UNIVERSITI PUTRA MALAYSIA

ENZYMATIC SYNTHESIS, CHARACTERIZATION AND ANALYTICAL APPLICATIONS OF FATTY HYDRAZIDES FROM PALM OIL

SHARIFAH MOHAMAD

FS 2008 34

ENZYMATIC SYNTHESIS, CHARACTERIZATION AND ANALYTICAL APPLICATIONS OF FATTY HYDRAZIDES FROM PALM OIL

By

SHARIFAH MOHAMAD

Thesis Submitted to the School of Graduate Studies, Universiti Putra Malayisa, in Fulfilment of the Requirements for the Degree of Doctor of Philosophy

May 2008

Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfilment of the requirement for the degree of Doctor of Philosophy

ENZYMATIC SYNTHESIS, CHARACTERIZATION AND ANALYTICAL

APPLICATIONS OF FATTY HYDRAZIDES FROM PALM OIL

By

SHARIFAH MOHAMAD

May 2008

Chair: Professor Dato’ Wan Md. Zin Wan Yunus, PhD

Faculty: Faculty of Science

Hydrazides with the general formula R-CO-NHNH2, have received a lot of attention due

to their applications in biological, organic synthesis and analytical chemistry fields. The

preparation of fatty hydrazides from palm oil has been developed in this research. Fatty

hydrazides (FH) and fatty phenyl hydrazides (FPH) were successfully synthesized from

palm oil as a raw material by one-step lipase catalyzed reaction. FTIR and CHN

elemental analyses were carried out to determine the presence of the hydrazides in the

products. The method offers several advantages such as renewable and abundant of raw

materials, simple reaction procedure and high yield of products.

The application of the products as a reagent was carried out based on the ability of the

FH and FPH to form complexes with some metal ions. FH was successfully used as an

extractant for extraction and separation of copper(II). The separation of Cu(II) is

ii

possible from other metal ions such as Co(II), Cr(VI), Ni(II), Zn(II) and Fe(III) at pH

5.5-6.5. A preconcentration method was proposed for the determination of Cu(II) in

water samples. It was shown that the extraction from aqueous phase containing Cu(II)

with organic phase containing FH and then stripping the organic phase with 2 M of HCl

solution give a solution of Cu(II) 10 fold in concentrations.

FH was also successfully used as an extractant for extraction and separation of Mo(VI).

This metal successfully separated from other metal ions such as Ni(II), Co(II), Al(III),

Fe(III) and Mn(II). Quantitative stripping of Mo(VI) ion from the organic phase can be

carried out using 2 M ammonium hydroxide. This proposed method was applied for the

recovery of Mo(VI) from synthetic mixture and the results showed that more than 90%

recovery of Mo(VI) is achieved and the metal ion solution is free from the impurities.

FH and FPH were also evaluated as an extractant for extraction and separation of

gold(III) from chloride media. This metal was extracted quantitatively from hydrochloric

media at 0.001-0.1 M into the organic phase. Gold(III) was successfully separated from

Cu(II), Ni(II), Zn(II), Co(II) and Fe(III) by using FH or FPH. Quantitative stripping of

gold(III) from the organic phase can be carried out using 2.0 M thiourea in 1.0 M HCl

solution. The extraction and separation of gold(III) by FH or FPH was applied to

separate and recover pure gold(III) from synthetic mixtures.

iii

FPH synthesized from palm olein was also successfully used as a new reagent for the

determination of V(V) by spetrophotometric method. The method is based on the

colored complex of vanadium(V)-FPH. The metal ion forms dark brown colored

complex which has an absorption maximum at 405 nm. Beer’s law is valid over the

concentration range of 0.2-20 mg/L and the limit of detection of this method is

0.01 mg/L. The characteristics of this developed method are simple, good selectivity,

high sensitivity and rapid.

iv

Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai memenuhi keperluan untuk ijazah Doktor Falsafah

SINTESIS BERENZIM, PENCIRIAN DAN PENGGUNAAN ANALISIS HIDRAZIDA LEMAK DARIPADA MINYAK KELAPA SAWIT

Oleh

SHARIFAH MOHAMAD

Mei 2008

Pengerusi: Professor Dato’ Wan Md Zin Wan Yunus, PhD

Fakulti: Fakulti Sains

Hidrazida dengan formula umumnya R-CO-NHNH2, mendapat banyak perhatian kerana

pengunaannya dalam bidang biologi, sintesis organik dan kimia analisis. Penyediaan

hidrazida lemak daripada minyak kelapa sawit telah dimajukan di dalam penyelidikan

ini. Hidrazida lemak (FH) dan hidrazida fenil lemak (FPH) telah berjaya disintesiskan

daripada minyak kelapa sawit sebagai bahan asas dengan tindakbalas satu peringkat

menggunakan enzim lipase sebagai mangkin. Analisis FTIR dan analisis unsur CHN

telah dijalankan untuk menentukan kehadiran kumpulan hidrazida di dalam produk.

Kaedah ini mempunyai beberapa kelebihan seperti bahan asas yang berterusan dan amat

banyak, prosedur tindak balas yang mudah dan hasil produk yang tinggi.

v

Penggunaan produk sebagai reagen telah dijalankan berasaskan kebolehan FH dan FPH

membentuk kompleks dengan beberapa ion logam. FH telah berjaya digunakan sebagai

pengekstrak untuk pengekstrakan dan pemisahan kuprum(II). Pemisahan kuprum(II)

adalah mungkin dari ion logam yang lain seperti Co(II), Cr(VI), Ni(II), Zn(II) dan

Fe(III) pada pH 5.5-6.5. Kaedah pra-pemekatan dicadangkan untuk penentuan Cu(II) di

dalam sampel air. Ini telah ditunjukkan dengan pengekstrakan fasa akueus yang

mengandungi Cu(II) dengan fasa organik yang mengandungi FH dan penanggalan

daripada fasa organik dengan larutan 2 M HCl memberikan pemekatan larutan Cu(II)

sebanyak 10 kali ganda.

FH juga telah berjaya digunakan sebagai pengekstrak untuk pengekstrakan dan

pemisahan Mo(VI). Logam ini telah berjaya dipisahkan dari ion logam seperti Ni(II),

Co(II), Al(III), Fe(III) dan Mn(II). Penanggalan kuantitatif bagi ion Mo(VI) dari fasa

organik boleh dijalankan dengan menggunakan larutan 2 M ammonia. Kaedah ini

diaplikasi untuk perolehan semula Mo(VI) dari campuran sintetik dan keputusan

menunjukkan lebih dari 90% perolehan semula Mo(VI) telah dicapai and larutan ion

logam tersebut bebas dari ketidaktulenan.

FH dan FPH telah berjaya digunakan sebagai pengekstrak untuk pengekstrakan dan

pemisahan emas(III) dari medium klorida. Logam tersebut telah diekstrak secara

kuantitatif dari medium hidroklorik pada 0.001-0.1 M ke dalam fasa organik. Emas(III)

telah berjaya dipisahkan dari Cu(II), Ni(II), Zn(II), Co(II) dan Fe(III) dengan

mengunakan FH atau FPH. Penanggalan kuantitatif bagi emas(III) dari fasa organik

boleh dijalankan dengan menggunakan larutan 2.0 M thiourea di dalam larutan HCl

vi

1.0 M. Pengekstrakan dan pemisahan emas(III) dengan FH atau FPH telah digunakan

untuk pisah dan peroleh semula emas(III) tulen dari campuran sintetik.

FPH disintesis dari olein minyak sawit telah berjaya digunakan sebagai reagen baru

dalam penentuan V(V) dengan kaedah spektrofotometrik. Kaedah ini beasaskan warna

bagi kompleks vanadium(V)-FPH. Ion logam tersebut membentuk kompleks berwarna

perang kehitaman dengan penyerapan maksimum pada 405 nm. Hukum Beer adalah sah

di dalam julat kepekatan 0.2-20 mg/L dan had pengesanan bagi kaedah ini adalah

0.01 mg/L. Ciri kaedah yang dimajukan ini adalah mudah, keselektifan yang bagus,

kesensitifan yang tinggi dan cepat.

vii

ACKNOWLEDGEMENTS

In the name of Allah, Most Gracious, Most Merciful. Praise be to Allah, the Cherisher

and Sustainer of the worlds. May the blessings and peace of Allah be upon our prophet

Muhammad ibn Abdullah (peace be upon him), upon his families and upon his

companions.

I wish to express my gratitude to my supervisor, Prof. Dato’ Dr Wan Md Zin Wan

Yunus. This thesis would not have been realized without his support, encouragement,

guidance and supervision throughout this work. I am also grateful to Prof Dr Md Jelas

Haron and Assoc. Prof Dr Mohamad Zaki Abd Rahman for their capacities as members

of Supervisory Committee. Thank you for the comments and suggestions. To the staff of

Chemistry Department thank you for their help and co-operation. A word of thanks is

also to Prof Dr Mhd Radzi Abas, University of Malaya for his advice and discussions.

My sincere thanks are to all my friends for the needed chats we had every day during our

lunch breaks, which took my mind off work. This thesis may not be completed without

your help and friendship, you are all great friends.

And to my family, thank you for supporting me throughout my long education. And

special thanks with heart full of love to my beloved sister, Sheela for her love,

encouragement, patience, understanding and moral support.

viii

I certify that an Examination Committee has met on date of viva voce to conduct the final examination of Sharifah Mohamad on her Doctor of Philosophy thesis entitled “Enzymatic Synthesis Characterization and Analytical Application of Fatty Hydrazides from Palm Oil” in accordance with Universiti Pertanian Malaysia (Higher Degree) Act 1980 and Universiti Pertanian Malaysia (Higher Degree) Regulations 1981. The Committee recommends that the student be awarded the Doctor of Philosophy. Members of the Examination Committee were as follows: Chairman, PhD Professor Department of Chemistry Faculty of Science Universiti Putra Malaysia (Chairman) Examiner 1, PhD Associate Professor Department of Chemistry Faculty of Science Universiti Putra Malaysia (Internal Examiner) Examiner 2, PhD Department of Chemistry Faculty of Science Universiti Putra Malaysia (Internal Examiner) External Examiner, PhD Professor Department of Chemistry Faculty of Science University Kebangsaan Malaysia (External Examiner) HASANAH MOHD. GHAZALI, PhD Professor and Deputy Dean School of Graduate Studies

Universiti Putra Malaysia

Date:

ix

This thesis was submitted to the Senate of Universiti Putra Malaysia and has been accepted as fulfilment for the requirement for the degree of Doctor of Philosophy. The members of the Supervisory Committee were as follows: Wan Md. Zin Wan Yunus, PhD Professor Department of Chemistry Faculty of Science Universiti Putra Malaysia (Chairman)

Md. Jelas Haron, PhD Professor Department of Chemistry Faculty of Science Universiti Putra Malaysia (Member)

Mohamad Zaki Abd Rahman, PhD Associate Professor Department of Chemistry Faculty of Science Universiti Putra Malaysia (Member)

AINI IDERIS, PhD Professor and Dean School of Graduate Studies Universiti Putra Malaysia Date: 10 July 2008

x

DECLARATION

I declare that the thesis is my original work expert for quotations and citations which have been duly acknowledgment. I also declare that it has not been previously, and is not concurrently, submitted for any other degree at Universiti Putra Malaysia or at any other degree at Universiti Putra Malaysia or at any other institution. SHARIFAH MOHAMAD

Date: 18 June 2008

xi

LIST OF TABLES

Table

2.1

2.2

2.3

2.4

3.1

3.2

3.3

3.4

3.5

4.1

4.2

4.3

4.4

4.5

4.6

4.7

4.8

4.9

Physical and chemical properties of palm oil and palm kernel olein Fatty acid composition (%) in palm oil and palm kernel olein Review of some extractants for copper separation and preconcentration Review of some spectrophotometric reagents for determination of V(V) Composition of synthetic mixture solutions containing Mo(VI) Composition of synthetic mixture solutions containing gold(III) Experimental conditions for determination PPH-vanadium(V) ratio (Job’s method) Experimental conditions for determination PPH-vanadium(V) ratio (slope ratio method) Composition of synthetic mixture solutions containing V(V) Optimum conditions of hydrazinolysis of palm oils Percentage of conversion of various fraction of palm oil to fatty hydrazide The elemental analysis data of FH Spectral assignments of palm olein, fatty hydrazide and palmityl hydrazide Optimum conditions of hydrazinolysis of palm oils Percentage of conversion of various fraction of palm oil to fatty phenyl hydrazide The elemental analysis data of FPH Spectral assignments of palm olein, fatty phenyl hydrazide and palmityl phenyl hydrazide Separation factor of separation of Cu(II) from other metal ions

Page

23

25

32

42

61

65

68

68

69

78

78

81

82

90

90

92

94

107

xii

4.10

4.11

4.12

4.13

4.14

4.15

4.16

4.17

4.18

4.19

4.20

4.21

4.22

4.23

4.24

4.25

4.26

4.27

4.28

Effect of foreign ions (500 mg/L) on the extractant of Cu(II) ion (10 mg/L) from the aqueous phase Effect of stripping agents on recovery of Cu(II) The application of the preconcentration procedure on water samples (n=5) Percentage of extraction of Mo(VI) in the presence of Ni(II), Co(II), Al(III), Mn(II) or Fe(III) from HCl solutions Effect of anions on extraction of Mo(VI) Stripping agent efficiency Effect of alkaline solutions concentration on stripping of Mo(VI) Recovery of pure molybdenum(VI) from synthetic mixture Binary separation of gold(III) from base metals by FH Binary separation of gold(III) from base metals by FPH Effect of the presence of chloride salts on the extraction of gold(III) by FH and FPH Stripping agent efficiency Recovery of gold(III) from synthetic mixture solutions by using FH Recovery of gold(III) from synthetic mixture solutions by using FPH The effect of foreign ions on the determination of 1 mg/L of vanadium(V) Molar absorptivity, specific absorptivity and Sandell sensitivity for FPH and PPH Limit of detection of vanadium(V) using FPH and PPH Determination of vanadium(V) in some synthetic mixture solutions Recovery of vanadium(V) in water samples

108

110

110

119

120

122

122

123

133

134

136

139

140

141

147

154

155

156

157

xiii

LIST OF FIGURES

Figure

2.1

2.2

4.1

4.2

4.3

4.4

4.5

4.6

4.7

4.8

4.9

4.10

4.11

4.12

4.13

4.14

4.15

4.16

4.17

Biotransformation of an ester in organic solvents World production and uses of oils and fats (1998, in million tones) Effect of temperature on hydrazinolysis of FH by various lipases Effect of various solvent on hydrazinolysis of FH Effect of reaction period on hydrazinolysis of FH Effect of catalyst loading on hydrazinolysis of FH Effect of mole ratio of hydrazine hydrate on hydrazinolysis of FH Possible reaction for the biosynthesis of FH in the presence of water FTIR spectra of (a): Palm olein (b) FH from palm olein FTIR spectra of (a): Palmityl hydrazide (PH) (b) FH from palm olein Effect of temperature on hydrazinolysis of FPH by various lipases Effect of solvent on hydrazinolysis of FPH Effect of reaction period on hydrazinolysis of FPH Effect of catalyst loading on hydrazinolysis of FPH Effect of mole ratio of phenylhydrazine on hydrazinolysis of FPH Possible reaction for the biosynthesis of FPH in anhydrous medium FTIR spectra of (a) Palm Olein, and (b) FPH from palm olein

FTIR spectra of (a) Palmityl phenyl hydrazide (PPH) and (b) FPH from palm olein 1H NMR spectra of FPH from palm olein

Page

13

20

71

73

74

76

77

80

83

83

85

86

87

88

89

91

94

95

96

xiv

4.18

4.19

4.20

4.21

4.22

4.23

4.24

4.25

4.26

4.27

4.28

4.29

4.30

4.31

4.32

4.33

4.34

4.35

4.36

4.37

4.38

4.39

1H NMR spectra of palmityl phenyl hydrazide (PPH)

Effect of pH on extraction of Cu(II) by FH and FPH Effect of pH on extraction of Ni(II) by FH and FPH Effect of pH on extraction of Zn(II) by FH and FPH Effect of pH on extraction of Co(II) by FH and FPH Effect of pH on extraction of Fe(III) by FH and FPH Effect of pH on extraction of Mn(II) by FH and FPH Distribution ratio for Cu(II), Ni(II), Zn(II), Co(II), Mn(II) and Fe(III) with FH Distribution ratio for Cu(II), Ni(II), Zn(II), Co(II), Mn(II) and Fe(III) with FPH

Effect of solvent on the extraction of copper(II) by FH Effect of pH on the extraction of Cu(II) Effect of shaking period on the extraction of Cu(II) Plot of log D vs log [FH] Effect of the A/O ratio on the extraction of Cu(II) with FH in toluene Extraction of molybdenum(VI) using FH and FPH Effect of solvent on the extraction of Mo(VI) Effect of shaking period on the extraction of Mo(VI) Effect of HCl concentration on the extraction of Mo(VI) Effect of the concentration of FH on the extraction of Mo(VI) Effect of HCl concentration on the extraction of metal ions by FH Extraction of gold(III) by using FH and FPH Effect of solvent on the extraction of gold(III)

96

98

98

99

99

100

100

101

101

103

104

105

106

109

112

113

114

116

117

118

125

126

xv

4.40

4.41

4.42

4.43

4.44

4.45

4.46

4.47

4.48

4.49

4.50

4.51

4.52

4.53

4.54

4.55

4.56

4.57

4.58

Effect of shaking period on the extraction of gold(III) ion

Effect of HCl concentration on the extraction of gold(III) by using FH and FPH Effect of the concentration of FH on the extraction of gold(III) Effect of the concentration of FPH on the extraction of gold(III) Effect of HCl concentration on the extraction of metal ions by FH Effect of HCl concentration on the extraction of metal ions by FPH Effect of anions on the extraction of gold(III) by FH Effect of anions on the extraction of gold(III) by FPH Absorption spectra (a) absorption spectra of FPH (b) absorption spectra of FPH-V(V) complex Effect of solvent on the absorbance of V(V)-FPH system Effect of acid on the absorbance of V(V)-FPH system Effect of nitric acid concentration on the absorbance of V(V)-FPH system Effect of FPH concentration on the absorbance of V(V)-FPH system Absorption spectra of FPH-V(V), FPH-Fe(III) and FPH-Cr(VI) complex V(V) complex stoichiometry determination with PPH at 405 nm (Job’s method) V(V) complex stoichiometry determination with PPH at 405 nm (Slope ratio method) Response curve of FPH towards different concentration of V(V) Response curve of PPH towards different concentration of V(V) Calibration curve of complex FPH-vanadium(V)

127

129

130

130

131

132

137

138

142

143

144

144

145

148

149

150

151

151

153

xvi

4.59

4.60

Calibration curve of complex PPH-vanadium(V) Reproducibility study in FPH response towards V(V)

153

155

xvii

xviii

LIST OF ABBREVIATIONS

DAGs

DBC

DEHPA

FAAS

FFAs

FH

FPH

FTIR

ICP-AES

MAGs

NMR

PH

PPH

TAG

TAGS

TBP

TBPO

diacylglycerols di-butylcarbitol di-2-ethylhexylphosphoric acid flame atomic absorption spectroscopy free fatty acids fatty hydrazide fatty phenyl hydrazide fourier transforms infrared spectroscopy inductively coupled plasma-atomic emission spectroscopy monoacylglycerols nuclear magnetic resonance palmityl hydrazide palmityl phenyl hydrazide triacylglycerol triacylglycerols tributyl phosphate tributyl phosphine oxide

TABLE OF CONTENTS

Page ABSTRACT ABSTRAK ACKNOWLEDGEMENTS APPROVAL DECLARATION LIST OF TABLES LIST OF FIGURES LIST OF ABBREVIATIONS

ii v

vii ix xi xii xiv

xviii

CHAPTER 1

2

INTRODUCTION 1.1 Background of study 1.1.1 Hydrazide 1.1.2 Application of hydrazide in analytical chemistry 1.2 Objectives of study LITERATURE REVIEW 2.1 Synthesis of hydrazide 2.2 Lipases 2.2.1 Lipase-catalyzed synthesis of carboxylic amides 2.2.2 Lipases in fat and oleochemical industry 2.3 Fats and oils as oleochemical raw materials 2.3.1 Palm oils 2.3.2 Characteristics of palm oil 2.3.3 Chemical composition of palm oil 2.3.4 Current status of the palm oil industry 2.4 Organic reagent for analytical application 2.5 Solvent extraction 2.5.1 Preconcentration and separation of copper(II) by solvent extraction 2.5.2 Recovery and purification of molybdenum(VI) by solvent extraction 2.5.3 Recovery and purification of gold by solvent extraction 2.6 Spectrophotometric method 2.6.1 Spectrophotometric method for determination of vanadium

1 1 1 4 6

8 8 11 14 16 19 20 21 23 25 27 28 30

32

34 37 39

3

MATERIALS AND METHODS 3.1 Materials and equipments 3.1.1 Materials and equipments for synthesis of FH and FPH 3.1.2 Materials and equipments for application studies 3.2 Synthesis of FH 3.2.1 General procedure for enzymatic synthesis of FH 3.2.2 Separation and purification of FH 3.3 Optimization study of FH synthesis 3.3.1 Screening of lipase 3.3.2 Effect of solvent 3.3.3 Effect of reaction period 3.3.4 Effect of catalyst loading 3.3.5 Effect of mole ratio of hydrazine 3.4 Synthesis of fatty phenylhydrazide (FPH) 3.4.1 General procedure for enzymatic synthesis of FPH 3.4.2 Separation and purification of FPH 3.5 Optimization study of FPH synthesis 3.5.1 Screening of lipase 3.5.2 Effect of solvent 3.5.3 Effect of reaction period 3.5.4 Effect of catalyst loading 3.5.5 Effect of mole ratio of phenyl hydrazine 3.6 Characterization of FH and FPH 3.6.1 Elemental analysis 3.6.2 Fourier Transform Infrared (FTIR) spectroscopy 3.6.3 1H Nuclear Magnetic Resonance (NMR) 3.7 Metal ion extraction by solvent extraction 3.7.1 Effect of pH on metal ion extraction by using FH and FPH 3.8 Preconcentration and separation of copper(II) ion by using FH 3.8.1 Effect of solvent on extraction of copper(II) ion 3.8.2 Effect of pH on the extraction of copper(II) ion 3.8.3 Effect of shaking period 3.8.4 Effect of FH concentration on the extraction of copper(II) ion 3.8.5 Separation of copper(II) from other metal ions 3.8.6 Effect of foreign ions on extraction of copper(II) ion 3.8.7 Effect of aqueous to organic phase ratio 3.8.8 Effect of different acids of various concentration stripping of copper(II) from organic phase 3.8.9 Preconcentration of copper(II) ion from water samples using optimum conditions of extraction and stripping 3.9 Extraction and separation of molybdenum(VI) using FH 3.9.1 Effect of solvent on the extraction of molybdenum (VI) ion

44 44 44 45 48 48 48 48 48 49 49 49 50 50 50 50 51 51 51 51 52 52 52 52 53 53 53 53

54

54 55 55 55

56 56 56 57

57

58 58

4

3.9.2 Effect of HCl concentration on the extraction of molybdenum(VI) ion 3.9.3 Effect of shaking period 3.9.4 Effect of FH concentration 3.9.5 Separation of molybdenum(VI) from associated metals 3.9.6 Effect of anions on the extraction of molybdenum(VI) 3.9.7 Stripping study 3.9.8 Recovery of molybdenum(VI) from synthetic solutions 3.10 Extraction and separation of gold(III) using FH and FPH 3.10.1 Effect of solvent on the extraction of gold(III) 3.10.2 Effect of HCl concentration on the extraction of gold(III) ion 3.10.3 Effect of shaking period 3.10.4 Effect of extractant concentration 3.10.5 Separation of gold(III) from associated metals 3.10.6 Effect of the presence of chloride salts on the extraction of gold(III) 3.10.7 Effect of anions on the extraction of gold(III) 3.10.8 Stripping agent studies 3.10.9 Recovery of gold(III) from synthetic solutions 3.11 Spectrophotometric studies for determination of vanadium (V) using FPH 3.11.1 Absorption of spectra 3.11.2 Effect solvent 3.11.3 Effect of acidity 3.11.4 Effect of concentration of FPH 3.11.5 Effect of foreign ions 3.11.6 Determination of molar ratio of FPH-vanadium(V) complexes 3.11.7 Dynamic range and sensitivity 3.11.8 Reproducibility of the method 3.11.9 Determination of vanadium(V) from synthetic solutions and water samples RESULTS AND DISCUSSION 4.1 Optimization of FH synthesis 4.1.1 Effect of various lipases on the biosynthesis of FH 4.1.2 Effect of solvent on biosynthesis of FH 4.1.3 Effect of reaction period on biosynthesis of FH 4.1.4 Effect of catalyst loading on biosynthesis of FH 4.1.5 Effect of mole ratio of hydrazine hydrate on biosynthesis of FH 4.1.6 Hydrazinolysis of various products of palm oil 4.1.7 Proposed reaction involved in the biosynthesis of FH

58

59 59 59

60 60 61 62 62 62

62 63 63 63

64 64 65 65

65 66 66 66 67 67

68 69 69

70 70 70 72 74 75 76

77 79

4.2 Characterization of FH 4.2.1 Elemental analysis of FH 4.2.2 FTIR spectra of FH 4.3 Optimization of FPH synthesis 4.3.1 Effect of various lipases on the biosynthesis of FPH 4.3.2 Effect of solvent on biosynthesis of FPH 4.3.3 Effect of reaction period on biosynthesis of FPH 4.3.4 Effect of catalyst loading on biosynthesis of FPH 4.3.5 Effect of mole ratio of phenylhydrazine on biosynthesis of FPH 4.3.6 Hydrazinolysis of various products of palm oil 4.3.7 Proposed reaction involved in biosynthesis of FPH 4.4 Characterization of FPH 4.4.1 Elemental analysis of FPH 4.4.2 FTIR spectra of FPH 4.4.3 1H NMR of FPH 4.5 Solvent extraction of metals by FH and FPH 4.5.1 Effect of pH on various metal ion extraction 4.6 Preconcentration and separation of copper(II) by FH 4.6.1 Effect of solvent on the extraction of Cu(II) 4.6.2 Effect of pH on the extraction of Cu(II) 4.6.3 Effect of shaking period on the extraction of Cu(II) 4.6.4 Effect of FH concentration on the extraction of Cu(II) 4.6.5 Separation of Cu(II) from other metals 4.6.6 Effect of foreign ions on the extraction of Cu(II) 4.6.7 Effect of aqueous to organic phase ratio 4.6.8 Effect of stripping agent for preconcentration of Cu 4.6.9 Preconcentration of Cu in water samples 4.7 Extraction and separation of molybdenum(VI) using FH 4.7.1 Extraction of molybdenum(VI) ion by using FH and FPH 4.7.2 Effect of solvent on the extraction of molybdenum(VI) 4.7.3 Effect of shaking period 4.7.4 Effect of HCl concentration on the extraction of molybdenum(VI) 4.7.5 Effect of FH concentration on the extraction of molybdenum(VI) 4.7.6 HCl concentration effect on extraction of various metal ions 4.7.7 Separation of molybdenum(VI) from associated metals 4.7.8 Effect of anions on the extraction of molybdenum(VI) 4.7.9 Stripping agent studies 4.7.10 Recovery of molybdenum(VI) from synthetic solutions

80 80 81 84 84 85 87 88 89

90 91 92 92 93 95 97 97 102 102 103 105 105 106 107 108 109 110 111 112

113 114 114

116

117

118 119 121 123

5

4.8 Extraction and separation of gold(III) using FH and FPH 4.8.1 Extraction of gold(III) ion by using FH and FPH 4.8.2 Effect of solvent on the extraction of gold(III) 4.8.3 Effect of shaking period on gold(III) extraction 4.8.4 Effect of HCl concentration on the extraction of gold(III) 4.8.5 Effect of FH concentration on the extraction of gold(III) 4.8.6 HCl concentration effect on extraction of various metal ions 4.8.7 Separation of gold(III) from associated metals 4.8.8 Effect of the presence of chloride salts on the extraction of gold(III) 4.8.9 Effect anions on the extraction of gold(III) 4.8.10 Stripping agent studies 4.8.11 Recovery of gold(III) from synthetic solutions 4.9 Spectrophotometric studies for determination of vanadium (V) using FPH 4.9.1 Absorption spectra 4.9.2 Effect of solvent 4.9.3 Effect of acidity 4.9.4 Effect of concentration of FPH 4.9.5 Effect of foreign ions 4.9.6 Molar ratio of PPH-vanadium(V) complex 4.9.7 Dynamic range and sensitivity 4.9.8 Limit of detection (LOD) 4.9.9 Reproducibility of the method 4.9.10 Determination of vanadium in synthetic solutions 4.9.11 Application: Analysis of vanadium in spiked water samples CONCLUSIONS 5.1 Recommendations for further studies

124 125 126 126 127

129

131

132 134

136 138 139 141

142 143 143 145 146 148 150 154 155 156 156

158 160

REFERENCES APPENDICES BIODATA OF THE STUDENT LIST OF PUBLICATIONS

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CHAPTER I

INTRODUCTION

1.1 Background of study

1.1.1 Hydrazide

Hydrazides with the general formula RCONHNH2 have been thoroughly studied as

ligands with non-bonded lone pair of electrons on the nitrogen of amino group and

oxygen of the carbonyl group. It may coordinate with many metal ions monodentate or

multidentate (Gudasi et al., 2007; Becher et al., 2006; Zhang et al., 2006). The

molecular structure of chelate-forming reagents must contain at least two donor atoms

capable of bonding to the same metal atom. The complex formation with hydrazide

ligands in aqueous solution demonstrated clearly that two atoms (O, N) bonding modes

of the ligands are accessible to metal ions (Bontchev et al., 1981; Ahmed and Chaudhuri,

1971).

The formation of metal complexes plays an important role in the enhancement of their

biological activity (Singh et al., 2000). In recent years, hydrazides have received a lot of

attention due to their biological activity as tuberculostatic (Yadav et al., 2005),

antibacterial agent (Malhotra et al., 1992), antitumor agent (Dodoff et al., 1994) and

anticancer agent (Zhang et al., 2004). In addition, hydrazides have received considerable

attention in analytical chemistry chelating reagents for metal extraction and

determination (Jal et al., 2001; Ahmed and Banoo, 1999; Liu et al., 1999).