# New Bioactive Oleanane Type Compounds fromCoriandrum sativum Linn.

Authors: Muhammad Iqbal; Muhammad Bilal; Rubina Iqbal; Muhammad Akram; Imam Bakhsh Baloch; Musa Kaleem Baloch
Journal: Journal of Chemistry
 (2014)

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## Abstract

Five (1–5) new bioactive oleanane type triterpenoids have been isolated from ethyl acetate soluble fraction of ethanolic extract of Coriandrum sativum Linn. of Umbelliferae family. Ethanolic extract of the whole plant was fractionated in organic solvents. Ethyl acetate fraction was subjected to column chromatography on HPLC RP-18 to get 1-oxo-11β,21β-dihydroxy-oleanane (1), 1-oxo-11β-hydroxy-21β-O-acetyloleanane (2), 1-oxo-11β-hydroxy-21β-O-angeloyloleanane (3), 1-oxo-11β-O-angeloyl-21β-O-acetyloleanane (4), and 1-oxo-11β,21β-O-dibenzoyloleanane (5). The structures were elucidated after analysis of spectroscopic data, UV, IR, NMR (1H, 13C, 1D, and 2D), and mass measurements. Suspension in water of crude ethyl acetate extract was employed to treat sheep with ringworm disease. All isolated compounds (1–5) displayed excellent activity in terms of inhibition zones, MICs, MBCs, and MFCs against both bacteria and fungi. Ethyl acetate extract showed excellent antiringworm activity in sheep.


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## Body

## 1. Introduction

Coriandrum sativum Linn. of family Umbelliferae is an annual herbaceous plant and is cultivated all over the world for its use not only in the indigenous medicines but also as one of the ingredients of all spicy foods especially of Pakistan and India. The plant is a rich source of essential oil and many of the researchers have almost concentrated on extraction, composition, biological activities, and use against various diseases of its crude extracts and essential oils.C. sativum is an important medicinal plant used against a number of diseases. Nair et al. have indicated the antiarthritic activity of hydroalcoholic extract ofC. sativum [1]. Aqueous extract ofC. sativumseed has anxiolytic effect [2]. Ethanolic extract of the species has been reported to display anti-inflammatory and analgesic activity [3]. Its ethanolic extract possesses antitumor activity [4, 5]. Extracts ofC. sativum show diuretic and cholesterol lowering activity [6]. The extract of the plant has been used against diabetes mellitus [7]. Essential oil from seeds ofC. sativum has been used as antioxidant and antifungal agent [8]. Essential oil of the species has been used as wound healing medicine [9]. Naik et al. reported isolation of two pentacyclic oleanane type triterpenoids, coriandinonediol and its acetyl derivative, from the seeds of the species [10]. Dharmalingam and Nazni have shown the presence of alkaloids, steroids, saponins, tannins, and glycosides in the flowers ofC. sativum Linn. [11].Up till now, all of the activities ofC. sativum Linn. such as radical scavengers, antioxidants, anti-inflammatory, antianxiety, antibacterial, and antifungal have been associated with its essential oils and glycolipids [12–18]. Many other species of the Umbelliferae are rich in terpenes and terpenoids [19]; however, no proper attention has been paid toCoriandrum sativum Linn. for isolation of new compounds. These aspects prompted us to explore the locally cultivated species ofC. sativumLinn. for phytochemical constituents. In the present work, whole plant ofC. sativumLinn. was investigated for phytochemicals and resulted in the isolation of five (1–5) new oleanane type triterpenoids.


## 2. Material and Methods

### 2.1. Plant Material

C. sativum Linn. was grown in the botanical garden, Rakh Bibi Campus, Gomal University, Dera Ismail Khan, Khyber Pakhtunkhwa, Pakistan. The plants were identified by Professor Hamidullah Khan, Head of Pharmacognosy Department, Faculty of Pharmacy, Gomal University, Dera Ismail Khan. A specimen number CS 25 was retained in the herbarium. The seeds, the leaves, and whole parts of the plant were collected at appropriate time and investigated for their essential oil, bioactive constituents, and bioactivity.


### 2.2. Extraction

The whole dried plant material (roots, stems, leaves, and seeds) ofC. sativum Linn. was powdered using grinding machine. The grinded material (5000 g) was extracted with MeOH (7.5 L × 3) at room temperature. Methanolic solution was filtered using Whatman number 1 filter paper and concentrated at reduced temperature and pressure to get dark brown extract 500 g.


### 2.3. Isolation of Compounds

The methanol extract (480 g) was suspended in water (1000 mL) and reextracted with n-hexane (3 × 300 mL) chloroform (3 × 250 mL), ethyl acetate (3 × 250 mL), andn-butanol (3 × 50 mL). Each fraction was dried over anhydrous sodium sulphate and evaporated to dryness to yield n-hexane fraction (122.4 g, 25.5%), chloroform fraction (134.3 g, 27.98%), ethyl acetate fraction (124.3 g, 25.89%), n-butanol fraction (72.3 g, 15.06%), and aqueous fraction (25.4 g, 5.29%).The ethyl acetate soluble fraction (20 g) was subjected to chromatography on silica gel column (192 × 5.4 cm). It was eluted with petroleum ether/ethyl acetate mixture (6/4, 25000 mL) with flow rate of 2 mL/min to yield twenty (1–20) fractions each 250 mL. Fractions 3–5, containing identical constituents (on TLC similarities), were combined, evaporated to dryness in vacuo to afford a crude mixture (1.4 g), and subsequently purified by semipreparative HPLC using 15% water: acetonitrile as eluent, yielding1 (7 mg) and an inseparable mixture of2 and3 (11 mg). Fractions 11–13 after combination (1.2 g) were subjected to rechromatography on the same column of silica gel. The column was eluted using increasing concentrations of ethyl acetate in petroleum ether to end up with nine (1.1–1.9) pooled fractions.Fractions 1.4 and 1.5, based on TLC behavior, were combined together (1.3 g) and chromatographed over a silica gel column. The column was eluted using chloroform, 5% methanol in chloroform, and 10% methanol in chloroform, successively. The 5% methanol in chloroform elution resulted in three (1.4.1–1.4.3) fractions. Repeated column chromatography of these fractions resulted in the isolation of compounds4 (23 mg) and5 (15 mg). Fractions 6–8 (1.7 g) were combined and chromatographed over silica gel column using chloroform as eluent and the polarity was increased by methanol. Ten (6.1–6.10) fractions were collected. Fraction 6.1.4–6.1.7 (350 mg) was applied to a reversed-phase preparative HPLC on a LiChrosorb RP-18 column (7 μm, 2 × 30 cm i.d, Merck) and eluted with MeOH-H2O (80/20) at the rate of 10 mL/min (tR=7.5 min) to afford pure compounds (1) (31.3 mg), (tR=7.9 min) (2), (tR=8.1 min) (25.3 mg), (tR=8.4 min) (3) (31.3 mg), (tR=8.7 min) (4) (15.3 mg), and (tR=9.3 min) (5) (37.3 mg), respectively.Compound
1. White solid needles, crystals, m.p. 255–258°C; [α]D25+5.43 (c 1.4, CHCl3); UV (MeOH) λmax⁡  (log⁡⁡ε) 216 (1.56) nm; IR (dry) νmax⁡ 3532, 3404, 2940, 2870, 1620, 1450, 1250, 760 cm−1; 1H-NMR (C5D5N, 300 MHz) δ: 2.22 (2H, ddd,J= 13, 5.2, 3.5 Hz, H-2), 1.97 (2H, dd,J= 13, 5.2 Hz, H-3), 1.83 (1H, d,J= 11.5 Hz, H-5), 2.17 (2H, dd,J= 14, 6.5 Hz, H-6), 2.89 (1H, ddd,J = 16.5, 12.5, 6.5 Hz, H-7α), 2.74 (1H, dd,J = 16.5, 5.0 Hz, H-7β), 1.64 (1H, dd,J = 14, 6.5 Hz, H-9), 3.29 (1H, ddd,J = 14, 6.5, 4.2 Hz, H-11), 1.88 (2H, ddd,J = 14.1, 4.5, 12.2 Hz, H-12), 2.01 (1H, dd,J = 12.2, 4.5 Hz, H-13), 2.44 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-15α), 1.46 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15β), 2.53 (1H, ddd,J= 13.5, 13.5, 3.0 Hz, H-16α), 1.56 (1H, ddd,J= 13.5, 13.5, 3.0 Hz, H-16β), 1.78 (1H, dddJ = 12.5, 12.5, 3.5 Hz, H-18), 1.71 (1H, ddd,J = 14.5, 11.5, 4.0 Hz, H-19α), 1.17 (1H, dd,J = 14.5, 4.0 Hz, H-19β), 3.46 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-21), 2.91 (1H, ddd,J = 13.5, 13.5, 2.5 Hz, H-22α), 1.91 (1H, dd,J = 13.5, 13.5 Hz, H-22β), 1.66 (3H, s, H-23), 1.07 (3H, s, H-24), 1.13 (3H, s, H-25), 1.16 (3H, s, H-26), 1.55 (3H, s, H-27), 1.17 (3H, s, H-28), 1.38 (3H, s, H-29), 0.97 (3H, s, H-30); 13C-NMR (C5D5N, 75 MHz) δ: 214.7 (C-1), 68.2 (C-2), 47.9 (C-3), 45.7 (C-4), 56.7 (C-5), 19.5 (C-6), 35.5 (C-7), 40.4 (C-8), 50.6 (C-9), 38.8 (C-10), 71.4 (C-11), 43.7 (C-12), 51.3 (C-13), 42.1 (C-14), 29.1 (C-15), 27.9 (C-16), 46.5 (C-17), 44.6 (C-18), 27.1 (C-19), 35.7 (C-20), 72.8 (C-21), 42.9 (C-22), 24.1 (C-23), 23.5 (C-24), 17.5 (C-25), 17.4 (C-26), 24.9 (C-27), 25.1 (C-28), 28.8 (C-29), 24.6 (C-30). EI-MS:m/z = 458 (M+, 3, C30H50O3), 440 (12, M+-H2O), 430 (27, M+-CO), 422 (15, M+-2H2O), 291 (10, C19H31O2+), 263 (11, C18H31O+), 249 (23, C16H25O2+), 207 (31, C14H23O+), 189 (43, C14H21+), 135 (42, C9H11O+), 109 (34, C7H9O+), 95 (44, C7H11+); HR-EI-MS:m/z = 458.7162, calcd. for C30H50O3; 458.7158 observed.Compound
2. White solid, crystals, m.p. 291–293°C; [α]D25 + 4.43 (c1.4, CHCl3); UV (MeOH) λmax⁡  (log⁡⁡ε) 247 (1.7) nm; IR (dry) νmax⁡ 3436, 2965, 2879, 1728, 1712, 1458, 1266, 779 cm−1; 1H-NMR (C5D5N, 300 MHz) δ: 1.88 (2H, ddd,J = 13, 5.2, 3.5 Hz, H-2), 2.24 (2H, dd,J = 5, 12 Hz, H-3), 1.72 (1H, d = 11.5 Hz, H-5), 2.19 (2H, dd,J = 6.5, 14 Hz, H-6), 2.87 (1H, ddd,J = 16.5, 12.5, 6.5 Hz, H-7α), 2.71 (1H, dd, 16.5, 5.0 H-7β), 1.65 (1H, dd,J = 14, 6.5 Hz, H-9), 4.23 (1H, ddd,J = 3.0, 13.5, 13.5 Hz, H-11), 5.79 (2H, ddd,J = 14.1, 4.5, 12.2 Hz, H-12), 2.02 (1H, dd,J = 12.2, 4.5 Hz, H-13), 2.43 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-15α), 1.47 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15β), 2.55 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16α), 2.57 (ddd,J = 13.5, 13.5, 3.0 Hz, H-16β), 1.77 (1H, ddd,J = 12.5, 12.5, 3.5 Hz, H-18), 1.70 (1H, ddd,J = 14.5, 11.5, 4.0 Hz, H-19α), 1.16 (1H, dd,J = 14.5, 4.0 Hz, H-19β), 4.46 (1H, ddd,J = 13.5, 13.5, 3.0, Hz, H-21), 2.90 (1H, ddd,J = 13.5, 13.5, 2.5 Hz, 22α), 1.92 (1H, dd,J = 13.5, 13.5 Hz, H-22β), 1.67 (3H, s, H-23), 1.08 (3H, s, H-24), 1.12 (3H, s, H-25), 1.17 (3H, s, H-26), 1.56 (3H, s, H-27), 1.15 (3H, s, H-28), 1.37 (3H, s, H-29), 0.98 (3H, s, H-30), Ac: 2.29 (3H, s, CH3-CO); 13C-NMR (C5D5N, 75 MHz) δ: 214.5 (C-1), 67.9 (C-2), 47.8 (C-3), 45.5 (C-4), 56.6 (C-5), 19.6 (C-6), 35.4 (C-7), 40.3 (C-8), 50.6 (C-9), 38.8 (C-10), 77.4 (C-11), 44.6 (C-12), 54.4 (C-13), 42.4 (C-14), 29.0 (C-15), 28.0 (C-16), 46.6 (C-17), 44.8 (C-18), 27.3 (C-19), 35.5 (C-20), 78.8 (C-21), 43.0 (C-22), 24.2 (C-23), 23.7 (C-24), 17.4 (C-25), 17.3 (C-26), 24.7 (C-27), 25.5 (C-28), 28.7 (C-29), 24.8 (C-30), Ac: 176.2 (CH3–CO), 22.3 (CH3–CO); EI-MS:m/z = 500 (M+, 3, C32H52O4), 482 (15, M+-H2O), 472 (21, M+-CO), 440 (15, M+-MeCO2H), 291 (10, C19H31O2+), 263 (11, C18H31O+), 249 (23, C16H25O2+), 207 (31, C14H23O+), (100, C5H8O2+), 189 (43, C14H21+), 135 (42, C9H11O+), 109 (34, C7H9O+), 95 (44, C7H11+), 43 (46); HR-EI-MS:m/z = 500.7529, calcd. for C32H52O4; 500.7515 observed.Compound
3. White solid, crystalline, m.p. 258–260°C; [α]D25 + 5.43 (c1.4, CHCl3); UV (MeOH) λmax⁡  (log⁡⁡ε) 308 (4.28) and 214 (4.26), 267 (3.7) nm; IR (dry) νmax⁡ 3348 (OH), 2947, 2877, 1723 (C=C–CO2), 1707 (CO), 1599, 1534 (C=C), 1451, 1256, 767 cm−1;1H-NMR (C5D5N, 300 MHz) δ: 1.87 (2H, ddd,J = 13, 5.2, 3.5 Hz, H-2), 2.25 (2H, dd,J = 4.5, 12 Hz, H-3), 1.74 (1H, d,J = 11.5 Hz, H-5), 2.17 (2H, dd,J = 6.5, 14 Hz, H-6), 2.87 (1H, ddd,J = 16.5, 12.5, 6.5 Hz, H-7α), 2.69 (1H, dd,J = 16.5, 5.0 Hz, H-7β), 1.66 (1H, dd,J = 14, 6.5 Hz, H-9), 4.47 (1H, ddd,J = 3.0, 13.5, 13.5 Hz, H-11), 5.77 (2H, ddd,J = 14.1, 4.5, 12.2 Hz, H-12), 2.05 (1H, dd,J = 12.2, 4.5 Hz, H-13), 2.44 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-15α), 1.46 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15β), 2.52 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16α), 1.56 (1H, ddd, 13.5, 13.5, 3.0 Hz, H-16β), 1.78 (1H, ddd,J = 12.5, 12.5, 3.5 Hz, H-18), 1.71 (1H, ddd,J = 14.5, 11.5, 4.0 Hz, H-19α), 1.19 (1H, dd,J = 14.5, 4.0 Hz, H-19β), 4.57 (1H, ddd,J= 13.5, 13.5, 3.0 Hz, H-2), 2.93 (1H, ddd,J = 13.5, 13.5, 2.5 Hz, H-22α), 1.95 (1H, dd,J = 13.5, 13.5 Hz, H-22α), 1.95 (1H, dd,J = 13.5, 13.5 Hz, H-22β), 1.65 (3H, s, H-23), 1.09 (3H, s, H-24), 1.09 (3H, s, H-25), 1.18 (3H, s, H-26), 1.58 (3H, s, H-27), 1.14 (3H, s, H-28), 1.36 (3H, s, H-29), 0.97s (3H, s, H-30), Ang: 6.13 (1H, qq,J = 6.6 Hz, H-3), 2.01 (3H, dq,J = 6.5, 2.5 Hz, H-4), 1.93 (3H, dqm,J = 6.5, 2.2 Hz, H-5); 13C-NMR (C5D5N, 75 MHz) δ: 214.2 (C-1), 68.4 (C-2), 47.7 (C-3), 45.8 (C-4), 56.9 (C-5), 19.4 (C-6), 35.5 (C-7), 40.3 (C-8), 50.6 (C-9), 38.7 (C-10), 78.1 (C-11), 44.6 (C-12), 54.8 (C-13), 42.3 (C-14), 29.2 (C-15), 28.4 (C-16), 46.7 (C-17), 44.5 (C-18), 27.8 (C-19), 35.4 (C-20), 78.8 (21), 43.5 (C-22), 24.0 (C-23), 23.8 (C-24), 17.6 (C-25), 17.3 (C-26), 24.8 (C-27), 25.6 (C-28), 28.6 (C-29), 24.9 (C-30), Ang: 168.3 (C-1), 128.0 (C-2), 137.3 (C-3), 15.9 (C-4), 20.6 (C-5); EI-MS:m/z = 540 (M+, 3, C35H56O4), 518 (12, M+-H2O), 512 (27, M+-CO), 440 (15, M+-C5H8O2), 291 (10, C19H31O2+), 263 (11, C18H31O+), 249 (23, C16H25O2+), 207 (31, C14H23O+), (100, C5H8O2+), 189 (43, C14H21+), 135 (42, C9H11O+), 109 (34, C7H9O+), 95 (44, C7H11+), 83 (45), 82 (34); HR-EI-MS:m/z = 540.8167, calcd. for C35H56O4; 540.8143 observed.Compound
4. White solid, amorphous, m.p. 267-268°C; [α]D25 + 6.43 (c1.4, CHCl3); UV (MeOH) λmax⁡ (log ε) 347 (4.55), 284 (4.56), 253 (4.22) 234 (2.7) nm; IR (dry) νmax⁡ 2955, 2885, 1738, 1690, 1635, 1593, 1465, 1260, 926, 807, 764 cm−1;1H-NMR (C5D5N, 300 MHz) δ: 1.86 (2H, ddd,J = 13, 5.2, 3.5 Hz, H-2), 2.24 (2H, dd,J = 4.5, 12 Hz, H-3), 1.73 (1H, d,J = 11.5 Hz, H-5), 2.18 (2H, dd,J= 6.5, 14 Hz, H-6), 2.84 (1H, ddd,J = 16.5, 12.5, 6.5 Hz, H-7α), 2.68 (1H, dd, 16.5, 5.0 Hz, H-7β), 1.62 (1H, dd,J = 14, 6.5 Hz, H-9), 4.29 (1H, ddd,J = 3.0, 13.5, 13.5 Hz, H-11), 5.77 (2H, ddd,J = 14.1, 4.5, 12.2 Hz, H-12), 2.06 (1H, dd,J = 12.2, 4.5 Hz, H-13), 2.45 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-15α), 1.45 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15α), 1.45 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15β), 2.51 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16α), 1.55 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16α), 1.55 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16β), 1.79 (1H, ddd,J = 12.5, 12.5, 3.5 Hz, H-18), 1.72 (1H, ddd,J = 14.5, 11.5, 4.0 Hz, H-19α), 1.18 (1H, dd,J = 14.5, 4.0 Hz, H-19β), 4.29 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-21), 2.92 (1H, ddd,J = 13.5, 13.5, 2.5 Hz, H-22α), 2.89 (1H, dd,J = 13.5, 13.5 Hz, H-22β), 1.66 (3H, s, H-23), 1.09 (3H, s, H-24), 1.08 (3H, s, H-25), 1.15 (3H, s, H-26), 1.61 (3H, s, H-27), 1.18 (3H, s, H-28), 1.39 (3H, s, H-29), 0.96 (3H, s, H-30), Ang: 6.13 (1H, qq,J = 6.6 Hz, H-3), 2.01 (3H, dq,J = 6.5, 2.5 Hz, H-4), 1.93 (3H, dqm,J = 6.5, 2.2 Hz, H-5), Ac: 2.29 (3H, s, CH3–CO); 13C-NMR (C5D5N, 75 MHz) δ: 214.8 (C-1), 68.3 (C-2), 47.6 (C-3), 45.9 (C-4), 56.6 (C-5), 19.3 (C-6), 35.5 (C-7), 40.2 (C-8), 50.6 (C-9), 38.8 (C-10), 77.9 (C-11), 44.5 (C-12), 54.7 (C-13), 42.7 (C-14), 29.3 (C-15), 27.9 (C-16), 46.4 (C-17), 44.6 (C-18), 27.6 (C-19), 35.5 (C-20), 81.8 (C-21), 43.6 (C-22), 24.1 (C-23), 23.8 (C-24), 17.5 (C-25), 17.1 (C-26), 24.8 (C-27), 25.7 (C-28), 28.8 (C-29), 24.6 (C-30), Ang: 168.3 (C-1), 128.0 (C-2), 137.3 (C-3), 15.9 (C-4), 20.6 (C-5); Ac: 176.2 (CH3–CO), 22.3 (CH3–CO); EI-MS:m/z = 582 (M+, 3, C37H58O5), 564 (12, M+-H2O), 554 (27, M+-CO), 522 (15, M+-MeCO2H), 464 (15, M+-C5H8O2), 291 (10, C19H31O2+), 263 (11, C18H31O+), 249 (23, C16H25O2+), 207 (31, C14H23O+), (100, C5H8O2+), 189 (43, C14H21+), 135 (42, C9H11O+), 109 (34, C7H9O+), 95 (44, C7H11+), 83 (45), 82 (56), 43 (78); HR-EI-MS:m/z = 582.8334, calcd. for C37H58O5; 582.8343, observed.Compound
5. White solid, crystals, m.p. 264–268°C; [α]D25 + 5.43 (c1.4, CHCl3); UV (MeOH) λmax⁡ (log ε) 349 (4.55), 286 (4.56), 254 (4.22), 247 (1.7) nm; IR (dry) νmax⁡ 3133, 2940, 2870, 1745, 1620, 1527, 1450, 1250, 923, 806, 791, 760 cm−1;1H-NMR (C5D5N, 300 MHz) δ: 1.87 (2H, ddd,J = 13, 5.2, 3.5 Hz, H-2), 4.35 (2H, dd,J = 4.5, 12 Hz, H-3), 1.76 (1H, d,J = 11.5 Hz, H-5), 2.17 (1H, dd,J = 6.5, 14 Hz, H-6), 2.83 (1H, ddd,J = 16.5, 12.5, 6.5 Hz, H-7α), 2.69 (1H, dd,J = 16.5, 5.0 Hz, H-7β), 1.63 (1H, dd,J = 14, 6.5 Hz, H-9), 4.29 (1H, ddd,J = 3.0, 13.5, 13.5 Hz, H-11), 5.78 (2H, ddd,J = 14.1, 4.5, 12.2 Hz, H-12), 2.07 (1H, dd,J = 12.2, 4.5 Hz, H-13), 2.44 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-15α), 1.46 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15β), 2.54 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16α), 1.56 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16β), 1.78 (1H, ddd,J = 12.5, 12.5, 3.5 Hz, H-18), 1.73 (1H, ddd,J = 14.5, 11.5, 4.0 Hz, H-19α), 1.17 (1H, dd,J = 14.5, 4.0 Hz, H-19β), 4.46 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-21), 2.97 (1H, ddd,J = 13.5, 13.5, 2.5 Hz, H-22α), 1.90 (1H, dd,J = 13.5, 13.5 Hz, H-22β), 1.65 (3H, s, H-23), 1.10 (3H, s, H-24), 1.14 (3H, s, H-25), 1.19 (3H, s, H-26), 1.60 (3H, s, H-27), 1.17 (3H, s, H-28), 1.37 (3H, s, H-29), 0.99 (3H, s, H-30), 2 × Bz: 8.04 (1H, d,J = 7.2 Hz, H-2), 7.41 (1H, t,J = 7.2 Hz, H-3), 7.53 (1H, dt,J = 7.2, 2.5 Hz, H-4), 7.41 (1H, t,J = 7.2 Hz, H-5), 8.04 (1H, d,J = 7.2 Hz, H-6); 13C-NMR (C5D5N, 75 MHz) δ: 214.9 (C-1), 68.5 (C-2), 47.5 (C-3), 46.1 (C-4), 56.7 (C-5), 19.8 (C-6), 35.6 (C-7), 40.3 (C-8), 50.6 (C-9), 38.9 (C-10), 77.8 (C-11), 44.6 (C-12), 54.5 (C-13), 42.6 (C-14), 29.1 (C-15), 28.3 (C-16), 46.5 (C-17), 44.9 (C-18), 27.5 (C-19), 35.5 (C-20), 81.8 (C-21), 44.1 (C-22), 24.2 (C-23), 23.5 (C-24), 17.4 (C-25), 17.4 (C-26), 24.9 (C-27), 25.5 (C-28), 28.9 (C-29), 24.8 (C-30), 2 × Bz: 164.6 (C-1), 129.7 (C-2), 130.0 (C-3), 128.4 (C-4), 133.7 (C-5), 128.4 (C-6), 130.0 (C-7); EI-MS:m/z = 666 (M+, 3, C44H54O5), 440 (12, M+-H2O), 430 (27, M+-CO), 422 (15, M+-2H2O), 291 (10, C19H31O2+), 263 (11, C18H31O+), 249 (23, C16H25O2+), 207 (31, C14H23O+), 189 (43, C14H21+), 135 (42, C9H11O+), 105 (34, C7H5O+), 109 (34, C7H9O+), 95 (44, C7H11+), 77 (23, C6H5+), 65 (23, C5H5+); HR-EI-MS:m/z = 666.9283, calcd. for C44H54O5; 666.9273 observed.


### 2.4. Antiringworm Activity of Ethyl Acetate Extract ofCoriandrum sativumLinn.

In this study five sheep infected with ringworm disease were selected. For each infected sheep, ethyl acetate extract (10 gram) was suspended in preboiled water (10 Liter). Each of the infected sheep was given bath with the suspension after every eight hours continually for four days. On the fourth day, four of the infected sheep recovered from ringworms’ infections, while the fifth test sheep took a longer time (10 days) along with vitamin A injection medication.


### 2.5. Antimicrobial Activities of Compounds 1–5

Compounds1–5 were tested for their antibacterial and antifungal activities in terms of inhibition zones, MICs, MBCs, and MFCs using standard procedures [8, 12]. The results of antimicrobial activity of compounds1–5 are displayed in Tables 1, 2, 3, 4, 5, and 6. Briefly, LB medium 20 mL was poured in petri plates and after solidification standard inoculum (100 μL) bacteria/fungi concentration 107 CFU/mL suspension was poured and dried for 5 min. DMSO was used as negative control while standard drugs Miconazole and Imipenem (10 μg/disc) were used as positive control.Table 1
Inhibition zones (mm) of1–5 isolated from C. sativum against bacteria.
Comp./organism
1
2
3
4
5
Std*
E. coli
7.1 ± 1.2
22 ± 2
17 ± 2
8 ± 2
8 ± 2
28 ± 1.2
S.  aureus
3.6 ± 1.2
23 ± 2
16 ± 2
8 ± 2
18 ± 2
31 ± 1.2
P. mirabilis
2.4 ± 1.2
22 ± 2
19 ± 2
21 ± 2
10 ± 2
29 ± 1.2
P. aeruginosa
2.2 ± 1.2
29 ± 2
21 ± 2
29 ± 2
19 ± 2
33 ± 1.2
B. cereus
2.2 ± 1.2
23 ± 2
22 ± 2
8 ± 2
8 ± 2
27 ± 1.2
K. pneumonia
0
0
17 ± 2
8 ± 2
8 ± 2
29 ± 1.2
M. luteus
0
0
8 ± 2
8 ± 2
17 ± 2
37 ± 1.2
E. cloacae
0
0
0
0
0
34 ± 1.2
*Imipenem was used as standard.Table 2
Inhibition zones (mm) of1–5isolated from C. sativum against fungi.
Comp./organism
1
2
3
4
5
Std*
T. rubrum
9.1 ± 2
7 ± 2
3 ± 2
2 ± 2
1 ± 2
29 ± 2
C. albicans
7.6 ± 2
5 ± 2
3 ± 2
2 ± 2
2 ± 2
31 ± 2
M. audouinii
5.4 ± 2
6 ± 2
2 ± 2
2 ± 2
1 ± 2
24 ± 2
C. neoformans
6.2 ± 2
6 ± 2
2 ± 2
3 ± 2
1 ± 2
26 ± 2
T. mentagrophytes
9.0 ± 2
5 ± 2
3 ± 2
2 ± 2
1 ± 2
29 ± 2
E. floccosum
6.3 ± 2
6 ± 2
3 ± 2
1 ± 2
2 ± 2
27 ± 2
M. canis
7.6 ± 2
3 ± 2
3 ± 2
3 ± 2
1 ± 2
28 ± 2
A. niger
2.3 ± 2
0
3 ± 2
3 ± 2
1 ± 2
28 ± 2
*Imipenem was used as standard.Table 3
MICs (µg/mL) of 1–5isolated from C. sativum against bacterial strains.
Comp.
E. coli
P. mirabilis
P. aeruginosa
M. luteus
E. cloacae
K. pneumonia
B. cereus
S.  aureus
1
4 ± 0.5
8 ± 1.5
8 ± 1.5
4 ± 0.5
3 ± 0.5
4 ± 0.5
4 ± 0.5
4 ± 0.5
2
4 ± 0.5
8 ± 1.5
8 ± 1.5
4 ± 0.5
4 ± 0.5
8 ± 1.5
8 ± 1.5
4 ± 0.5
3
4 ± 0.5
16 ± 2.0
8 ± 1.5
8 ± 1.5
5 ± 0.5
16 ± 1.5
16 ± 1.5
4 ± 0.5
4
8 ± 0.5
16 ± 1.5
8 ± 1.0
5 ± 0.5
4 ± 0.5
16 ± 1.5
4 ± 0.5
4 ± 0.5
5
64 ± 2.0
32 ± 2.0
32 ± 2.0
4 ± 0.5
4 ± 0.5
64 ± 2.0
64 ± 2.0
64 ± 2.0Table 4
MICs (µg/mL) of 1–5 from C. sativum against fungal strains.
Comp./organism
C. albicans
C. neoformans
M. audouinii
A. niger
T. mentagrophytes
E. floccosum
M. canis
T. rubrum
1
4 ± 0.5
4 ± 0.5
8 ± 0.5
0
8 ± 0.5
16 ± 0.5
0
16 ± 1.5
2
4 ± 0.5
8 ± 1.5
16 ± 1.5
0
16 ± 0.5
16 ± 0.5
0
16 ± 1.5
3
4 ± 0.5
8 ± 1.5
16 ± 1.5
0
16 ± 0.5
32 ± 0.5
16 ± 1.5
32 ± 2.0
4
8 ± 1.5
8 ± 1.5
16 ± 1.5
0
64 ± 2.5
32 ± 1.5
16 ± 1.5
16 ± 1.5
5
16 ± 1.5
64 ± 2.0
16 ± 1.5
0
32 ± 1.5
32 ± 2.0
16 ± 1.5
8 ± 1.5Table 5
MBCs (µg/mL) of 1–5 from C. sativum against bacterial strains.
Comp./organism
E. coli
P. mirabilis
P. aeruginosa
M. luteus
E. cloacae
K. pneumoniae
B. cereus
S.  aureus
1
8 ± 1.5
16 ± 1.5
16 ± 1.5
8 ± 1.5
0
8 ± 1.5
9 ± 1.5
8  ± 1.5
2
8 ± 1.5
16 ± 1.5
16 ± 1.5
9 ± 1.5
0
16 ± 1.5
16 ± 1.5
8 ± 1.5
3
8 ± 1.5
33 ± 1.5
16 ± 1.5
16 ± 1.5
0
32 ± 1.5
33 ± 1.5
8 ± 1.5
4
17 ± 1.5
32 ± 1.5
16 ± 1.5
0
0
33 ± 1.5
9 ± 1.5
9 ± 1.5
5
12 ± 1.5
65 ± 1.5
15 ± 1.5
0
0
12 ± 1.5
12 ± 1.5
12 ± 1.5Table 6
MFCs (µg/mL) of 1–5 from C. sativum seeds against fungal strains.
Comp.
C. albicans
C. neoformans
M. audouinii
A. niger
T. mentagrophytes
E. floccosum
M. canis
T. rubrum
1
9 ± 1.5
9 ± 1.5
17 ± 1.5
0
17 ± 1.5
31 ± 1.5
0
31 ± 1.5
2
9 ± 1.5
16 ± 1.5
31 ± 1.5
0
31
31
0
31 ± 1.5
3
9 ± 1.5
16 ± 1.5
31 ± 1.5
0
31 ± 1.5
63 ± 1.5
32 ± 1.5
63 ± 1.5
4
16 ± 1.5
16 ± 1.5
31 ± 1.5
0
122 ± 1.5
61 ± 1.5
3 ± 1.52
31 ± 1.5
5
32 ± 1.5
112 ± 1.5
31 ± 1.5
0
62 ± 1.5
61 ± 1.5
32 ± 1.5
16 ± 1.5


## 2.1. Plant Material

C. sativum Linn. was grown in the botanical garden, Rakh Bibi Campus, Gomal University, Dera Ismail Khan, Khyber Pakhtunkhwa, Pakistan. The plants were identified by Professor Hamidullah Khan, Head of Pharmacognosy Department, Faculty of Pharmacy, Gomal University, Dera Ismail Khan. A specimen number CS 25 was retained in the herbarium. The seeds, the leaves, and whole parts of the plant were collected at appropriate time and investigated for their essential oil, bioactive constituents, and bioactivity.


## 2.2. Extraction

The whole dried plant material (roots, stems, leaves, and seeds) ofC. sativum Linn. was powdered using grinding machine. The grinded material (5000 g) was extracted with MeOH (7.5 L × 3) at room temperature. Methanolic solution was filtered using Whatman number 1 filter paper and concentrated at reduced temperature and pressure to get dark brown extract 500 g.


## 2.3. Isolation of Compounds

The methanol extract (480 g) was suspended in water (1000 mL) and reextracted with n-hexane (3 × 300 mL) chloroform (3 × 250 mL), ethyl acetate (3 × 250 mL), andn-butanol (3 × 50 mL). Each fraction was dried over anhydrous sodium sulphate and evaporated to dryness to yield n-hexane fraction (122.4 g, 25.5%), chloroform fraction (134.3 g, 27.98%), ethyl acetate fraction (124.3 g, 25.89%), n-butanol fraction (72.3 g, 15.06%), and aqueous fraction (25.4 g, 5.29%).The ethyl acetate soluble fraction (20 g) was subjected to chromatography on silica gel column (192 × 5.4 cm). It was eluted with petroleum ether/ethyl acetate mixture (6/4, 25000 mL) with flow rate of 2 mL/min to yield twenty (1–20) fractions each 250 mL. Fractions 3–5, containing identical constituents (on TLC similarities), were combined, evaporated to dryness in vacuo to afford a crude mixture (1.4 g), and subsequently purified by semipreparative HPLC using 15% water: acetonitrile as eluent, yielding1 (7 mg) and an inseparable mixture of2 and3 (11 mg). Fractions 11–13 after combination (1.2 g) were subjected to rechromatography on the same column of silica gel. The column was eluted using increasing concentrations of ethyl acetate in petroleum ether to end up with nine (1.1–1.9) pooled fractions.Fractions 1.4 and 1.5, based on TLC behavior, were combined together (1.3 g) and chromatographed over a silica gel column. The column was eluted using chloroform, 5% methanol in chloroform, and 10% methanol in chloroform, successively. The 5% methanol in chloroform elution resulted in three (1.4.1–1.4.3) fractions. Repeated column chromatography of these fractions resulted in the isolation of compounds4 (23 mg) and5 (15 mg). Fractions 6–8 (1.7 g) were combined and chromatographed over silica gel column using chloroform as eluent and the polarity was increased by methanol. Ten (6.1–6.10) fractions were collected. Fraction 6.1.4–6.1.7 (350 mg) was applied to a reversed-phase preparative HPLC on a LiChrosorb RP-18 column (7 μm, 2 × 30 cm i.d, Merck) and eluted with MeOH-H2O (80/20) at the rate of 10 mL/min (tR=7.5 min) to afford pure compounds (1) (31.3 mg), (tR=7.9 min) (2), (tR=8.1 min) (25.3 mg), (tR=8.4 min) (3) (31.3 mg), (tR=8.7 min) (4) (15.3 mg), and (tR=9.3 min) (5) (37.3 mg), respectively.Compound
1. White solid needles, crystals, m.p. 255–258°C; [α]D25+5.43 (c 1.4, CHCl3); UV (MeOH) λmax⁡  (log⁡⁡ε) 216 (1.56) nm; IR (dry) νmax⁡ 3532, 3404, 2940, 2870, 1620, 1450, 1250, 760 cm−1; 1H-NMR (C5D5N, 300 MHz) δ: 2.22 (2H, ddd,J= 13, 5.2, 3.5 Hz, H-2), 1.97 (2H, dd,J= 13, 5.2 Hz, H-3), 1.83 (1H, d,J= 11.5 Hz, H-5), 2.17 (2H, dd,J= 14, 6.5 Hz, H-6), 2.89 (1H, ddd,J = 16.5, 12.5, 6.5 Hz, H-7α), 2.74 (1H, dd,J = 16.5, 5.0 Hz, H-7β), 1.64 (1H, dd,J = 14, 6.5 Hz, H-9), 3.29 (1H, ddd,J = 14, 6.5, 4.2 Hz, H-11), 1.88 (2H, ddd,J = 14.1, 4.5, 12.2 Hz, H-12), 2.01 (1H, dd,J = 12.2, 4.5 Hz, H-13), 2.44 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-15α), 1.46 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15β), 2.53 (1H, ddd,J= 13.5, 13.5, 3.0 Hz, H-16α), 1.56 (1H, ddd,J= 13.5, 13.5, 3.0 Hz, H-16β), 1.78 (1H, dddJ = 12.5, 12.5, 3.5 Hz, H-18), 1.71 (1H, ddd,J = 14.5, 11.5, 4.0 Hz, H-19α), 1.17 (1H, dd,J = 14.5, 4.0 Hz, H-19β), 3.46 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-21), 2.91 (1H, ddd,J = 13.5, 13.5, 2.5 Hz, H-22α), 1.91 (1H, dd,J = 13.5, 13.5 Hz, H-22β), 1.66 (3H, s, H-23), 1.07 (3H, s, H-24), 1.13 (3H, s, H-25), 1.16 (3H, s, H-26), 1.55 (3H, s, H-27), 1.17 (3H, s, H-28), 1.38 (3H, s, H-29), 0.97 (3H, s, H-30); 13C-NMR (C5D5N, 75 MHz) δ: 214.7 (C-1), 68.2 (C-2), 47.9 (C-3), 45.7 (C-4), 56.7 (C-5), 19.5 (C-6), 35.5 (C-7), 40.4 (C-8), 50.6 (C-9), 38.8 (C-10), 71.4 (C-11), 43.7 (C-12), 51.3 (C-13), 42.1 (C-14), 29.1 (C-15), 27.9 (C-16), 46.5 (C-17), 44.6 (C-18), 27.1 (C-19), 35.7 (C-20), 72.8 (C-21), 42.9 (C-22), 24.1 (C-23), 23.5 (C-24), 17.5 (C-25), 17.4 (C-26), 24.9 (C-27), 25.1 (C-28), 28.8 (C-29), 24.6 (C-30). EI-MS:m/z = 458 (M+, 3, C30H50O3), 440 (12, M+-H2O), 430 (27, M+-CO), 422 (15, M+-2H2O), 291 (10, C19H31O2+), 263 (11, C18H31O+), 249 (23, C16H25O2+), 207 (31, C14H23O+), 189 (43, C14H21+), 135 (42, C9H11O+), 109 (34, C7H9O+), 95 (44, C7H11+); HR-EI-MS:m/z = 458.7162, calcd. for C30H50O3; 458.7158 observed.Compound
2. White solid, crystals, m.p. 291–293°C; [α]D25 + 4.43 (c1.4, CHCl3); UV (MeOH) λmax⁡  (log⁡⁡ε) 247 (1.7) nm; IR (dry) νmax⁡ 3436, 2965, 2879, 1728, 1712, 1458, 1266, 779 cm−1; 1H-NMR (C5D5N, 300 MHz) δ: 1.88 (2H, ddd,J = 13, 5.2, 3.5 Hz, H-2), 2.24 (2H, dd,J = 5, 12 Hz, H-3), 1.72 (1H, d = 11.5 Hz, H-5), 2.19 (2H, dd,J = 6.5, 14 Hz, H-6), 2.87 (1H, ddd,J = 16.5, 12.5, 6.5 Hz, H-7α), 2.71 (1H, dd, 16.5, 5.0 H-7β), 1.65 (1H, dd,J = 14, 6.5 Hz, H-9), 4.23 (1H, ddd,J = 3.0, 13.5, 13.5 Hz, H-11), 5.79 (2H, ddd,J = 14.1, 4.5, 12.2 Hz, H-12), 2.02 (1H, dd,J = 12.2, 4.5 Hz, H-13), 2.43 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-15α), 1.47 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15β), 2.55 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16α), 2.57 (ddd,J = 13.5, 13.5, 3.0 Hz, H-16β), 1.77 (1H, ddd,J = 12.5, 12.5, 3.5 Hz, H-18), 1.70 (1H, ddd,J = 14.5, 11.5, 4.0 Hz, H-19α), 1.16 (1H, dd,J = 14.5, 4.0 Hz, H-19β), 4.46 (1H, ddd,J = 13.5, 13.5, 3.0, Hz, H-21), 2.90 (1H, ddd,J = 13.5, 13.5, 2.5 Hz, 22α), 1.92 (1H, dd,J = 13.5, 13.5 Hz, H-22β), 1.67 (3H, s, H-23), 1.08 (3H, s, H-24), 1.12 (3H, s, H-25), 1.17 (3H, s, H-26), 1.56 (3H, s, H-27), 1.15 (3H, s, H-28), 1.37 (3H, s, H-29), 0.98 (3H, s, H-30), Ac: 2.29 (3H, s, CH3-CO); 13C-NMR (C5D5N, 75 MHz) δ: 214.5 (C-1), 67.9 (C-2), 47.8 (C-3), 45.5 (C-4), 56.6 (C-5), 19.6 (C-6), 35.4 (C-7), 40.3 (C-8), 50.6 (C-9), 38.8 (C-10), 77.4 (C-11), 44.6 (C-12), 54.4 (C-13), 42.4 (C-14), 29.0 (C-15), 28.0 (C-16), 46.6 (C-17), 44.8 (C-18), 27.3 (C-19), 35.5 (C-20), 78.8 (C-21), 43.0 (C-22), 24.2 (C-23), 23.7 (C-24), 17.4 (C-25), 17.3 (C-26), 24.7 (C-27), 25.5 (C-28), 28.7 (C-29), 24.8 (C-30), Ac: 176.2 (CH3–CO), 22.3 (CH3–CO); EI-MS:m/z = 500 (M+, 3, C32H52O4), 482 (15, M+-H2O), 472 (21, M+-CO), 440 (15, M+-MeCO2H), 291 (10, C19H31O2+), 263 (11, C18H31O+), 249 (23, C16H25O2+), 207 (31, C14H23O+), (100, C5H8O2+), 189 (43, C14H21+), 135 (42, C9H11O+), 109 (34, C7H9O+), 95 (44, C7H11+), 43 (46); HR-EI-MS:m/z = 500.7529, calcd. for C32H52O4; 500.7515 observed.Compound
3. White solid, crystalline, m.p. 258–260°C; [α]D25 + 5.43 (c1.4, CHCl3); UV (MeOH) λmax⁡  (log⁡⁡ε) 308 (4.28) and 214 (4.26), 267 (3.7) nm; IR (dry) νmax⁡ 3348 (OH), 2947, 2877, 1723 (C=C–CO2), 1707 (CO), 1599, 1534 (C=C), 1451, 1256, 767 cm−1;1H-NMR (C5D5N, 300 MHz) δ: 1.87 (2H, ddd,J = 13, 5.2, 3.5 Hz, H-2), 2.25 (2H, dd,J = 4.5, 12 Hz, H-3), 1.74 (1H, d,J = 11.5 Hz, H-5), 2.17 (2H, dd,J = 6.5, 14 Hz, H-6), 2.87 (1H, ddd,J = 16.5, 12.5, 6.5 Hz, H-7α), 2.69 (1H, dd,J = 16.5, 5.0 Hz, H-7β), 1.66 (1H, dd,J = 14, 6.5 Hz, H-9), 4.47 (1H, ddd,J = 3.0, 13.5, 13.5 Hz, H-11), 5.77 (2H, ddd,J = 14.1, 4.5, 12.2 Hz, H-12), 2.05 (1H, dd,J = 12.2, 4.5 Hz, H-13), 2.44 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-15α), 1.46 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15β), 2.52 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16α), 1.56 (1H, ddd, 13.5, 13.5, 3.0 Hz, H-16β), 1.78 (1H, ddd,J = 12.5, 12.5, 3.5 Hz, H-18), 1.71 (1H, ddd,J = 14.5, 11.5, 4.0 Hz, H-19α), 1.19 (1H, dd,J = 14.5, 4.0 Hz, H-19β), 4.57 (1H, ddd,J= 13.5, 13.5, 3.0 Hz, H-2), 2.93 (1H, ddd,J = 13.5, 13.5, 2.5 Hz, H-22α), 1.95 (1H, dd,J = 13.5, 13.5 Hz, H-22α), 1.95 (1H, dd,J = 13.5, 13.5 Hz, H-22β), 1.65 (3H, s, H-23), 1.09 (3H, s, H-24), 1.09 (3H, s, H-25), 1.18 (3H, s, H-26), 1.58 (3H, s, H-27), 1.14 (3H, s, H-28), 1.36 (3H, s, H-29), 0.97s (3H, s, H-30), Ang: 6.13 (1H, qq,J = 6.6 Hz, H-3), 2.01 (3H, dq,J = 6.5, 2.5 Hz, H-4), 1.93 (3H, dqm,J = 6.5, 2.2 Hz, H-5); 13C-NMR (C5D5N, 75 MHz) δ: 214.2 (C-1), 68.4 (C-2), 47.7 (C-3), 45.8 (C-4), 56.9 (C-5), 19.4 (C-6), 35.5 (C-7), 40.3 (C-8), 50.6 (C-9), 38.7 (C-10), 78.1 (C-11), 44.6 (C-12), 54.8 (C-13), 42.3 (C-14), 29.2 (C-15), 28.4 (C-16), 46.7 (C-17), 44.5 (C-18), 27.8 (C-19), 35.4 (C-20), 78.8 (21), 43.5 (C-22), 24.0 (C-23), 23.8 (C-24), 17.6 (C-25), 17.3 (C-26), 24.8 (C-27), 25.6 (C-28), 28.6 (C-29), 24.9 (C-30), Ang: 168.3 (C-1), 128.0 (C-2), 137.3 (C-3), 15.9 (C-4), 20.6 (C-5); EI-MS:m/z = 540 (M+, 3, C35H56O4), 518 (12, M+-H2O), 512 (27, M+-CO), 440 (15, M+-C5H8O2), 291 (10, C19H31O2+), 263 (11, C18H31O+), 249 (23, C16H25O2+), 207 (31, C14H23O+), (100, C5H8O2+), 189 (43, C14H21+), 135 (42, C9H11O+), 109 (34, C7H9O+), 95 (44, C7H11+), 83 (45), 82 (34); HR-EI-MS:m/z = 540.8167, calcd. for C35H56O4; 540.8143 observed.Compound
4. White solid, amorphous, m.p. 267-268°C; [α]D25 + 6.43 (c1.4, CHCl3); UV (MeOH) λmax⁡ (log ε) 347 (4.55), 284 (4.56), 253 (4.22) 234 (2.7) nm; IR (dry) νmax⁡ 2955, 2885, 1738, 1690, 1635, 1593, 1465, 1260, 926, 807, 764 cm−1;1H-NMR (C5D5N, 300 MHz) δ: 1.86 (2H, ddd,J = 13, 5.2, 3.5 Hz, H-2), 2.24 (2H, dd,J = 4.5, 12 Hz, H-3), 1.73 (1H, d,J = 11.5 Hz, H-5), 2.18 (2H, dd,J= 6.5, 14 Hz, H-6), 2.84 (1H, ddd,J = 16.5, 12.5, 6.5 Hz, H-7α), 2.68 (1H, dd, 16.5, 5.0 Hz, H-7β), 1.62 (1H, dd,J = 14, 6.5 Hz, H-9), 4.29 (1H, ddd,J = 3.0, 13.5, 13.5 Hz, H-11), 5.77 (2H, ddd,J = 14.1, 4.5, 12.2 Hz, H-12), 2.06 (1H, dd,J = 12.2, 4.5 Hz, H-13), 2.45 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-15α), 1.45 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15α), 1.45 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15β), 2.51 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16α), 1.55 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16α), 1.55 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16β), 1.79 (1H, ddd,J = 12.5, 12.5, 3.5 Hz, H-18), 1.72 (1H, ddd,J = 14.5, 11.5, 4.0 Hz, H-19α), 1.18 (1H, dd,J = 14.5, 4.0 Hz, H-19β), 4.29 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-21), 2.92 (1H, ddd,J = 13.5, 13.5, 2.5 Hz, H-22α), 2.89 (1H, dd,J = 13.5, 13.5 Hz, H-22β), 1.66 (3H, s, H-23), 1.09 (3H, s, H-24), 1.08 (3H, s, H-25), 1.15 (3H, s, H-26), 1.61 (3H, s, H-27), 1.18 (3H, s, H-28), 1.39 (3H, s, H-29), 0.96 (3H, s, H-30), Ang: 6.13 (1H, qq,J = 6.6 Hz, H-3), 2.01 (3H, dq,J = 6.5, 2.5 Hz, H-4), 1.93 (3H, dqm,J = 6.5, 2.2 Hz, H-5), Ac: 2.29 (3H, s, CH3–CO); 13C-NMR (C5D5N, 75 MHz) δ: 214.8 (C-1), 68.3 (C-2), 47.6 (C-3), 45.9 (C-4), 56.6 (C-5), 19.3 (C-6), 35.5 (C-7), 40.2 (C-8), 50.6 (C-9), 38.8 (C-10), 77.9 (C-11), 44.5 (C-12), 54.7 (C-13), 42.7 (C-14), 29.3 (C-15), 27.9 (C-16), 46.4 (C-17), 44.6 (C-18), 27.6 (C-19), 35.5 (C-20), 81.8 (C-21), 43.6 (C-22), 24.1 (C-23), 23.8 (C-24), 17.5 (C-25), 17.1 (C-26), 24.8 (C-27), 25.7 (C-28), 28.8 (C-29), 24.6 (C-30), Ang: 168.3 (C-1), 128.0 (C-2), 137.3 (C-3), 15.9 (C-4), 20.6 (C-5); Ac: 176.2 (CH3–CO), 22.3 (CH3–CO); EI-MS:m/z = 582 (M+, 3, C37H58O5), 564 (12, M+-H2O), 554 (27, M+-CO), 522 (15, M+-MeCO2H), 464 (15, M+-C5H8O2), 291 (10, C19H31O2+), 263 (11, C18H31O+), 249 (23, C16H25O2+), 207 (31, C14H23O+), (100, C5H8O2+), 189 (43, C14H21+), 135 (42, C9H11O+), 109 (34, C7H9O+), 95 (44, C7H11+), 83 (45), 82 (56), 43 (78); HR-EI-MS:m/z = 582.8334, calcd. for C37H58O5; 582.8343, observed.Compound
5. White solid, crystals, m.p. 264–268°C; [α]D25 + 5.43 (c1.4, CHCl3); UV (MeOH) λmax⁡ (log ε) 349 (4.55), 286 (4.56), 254 (4.22), 247 (1.7) nm; IR (dry) νmax⁡ 3133, 2940, 2870, 1745, 1620, 1527, 1450, 1250, 923, 806, 791, 760 cm−1;1H-NMR (C5D5N, 300 MHz) δ: 1.87 (2H, ddd,J = 13, 5.2, 3.5 Hz, H-2), 4.35 (2H, dd,J = 4.5, 12 Hz, H-3), 1.76 (1H, d,J = 11.5 Hz, H-5), 2.17 (1H, dd,J = 6.5, 14 Hz, H-6), 2.83 (1H, ddd,J = 16.5, 12.5, 6.5 Hz, H-7α), 2.69 (1H, dd,J = 16.5, 5.0 Hz, H-7β), 1.63 (1H, dd,J = 14, 6.5 Hz, H-9), 4.29 (1H, ddd,J = 3.0, 13.5, 13.5 Hz, H-11), 5.78 (2H, ddd,J = 14.1, 4.5, 12.2 Hz, H-12), 2.07 (1H, dd,J = 12.2, 4.5 Hz, H-13), 2.44 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-15α), 1.46 (1H, ddd,J = 13.5, 13.5, 3.1 Hz, H-15β), 2.54 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16α), 1.56 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-16β), 1.78 (1H, ddd,J = 12.5, 12.5, 3.5 Hz, H-18), 1.73 (1H, ddd,J = 14.5, 11.5, 4.0 Hz, H-19α), 1.17 (1H, dd,J = 14.5, 4.0 Hz, H-19β), 4.46 (1H, ddd,J = 13.5, 13.5, 3.0 Hz, H-21), 2.97 (1H, ddd,J = 13.5, 13.5, 2.5 Hz, H-22α), 1.90 (1H, dd,J = 13.5, 13.5 Hz, H-22β), 1.65 (3H, s, H-23), 1.10 (3H, s, H-24), 1.14 (3H, s, H-25), 1.19 (3H, s, H-26), 1.60 (3H, s, H-27), 1.17 (3H, s, H-28), 1.37 (3H, s, H-29), 0.99 (3H, s, H-30), 2 × Bz: 8.04 (1H, d,J = 7.2 Hz, H-2), 7.41 (1H, t,J = 7.2 Hz, H-3), 7.53 (1H, dt,J = 7.2, 2.5 Hz, H-4), 7.41 (1H, t,J = 7.2 Hz, H-5), 8.04 (1H, d,J = 7.2 Hz, H-6); 13C-NMR (C5D5N, 75 MHz) δ: 214.9 (C-1), 68.5 (C-2), 47.5 (C-3), 46.1 (C-4), 56.7 (C-5), 19.8 (C-6), 35.6 (C-7), 40.3 (C-8), 50.6 (C-9), 38.9 (C-10), 77.8 (C-11), 44.6 (C-12), 54.5 (C-13), 42.6 (C-14), 29.1 (C-15), 28.3 (C-16), 46.5 (C-17), 44.9 (C-18), 27.5 (C-19), 35.5 (C-20), 81.8 (C-21), 44.1 (C-22), 24.2 (C-23), 23.5 (C-24), 17.4 (C-25), 17.4 (C-26), 24.9 (C-27), 25.5 (C-28), 28.9 (C-29), 24.8 (C-30), 2 × Bz: 164.6 (C-1), 129.7 (C-2), 130.0 (C-3), 128.4 (C-4), 133.7 (C-5), 128.4 (C-6), 130.0 (C-7); EI-MS:m/z = 666 (M+, 3, C44H54O5), 440 (12, M+-H2O), 430 (27, M+-CO), 422 (15, M+-2H2O), 291 (10, C19H31O2+), 263 (11, C18H31O+), 249 (23, C16H25O2+), 207 (31, C14H23O+), 189 (43, C14H21+), 135 (42, C9H11O+), 105 (34, C7H5O+), 109 (34, C7H9O+), 95 (44, C7H11+), 77 (23, C6H5+), 65 (23, C5H5+); HR-EI-MS:m/z = 666.9283, calcd. for C44H54O5; 666.9273 observed.


## 2.4. Antiringworm Activity of Ethyl Acetate Extract ofCoriandrum sativumLinn.

In this study five sheep infected with ringworm disease were selected. For each infected sheep, ethyl acetate extract (10 gram) was suspended in preboiled water (10 Liter). Each of the infected sheep was given bath with the suspension after every eight hours continually for four days. On the fourth day, four of the infected sheep recovered from ringworms’ infections, while the fifth test sheep took a longer time (10 days) along with vitamin A injection medication.


## 2.5. Antimicrobial Activities of Compounds 1–5

Compounds1–5 were tested for their antibacterial and antifungal activities in terms of inhibition zones, MICs, MBCs, and MFCs using standard procedures [8, 12]. The results of antimicrobial activity of compounds1–5 are displayed in Tables 1, 2, 3, 4, 5, and 6. Briefly, LB medium 20 mL was poured in petri plates and after solidification standard inoculum (100 μL) bacteria/fungi concentration 107 CFU/mL suspension was poured and dried for 5 min. DMSO was used as negative control while standard drugs Miconazole and Imipenem (10 μg/disc) were used as positive control.Table 1
Inhibition zones (mm) of1–5 isolated from C. sativum against bacteria.
Comp./organism
1
2
3
4
5
Std*
E. coli
7.1 ± 1.2
22 ± 2
17 ± 2
8 ± 2
8 ± 2
28 ± 1.2
S.  aureus
3.6 ± 1.2
23 ± 2
16 ± 2
8 ± 2
18 ± 2
31 ± 1.2
P. mirabilis
2.4 ± 1.2
22 ± 2
19 ± 2
21 ± 2
10 ± 2
29 ± 1.2
P. aeruginosa
2.2 ± 1.2
29 ± 2
21 ± 2
29 ± 2
19 ± 2
33 ± 1.2
B. cereus
2.2 ± 1.2
23 ± 2
22 ± 2
8 ± 2
8 ± 2
27 ± 1.2
K. pneumonia
0
0
17 ± 2
8 ± 2
8 ± 2
29 ± 1.2
M. luteus
0
0
8 ± 2
8 ± 2
17 ± 2
37 ± 1.2
E. cloacae
0
0
0
0
0
34 ± 1.2
*Imipenem was used as standard.Table 2
Inhibition zones (mm) of1–5isolated from C. sativum against fungi.
Comp./organism
1
2
3
4
5
Std*
T. rubrum
9.1 ± 2
7 ± 2
3 ± 2
2 ± 2
1 ± 2
29 ± 2
C. albicans
7.6 ± 2
5 ± 2
3 ± 2
2 ± 2
2 ± 2
31 ± 2
M. audouinii
5.4 ± 2
6 ± 2
2 ± 2
2 ± 2
1 ± 2
24 ± 2
C. neoformans
6.2 ± 2
6 ± 2
2 ± 2
3 ± 2
1 ± 2
26 ± 2
T. mentagrophytes
9.0 ± 2
5 ± 2
3 ± 2
2 ± 2
1 ± 2
29 ± 2
E. floccosum
6.3 ± 2
6 ± 2
3 ± 2
1 ± 2
2 ± 2
27 ± 2
M. canis
7.6 ± 2
3 ± 2
3 ± 2
3 ± 2
1 ± 2
28 ± 2
A. niger
2.3 ± 2
0
3 ± 2
3 ± 2
1 ± 2
28 ± 2
*Imipenem was used as standard.Table 3
MICs (µg/mL) of 1–5isolated from C. sativum against bacterial strains.
Comp.
E. coli
P. mirabilis
P. aeruginosa
M. luteus
E. cloacae
K. pneumonia
B. cereus
S.  aureus
1
4 ± 0.5
8 ± 1.5
8 ± 1.5
4 ± 0.5
3 ± 0.5
4 ± 0.5
4 ± 0.5
4 ± 0.5
2
4 ± 0.5
8 ± 1.5
8 ± 1.5
4 ± 0.5
4 ± 0.5
8 ± 1.5
8 ± 1.5
4 ± 0.5
3
4 ± 0.5
16 ± 2.0
8 ± 1.5
8 ± 1.5
5 ± 0.5
16 ± 1.5
16 ± 1.5
4 ± 0.5
4
8 ± 0.5
16 ± 1.5
8 ± 1.0
5 ± 0.5
4 ± 0.5
16 ± 1.5
4 ± 0.5
4 ± 0.5
5
64 ± 2.0
32 ± 2.0
32 ± 2.0
4 ± 0.5
4 ± 0.5
64 ± 2.0
64 ± 2.0
64 ± 2.0Table 4
MICs (µg/mL) of 1–5 from C. sativum against fungal strains.
Comp./organism
C. albicans
C. neoformans
M. audouinii
A. niger
T. mentagrophytes
E. floccosum
M. canis
T. rubrum
1
4 ± 0.5
4 ± 0.5
8 ± 0.5
0
8 ± 0.5
16 ± 0.5
0
16 ± 1.5
2
4 ± 0.5
8 ± 1.5
16 ± 1.5
0
16 ± 0.5
16 ± 0.5
0
16 ± 1.5
3
4 ± 0.5
8 ± 1.5
16 ± 1.5
0
16 ± 0.5
32 ± 0.5
16 ± 1.5
32 ± 2.0
4
8 ± 1.5
8 ± 1.5
16 ± 1.5
0
64 ± 2.5
32 ± 1.5
16 ± 1.5
16 ± 1.5
5
16 ± 1.5
64 ± 2.0
16 ± 1.5
0
32 ± 1.5
32 ± 2.0
16 ± 1.5
8 ± 1.5Table 5
MBCs (µg/mL) of 1–5 from C. sativum against bacterial strains.
Comp./organism
E. coli
P. mirabilis
P. aeruginosa
M. luteus
E. cloacae
K. pneumoniae
B. cereus
S.  aureus
1
8 ± 1.5
16 ± 1.5
16 ± 1.5
8 ± 1.5
0
8 ± 1.5
9 ± 1.5
8  ± 1.5
2
8 ± 1.5
16 ± 1.5
16 ± 1.5
9 ± 1.5
0
16 ± 1.5
16 ± 1.5
8 ± 1.5
3
8 ± 1.5
33 ± 1.5
16 ± 1.5
16 ± 1.5
0
32 ± 1.5
33 ± 1.5
8 ± 1.5
4
17 ± 1.5
32 ± 1.5
16 ± 1.5
0
0
33 ± 1.5
9 ± 1.5
9 ± 1.5
5
12 ± 1.5
65 ± 1.5
15 ± 1.5
0
0
12 ± 1.5
12 ± 1.5
12 ± 1.5Table 6
MFCs (µg/mL) of 1–5 from C. sativum seeds against fungal strains.
Comp.
C. albicans
C. neoformans
M. audouinii
A. niger
T. mentagrophytes
E. floccosum
M. canis
T. rubrum
1
9 ± 1.5
9 ± 1.5
17 ± 1.5
0
17 ± 1.5
31 ± 1.5
0
31 ± 1.5
2
9 ± 1.5
16 ± 1.5
31 ± 1.5
0
31
31
0
31 ± 1.5
3
9 ± 1.5
16 ± 1.5
31 ± 1.5
0
31 ± 1.5
63 ± 1.5
32 ± 1.5
63 ± 1.5
4
16 ± 1.5
16 ± 1.5
31 ± 1.5
0
122 ± 1.5
61 ± 1.5
3 ± 1.52
31 ± 1.5
5
32 ± 1.5
112 ± 1.5
31 ± 1.5
0
62 ± 1.5
61 ± 1.5
32 ± 1.5
16 ± 1.5


## 3. Results and Discussion

The methanolic extract of the whole plant was suspended in water and reextracted with various organic solvents. Ethyl acetate soluble portion was subjected to column chromatography on silica gel; elution was carried out with mixture of petroleum ether/ethyl acetate to yield eight (1–8) fractions. The active fractions 3-4 containing more than two components were subjected to separation on the same silica gel column and eluted with petroleum ether/acetone (7/3) to get semipure triterpenes. Further purification was carried out on preparative HPLC and the column was eluted with MeOH-H2O (80/20) to get compounds (1) (31.3 mg), (tR = 7.9 min) (2), (tR = 8.1 min) (25.5 mg), (tR = 8.4 min) (3) (31.7 mg), (tR = 8.7 min) (4) (15.8 mg), and (tR = 9.3 min) (5) (37.6 mg).Compound1 was obtained as solid needles, m.p. 255–258°C, [α] +5.43°, and displayed an [M]+ ion peak atm/z = 458 in HR-EIMS for [C30H50O3]+. EIMS gave peaks atm/z = 430 [M-28-CO]+, 440 [M-18-H2O]+, andm/z = 412 [M-36-2H2O]+. In IR spectrum, bands were at 1689 cm−1 (ketone) and 3404, 3532 (OH) cm−1. Its UV spectrum displayed an absorption band at 216 nm (1.56) for an isolated cyclic ketone [20]. Compound1 in its 1H-NMR displayed eight singlets (three protons each) at δ 0.97, 1.07, 1.13, 1.16, 1.17, 1.55, 1.66, and 1.38. These singlets were attributed to eight methyl groups present in the basic skeleton of the molecule. Out of these four were gem-methyl. Two methyl groups resonating at δ 1.66, 1.07 (Me-23, Me-24) were gem-methyl present at C-4, other two gem-methyl resonating at 1.38, 0.97 (Me-29, Me-30) were at C-20 and were identified by HMBC (Figure 2). In the same spectrum, there were nine methylene groups that appeared as a complex multiplicity. Peaks at δ 2.22 ddd (J = 13, 5.2, 3.5 Hz) and 1.97 dd (J = 13, 5.2 Hz) were assigned to CH2-2 and CH2-3. Similarly, other peaks integrated for two protons were at δ 1.83 d (J = 11.5 Hz) and 2.17 dd (J = 14, 6.5 Hz) for CH2-6 and CH2-7 and peaks at δ 2.44 ddd (J = 13.5, 13.5, 3.0 Hz), 1.46 ddd (J = 13.5, 13.5, 3.1 Hz), 2.53 ddd (J = 13.5, 13.5, 3.0 Hz), and 1.56 ddd (J = 13.5, 13.5, 3.0 Hz) were assigned to αβ methylene protons of C-15,16. In addition, there was another set of methylene protons that resonated at δ 1.88 ddd (J = 14.1, 4.5, 12.2 Hz, CH2-12), δ 1.71 ddd (J = 14.5, 11.5, 4.0 Hz, H-19α), δ 1.17 dd (J = 14.5, 4.0 Hz, H-19β), δ 2.91 ddd (J = 13.5, 13.5, 2.5 Hz, H-22α), and δ 1.91 dd (J = 13.5, 13.5 Hz, H-22β). 1H-NMR spectrum of compound1 revealed six methine protons. Out of these, two were in the middle region of NMR (δ 3.29 ddd (J = 14, 6.5, 4.2 Hz) and δ 3.46 ddd (J = 13.5, 13.5, 3.0 Hz)) for hydroxymethine moieties at C-11 and C-21 by NOE irradiation. Remaining four methine protons were identified by peaks at δ 1.83 d (J = 11.5 Hz), 1.64 dd (J = 14, 6.5 Hz), 2.01 dd (J = 12.2, 4.5 Hz), and 1.78 ddd (J = 12.5, 12.5, 3.5 Hz). 13C-NMR of compound1 displayed 30 peaks for thirty carbons. In 13C-NMR spectrum, peak appeared at δ 214.7 due to carbonyl group and its position was located with the help of HMBC experiments (Figure 2). Various protons bearing fragments were identified with the help of COSY-45° and HOHAHA experiments (Figure 2). These fragments were connected with each other with the help of HMBC technique (Figure 2).The BB and DEPT13C-NMR spectrum of1 showed 30 carbon signals, 8Me, 9CH2, 6CH, and 7C. The relative stereochemistry of OH groups at C-11 and C-21 in1 was deduced by NOESY spectrum (Figure 3). In this spectrum, interactions were observed among α-oriented H-8, H-13, H-18, Me-23, and Me-27. In the light of these experimental results, it was concluded that compound1 has basic skeleton of triterpene as reported earlier [21–23]. Hence, structure for1 was confirmed as 1-oxo-11β, 21β-dihydroxy-oleanane (Figure 1).Figure 1
Presentation of compounds1–5.Figure 2
COSY and HOHAHA; HMBC interaction in compounds1–5.Figure 3
NOESY interaction in compounds1–5.Compound2 was acetyl derivative of1 as in HR-EIMS molecular ion peak was atm/z = 500.7515 for C32H52O4. 1H-NMR spectrum of2 was very close to1 except for presence of an extra peak at δ 2.29 for three protons. Presence of acetyl moiety in2 was proved by 13C-NMR spectrum. In this spectrum, in addition to other peaks, there were two more peaks present at δ 176.2 and δ 22.3 which were assigned to an acetyl group. Position of the acetyl group was located at C-21 with the help of HMBC and NOE experiments (Figures 2 and 3). The DEPT 13C-NMR showed 32 carbon signals, 9Me, 9CH2, 6CH, and 8C. Hence, the proposed structure of2 was 1-oxo-11β-hydroxy-21β-O-acetyloleanane (Figure 1).Compound3 was obtained as white solid, crystalline, m.p. 258–260°C, [α] +5.43°. In UV spectrum displayed λmax⁡ at 308 (4.28) and 214 (4.26); 267 (3.7) nm indicated presence of α,β-unsaturated ester as a chromophore. Compound3 showed molecular ion [M+] peakm/z = 540.8143 in HR-EIMS corresponding to C35H56O4. Therefore, compound3 was suggested as an angeloyl derivative of1. 13C-NMR spectra of3 were similar to1 but there were peaks for angelate moiety: H-3 (δ 6.13, 1H, qq,J = 6.6 Hz), CH3-4 (δ 2.01, 3H, dq,J = 6.5, 2.5 Hz), and CH3-5 at δ 1.93 [3H, dqm,J = 6.5, 2.2 Hz]; δ 168.3 (C-1), 128.0 (C-2), 137.3 (C-3), 15.9 (C-4), and δ 20.6 (C-5). Therefore,3 was considered as angeloyl derivative of1. Position of angelate was inferred at C-21 by HMBC and NOE interaction between H-21 and an angeloyl proton H-3. The DEPT 13C-NMR spectrum of3 showed 35 carbon signals, 10Me, 9CH2, 7CH, and 9C. On the basis of these results, structure of3 was concluded as 1-oxo-11β-hydroxy-21β-O-angeloyloleanane.Compound4is colorless, amorphous powder, m.p. 267-268°C, [α] +6.43°; UV spectrum showed compound is UV active by displaying λmax⁡ at 347 (4.55), 284 (4.56), 253 (4.22), and 234 (2.7) nm typical for angelic moiety. IR spectrum showed absence of OH groups in the compound. There were peaks at νmax⁡ 1723 and 1706 cm−1 for ester carbonyl groups. Molecular formula C37H58O5 of compound4 was established with the help of HR-EIMS, displaying molecular ion peak atm/z = 582. In EIMS, fragmentation peak appeared atm/z = 522 [M+-60] indicating presence of an acetate. Another peak atm/z = 482 was due to loss of angelic acid from the parent molecule. Therefore, it was considered as diester of acetic acid and angelic acid of1.NMR (1H- and 13C-) spectra of4 were almost same as3 except for presence of an extra acetyl moiety. It was also proved by DEPT 13C-NMR spectrum which showed 37 carbon signals, 11Me, 9CH2, 7CH, and 10C. Position of the acetate moiety was deduced at C-21 by HMBC and NOE interaction. Thus, structure of 4 was concluded as 1-oxo-11β-O-acetyl-21β-O-angeloyloleanane.Fractions 6–8 were combined and chromatographed over silica gel column. Active fraction was subjected to a reversed-phase preparative HPLC and afforded a pure compound5 (tR = 9.3 min). It was white solid, m.p. 264–268°C, optically active and displayed [α] +5.43° in polarimeter. Presence of chromophore was revealed by displaying λmax⁡ peaks at 349 (4.55), 286 (4.56), 254 (4.22), and 247 (1.7) nm in UV. Presence of aromatic moiety was indicated by IR spectrum. Peak at νmax⁡ 3133 (Ar–H stretching) and peaks at 806, 791, and 760 cm−1 suggested monosubstituted benzene ring. Presence of the ester carbonyl was proved by peaks at 1745 and 1725 cm−1. Molecular composition C44H58O5 of5 was proposed by HR positive ion FAB-MS in which it indicated an [M]+ ion peak atm/z = 667. In EIMS, fragmentation pattern revealed presence of two benzoic acid species by displaying peaks atm/z = 544 [M-122-benzoic acid]+ and 422 [M-2 × 122-2 × benzoic acid]+. The 1H, 13C-NMR spectra of 5 were close to1 except for presence of (double) peaks in aromatic region at δ 8.04 (2H, d,J = 7.2 Hz), 7.41 (2H, t,J = 7.2 Hz), 7.53 (1H, dt,J = 7.2, 2.5 Hz) δ 164.6 [2 × CO s]), 2 × 5CH (δ 129.7 d, 130.0 d, 130.0 d, 128.4 d, 128.4 d), and 2 × C (133.7). Therefore, it was suggested as diester of benzoic acid of1(Figure 1). Positions of the ester moieties were deduced by HMBC interactions. The H-11 (δ 4.29) displayed interaction with carbonyl carbon of ester (δ 164.6); similarly, H-21(δ 4.46) displayed interaction with other carbonyl (δ 164.4) in HMBC spectrum (Figure 2). Therefore, ester moieties were suggested at C-11 and C-21, respectively.On this basis, compound5 was declared as 1-oxo-11β,21β-O-dibenzoyloleanane. The DEPT 13C-NMR spectrum of5 showed 44 carbon signals, 8Me, 9CH2, 16CH, and 11C. On this basis, compound5 was declared as 1-oxo-11β,21β-O-dibenzoyloleanane. According to our knowledge, compounds1–5 have not been reported earlier from plant kingdom and hence can be declared as new entities.Ringworm (Club Lamb Fungus) disease in sheep is common and is a matter of intense concern all over the world. The disease once its symptoms appear is not restricted to sheep hosts but is fast transmitted to other animals and human beings who rear up the infected animals. Ethyl acetate extract of whole plant ofCoriandrum sativum Linn. was effectively employed to heal ringworm disease in sheep.Isolated compounds displayed activity against used microbes. Compounds2 and4 were most active againstP. aeruginosawith inhibition zones (29 mm), whereas standard (Imipenem) displayed 31 mm. Compounds1–5 displayed no activity againstE. coli. Compounds2 and3 were found to be also inactive againstM. luteus and K. pneumoniabut displayed activity againstE. coli(inhibition zones 22 and 17 mm) compared to standard (33 mm). Compounds2–4 also displayed moderate activity againstP. mirabilis, P. aeruginosa, andB. cereus. Same trend of biological activities of compounds1–5 isolated fromC. sativum was found in terms of MICs against used microbes.


## 4. Conclusions

The ethyl acetate extract exhibited excellent antiringworms activity and antibacterial activity. We have for the first time isolated oleanane type compounds (1–5) fromC. sativum Linn. This study reveals that these oleananes could be potential bioactive compounds that will be useful for the development of new tools for the control of diseases.



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*Source: 396436-2014-11-18.xml*