**3. Result and discussion**

#### **3.1 Isolation and separation**

A brownish yellow gel of CocoPLs was obtained from dried coconut meat (6.86 × 10 −2%, w/w) (**Figure 1**). The result was confirmed by the appearance similarity of the CocoPLs from the previous research [20, 21]. The CocoPLs was then subjected to separation to obtain CocoPEs.

In the separation process using vacuum column chromatography, CocoPLs was eluted continuously using chloroform:methanol (9:1, v/v). Each fraction of 10 mL eluent was collected and subjected to identification. As much as 520 fractions were obtained to elute CocoPEs from the CocoPLs samples completely. Identification by

**185**

**Figure 2.** *CocoPEs.*

*Coconut Phospholipid Species: Isolation, Characterization and Application as Drug Delivery…*

TLC using 10% H2SO4 and ninhydrin spotting agent [23] resulted in that CocoPEs

the eluent that resulted in dark brown CocoPEs gel (9.8 × 10−3%, w/w of dried

The fraction contained CocoPEs were then combined and evaporated to remove

The functional groups identification of CocoPLs and CocoPEs was conducted by FTIR spectra analysis. The FTIR spectra of both CocoPLs and CocoPEs were displayed on **Figure 3**. To analyze further the spectra were scrutinized using a decon-

The absorption data obtained from both FTIR spectra and deconvolution analysis were compared (see **Table 1**) to the specific infrared absorption area for phospholipids proposed by Stuart [25] and Hudiyanti et al. [20, 21]. The presence of a typical spectrum of phospholipids was clearly revealed. Significant differences between CocoPLs and CocoPEs spectra was disclosed by the typical absorption of choline and ethanolamine groups on both spectra of CocoPLs and CocoPEs. The

try stretching; were not present on the CocoPEs spectra. The typical absorption that indicate the presence of ethanolamine species by N-H vibration absorptions was displayed on CocoPLs and CocoPEs spectra. This evident indicated that CocoPLs contained choline and ethanolamine species while CocoPEs did not contain choline

asymmetric bending and (CH3)3N+

and 1800–700 cm<sup>−</sup><sup>1</sup>

as

asymme-

*DOI: http://dx.doi.org/10.5772/intechopen.88176*

**3.2 Functional groups identification**

choline group absorptions; (CH3)3N+

coconut meat) (**Figure 2**).

presented in **Figure 4**.

were present in the 105th to the 520th fraction.

volution program [21, 24], at wavenumbers 3500–2800 cm<sup>−</sup><sup>1</sup>

**Figure 1.** *CocoPLs.*

*Coconut Phospholipid Species: Isolation, Characterization and Application as Drug Delivery… DOI: http://dx.doi.org/10.5772/intechopen.88176*

TLC using 10% H2SO4 and ninhydrin spotting agent [23] resulted in that CocoPEs were present in the 105th to the 520th fraction.

The fraction contained CocoPEs were then combined and evaporated to remove the eluent that resulted in dark brown CocoPEs gel (9.8 × 10−3%, w/w of dried coconut meat) (**Figure 2**).

#### **3.2 Functional groups identification**

*Nano- and Microencapsulation - Techniques and Applications*

*EE* = \_

and *Ct* is the unencapsulated VC concentration.

**3. Result and discussion**

**3.1 Isolation and separation**

to separation to obtain CocoPEs.

coconut liposome was determined based on Eqs. (1) and (2):

In addition we used CocoPLs as comparison. The encapsulation efficiency of VC in

*C*<sup>0</sup> − *Ct C*<sup>0</sup>

where *EE* is the encapsulation efficiency*; C0* is the initial concentration of VC;

A brownish yellow gel of CocoPLs was obtained from dried coconut meat (6.86 × 10 −2%, w/w) (**Figure 1**). The result was confirmed by the appearance similarity of the CocoPLs from the previous research [20, 21]. The CocoPLs was then subjected

In the separation process using vacuum column chromatography, CocoPLs was eluted continuously using chloroform:methanol (9:1, v/v). Each fraction of 10 mL eluent was collected and subjected to identification. As much as 520 fractions were obtained to elute CocoPEs from the CocoPLs samples completely. Identification by

× 100% (1)

*Ct* = *Cliposome*<sup>+</sup>*VC* − *Cempty liposome* (2)

**184**

**Figure 1.** *CocoPLs.*

The functional groups identification of CocoPLs and CocoPEs was conducted by FTIR spectra analysis. The FTIR spectra of both CocoPLs and CocoPEs were displayed on **Figure 3**. To analyze further the spectra were scrutinized using a deconvolution program [21, 24], at wavenumbers 3500–2800 cm<sup>−</sup><sup>1</sup> and 1800–700 cm<sup>−</sup><sup>1</sup> as presented in **Figure 4**.

The absorption data obtained from both FTIR spectra and deconvolution analysis were compared (see **Table 1**) to the specific infrared absorption area for phospholipids proposed by Stuart [25] and Hudiyanti et al. [20, 21]. The presence of a typical spectrum of phospholipids was clearly revealed. Significant differences between CocoPLs and CocoPEs spectra was disclosed by the typical absorption of choline and ethanolamine groups on both spectra of CocoPLs and CocoPEs. The choline group absorptions; (CH3)3N+ asymmetric bending and (CH3)3N+ asymmetry stretching; were not present on the CocoPEs spectra. The typical absorption that indicate the presence of ethanolamine species by N-H vibration absorptions was displayed on CocoPLs and CocoPEs spectra. This evident indicated that CocoPLs contained choline and ethanolamine species while CocoPEs did not contain choline

**Figure 2.** *CocoPEs.*

**Figure 3.** *CocoPLs and CocoPEs absorption spectra.*

**Figure 4.**

*Deconvolution results: (a) CocoPLs at wavenumbers 1800–700 cm<sup>−</sup><sup>1</sup> ; (b) CocoPLs at wavenumbers 3500– 2800 cm<sup>−</sup><sup>1</sup> ; (c) CocoPEs at wavenumbers 1800–700 cm<sup>−</sup><sup>1</sup> ; (d) CocoPEs at wavenumbers 3500–2800 cm<sup>−</sup><sup>1</sup> .*

species. From The FTIR spectra point of view this data disclosed that the CocoPEs separation from CocoPLs was successful.

## **3.3 Characterization of fatty acyl chains**

The fatty acyl chains content of CocoPLs and CocoPEs was analyzed by GC-MS. The CocoPLs chromatogram was presented on **Figure 5**. A total of nine

**187**

*CocoPEs.*

**Table 1.**

*Coconut Phospholipid Species: Isolation, Characterization and Application as Drug Delivery…*

**CocoPLs (cm<sup>−</sup><sup>1</sup> )**

1. **N-H vibration 3471 3394 3379 3403 3373** 2. =C-H stretching 3010 — — 3001 3002

**CocoPEs (cm<sup>−</sup><sup>1</sup> )**

2956 — — 2958 2956

2920 2924 2924 2923 2919

2870 — — 2885 2890

2850 2854 2854 2850 2848

1730 1735 1735 1738 1739

**1485 — — 1493 —**

1460 1458 1458 1461 1464

1405 — — — —

1378 1373 1373 1376 1378

1228 1226 1242 1225 1222

1170 1165 — 1150 1165

1085 — 1080 1106 1107

1070 1072 — 1071 1070

**972 — — 973 —**

820 817 — 813 819

18. C-O-P stretching 1047 — — 1020 1003

21. CH2 rocking 730, 720, 718 717 725 714 713 *Bold entries represented the typical absorption of choline and ethanolamine groups on both spectra of CocoPLs and* 

1400–1200 — — 1333 1266

— — — —

**CocoPLs Deconvolution (cm<sup>−</sup><sup>1</sup> )**

**CocoPEs Deconvolution (cm<sup>−</sup><sup>1</sup> )**

*DOI: http://dx.doi.org/10.5772/intechopen.88176*

**20, 21, 25] (cm<sup>−</sup><sup>1</sup> )**

1472, 1468, 1463

**No. Absorption type References [15,** 

3. CH3 asymmetric stretching

4. CH2 asymmetric stretching

5. CH3 symmetric stretching

6. CH2 symmetric stretching

7. C=O stretching, sn-1 chain transconformation

8. **(CH3)3N+**

10. CH3 asymmetric bending

11. (CH3)3N+

12. CH3 symmetric bending

13. CH3 rocking

15. CO-O-C

17. CO-O-C

19. **(CH3)3N+**

20. P-O asymmetric stretching

14. PO2

16. PO2

**asymmetric bending**

symmetric bending

ribbon progression

asymmetric stretching

symmetric stretching

**asymmetric stretching**

*Typical Absorption of CocoPLs and CocoPEs functional groups.*

<sup>−</sup> symmetric stretching

<sup>−</sup>asymmetric stretching

9. CH2 scissoring 1473,


*Coconut Phospholipid Species: Isolation, Characterization and Application as Drug Delivery… DOI: http://dx.doi.org/10.5772/intechopen.88176*

#### *Bold entries represented the typical absorption of choline and ethanolamine groups on both spectra of CocoPLs and CocoPEs.*

#### **Table 1.**

*Typical Absorption of CocoPLs and CocoPEs functional groups.*

*Nano- and Microencapsulation - Techniques and Applications*

species. From The FTIR spectra point of view this data disclosed that the CocoPEs

*; (b) CocoPLs at wavenumbers 3500–*

*.*

*; (d) CocoPEs at wavenumbers 3500–2800 cm<sup>−</sup><sup>1</sup>*

The fatty acyl chains content of CocoPLs and CocoPEs was analyzed by GC-MS. The CocoPLs chromatogram was presented on **Figure 5**. A total of nine

separation from CocoPLs was successful.

*Deconvolution results: (a) CocoPLs at wavenumbers 1800–700 cm<sup>−</sup><sup>1</sup>*

*; (c) CocoPEs at wavenumbers 1800–700 cm<sup>−</sup><sup>1</sup>*

**3.3 Characterization of fatty acyl chains**

**186**

**Figure 4.**

*2800 cm<sup>−</sup><sup>1</sup>*

**Figure 3.**

*CocoPLs and CocoPEs absorption spectra.*

#### *Nano- and Microencapsulation - Techniques and Applications*

**Figure 5.** *CocoPLs chromatogram.*


#### **Table 2.**

*The fatty acyl chains of CocoPLs.*

peaks was recognized. Seven peaks were with abundance above 1%. The chromatogram suggested that there were at least 9 types of fatty acyl chains present on the CocoPLs. The MS reading revealed the identity of these fatty acyl chains. Three fatty acyl chains worth mentioning with the abundance more than 10%, i.e., lauric acid, palmitic acid and oleic acid which were indicated by peak number 3 (abundance of 11.31%); peak number 5 (15.26%); and peak number 7 (55.18%). The result was in agreement with previous research [15, 20, 21]. The seven fatty acyl chains recognized in CocoPLs was displayed on **Table 2**.

The chromatogram of CocoPEs was disclosed on **Figure 6**. The resulting chromatogram exposed the presence of five peaks with abundance above 1% which suggested the presence of five types of fatty acyl chains in the CocoPEs. Three of them had great abundance i.e. capric, linoleic and oleic acids as indicated by peak number 2, 3 and 4 and with abundance of 17.09%, 43.17% and 31.88% respectively. The MS reading of fatty acyl chains content in the CocoPEs was tabulated on **Table 3**.

**Tables 2** and **3** revealed differences to some extent in fatty acyl chains composition between CocoPLs and CocoPEs. CocoPLs had more variation in fatty acyl chains type compared to CocoPEs. This fact plausible considering that CocoPEs was obtained from the separation of CocoPLs. The separation was mainly based on the common head group namely ethanolamine that reflected on the polarity of the separated CocoPEs molecules hence the choice of the separation eluent. More over fatty acyl chains profile were closely related to the position of phospholipid species in the bio-membrane bilayer [26–28]. Phosphatidylethanolamine (PE) species

**189**

**Table 3**.

**Table 3.**

**Figure 6.**

*CocoPEs chromatogram.*

**3.4 Parent ion screening**

*The Fatty acyl chains of CocoPEs.*

**3.5 Phase behavior**

phase behavior and other properties as well.

*Coconut Phospholipid Species: Isolation, Characterization and Application as Drug Delivery…*

generally would be positioned in the inner leaflet of bilayer due to their molecular geometry, i.e. cylinder [2]. The PE species molecular shape was supported by more abundance composition of unsaturated fatty acyl chains in the CocoPEs extract,

**Peak number tR (min) Fatty acyl chains Area (%)** 2. 38.566 Capric acid, C10:0 (decanoic acid) 17.09 3. 42.041 Linoleic acid, C18:2 (9(Z),12(Z)-octadecadienoic acid) 43.17 4. 42.198 Oleic acid, C18:1 (9(Z)-octadecenoic acid) 31.88 5. 42.555 Stearic acid, C18: 0 (octadecanoic acid) 5.93 8. 46.186 Arachidic acid, C20:0 (eicosanoic acid) 1.04

Based on the fatty acyl chains of the CocoPEs we conducted parent ion screening using LCMSMS. The CocoPEs parent ion spectrogram was presented on **Figure 7**. The spectrogram gave us a representation of the molecular species composing CocoPEs extract. At least 11 molecular species of CocoPEs were found. The CocoPEs molecular species was tabulated on **Table 4**. The molecular species was predicted based on the head group and combination of two fatty acyl chains for the nonpolar part of CocoPEs species. These similar species would govern the CocoPEs

Every phospholipid species has unique phase behavior that related to their molecular structure and phase behavior. The phase behavior of CocoPLs and CocoPEs were investigated by thermal analysis using DSC. The thermogram for CocoPLs, **Figure 8**, exhibited a small peak at 28.85°C and larger peak at 83.95°C. These peaks indicated that CocoPLs underwent phase changes as temperature changes. A pre-transition process from planar-shaped gel (Lb′) to the rippling phase (Pb′) was at a temperature of 28.85°C (Tp), then proceed with the

*DOI: http://dx.doi.org/10.5772/intechopen.88176*

*Coconut Phospholipid Species: Isolation, Characterization and Application as Drug Delivery… DOI: http://dx.doi.org/10.5772/intechopen.88176*

**Figure 6.** *CocoPEs chromatogram.*

*Nano- and Microencapsulation - Techniques and Applications*

peaks was recognized. Seven peaks were with abundance above 1%. The chromatogram suggested that there were at least 9 types of fatty acyl chains present on the CocoPLs. The MS reading revealed the identity of these fatty acyl chains. Three fatty acyl chains worth mentioning with the abundance more than 10%, i.e., lauric acid, palmitic acid and oleic acid which were indicated by peak number 3 (abundance of 11.31%); peak number 5 (15.26%); and peak number 7 (55.18%). The result was in agreement with previous research [15, 20, 21]. The seven fatty acyl chains

**Peak number tR (min) Fatty acyl chains Area (%)** 3. 29.164 Lauric acid, C12:0 (dodecanoic acid) 11.31 4. 34.037 Myristic acid, C14:0 (tetradecanoic acid) 5.71 5. 38.497 Palmitic acid, C16:0 (hexadecanoic acid) 15.26 6. 41.872 Linoleic acid, C18:2 (9(Z),12(Z)-octadecadienoic acid) 6.00 7. 42.117 Oleic acid, C18:1 (9(Z)-octadecenoic acid) 55.18 8. 42.482 Stearic acid, C18:0 (octadecanoic acid) 3.97 9. 52.794 Lignoceric acid, C24:0 (tetracosanoic acid) 1.49

The chromatogram of CocoPEs was disclosed on **Figure 6**. The resulting chromatogram exposed the presence of five peaks with abundance above 1% which suggested the presence of five types of fatty acyl chains in the CocoPEs. Three of them had great abundance i.e. capric, linoleic and oleic acids as indicated by peak number 2, 3 and 4 and with abundance of 17.09%, 43.17% and 31.88% respectively. The MS reading of fatty acyl chains content in the CocoPEs was tabulated

**Tables 2** and **3** revealed differences to some extent in fatty acyl chains composi-

tion between CocoPLs and CocoPEs. CocoPLs had more variation in fatty acyl chains type compared to CocoPEs. This fact plausible considering that CocoPEs was obtained from the separation of CocoPLs. The separation was mainly based on the common head group namely ethanolamine that reflected on the polarity of the separated CocoPEs molecules hence the choice of the separation eluent. More over fatty acyl chains profile were closely related to the position of phospholipid species in the bio-membrane bilayer [26–28]. Phosphatidylethanolamine (PE) species

recognized in CocoPLs was displayed on **Table 2**.

**188**

on **Table 3**.

**Figure 5.**

**Table 2.**

*The fatty acyl chains of CocoPLs.*

*CocoPLs chromatogram.*


## **Table 3.**

*The Fatty acyl chains of CocoPEs.*

generally would be positioned in the inner leaflet of bilayer due to their molecular geometry, i.e. cylinder [2]. The PE species molecular shape was supported by more abundance composition of unsaturated fatty acyl chains in the CocoPEs extract, **Table 3**.

## **3.4 Parent ion screening**

Based on the fatty acyl chains of the CocoPEs we conducted parent ion screening using LCMSMS. The CocoPEs parent ion spectrogram was presented on **Figure 7**. The spectrogram gave us a representation of the molecular species composing CocoPEs extract. At least 11 molecular species of CocoPEs were found. The CocoPEs molecular species was tabulated on **Table 4**. The molecular species was predicted based on the head group and combination of two fatty acyl chains for the nonpolar part of CocoPEs species. These similar species would govern the CocoPEs phase behavior and other properties as well.

#### **3.5 Phase behavior**

Every phospholipid species has unique phase behavior that related to their molecular structure and phase behavior. The phase behavior of CocoPLs and CocoPEs were investigated by thermal analysis using DSC. The thermogram for CocoPLs, **Figure 8**, exhibited a small peak at 28.85°C and larger peak at 83.95°C. These peaks indicated that CocoPLs underwent phase changes as temperature changes. A pre-transition process from planar-shaped gel (Lb′) to the rippling phase (Pb′) was at a temperature of 28.85°C (Tp), then proceed with the

**Figure 7.**

*CocoPEs spectrogram.*


**191**

**Figure 9.**

*Thermal analysis of CocoPEs.*

**Figure 8.**

*Thermal analysis of CocoPLs.*

*Coconut Phospholipid Species: Isolation, Characterization and Application as Drug Delivery…*

main transition from gel (Lb′) to the liquid crystal phase (La) at a temperature of 83.95°C (Tm) [29–31]. Tp and Tm were the pre-transition and melting temperature

Different phase behavior of CocoPEs was exhibited in **Figure 9**. The thermogram for CocoPEs was more complex than CocoPLs indicated that CocoPEs had more complex phase transition than CocoPLs. CocoPEs displayed pre-transition process from planar-shaped gel (Lb′) to a rippling phase (Pb′) at a temperature of 25.29°C (Tp), followed by a major transition from gel (Lb′) to liquid crystal phase (La) at a temperature of 32.62°C (Tm), and then a transition from the liquid crystal phase (La) to hexagonal phase (H) at a temperature of 65.53°C (Th) [32]. The

*DOI: http://dx.doi.org/10.5772/intechopen.88176*

correspondingly.

#### **Table 4.**

*CocoPEs molecular species prediction.*

*Coconut Phospholipid Species: Isolation, Characterization and Application as Drug Delivery… DOI: http://dx.doi.org/10.5772/intechopen.88176*

main transition from gel (Lb′) to the liquid crystal phase (La) at a temperature of 83.95°C (Tm) [29–31]. Tp and Tm were the pre-transition and melting temperature correspondingly.

Different phase behavior of CocoPEs was exhibited in **Figure 9**. The thermogram for CocoPEs was more complex than CocoPLs indicated that CocoPEs had more complex phase transition than CocoPLs. CocoPEs displayed pre-transition process from planar-shaped gel (Lb′) to a rippling phase (Pb′) at a temperature of 25.29°C (Tp), followed by a major transition from gel (Lb′) to liquid crystal phase (La) at a temperature of 32.62°C (Tm), and then a transition from the liquid crystal phase (La) to hexagonal phase (H) at a temperature of 65.53°C (Th) [32]. The

**Figure 8.** *Thermal analysis of CocoPLs.*

*Nano- and Microencapsulation - Techniques and Applications*

**No. m/z (M-H) Molecular weight CocoPEs molecular species**

1. 554 555 Ethanolamine Capric acid

2. 662 663 Ethanolamine Capric acid

3. 664 665 Ethanolamine Capric acid

4. 666 667 Ethanolamine Capric acid

5. 694 695 Ethanolamine Capric acid

6. 770 771 Ethanolamine Linoleic acid

7. 774 775 Ethanolamine Oleic acid

8. 776 777 Ethanolamine Oleic acid

9. 802 803 Ethanolamine Linoleic acid

10. 806 807 Ethanolamine Stearic acid

11. 834 835 Ethanolamine Arachidic acid

**Head group Fatty acyl chains**

Capric acid

Linoleic acid

Oleic acid

Stearic acid

Arachidic acid

Linoleic acid

Oleic acid

Stearic acid

Arachidic acid

Arachidic acid

Arachidic acid

**190**

**Table 4.**

**Figure 7.**

*CocoPEs spectrogram.*

*CocoPEs molecular species prediction.*

**Figure 9.** *Thermal analysis of CocoPEs.*

hexagonal phase formation was consistent to cylindrical molecular shape attributed to CocoPEs. The CocoPEs gradual change of phase was estimated because of the similar molecular species composing CocoPEs.

The phase behavior of CocoPEs dan CocoPLs above indicated that they were both had complex self-assembly structures which would be beneficial for future applications [2].

#### **3.6 Encapsulation of vitamin C in coconut (CocoPLs and CocoPEs) liposomes**

Phospholipids has long been known as drug delivery substance due to their liposome forming ability. Liposome was a spherical aggregation structure with bilayer phospholipid as its shell surrounding aqueous core. This unique structure was especially a perfect vehicle for delivering hydrophilic and hydrophobic drugs with storage and controlled release purposes. In this paper as a preliminary study for further application of coconut phospholipid as drug delivery material we used vitamin C as a hydrophilic drug model to be encapsulated in coconut liposomes. Vitamin C was a hydrophilic drug and would be encapsulated inside the aqueous core of liposome. The study lead to that encapsulation efficiency of vitamin C in CocoPEs were higher than CocoPLs i.e. 94.44% and 92.40% respectively, **Figure 10**.

In relation to their application as drug delivery, liposomes were usually made from phospholipid and a small amount of cholesterol. Cholesterol was added to the liposome membrane to control liposome rigidity and penetrability [33]. Therefore to explore the effect of cholesterol on the encapsulation efficiency of coconut liposomes we also prepared coconut liposomes with several different concentration of cholesterol namely 10%, 20%, 30% and 40%. The encapsulation efficiency of the liposomes were presented on **Figure 10**. The results suggested that addition of cholesterol up to 40% in the liposome's membrane reduced the encapsulation efficiency of CocoPEs and CocoPLs liposomes. Furthermore CocoPEs liposomes demonstrated slighter reduction than CocoPLs liposomes. The encapsulation efficiency of CocoPEs diminished gradually as the cholesterol concentration increased while for CocoPLs liposomes the decline was arbitrary. Addition up to 30% of cholesterol only reduced the CocoPEs encapsulation efficiency to around 80% while CocoPLs was as low as 52%. Cholesterol effect on the encapsulation efficiency of CocoPEs

**Figure 10.** *Encapsulation efficiency of CocoPLs and CocoPEs liposomes with cholesterol composition variation.*

**193**

*Coconut Phospholipid Species: Isolation, Characterization and Application as Drug Delivery…*

liposomes more consistent than CocoPLs. We suspected it was due to the molecular composition of the phospholipid in the membrane. The molecular composition was represented by the composition of functional group and fatty acyl chains in the CocoPEs and CocoPLs, **Tables 1–3**. In the liposome membrane cholesterol interacted with CocoPEs and CocoPLs through their functional groups and fatty acyl chains. Cholesterol with small hydrophilic head group i.e., –OH and big and rigid hydrophobic steroid ring would interact better with small head group phospholipid species like CocoPEs than CocoPLs which had big spherical choline group and possibly other head groups as well. The composition of fatty acyl with double bonds also suspected would give more room for cholesterol hydrophobic moiety. The fatty acyl chains would assume "kink" structure at the double bond position [34, 35] and allocate more space hence more comfortable for cholesterol to integrate. With smaller number of fatty acyl chains type and higher concentration of double bond made cholesterol effect became more systematic in the CocoPEs liposome membrane. The data gave an insight about the application of CocoPEs as encapsulation material. CocoPEs was a good candidate for encapsulation hydrophilic material.

A total of (9.8 × 10−3%, w/w) of coconut phosphatidylethanolamine species (CocoPEs) was isolated from dried coconut meat. The CocoPEs were obtained in the form of a dark brownish gel. Parent ion screening by LCMSMS revealed that 15 species were found in CocoPEs. Characterization of fatty acyl chains by GCMS resulted in that the hydrophobic part of the species were comprised of capric, linoleic, oleic, stearic and arachidic acyl chains. Phase behavior analysis using DSC obtained at least four different phases on CocoPEs i.e. planar-shape gel phase, rippling phase, liquid crystal phase and hexagonal phase. Each phase change occurred at a particular temperature. The pre-transition temperature (Tp) was from planar-shaped gel to rippling phase at 25.29°C, the melting temperature (Tm) for major transition from gel to liquid crystal at 32.62°C, and the hexagonal phase formation from liquid crystal (Th) at 65.53°C. CocoPEs liposome had high encapsulation efficiency. The presence of cholesterol in the membrane liposome up to 30% did not change much of their encapsulation efficiency. The encapsulation efficiencies were above 80%. Meanwhile coconut phospholipids (CocoPLs) had them above 90% but then decrease irregularly to 52% at 0% and 30% choles-

DH and KA would like to express their gratitude of financial support by DIPA

The authors declare that there is no conflict of interests regarding the publica-

*DOI: http://dx.doi.org/10.5772/intechopen.88176*

**4. Conclusion**

terol respectively.

**Acknowledgements**

**Conflict of interest**

tion of this chapter.

Selain APBN FSM UNDIP Riset Madya, 2018.

*Coconut Phospholipid Species: Isolation, Characterization and Application as Drug Delivery… DOI: http://dx.doi.org/10.5772/intechopen.88176*

liposomes more consistent than CocoPLs. We suspected it was due to the molecular composition of the phospholipid in the membrane. The molecular composition was represented by the composition of functional group and fatty acyl chains in the CocoPEs and CocoPLs, **Tables 1–3**. In the liposome membrane cholesterol interacted with CocoPEs and CocoPLs through their functional groups and fatty acyl chains. Cholesterol with small hydrophilic head group i.e., –OH and big and rigid hydrophobic steroid ring would interact better with small head group phospholipid species like CocoPEs than CocoPLs which had big spherical choline group and possibly other head groups as well. The composition of fatty acyl with double bonds also suspected would give more room for cholesterol hydrophobic moiety. The fatty acyl chains would assume "kink" structure at the double bond position [34, 35] and allocate more space hence more comfortable for cholesterol to integrate. With smaller number of fatty acyl chains type and higher concentration of double bond made cholesterol effect became more systematic in the CocoPEs liposome membrane. The data gave an insight about the application of CocoPEs as encapsulation material. CocoPEs was a good candidate for encapsulation hydrophilic material.
