

DarkDark

BrightDull

Dry powder Brown Violet Light BrightDry powder rubbed on filter paper

Cane colour Yellowish white Light Dull

Powder treated with 11MNaOH in methanol

Yellowishbrown

Blackish brown Light . Bright

Powder treated with 1N NaOH in water

Yellow Orange Dark Bright

Powder treated with 50% H2SO4

Grey Black Dark Dull

Powder treated with 50% H N O 3

Whitishyellow

Black Dark Dull

Powder mounted in nitrocellulose in amylacetate

Cream Golden Light Bright

Powder treated with 1N NaOH in water, dried then mounted in nitrocellulose in amylacetate

Reddishbrown

Pale green Light Dull

Powder treated with 1N NaOH in methanol, dried and then mounted in nitrocellulosein amylc.,cetate

Brown Greenish brown

r

Light . Bright

Powder treated with 1N NCI, dried and then mounted in nitrocellulose in amylacetate

Dark brown Pale yellow Light Dull

Alcoholic extract f

Light yellow Turbid yellowish white

Light Bright1,

Aqueous extract pH-6

pH-7 pH-9

Light yellowLight yellow

Yellow

Greenish yellow Yellow Yellow

DarkDarkDark

DullDullDull

Pharmaco-anatomy ■ Laboratory, Department o f Botany, Faculty o f Science, Jamla Hamdard

Tahle.'i a n d G raphs

TabIe-11: Reactions of the stem bark povi/der of Alstonia scholaris with different chemical reagents:

Acid /Chemical reagents Observations

Cone. Sulphuric acid (8 8%) Reddish black

Cone. Hydrochloric acid Yellowish brown

Cone. Nitric acid Orange

Glacial acetic acid Pale yellow

Picric acid Oily yellow

Iodine solution Light red

Ferric chloride solution (aqueous) » Golden yellow

Sodium hydroxide solution (aqueous 5%) Light yellow

Potassium hydroxide solution (aqueous 5%) Reddish yellow

Powder as such...................................... - ...................................................................................... . ...

Brown

Pharmaco-anatomy Laboratory, Department o f Botany, faculty o f StJience. Jamia Harndard '

Ta hle.'i and Graphs

TabIe-12: Thin layer chromatographic studies of the different extracts of the stem bark of Alstonia scholaris:

Extracts Solvent system Spray reagent/ treatment

No., of spots R. F, Values

Hexane Hexane (90): ethylacetate (10 )

Exposed to h Vapours

4 0.11. 0.42, 0.66, 0.92

Petroleum ether (60- 80°C)

Hexane (90); ethylacetate (10 )

Exposed to h Vapours

3 0.*16, 0.45, 0.79

Benzene TEF Exposed to I2 Vapours

6 0.33, 0.43, 0.50, 0.61, 0.71, 0.80

Chloroform TEF Exposed to I2

Vapours2 0.12, 0.55

Acetone TEF Exposed to I2

Vapours2 0.25, 0.41

ft

Methanol n-Butenol (6): acetic acid (1 ); water (2 )

Exposed to I2

Vapours5 0,13, 0.21,

0.46, 0.51, 0.71

Ethanol n-Butenoi (6): acetic acid (1 );water (2 )

Exposed to I2

Vapours4 0.25, 0.47,

0^5, 0.79

Water n-Butenol (6); acetic acid (1 ):water (2 )

Exposed to I2

Vapours3 0.27, 0.52,

0.63

Phannaco-anatomy Laboratory, Department of Botany, Faculty o f Science, Janaia Hamdard

U k s m O r a p k

TaWe-IS: Eiemental ansifsis of llie stem barl powder of ilstom sctolaris d o ig Janoar|» 2fi81 to December 2IS1

Jaa Feb. la r. Apr. l8 f . Jim. k i , Dec.

2.S0

±0742

2,62

±0.03?

2.64

±0.1149

2.64

+0.068

2,88 2,?4 2.71

±ai3 milm

±0.06

11

±0,02 ±0,055

2,oe

i0.065

Pliospfiorus 0,8±0,04'

0,82 0,83

±0.012

0,83

±0,014

0,83

±0,007 ±0,013 ±0.012

0.S4

±0,012

0,04

±0,0

0,S4

±0,003

o,eo±o.ooe

Potassiyinf/o) 3.80

±0.080 ±0.044

3,87

±0.035

3.82

±0.051

3,81

±0,048 ±0,026

3,8

±0,012

3,82 I 3.86 3,85 3,87

±0,041 ±0,018 i ±0,021 ±0,03

3,85

±0.013

■Su|taf/2 ,047 .047 .047

±0.008 +0.002

.047

±0.007

.048

±0.001 ±0.00'

,046 0.043 .047 I ,042 ,047 ,047

±0.00 ±0.005 ±0,001 ±0.00 ±0,002 +0.002

t im % 3.37

+0.025

3.34

+0,040

3,37

±0.015

3,37

±0,027

3.38

+0,046

3.38

+0,008

3.39

+0,013

3,41 3.42 ! 3,40

+0,0101 +J.03i

3,36

±0,018 +0,027

3,4'

±0,020

Iron %' 0,65

+0,02

,65

+0,023

0.65

+0.030

0,6

+0,002

0,63 0.63 0,7

±0,0411 ±0,0251 +0,028

0,72 0,681 I

±0,080 i +0,007 i +0.053

0,6?

+0.C3 +0.03

s!, V

iam aco -ana to m y L a k a to r ) ; Departinent of Botany, Faculty of Science, lamia Hamdard

M k ssiid G rsp k

IaWe-14: Pfeliminarf pfiftocliemical screeiiing for l e delecioo of cfiemica! constityenfe from diferent extracts of l e $lm MofMoniascMm

TestsHexaneextract

Petetler{6 i# C )a t e t

Bsffisae eitrast '

ChorofDSextract

MetoReextracl

le la o sextract

la te rextract

Redpcini

sugar.

+ « ii

■f

Tannins - - - - -[

■Aka olds +I1 it 1* A

iSap'onins ' ■

j j i i

■ - :

Steros + + - X11

4! I

Proteins - ^ 1 ■1 i

t 1 +! i

Flawnoids * +T- - - -4* + 1 4

j

Resifis ■ ■ + + 1 T1

+ 1i

Gfcosides + i! 5+ I

i i

1

1

+ 1

1

'liaimaco-anatoiny Laboraioiy, Department o f Botany, Faculty o f Science, Jamia Hamdard

Tables and Graphs

Table-15: Qyantitative microscopy of the stem bark of Balanites aegyptiaca

S, No. individual ceils Length (in [im) ±SD (Rantje)

Width (in |im) ±SD fRangel .

1 . Cork cells 26.4+1.16

(15.3-36.2)

19.3±6.21

(13.4-23.5)

2 . Phelloderm cells 26,8±2,19

(21.8-32.5)

16.3±3.97

(13.3-23.5)

3. Ray ceils 78,5±3.52

(54,5-98.3)

27.5+5.31

(23.5-39.3)

4. Phloem fibres 198±11.38

(172-253)

22.3±2.76

(15.6-28,7)

5. Stone cells 95.5+3.38

(76.0-115.8)

78.0±1.53

(62.2-89,6)


"Si:#?-' Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Seience, Jamia Hamdar.d

Tables and Graphs

Tal3le-16: Preliminary evaluation of the stem bark of Balanites aegyptiaca

Organoleptic charactersColour GreyOdour PungentTaste BitterTexture RoughPhysico-chemica! constants (% w/w|Loss in weight on drying at 105°C 55.50Solid content 65.30Extractive values (% w/w)Water soluble matters 7.77 IAlcohol soluble mattersCrude fibers 16.00pH values

pH of 1 % aqueous solution 5.34pH of 10% aqueous solution 4.97Quantitative estimation of total phenolics, tannins, and saponins (% w/w)Total phenolics 3,19Tannins 0.87Saponins 0.50

Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard '

Tables and Graphs

Table-17: Histociietnicai *tests of stem bark of Balanites aegyptiaca

Reactions Colour Observations Inferences


Dark bluish patches appeared

Starch present

Section placed in a drop of H2SO4

Yellow colour appeared Saponins present.

Section placed in 10% FeCl3+ a little Na2 CO3

Bluish colour appeared Tannins present

Section placed in a drop of phluroglucinol them a drop of HCI


Lignins present

Section placed inDragendorff's reagent

An orange colour appeared Alkaloids present


A strong yellow colourappeared

Flavonoids present

II



Calcium oxalate present


Yelloish brown colourappeared

Proteins present

*Colour reactions mentioned are on the basis of average observation recorded

Pharmaco-anatomy Laboratoiy, Department o f Botany, Faculty o f Science, Jamia Hamdard

Tables and Graphs

Tal3!e-18: Ash Values of the stem bark of Balanites aegyptiaca

S. No. Determinants Average values of ash (%w/w)

1. Total ash 12.16

,2. Water soluble ash 7.59

3. Acid insoluble ash 2.34

4. Sulphated ash 18.21


Tab!e-19: Successive extractive values of the stem bark of Balanites aegyptiaca

S. No. SolveEits Colour , Average values of extractives (%w/w)

1 Petroleum ether (60- 80°C)

Off white 3.61

2. Ether Cane colour 1.803. * Chloroform Cream 1.604. Benzene Silk colour 3.015. Ethanol 95% Pale yellow 5,606. Chloroform water Light yellow 4,80


Phanmco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

Tables and Graphs

Table-20: Fiiiorescerice analysis of the stem iaark of Balanites aegyptiaca

Materials I Treatments Colour in clay iiglit

Observation under UV lightModifying colour Colour

QualityDegree of Radiance

Entire dry barkC) Upp.er surfaceD) Lower surface

GrayCane colour

Iron gray Yellow

LightDark

BrightBright

Dry powder Brown Blackish brown Light DullDry powder rubbed on filter paper

Brown Cream Light Bright

Powder,treated with 1 N NaOH in methanol

Light yellow Blue Dark Bright


Whitish yellow Blue bark Bright


Reddishyellow

Black Dark Dull

Powder treated with 50% HNO3

Pale yellow Black Dark Dull

Powder mounted in nitrocellulose inarnylacetate

Dark gray Gray Light Bright

Powder treated with 1N NaOH in water, dried then mounted in nitrocellulose in arnylacetate

Gray Pale yellow Light Bright

Powder treated with 1N NaOH in methanol, dried and then mounted in nitrocellulose in amylacetat

Yellow Golden Dark Bright

Powder treated with 1N MCI, dried and then mounted in nitrocellulose in arnylacetate

Brown Gray Light iDull ...

Alcoholic extract Light yellow Turbid yellowish white

Light Bright

Aqueous extract pH-5 Light yellow Greenish yellow Light DullpH-7 Light yellow Yellow Dark BrightpH-9 Yellow Yellow Dark Dull


Tables and Graphs

Tabfe-21: Reactions of the stem bark powdered of Balanites aegyptiaca with different chemical reagents:


Cone. Sulphuric acid (98%) Dark black

Cone. Hydrochloric acid Light brown

Cone, Nitric acid Reddish yellow

Glacial acetic acid Light grey

Picric acid Chocolate colour

Iodine solution Light yellow

Ferric chloride solution (aqueous) Dark brown

Sodium hydroxide solution (aqueous 5%) Cream colour

Potassium hydroxide solution (aqueous 5%) Yellowish white

Powder as such Light brown

pharmaco-anatomy Laboratoiy, Department o f Botany, Faculty o f Science, Jamia Hamdard .

Tables and Graphs-

TabIe-22: Thin layer chromatographic studies of different extracts of the stem bark off Balanites aegyptiaca


No.of

spots

R. F. Values

Hexane Hexane (90); ethylacetate (10)

Exposed to I2 Vapours

3 0.38, 0.60, 0.72

Petroleum ether (60-80°C)

Hexane (90): ethylacetate (10)

Exposed to I2

Vapours4 0.26, 0.50, 0,69,

0.90

Benzene TEF Exposed to I2

Vapours2 0.48, 0.74


Vapours1 0.52


Vapours4 0.14, 0.41, 0,54,

0.76Methanol n-Butenol (6):

acetic acid (1); water (2 )

Exposed to I2

Vapours3 0,27, 0,36, 0,69

%

Ethanol n-Butenol (6):acetic? acid (1); water (2 )

Exposed to I2

Vapours3 0 .2 0 , 0,61, 0,80

Water n-Butenoi (6 ); acetic acid (1 ): water (2)

Exposed to la Vapours

2 0.43,0.73 .

____ -....... .


Mksmiurspk

Iable*23: Elemental aaalfsis of tlie stem bark poidered of Bafente ssgjfpim diirifig Janyarf 2111 to Oeceiber 281

Eleineols Jan. Fel). ia r. Apr. ia f. J®. Sep. Oot is i Oec.

litrogeRf/o' 5.64

+0.314

5,76

+0,171

511

+0J53

5.1

i 0,9?3 +0.075 +0.113

512

+0.112

5.83

+ a o 5i

0.86

+0,018

0.87

+0,015

0,87

i 0,012

0,87

+0,017

§ ,i

+0.015

9,87

+.016

0.8?

+0.015

0,87

+0.015

0.88

+0.010

,86

1015

0.87

+0 ,01^

Potassium.(% 2,59

+i.019

2,60

+_0,020

2.58

+0,010

2.64

+0.010

2.58

+0,082

2,57

+0.047 +0,005

2.61 2,66I^+0,016

2.58 2.62 2,58

+0,0651+0.0211+0.0151

Sii}ptar(%)

+0,00

,023+01.022

+0,002 +0.001

.025

+0,002

.024 .02

+0.002 i +0,005

.024 ,028 1.022

+0.0031+0.003 +0,00

Caiciy!n(%l ,90

+J,021

.88 ,85

+0,014'+0.0'

,93 1.86

+0.025 +0,019 +0,022

1.87

+0.022

1,88

2 +0,032

1,87 1,87 11,92

+0,016 +0,005! +0,015

:ron(%) 0,74 0,71 0,75

+0,0191+0,013

0,76 0.72 0.79

+0,015 +0,015 +0.032

0,73

+0,032

9,72

+0,019

0,75

+0,018

0,75 10,78 10.75i I

iO.023l+O,O32|+O,O12

Pliamaco-anatoniyLaboratoiy, Department of B otanyiaculty^ HamdaK

iSommirmn

Tesfe faaneextract

Petefter(6M0‘C)g *ac t

Benzeneextract

Cliioroforieitract

i

‘ te fe iie erfrast

i

Mkwsiextract Ij

ia te re m e !

R e id o g sugar i + - + ■f[

Tannins I ■f I - + T -

Aikaoids + - + 1 •f (T 1-

Saponins ■ - f t -

1!i

Steros + 1I + 1- + 1 1 !

Mm + + (T T !

* i 4-1 ‘ i IiFlavonoids + ■

1 __ _ _+ 11

i

+ +' ^ 1 {

Resins -

1iii1 - -

1

G fc o s ite f +i

!

I1

+j

+i

1 ^

ii

P k m ^o-ana to m y Laboratory, Department of Botaay, Faculty' of Science. Janiia HamdaK

Tables and Grapjis

TabIe-25: Quantitative microscopy of the stem bark of Bombax ceiba

S. No. Iridivicliiai cells Length (in |itTi| ±SD (Range)

Width (in |im| ±SD (Range)

1. Cork cells 48.8+7,74 24,2±3.44^

(38.0-60.8) (19.8-30.4)

2. Phelloderm cells 35.0±5.11 25.6±2.53

(26.6-41,8) (21.5-30.4)

3. Ray cells 62.5+12.51 31.0+2.534

(45.5-75,5) (28.5-35.7)

4. Phloem fibres 336+8.96 33.6+5.44

(320-350) (26.0-41,8)

5. Stone cells 57,6+5.94 40.1 ±5.24

(49,4-64.6) (31.7-49,4)


/I-:--SS.TOmaaWirfff,imiT.«rrr,-as-a-«

Pharmaco-anatoray Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

Tables and Graphs

Tabie-26: F*reiiminary evaiyafion of the stem bark of Bombax Ceiba

Organoleptic charactersColour IViashroom to greyOdour Not characteristicTaste AcridTexture RoughPhysico-chemical constants (% w/w|Loss in weight on drying at 79,70Solid content 57,53Extractive values (% w/w)Water soluble matters 17.35Alcohol soluble matters 16,08Crude fibers 18,33pH values c

pH of 1% aqueous solution 4,75pH of 10% aqueous solution 3.69Quantitative ©stimstion of total alkaloid, steroids and resins {% w/w)Total alkaloids 0.13Tannins 2,92Resins 0.08

......•■S' v-?:' Pharmaco-anatomy Laboratory, Depamnent o f Botany, Faculty o f ScienGe, Jamia J-Jamdard ,

Tables and Graphs

Table-27; Histochemica! *tests of the stem bark of Bombax Ceiba

Fleactions Colour Observations inferences


Dark bluish patchesappeared

Starch present

Section placed in a drop of H 2 S O 4

Bluish black to dark violet; no yellow colour or red colour appeared


Section placed in 10% FeCl3+ ,a little Naa CO 3

Bluish colour appeared f

Tannins present

Section placed in a drop ofphiuroglucinol then a drop of HCI

... .... ...... f .... ..... .... ... .............


Lignins present

Section placed in Dragendorff's reagent

An orange colour appeared Alkaloids present

Section placed in a drop of5% aqueous KOH solution

A strong yellow colour appeared

Flavonoids present


Yellowish patches not appeared

Calcium oxalate absent

Section placed in 5% tartaric acid in 95% ethanol for 2-3 days washed and then treatedwith iodine... .


Proteins present

^Colour reactions mentioned are on the basis of average observationrecorded

phanTiaco-anatomy Laboratory, Department o f Botsny, Facility o f Science, Jamia Hamdarci

7'ahles and Graphs

Table-28; Ash values of the stem bark of Bombax ceiba

S. No. Determinants Average values of ash f%w/w)

1, Total ash 20.522. Water soluble ash 4,333. Acid insoluble ash 3,064. Sulphated ash 14.95


Table"2&: Syecessive extractive values of the stem bark of Bombax ceiba

S. No. Solvents Colour Average values of' extractives (%w/w)

1. Petroleum ether (60~80°C) Yellow 1.262. Ether Light Yellow 2,613. Chioroform Mashroom 1,604. Benzene Cream 1.40S. Ethanol 95% Leaf brown 4.006. Chloroform water Suede colour 8.80


Pharmaco-anatomy Laboratoiy, Department o f Botany, Faculty' o f Science, Jamia Hamdard

Tables and Graphs

Table-30: Filoorescence analysis of the stem ijark of Bombax ceiba



QualityDegree

ofRadiance

Entire dry barkE) Upper surfaceF) Lower surface

Mashroom Golden brown

MashroomBrown

DarkPark

BrightBright

Dry powder Reddishbrown

Blackish brown Light Bright

Dry powder rubbed on filter paper

Pale brown Brown Dark Dull

Powder treated with 1N NaOH in methanol

Yellowishbrown

Black Light Bright


Grey Sky blue Light Dull


Dirty colour Blue Dark Bright


Yellow Brown Dark Bright

Powder mounted in nitrocellulose In amylacetate

Dark red Brownish black Light Dull <

Powder treated with 1N NaOH in water, dried then mounted in nitrocellulose inamylacetate

Reddishbrown

Grey Dark Dull


ReddishYellow(Orange)

Reddish brown Light Dull,

Powder treated with 1N HCl, dried and thenmounted in nitrocellulose inamylacetate

Reddishbrown

Blackish brown Light Bright

<

Alcoholic extract Reddishorange

Wild purple Light Bright

Aqueous extract pH-5 pH-7 pH~9

Raw silk Suede colourSuede colour

CreamGreenish white Greenish white

DarkLightLight

DullBrightBright

C&l)

t

Phamaco-anatomy Laboratoiy, Department o f Botany, Faculty o f Science, Jamia Hamdard

Tables and Graphs

Tabie-31: Reaction of the stem bark powder of Bombax ceiba with different chemical reagents:


Cone. Sulphuric acid (98%) Dark gray

Cone. Hydrochloric acid Yellow

Cone, Nitric acid Yellowish orange

Glacial acetic acid Dull brown

Picric acid-

Chocolate colour

Iodine solution Brown colour

Ferric chloride solution (aqueous) Black

Sodium hydroxide solution (aqueous 5%) Brown

Potassium hydroxide solution (aqueous 5%) Blackish brown

Powder as such Reddish brown

pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard ^

Tables and Graphs

Tal3le-32; Thin layer chromatograpfiic studies of the different extracts of the stem bark Bombax ceiba


No.of

spots

R. F. Values

Hexane Hexane (90):ethylacetate (1 0 )

Exposed to I2

Vapours3 0.41, 0.60, 0.72

Petroleum ether (60- 80°C)

Hexane (90):ethylacetate (1 0 )

Exposed to I2

Vapours4 0.27, 0.55, 0.77,

0.86

Benzene TEF Exposed to I2

Vapours2 0.41, 0.63


Vapours2 0.38, 0.54


Vapours2 0.43, 0.61

Methanol n-Butenol (6): acetic acid (1 );water (2)

Exposed to I2

Vapours3 0.47, 0.67, 6.85

Ethanol n-Butenol (6);acetic acid (1 ): water (2 )

Exposed to I2Vapours

2 0.16, 0.66

Water n-Butenol (6): acetic add (1 ) : water (2)

Exposed to I2

Vapours3 0.16, 0.38, 0.61


Tab!e*33: Etefnentai analysis o'f the s te i k t Powder of Sofulsi ee ia dorin§ Janyari* 2ii1 to Oeceoiiisr 2011

Eieinents Jan. Feb. ia r. Apr. ia f . Jus. M, 1 Sep. Oct. M Dec.

i!tro§0'H(% 1.77 1.75 171 16§ 187 157 153 168 185 185 185 173

iO.095 +0.074 +0.032 +0,089 i0.033 +0.069 +0.048 +0.084 +0,035 +0,035 +0.025 +0.068j

Phosphorus 0,87 0.88 0.86 0.86 0.83 0.85 0.83 0.84 0,86 0,85 0,85 0.83 1■(%) i0.010 +0.010 +0,024 +0.018 i0.027 ,+0.027

i1i0.011 1+0,010 +0,027

1+0,018

j1+ o ,o ie +0,011

Potassimnf/o) 4,60 4,62 4.63 4.60 4,65 4.56 4,55 4,60 4.62 4.58 i|4,57 14,58

iO.023 +0,021 +0.022 +0,038 ±0.027 +0,045 11

I +0,030(1i+0,046

i+0,021 i1 ±0.015i1i1

+0,015 ii i1

+0,02^

Sulphur % ,033i0. .034 ,032 .035 .031 ,034 1,033 ,035 ,037 ,034i ,035 ^,036

0021

+0,005 +0.005 +.001 +0,002 +0.003 i0.l)041fi1+0.002 i0,002

i+0.0021!

+0,0231

+0.008

Calciuin % 1 1 183 1.85 1,90 183 1,88 lie ?I 188 1,87 1,87I

187 182

+0.021 +0,014 +0,011 +0,025 +0,019 +0.022i

+0.0^2 i +0,032i i 1 1 i 1+0,016

!±0,022 +0,005 +0,015

ron(%) . 0.85 0,86'

,80 ,86 0,82 10,81 |0,85i

|0,85 |0,82 0,88 io ,e i 0.85

i0,032 +0,036 +J.042 +0,0451 '

1

+0,043 +0,061i

+0.053I

+0.032i

+0.023 +0,0321

j

+0,035!

+0,081Ij

• I'v

larraaco-anatomy Laboralor}', Department o f Botany, Facufe of Science, lam ia Hamdard

Tests Heianeextract

Fet8tfier(g8»i 'C je x t e t

BeRzeR8 ’ extract

d i o f o t a 'M ract

AeetoKgifest

ie la ^ is lextast

Warereitraet

R edm ingsyp ■f Xt +

t- ! ^

Tamiiis - + - - 1 +

Alkaloids + T 4.i - I 1

Sapopins - 11

' i ) 1 . j

Sterols I1 1 + 1 iI T

i

1 X ^ 1 + I1 { 1 ;1 . . . . . . 1 . . . . . 1

Proteins' ' I - - ■f + 1i[

Flawnoids f I + ' [ ,1 t - !+ 1 +ii)

Resins t 1 •1i!- - - - - - - - - - -i J.i i

1T j

1

Gfcosides + +1

\ 1Ii1t

!t + ■f

P'larmaco-anatoiny Laboratoi)’, Department o f Botany, hdtj of Science, laniia

Tables and Graphs

Tabte"35: Quantitative microscopy of the stem bark of Morus alba

s. No. indiviciual ceils Length (In |im)±SD (Range)

Width fin ^m)±SD (Range)

1. Cork cells 34.8±7.73

(26.4-49.4)f

22.9±5.53

(15.2-32.2)

2. Phelloderm cells 29.9+8.52

(19.8-41.8)

16,9±5.45^

(11.4-26.6)

3. Ray ceilsf

68.1+4,32

(62.0-73,0)

38.6+6,20

(29.3-45.2)

4. Phloem fibres 413+8,87

(350-380)

16,2+2,59'

(12,5-21,5)

5. Sieve tubes 184+18,5

(152-209)

29,2+4,66

(22,8-37.0)

6. Stone cells 35.5+7.52

(26.6-38,0)

21.5+3,05*

(19.0-26.8). .... ..................... .


Vf;, v-wr pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Haindard

'fables and Graphs

Table-36: Preliminary' evaluation of the stem bark of Moms alba

Organoleptic characters Colour Iron grayOdour PungentTaste AcridTexture RoughPhysico-chemical constants (% w/w)Loss in weight on drying at 105“c 33.04Solid content 59.53Extractive values (% w/w)Water soluble matters 18,37Alcohol soluble matters 15.41Crude fibers 23.00pH valuespH of 1 % aqueous solution 5.46pH of 10% aqueous solution 4.73Quantitative estimation of total ailcaloid, phenoiics arid resins {% w/w)Total alkaloids 0.07Phenolic compounds 2.13Resins 0.03

Pharrnaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jainia Hamdard

Tables and Graphs

Table-37: Histocfiemica! *tests of the stem bark of Uorus alba

Section placed in weakiodine solutionSection placed in a drop ofH2SO4

Section placed in 10% FeCl3+ a little Naa CO3

Colour ObservationsDark bluish patchesappearedBluish black to dark violet; no yellow colour or red colour appeared

InferencesStarch present

Bluish colour appeared

Section placed in a drop of phluroglucinol them a drop of HCl

Reddish-brown to rose-redcolour appeared


Tannins present

Lignins present

Section placed in Dragendorffs reagent

A Orange colour appeared Alkaloids present


A strong yellow colour appeared

Flavonoids present

Section placed in casticalkali-!- HCl




Calcium oxalate presentProteins present

*Colour reactions mentioned are on the basis of average observation recorded

phamaco-anatorny Laboratoiy, Department o f Botany, Faculty o f Science, Jamia Hamdard

Tables and Graphs

Table-38: Ash values of the stem foark of IWorus a!ba

S. No. Determirij'jiits

Total ashWater soluble ashAcid insoluble ashSulphated ash

Average values of ash_____ i% w /w j________

16.784.24

22.51


Table-39: Syccessive extractive values of the stem bark of Moms alba

S. No. Solvents Colour Average values of extractives (%w/w)

1. Petroleum ether (60- QO°C)

Pale cream 5.04

2. Ether Butter scotch 3,403. Chloroform Light yellow 2.594. Benzene Off white 4.205. Ethanol 95% Light yellow 12.046, Chloroform water Yellow f 9.60


pharraaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

Tables and Graphs

Tab!e-40: Fluorescence analysis of the stem bark of Uorus alba



QualityDegree

ofRadiance

Entire dry barkG) Upper surfaceH) Lower surface

Iron gray New Ivory

Mouse gray Yellow

LightLiqht

BrightBright

Dry powder Brown Blackish brown Light BrightDry powder rubbed on filter paper

Cream Brown Dark Dull

Powder treated with 1N NaOH in methanol

Reddishbrown

Black Light W


Gray Gray Dark Dull


Yellow Blue Light Dull4


Pale Yellow Smoke colour Dark Bright

Powder mounted in nitrocellulose inamyl acetate

Dark red Black bark ’ d"uIF ......

Powder treated with 1N NaOH in water, dried then mounted in nitrocellulose inamylacetate

Brown Brown Dark Brighf

i


Reddishyellow

Reddish brown Dark Dull

Powder treated with 1N HCI, dried and then mounted in nitrocellulose inamylacetate

Reddishbrown

Yellowish brown Dark Bright

Alcoholic extract Light yellow Golden brown Light DullAqueous extract pH-5 Golden brown Brown Light Dull

pH-7 Golden brown Brown Light Dull,pH*9 Brown Brown Dark Bright

I’harmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hamdard

Tables and Ctraphs

Tabfe-41: Reactions of the stem bark powder of ¥iorus aiba with different chemical reagents:

Acid/Ciiemicai reagents Observations

Cone, Sulphuric acid (98%) Bluish black

Cone. Hydrochloric acid Dull yellow

Cone. Nitric acid Chocklet colour

Glacial acetic acid Dull brown

Picric acid Yellowish brown

iodine solution Light reddish brown

Ferric chloride solution (aqueous).........-...

Blackish brown

Sodium hydroxide solution (aqueous 5%) Orange colour

Potassium hydroxide solution (aqueous 5%) Dark Orange colour

Powder as such Dark brown

Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jatnia Hamdard

7'ables atul Graphs

Tab!e-42: Thin layer ciirornatographic studies of the different extracts of the stern bark Moms alba

Extracts Solvent system Spray reagent/treatment

No.of

spots

R, F. Values

Hexane Hexane (90): athylacetate (10)

Exposed to I2

Vapours6 0,42, 0.59, 0.68,

0.75, 0.84, 0.94Petroleum ether (60- 80°C)

Hexane (90):athylacetate (10)


3 0.55, 0.67, 0,87

t

Benzene TEF Exposed to I2Vapours

2 0.50, 0.57

Chloroform TEF Exposed to I2Vapours

....3... 0.33, 0.41, 0.50

Acetone TEF Exposed to I2 Vapours

5 0.11, 0.18, 0.29, 0.35, 0.40

Methanol n-Butenol (6): acetic acid (1);water (2 )

Exposed to I2 Vapours

3 0.18, 0.62, 0.86

Ethanol n-Butenol (6): acetic acid (1): water (2)


2 0.41, 0.67

Water n-Butenol (6); acetic acid (1): Water (2)

Exposed to I2

Vapours ,. 2_. 0.21, 0.63

/I-..Phannaco-anatomy Laboratoiy, Department o f Botany, Faculty o f Science, Jaraia Hamciard

M isssridG njph

Tale43: Elemefital aRaifsis of tte stem liark powder of io fu s g la dyri!i| Januaii 2§i1 !§ D eee ik r 2911

Elenents Jan, Feb. la r. Apr.' I l f . inn. Jy. M§. Sep. Oct. M Bes.

fiitrogenf/o)- 173i0.

202

1.86+0.

059 052

1,e7+_0.

019

1,S9iO.

016

: 178+0.

010

: 1J2+0,

07?

1.88+0.

:050

I 1 M

012

170+0,

f 2

173+0.

088

187+

0,039

Phospfiorus

(%)

0.95ii).

018

0.96+0.

^015

0,95+0.

015

0.87+0,

016

0.95i0.

021

0.96+0.

020

0.85i0.'1018 018

! ;

0,95+0.

0221

o,8e+.o,

015

0.94+Q.

010

0.95+

10.012

Potassium % 5.27+0.{053

j

5.30+0,

018

5.27+0,

029

5.29i0,

029

5.27+0.

013

5.26+0.

007

5.31++

0.08

5.32+0.

011

5.30+0.

027

5,28+0,

012

5,28+0.:1010

5,28+

0,029

Syipter %) .056+0.:

002

0,57+0,

007

,055+0..

008'

.05610.

0031

.054+0,

005

.054+0.

002

,052+0,

001

.055+0,

002

.056+0,

007

,056+0,

003

,056+0.

0021

,058+

0.008I [j

Caciymf/o) 2,81+0.

oie.

2.91+i.

018

2.92+0,

008

2,93+0,

018

2.93+0.i005

2,92+0.

013

2,92+0,

028i

i

2,91+0.

008

2.93+0,

016

2,92+0.

005

2,8510. i 2,80iI0 1 i 0,010] ;

irofi(%j ■ .

A

0.59+0.

013

10.53+0,

031

0,55+0,

018

0.56+0,

0201jj

0.58+0.10.62+0.

035 03514

j0,63i0.

0351

0,61i0.

032A

0.65+0.

062

0,63+J.

021

0,64+0,!1022iIi

10,63+ i!i10.034

i fPliM aco-aiiatoniy Laboratory, Departoient o f B otanyjacu lty o f Scienc^^ Hamdan

I d k s

Table44: ?dmmrf pliifoclieiiiical screeninc o f le s t e in k k o f io i i is a la

Tests Hexanee itas t

P e te le r(6i-80“Cextract

Beozeneextract

C b rs fs ffiiixtrast

feetoneextnef

le la i iosstraet

ia ta rextract

Esdycifig sugari

..

I- i * +

Tannins - + ;1 + - 4-J 1 + 1Akaloids + - 1 ^ •f + + I

SaponifiS' - - - -ii * 1

Steros 4* i „

1 jjj ! i

t

!

1t

Proteins +!

' i "!i ■

I T

t 1

Fiawnoids + - + - i

!

+. 1 7 1

;

Resins 1 ^ 1 -i1

T

1 !t

!Glycosides - - + I + i + ■i'

1 ! 1

j: I'..T

PkruiacQ-aaatomy Laboratory, Department of Botany, Faculty of Science. Jamia HamdaK

Tables and Graphs

Tal3ie-4S: ‘'H and NMR Spectral Data of (AS^1)Position ^HNlMR ” c NMR

Alpha Beta1.31 dddd (3.42, 10,25,

6.35. 11.72)1.61dddd (7,83, 3.90, 3.42, 3.22)

39.61

2 1.76 rn 189 rn 26.62‘3 4.46 dd (5.86, 9.76) 80.964 - 33.73

» 145.186 5.12dd(3.91, 3.42) 124.337 1.82 d (12.70) 1.98 d (4.40) 33.73e _ 40.059 1.59 dd (3.90, 4.88) 55,41

1 0 - 3 6 , ^1 1 1.78 dd (4.88, 8.798) 2.36 dd (5,86, 4.88) 20,961 2 5.17 dd (3.42, 3,91) 121.6513 139.6214 _ 42,8315 1.15 ddd (5.86, 9.28,

4.39)1.37 ddd (6.34.

5,89, 7.32)27,45

16 1,15 ddd (2.93, 5.86, 13.18)

2.00 ddd (4.40, 4.88, 7.81)

39,99.

17 - 42.9818 . 2.36 d (5.86) 47.6819 1.52 ddd (5.86, 8.30,

11.23)47,99

20 - 150.8521 1.40 m 1.88 m 29.8622 4.09 dd (5,83,6.83) 76,35'23 0.96s 17,4724 0.99 s - 17.9925 0.83 s 15.98'26 0.79 s „ 16.4727 0.93 s 14.5028 0.87 - 19.2929 4.56 brs, 4.68 brs - 109.3330 1.68 brs 23.70

OAc 2.04 brs 21.23, 170.90

Coupling constants in Hertz are provided in parenthesis

|£S l)■ ^ 5 ^ Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jam ia Hamdard

Tables and Grapks

Tabie-46: and ” C NMR Spectral Data of (AS-2)13.

Position MR ^ ■' C NMR' Alpha Beta1 1.34 dddd(8.30. 11.72,

3.91, 12.25)1.72 m 39,31

2 1.64 m 1.87 m 26.693 3.26 dd (3.91, 9.27) 77,904 33.395 145.906 5.12 brs - 125.147 1.89 d (10.25) 1.98 d (.86) 34.528 - - 40.919 1.53 dd (4.89, 16,77) 55.9210 _ 36.2811 1.57 m 2.03 m 20.0212 '5.12 brs 122.4513 - - 140.3014 ~ 42.44'15 1.08 dd (5.37, 3.42,

8.79)1.29 ddd (3.42,

8.30, 11.72)27.66

16 1.62 ddd (6.35, 6.35, 7.81)

1.84 ddd (6.35, 3.90, 10.26)

39.57

17 - _ 43.7018 „ 2.23 d (6^83) 47.9619 1.69 ddd (6.83, 7.81,

6.36)“ 47.56

c

20 _ - 151.642 1 1.34 m 1.76 m 30.5722 - 3,20 dd(3.42, 6.83) 79.7323 0.96 s _ 32.6724 0.99 s - 17.5325 0.81 s - 15.2626 0.79 s _ _ 16.20.27 0.93 s - 14,7028 0.87 s - 19,0929 4,66 d (8.31), 4.71 d

(1 2 .2 )- 106,65

30 1 .6 8 s - 24.24


Pharmaco-anatomy Laboratory, Department o f Botany, Faculty o f Science, Jamia Hanidard

Tablea and Graphs

Table-47: and ’’^C MMR Spectral Data of Isostigmasterol (AS-3| and p-Sitosterol

Position NMR ” C N1 rAlpha Beta Isostigmasterol P-Sitoterol

1 1.41 dddd (16.12, 2.27 dddd 37.67 37.3315,13, 8.30.6.76) (13.67, 2.93,

1.95, 3.42)2 1 .8 6 m 170 m 32.09 31.633 3.52 brm

(w '^=10.9B)72.20 71,73

4 195 d (16.60) 2,30 d (5.83) 42,73 42,005 - 141.18 140,716 5.35 d (4.89) 122.08 121,167 1.99 d (11.23) 1.65 d (3.38) 30.08 31,96 ■8 1.16 dddd

(6.84, 3.91, 9.27, 6.84)

36.53 31,81

9 1.51 dddd (8.30, 6.84,4.89,7.81)

- 51,62 51,13

10 - 36.57 36.4311 2.20 m 1.49 m 21.48 21.0912 1.11 ddd (13.19,

6,83, 6,84)1.83 ddd(9.27,

3.42,4.89)40.19 39.79

13 - - 40.83 42.3714 1.15m - 57.18 56.7515 1.08 m 1.54 m 24.68 24.1516 1.67 m 1.49 m 28.61 28.7517 1.36 m 56.49 56.0218 0.69 brs - 12.38 ' 11.8419 1.01 brs 19.16 19.4620 - 2.02 36.92 36.072 1 0.96 d (7,33) 19.36 18.6822 5.14 dd (8.30,

6.84)- 129.71 33,95

23 5.18 dd (6.84, 8.30)

- 138.67 26.10

24 2.26 m - 50.56 45.8225 1.53 m - 29,27 29.5126 0.84 d (7.23) 19.29 19.7727 0.78 d (6.84) - 19.43 19.2128 1.13m 1.64 m 23.48 23.3129 0.82 d (7.32) - 12.25 11.04

Coupling constants in Hertz are provided in parentinesis


Tables and Graphs

Tabie-48: andand p-Sitosterol

13,€ NMR Spectral Data of p-Sitosteroi Glycoside (AS»5)

Position n¥ r NfmAlpha Beta P-Sitoterol

GlycosidejJ-Sitotprol

1 1.38 m 2.50 m 36.86 37.332 1.80 m 1.78 m 29.28 31.633 4.43 br m (w 1/2

18,5 Hz)- ■ 77.00 71.73

4 2,50 m 2.34 m 40.33 42.005 100,46 140.716 5.31 d (5,2) - 121.08 121.167 1,93 m 2.08 m 33.88 31,968 . 1.13 m 31,88 31,819 1.45 rn - ,49.64 51.1310 _ _ 36.22 36.4311 1.93 m 1,50 m 20,62 21.0912 1,13 m 1.80 m 38,34 39.7913 .. - 42.11 42.3714 1.15m - 56.21 56.75 ■15 1.06 m 1.80 m 23.88 24.1516 1.61 m 1,36 m 28.74 28.7517 1.45 m - 55.74 56.0218 0.64 m - 11,74 11,8419 0.95 brs - 19.10 19.4620 2,34 m 36.22 36.0721 0.90 d (6.0) 18.63 18.6622 1.78 m 1.10m 35.51 33.9523 1.25 m 1.80 m 25.49 26.1024 1.30 m 1.55 m 44.66 45.8225 1.73m - 27.81 29.5126 0.92 d (6.10) - 19.62 19.7727 0.80 d (6,6) - 18.95 19.2128 1.15m 1.42 m 23.63 23.3129 0.82 d (7,3) - 11.67 11.0T“r 4.90 brs - 100.84 -

2' 3.62 m - 76.74 -

3' 3.08 m - 73.474‘ 3.04 m - 70.09 -

5’ 4.22 d (5,6) - 76.746a ’ 4.88 brs - 61.10 -

6b’ 4.66 brs "


Pharmaco-anatomy Laboratoiy, Department o f Botany, Faculty o f Science, Janiia Hamdard ,

Tab Ies and Graphs

Tabie-49: NMR Spectral Data of Diglucodirharanoside |BA-3|c SC C 8C .. 5C C §C

1 103.61 r 102.70 1" 97.60 III 92.102 74.69 2' 74.35 2" 74.21 p Iff 74.113 72.91 3' 72.65 3" 72.45 3"' 72.004 70.98 4' 70.63 4" 70.14 4”’ 69.92a5 83.81 6' 76.84 5" 76.23 5'" 75.066 62.17 6' 60.98 6" 19,01 6"' 17.77



7'ahles and Graphs

Tab!e-50: NfViR Spectra! data of BC-1F^osition 5 Values of compound BC-1

1 40.022 27,443 79.024 38.875 55.356 18.347 35.618 40.889 50,4910 38.1111 20.961213 38.7514 42.8615 28.0016 34,3317 43.0118 48.3619 48.0020 150.9421 29.6822 37.2023 29.8924 18,0125 16.1126 . 16,00

. 27 15.3528 14.5629 109.3030 19.32


PharmacO'Einatomy Laboratory, Department oCBotany, Faculty o f Science, Jamia Hamdard

Tables and Graphs

Tabie-51: NMR SpectralAibaiupenoic acid (MA-2)

Data of Albanylacetate (fWA-1| and

Position ..................................... .......... MMRAliaanylacetate (fWA~1) Aibaiupenoic acid (MA-2)

1 40.8 39,032 36.83 27.393 80.99 79,04 39.71 37.055 55.31 55,356 18.27 18.287 31,26 34.348 38.51 40,699 47.69 50.51

10 37.73 37.2111 23.24 20.8512 124.36 27.99

139.66 38.7214 42.12 42.43 '15 28.75 27.2116 28.08 29,6917 33.76 56.35

,18 ^ 59.12 49.2819 39,68 46.9020 38,49 150.5221 32,91 30.5722 41.56 32.9023 28.11 29.1424 16,89 15.5625 16,73 16.1226 ■ 15.73 16.9827 23.63 15.3328 26.63 181,1129 17.49 109.730 21.32 19,37

OAc 171,9621.37



cooL.oQ.

T o t a l W a t e r - s o l u ble3 4 5

A c i d - i n s o lu b l e S u l p h a t e d

Fig. 1: P e r c e n t a g e o f T o t a l a s h . W a t e r - s o l u b l e a s h , A c id - in s o lu b le ash and S u lp h a te d ash o f the stem bark of A ls t o n ia sc ho laris

c0oL .oQl

Fig. 2: S u c c e s s iv e Extract ive va lues of air dried stem bark o1 A ls ton ia scho la ris

oo

s .

2 3 4

Total Water-soluble Acid-insoluble Sulphated

O.

Fig. 3: Percen tage of Total ash, W ater -so lub le ash, Ac id - inso lub le ash and Su lphated ash of the stem bark o1 Balan i tes aegypt iaca

16

1 4 -

12 -

10c«O 8

6 -

4 -

1 2 3 4 5 6 7

Pet. ether Ether Chloroform Benzene Ethanol Chloroform water

Fig. 4: Successive Extractive values of air dried stem bark of Balanites aegyptiaca

1 2 3 4 5

T o ta l W a t e r - s o l u b l e A c id - in s o fu b le S u lp h a t e d

Fig. 5: P e r c e n ta g e o f To ta l ash, W a te r -s o lu b le ash , Ac id - ins o lu b le ash and S u lp h a te d ash o f the stem bark of B o m b a x ce iba

cook .001

9

8

7

6

5 -

4 -

3 -

2-

1 -

1 2 3 4 5 6 7

Pet. e ther Ether Chloroform Benzene Ethanol Chloroform w ater

Fig. 6; Successive Extractive values of air dried stem bark of B om bax ceiba

Total2 3 4

Water -so lub le Acid- insoluble Sulphated

Fig. 7: Percentage of Total ash, Water-soluble ash, Acid-insolubie ash and Sulphated ash of stem bark of Morus alba

10-1

Cou<DQ. 6-

2^

1 2 3 4 5 8 7

Pet. ether Ether Chloroform Benzene Ethanol Chloroform wate

Fig. 8: S u cc e ss iv e Extractive values of air dr ied s tem bark of Mo r us alba

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