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Copolymers of vinylpyrrolidone with coumarin, umbelliferone, esculetin, osthole, ostholic acid, meranzin hydrate, isoimperatorin, oxypeucedanin hydrate, isooxypeucedanin, gosferol, pranchimgin, and marmesin have been synthesized and their structures have been studied.

From a methanolic extract of the roots ofPrangos ferulacea (L) Lindl., growing in the Nakhichevan ASSR, in addition to the compounds found previously, another six substances (I–VI) have been isolated, of which (I) and (VI) have been identified as, respectively, meransin hydrate monoacetate, found for the first time in nature, and umbelliferone: and in the study of a crystalline mixture obtained from the resin of the roots of the plant under investigation collected in Armenia, two coumarins — (VII) and (VIII) — have been isolated.

The O -dealkylation activity for 7-ethoxycoumarin of mouse small intestine was determined by recording the surface fluorescence of the umbelliferone formed in everted intestinal segments. A simple holding device suitable for commercially available cuvettes has been designed. After hypotonic shocking of the mucosal ccells the substrate was added in 0·1 M Tris buffer pH 7·9 causing a linear increase in fluorescence for more than 15 min. Pretreatment of mice with phenobarbital increased the activity several fold, the induction following a biphasic course. The maximal rate of O -dealkylation by the entire small intestine was calculated as 19 nmoles min −1 . In agreement with previous results obtained with homogenates of small intestine the highest activity was located at a distance of 8–12 cm from the pylorus. Inhibition experiments indicate the involvement of cytochrome P-450.

Endogenous growth substances were estimated byAvena coleoptile test in hypocotyls of cuttings ofPhaseolus vulgaris which were treated with Umbelliferone, a coumarin which has been found to have a stimulating effect on rooting. A similar growth promoting zone which corresponded chromatographically to indole-3-acetic acid, was found in both treated and control cuttings. A growth-inhibiting zone detected in control extracts was absent in UM treated cuttings. The results are discussed in relation to Umbelliferone effect on rooting stimulation.

Cultures of normal tobacco callus tissue 3 weeks after subculture contain free and bound forms of cinnamic, caffeic, and ferulic acids and bergapten, and bound umbelliferone, esculetin, and scopoletin. Cultures of tobacco crown gall tumor tissue lack cinnamic acid, umbelliferone, and bergapten; but they contain free as well as bound esculetin and scopoletin, and higher total levels of each of these coumarins than normal tissue. It is suggested that biosynthesis of coumarins in tumor cells has been diverted from the umbelliferone–furanocoumarin pathway in favor of esculetin and scopoletin.

From the roots ofFerula kopetdaghensis Eug. Kor. a new coumarin has been isolated which has been called kopetdaghin. On the basis of the hydrolysis products, the results of hydrogenation and acetylation, and spectroscopy, it has been established that kopetdaghin is an ether of 7-hydroxycoumarin and 1-(3-hydroxy-2,2,6-trimethylcyclohex-5-enyl)-3-methylpent-2-en-1-ol, i.e., an isomer of farnesiferol B.

The synthesis and properties (NMR, UV absorption, fluorescence emission spectra, and fluorescence quantum yield) of umbelliferone, 4-isopropylumbelliferone, and 4-carboxymethylumbelliferone are described. The optimal conditions for the formation of these umbelliferones from substituted malic acids and resorcinol have been determined. This forms the basis for a rapid and sensitive fluoremetric assay of enzymes catalyzing aldol condensation reactions involving acetyl coenzyme A.

Several esters were prepared as substrates for cholinesterase: the acetate, propionate and butyrate esters of N-methyl indoxyl, umbelliferone acetate, and the acetate and butyrate esters of 4-methyl umbelliferone. Comparison of these substrates with other fluorogenic esters: indoxyl acetate, indoxyl butyrate, resorufin acetate, B-carbonaphthoxycholine, and β-naphthyl acetate indicated that N-methyl indoxyl butyrate was the best substrate for pseudo cholinesterase. Analysis of as little as 5 × 10−6 units per ml. of cholinesterase can be performed by a direct initial reaction-rate method in 2–3 minutes with an accuracy and precision of about 1.5%.

Evidence was found for the existence in Lavandula officinalis Chaix of 2-glucosyloxy-4-methoxy- cis -cinnamic acid 1 , which yields 7-methoxy showed that D -glucose, L -phenylalanine, and cinnamic acid are common precursors of coumarin and herniarin, but that o - and p -hydroxycinnamic acids are selectively converted to coumarin and herniarin, respectively. Data obtained were consistent with the occurrence of herniarin biosynthesis according to the partial sequence: Cinnamic acid→ p -coumaric acid→ p -methoxycinnamic acid→2-glucosyloxy-4-methoxy- trans -cinnamic acid→-2-glucosyloxy-4-methoxy- cis -cinnamic acid. Umbellic acid (2,4-dihydroxy- trans -cinnamic acid) and umbelliferone were utilized to a minor degree. An improved preparation of umbelliferone-2- 14 C, and syntheses of umbellic acid-carboxy- 14 C, p -methoxycinnamic acid-α- 14 C, 1 and 2-glucosyloxy-4-methoxy- trans -cinnamic acid-α- 14 C are described.

1. Fluorescence changes of umbelliferone have been used as indicator for pH changes in the outer phase of the functional membrane of photosynthesis. In combination with the repetitive pulse technique it is possible to measure the pH change which is produced during one turn-over of the molecular machinery (Δ pH ≈ 10−4). The analysis of this primary pH changes gives insight in the primary ionic events in general. 2. The rise of the primary pH formation takes place in 20-30 ms. The decay is at 20°C of first order with k ≈ 0.7 s−1 or τ½ ≈ 1 s. 3. The rise of the pH formation corresponds to the decay of the electrical field across the membrane (pHout 7, pHin 7, isotonic solution). 4. The amplitude of the primary pH change indicates the ratio of the field driven K⊕ and H⊕ efflux across the membrane **. This has been proved by membrane active substances which are specific for K⊕ and H⊕ permeabilities respectively. 5. The ratio of the field driven K⊕ and H⊕ efflux depends on the pHin. At pHin 7 (pHout 7) the field drives 100% K⊕. With decreasing pHin the efflux shifts from K⊕ to H⊕ (at pHin 5, pHout 7 50% K⊕ and 50% H⊕). 6. The field driven K⊕ efflux causes the formation of the pH changes. The field driven H⊕ efflux causes no pH changes. 7. The K⊕ channel and the H⊕ channel and its regulation by pHin lead to an extended concept of the coupling of the primary events in photosynthesis. This is discussed in chapter II and fig. 4.

A study of growth substances in cuttings ofSalix viminalis L. andFicus carica L. has been made by extraction with methanol, paper and thin layer chromatography, ultraviolet, infrared and fluoresence spectroscopy andAvena coleoptile straight growth test. FromS. viminalis, p-hydroxybenzoic, protocatechuic, two hydroxycinnamic acids and catechol were isolated and their growth properties studied. The growth stimulating zone in the acidic biohistogram may be explained by the presence of the first two acids. In addition to the compounds characterized, syringic, vanillic andp-coumaric acids were isolated from the hydrolysate. FromF. carica, syringic, vanillic,p-coumaric acids and umbelliferone have been isolated and identified. The relevant growth-promoting activity in the acidic biohistogram seems not to be completely due to the presence of the acids mentioned, but no IAA has been found.

The biosynthesis of angular furanocoumarins from umbelliferone in Angelica archangelica and Heracleum lanatum has been shown by isotope studies to parallel that of linear furanocoumarins, via the dihydrofuranocoumarin columbianetin. Angelicin, the first angular furanocoumarin formed, is converted by hydroxylation and O-alkylation to a range of substituted angelicins. A detailed biosynthetic route to angular furanocoumarins, consistent with known stereochemistry, is proposed. By variations of this pathway, it is possible to account for the elaboration of pyranocoumarins, furanoquinoline alkaloids, furanochromones, and possibly other plant products which frequently co-occur with furanocoumarins.A separation of furanocoumarins by column absorption chromatography on silicic acid, and a synthesis of [2-14C]angelicin, are described.

Umbelliferone and a new monohydroxymonomethoxycoumarin have been isolated from Hydrangea macrophylla Ser. The new coumarin has been shown to be a derivative of daphnetin. It is present largely in a bound form in all parts of the plant and is especially abundant in the roots. Chromatographic characteristics of the compound as well as those of several known coumarins are included in this report.

We have isolated from Cephalanthus spathelliferus (Rubiaceae) five compounds: umbelliferone, skimmin, 7,4′-dimethylkaempferol,7,4′-dimethylaromadendrin and its 5-glucoside. The latter two are new compounds.

The results of a complete study of 8 substrates for acid and alkaline phosphatase indicated 7-hydroxycoumarin (umbelliferone) phosphate to be the best substrate for the analysis of these enzymes. Using this ester, from 10−6 to 2 × 10−2 units per ml. of alkaline phosphatase and 10−5 to 0.06 units per ml. of acid phosphatase can be determined with an accuracy and precision of about 1.5%. Samples of serum as small as 1 μ;1. can be assayed. Analysis is performed by a direct initial reaction rate method in 2–3 minutes.

1. 1. Data are presented which indicate that umbelliferone phosphate is a substrate for the K+-activated phosphatase which is found in preparations of (Na+ + K+)-activated ATPase (ATP phosphohydrolase, EC 3.6.1.3). 2. 2.|The hydrolysis of umbelliferone phosphate by this enzyme resembles that of p-nitrophenyl phosphate in that it requires Mg2+ and is activated by K+ and this activation is inhibited by ouabain. 3. 3. At low concentrations of K+, it is further activated by ATP in the presence of Na+ and this extra activation is inhibited by oligomycin. 4. 4. The fluorimetric assay method, which has previously been used in the determination of acid and alkaline phosphatases, is simple, convenient and highly sensitive.

14 C-Labelled 7-demethylsuberosin (DMS) was a precursor of linear furanocoumarins in Conium maculatum and Heracleum lanatum (Umbelliferae), and in Ruta graveolens (Rutaceae), but not in Coronilla glauca (Leguminosae). Trapping experiments with 14 C-umbelliferone over 3- to 24-hr periods revealed that DMS is rapidly metabolized, and that in short experiments its specific activity relative to the linear furano- coumarins′ is that of an intermediate. The specific activity of umbelliferone exhibited anomalies attributed to pool compartmentation. Analogous but less extensive experiments established that osthenol is an intermediate in angular furanocoumarin biosynthesis, but is much less rapidly metabolized than DMS. No significant incorporation into furanocoumarins of 14 C from labelled l -valine, l -leucine, acetate, 2,3-di-hydroxyisovaleric acid, senecioic acid, isopentenylpyrophosphate, or mevalonate could be demonstrated. A synthesis of [1- 14 C]2,3-dihydroxyisovaleric acid is described.

From the non-volatile, neutral fraction of Asa foedita (Umbelliferae) 3 isomeric compounds C24H30O4, designated farnesiferol A, B, and C, have been isolated. Farnesiferol A (I) is a mixed ether of umbelliferone with a sesquiterpene moiety of the bicyclofarnesylic type. It might arise biogenetically by oxidative cyclisation of the known umbelliprenine (XXXII).

Labelling experiments have shown that umbelliferone, and to a lesser degree p -coumaric acid, are precursors of bergapten, imperatorin, isopimpinellin and xanthotoxin in Pastinaca sativa L. C-2 and C-5 of mevalonate, which has been proposed as a precursor of two furan ring carbons and of dimethylallyl substituents, were both incorporated into furanocoumarins with low efficiency—poorer than that of acetate. Feeding experiments with [2- 14 C]5-methoxy-7-hydroxycoumarin provided no evidence that it is a major intermediate in bergapten formation, supporting the theory that carbon side-chain attachment precedes further oxygenation of umbelliferone in furanocoumarin biosynthesis.

The sodium salts of 4-hydroxycoumarin, 5-hydroxy[2′, 3′∶7, 6-furo]coumarin (bergaptol), 6-dyroxycoumarin, 7-hydroxycoumarin (umbelliferone) and 8-hydroxy[2′, 3′∶7, 6-furo]coumarin (santhotoxol) have been prepared and their IR and UV spectra determined. The position and intensity of the IR absorption bands of these salts (both in the crystalline state and in D2O solution) show that the ions of 7-hydroxycoumarin, 5-hydroxyfurocoumarin, and 5-hydroxycoumarin exist in a form intermediate between quinoid and coumarin, and the ions of 6-hydroxycoumarin and 8-hydroxyfurocoumarin exist as coumarins.

A number of umbelliferone derivatives with acetyl, benzoyl, carbethoxyl or carboxyl groups in the 3-position have been prepared of which 6 are new. The fluorescence exhibited by these has been studied and the results are discussed.

Phenolic components in the plant were investigated to give phyllodulcin, phyllodulcin-8-0-β-D-glucoside, hydrangenol, umbelliferone, p-hydroxybenzoic acid, protocatechuic acid, gallic acid, methyl chlorogenate, chlorogenic acid, kaempferol, quercetin, isoquercitrin, rutin and three new components, hydrangea glucosides A, B, and C. On the basis of chemical and spectral data hydrangea glucosides A (I), B (II), and C (III) were characterized to 2-[β-(4'-hydroxyphenyl)-β-hydroxyethyl]-6-hydroxybenzoic acid-6-0-β-D-glucopyranoside, 2-[β-(4'-hydroxyphenyl)-β-hydroxyethyl]-6-hydroxybenzoic acid-β-O-β-D-glucoside and p-hydroxybenzaldehyde-O-β-D-glucopyranoside, respectively.

Cytological observations in Vicia faba using asafoetida revealed that asafoetida could cause aberrations and be used as a mutagen. About 0.1% to 0.2%/ of natural mutations do occur in Vicia faba. Since water treatment caused some aberrations it may be presumed that certain chemicals which are present in seed coat of Vicia faba must be soluble in water and in turn acting as automutagenic substances. Coumarins cause mutations. Since asafoetida is having umbelliferone which is 7-hydroxy coumarin it must be acting as a mutagen. So asafoetida can be used as a cheaply available mutagen. But how far this mutagen can be used in boosting up Agricultural production needs further investigation.

14 C-labelled p -coumaric and o -coumaric acids, when administered to cut shoots of Hydrangea macrophylla Ser., were both converted with good efficiency to umbelliferone. No randomization of 14 C could be detected when the umbelliferone formed from o -coumaric acid was chemically degraded. Trapping experiments showed that both coumaric acids can be converted to umbelliferone via umbellic (2,4-dihydroxycinnamic) acid, which is itself an extremely efficient umbelliferone precursor when fed in labelled form. When cinnamic acid-β- 14 C was administered to H. macrophylla with non-radioactive o -coumaric acid, no 14 C could be detected in the recovered o -coumaric acid, although p -coumaric acid, umbelliferone, and hydrangetin (7-hydroxy-8-methoxycoumarin) were radioactive. The results establish the existence of a pathway: cinnamic acid → p -coumaric acid → umbellic acid → umbelliferone in Hydrangea , but the normal participation of o -coumaric acid in umbelliferone biosynthesis cannot be considered proved. The results of one experiment indicate that cinnamic and p -coumaric acids are also precursors of hydrangetin.

On the basis of chemical reactions and spectral data, the structures of the umbelliferone ethers of 3-oxo-1,2,5,9-tetramethyldecal-5-ylmethanol and of 3-hydroxy-1,2,5,9-tetramethyldecal-5-ylmethanol have been proposed for kamolone and kamolol, respectively.

The structure of the coumarin mogoltin isolated previously from the roots ofPeucedanum mogoltavicum Korov has been established on the basis of NMR, mass, and IR spectroscopy and the preparation of derivatives. It has been shown that mogoltin is the ether of umbelliferone and 6,7-dihydroxy-2,5,5,8a-tetramethyl-δ2-octahydro-1-naphthylmethanol with the equatorial arrangement of the hydroxy groups.

The roots ofFerula kopetdaghensis Eug. Kor. have yielded five coumarins — umbelliferone and four new compounds (kopetin, kopeodin, kopeolin, and kopeoside).

Umbelliferone (UM) stimulated rooting of Phaseolus vulgaris L. cuttings from 7-day-old seedlings at 50 ng/ml and 100 ng/ml, whereas at 150 ng/ml root formation was inhibited. Concentrations lower than 50 ng/ml showed no effect on rooting. Rooting stimulation caused by UM was not modified by 1 ng/ml indole-3-acetic (cid (IAA) or lower doses either when applied before or simultaneously, but an exuberant formation of roots occured in cuttings treated with 1 ng/ml IAA, after being dipped in UM (100 ng/ml) for 1 day. UM inhibited root grrowth, but did not affect the growth of cuttings.

1. The fluorescence characteristics of 3- and 7-hydroxycoumarin, and 7-hydroxy-and 7-methoxy-4-methylcoumarin, have been determined. 7-Hydroxycoumarin shows excited-state ionization from pH1 to 9. 2. A sensitive and specific fluorimetric method for the determination of 7-hydroxycoumarin (umbelliferone), and its application to liver homogenates and other tissue preparations, are described. 3. The enzymic hydroxylation of coumarin to 7-hydroxycoumarin has been studied by this method and the optimum conditions have been determined for rabbit-liver preparations. The enzymic activity was found in the microsomal fraction and required NADPH(2) and oxygen. Activity with NADH(2) was one-third of that with NADPH(2). 4. Addition of NADP was necessary for full activity of 10000g supernatant preparations of liver. Nicotinamide added during preparation preserved coenzymic activity in tissue stored at -12 degrees . Glucose 6-phosphate had no effect on the activity of stored or fresh tissue. 5. Inhibition occurred with p-chloromercuribenzoate, and with the usual inhibitors of the microsomal drug-metabolizing enzymes, SKF acid, SKF 525A, and Lilly 7132, but not with 2,2'-bipyridyl. 6. Liver homogenates from rabbit, guinea pig, coypu, cat and pigeon showed activity, but preparations of rat or mouse liver, and of locust fat bodies, did not hydroxylate coumarin to umbelliferone. The enzyme system was absent from rat-liver homogenates and microsomal preparations. Moreover, rat liver also contained inhibitors of the rabbit-liver coumarin-7-hydroxylase system and of the further metabolism of umbelliferone by guinea-pig liver. Guinea-pig-liver preparations hydroxylated coumarin to umbelliferone and then converted this product into its glucuronide. 7. The coumarin-7-hydroxylase activity of female rabbit liver was two to three times that of male rabbit liver.

The action of mercuric acetate in methyl alcoholic solution on (1) umbelliferone, (2) 4-methylumbelliferone and (3) 4: 7-dimethylcoumarin has been investigated. With umbelliferone addition at the double bond and substitution in positions 6 and 8 take place whereas in the case of its 4-methyl derivative position 8 alone is mercurated to yield a monoacetoxymercuri-compound. In regard to the last compound the reaction stops with addition at the double bond and mercuration of the 6th position, the position 8 escaping the attack of the reagent. It is concluded that owing to the influence of the substitutent groups the reactivity of the three active centres of the coumarin molecule are differently affected.

The metabolic fate of umbelliferone was studied by the quantitative analysis of urinary excrete of rabbits. The orally administered umbelliferone was converted into glucuronide (44.32%) and ethereal sulfate (19.10%) accompanying unchanged umbelliferone (20.23%). An extractor of high efficiency was designed to be employed for the continuous extraction of urinary metabolites.

Trapping experiments with [2-14C]umbelliferone and 2′,3′-dihydropsoralen (DHP) administered to Ruta graveolens yielded negative results over metabolic periods of 5–24 h. Specific radioactivities of psoralen were comparable to those previously observed, but those of DHP were about two orders of magnitude lower. These results do not support the theory (Z. Naturforsch. 25b, 700 (1970)) that DHP is the immediate precursor of psoralen in furanocoumarin biosynthesis, and it is proposed instead that DHP is not on the main pathway, but is in an equilibrium with psoralen that greatly favors dehydrogenation.

Tracer experiments involving the feeding of 14C-labelled cis-p-coumaric acid derivatives to intact Hydrangea macrophylla and Lavandula (Munstead) plants have provided strong evidence against the oxidative cyclization theories proposed for the biosynthesis of 7-oxygenated coumarins. In contrast, trans-p-coumaric acid and its β- d -glucoside were very well incorporated into umbelliferone in Hydrangea while trans-o-coumaric acid and its β- d -glucoside were very poorly incorporated. Evidence was obtained that “free” umbelliferone present in the plant extracts is almost entirely an artefact. The primary site of synthesis of umbelliferone appears to be the leaves.

The inhibitory effects of umbelliferone and a number of its alkyl and acyl derivatives on the growth of a variety of bacteria, yeasts and molds have been examined. Herniarin and similar alkyl ethers are more effective antimicrobial agents than umbelliferone.

Two umbelliferone-producing glucosides have been found in Hydrangea macrophylla leaf extracts. By chromatographic characterization and comparison with the synthetic compounds, they have been identified as skimmin (7-β- d -glucosyloxycoumarin) and cis-2,4-di-β- d -glucosyloxycinnamic acid. Skimmin is by far the major component. Both of these compounds are intermediates in a proposed scheme for umbelliferone biosynthesis consistent with the results of tracer studies. A new technique was developed involving thin-layer chromatography on cellulose powder which has made possible the separation of diglucosides, monoglucosides and phenolic aglycones. A rationale for the observed chemical properties of o-glucosyloxycinnamic acids is discussed.

Aus dem Lipoidanteil der Fruchte von Anethum graveolens (L.) und Coriandrum sativum (L.) konnten durch Atherextraktion und anschliesende Fraktionierung uber einer Aluminiumoxyd-Saule folgende Substanzen isoliert werden: Fructus Anethi: Ein Wachs (Zusammensetzung: C16-, C18-, C22-, C24-, C26- und C28-Sauren sowie C24-, C26-, C28-, C30- und C32-Alkohole), die Cumarine Bergapten und Umbelliprenin, eine nicht genau identifizierte Verbindung (Phytofluen?) sowie γ-Sitosterin. Fructus Coriandri: Eine nicht genau identifizierte Verbindung (Phytofluen?), γ-Sitosterin, Umbelliferon und Scopoletin. Some Lipid Containing Substances from Fruits of Anethum Graveolens L. and Coriandrum Sativum L. From the lipid part of fruits from anethum graveolens L. and coriandrum sativum L. the following substances could be isolated by ether extraction followed by fractionation on an aluminium oxide column: Fructus anethi: a wax (composition: C16-, C18-, C22-, C24-, C26- and C28-acids and C24-, C26-, C28-, C30- and C32- alcohols), the coumarins bergapten and umbelliprenin, an unidentified compound (phytofluen?) as well as γ-sitosterol. Fructus coriandri: an unidentified compound (phytofluen?), γ-sitosterol, umbelliferone and scopoletin.

Umbelliferone has been found to be a useful chemical marker in Hieracium. Its presence in nine of thirty taxa examined confirms the separation of the genus Pilosella previously proposed on morphological and genetical grounds.

The results of a complete study of 12 substrates for lipase indicated 4-methyl umbelliferone heptanoate to be the best substrate for the analysis of this enzyme. Using this ester, from 0.000020 to 0.066 units per ml. of porcine pancreas lipase can be determined with an accuracy and precision of about 1.5%. Analysis is performed by a direct initial reaction rate method in 2–3 minutes.