1H NMR Quantitative Determination of Chlorophyll a in the Spirulina Biomass

Dmitry V. Belykh,^a@ Elena N. Zainullina,^a and Ol’ga M. Startseva^b

^aInstitute of Chemistry of the Federal Research Center “Komi Scientific Center of the Ural Branch of the Russian Academy of Sciences”, 167000 Syktyvkar, Russia

^bFederal State Budgetary Educational Institution of Higher Education "Pitirim Sorokin Syktyvkar State University", 167000 Syktyvkar, Russia

^@Corresponding author E-mail: belykh-dv@mail.ru

DOI: 10.6060/mhc200607b
Macroheterocycles 2020 13 (3) 260-268

A method for determining the mass fraction of chlorophyll a in the spirulina biomass — the feedstock for the preparation of chlorophyll a derivatives — using ¹H NMR spectroscopy without preliminary separation of the extract components is proposed (the extract was obtained by exhaustive extraction with a mixture of chloroform and ethanol). Because of the relative lability of chlorophyll a for its determination the demetallation was performed under mild conditions (action on the extract by 10 % hydrochloric acid) with formation of more stable pheophytin a, which served as a derivate for the determination of chlorophyll a. The chlorophyll a mass fraction in the studied raw materials was calculated based on the amount of pheophytin a in the extract. The content of pheophytin a in the extract analyzed for calculating of the chlorophyll a mass fraction in the air-dry biomass of spirulina was determined by the internal standard method. The 4-nitrobenzaldehyde was used as an internal standard. The ratio of the amount of pheophytin a and 4-nitrobenzaldehyde added to the extract was determined by the signal integrated intensities of protons of the internal standard and protons in the meso-positions of the pheophytin a molecule. It was shown that, to increase the accuracy of determination, preliminary removal of a part of the lipid components by extraction with hexane is useful. The mass fraction of chlorophyll a in the raw material studied was ω(chl) = 0.81 ± 0.05 %. The result obtained corresponds to the data on the preparative yields of pheophytin a and methylpheophorbide a obtained from the same spirulina sample (the yields of these compounds can be used to judge the lower boundary of the content of chlorophyll a in the raw material). The proposed technique can be adapted for other types of raw materials containing chlorophyll a or its derivatives.

References:

Click here to expand or collapse this section

1. Imran M., Muhammad M., Kaleem Qureshi A.K., Khan M.A., Tariq M. Biosensors 2018, 8, 95.
https://doi.org/10.3390/bios8040095

2. Abrahamse H., Hamblin M.R. Biochem. J. 2016, 473(4), 347-364.
https://doi.org/10.1042/BJ20150942

3. Van Straten D., Mashayekhi V., de Bruijn H.S., Oliveira S., Robinson D.J. Cancers 2017, 9, 19.
https://doi.org/10.3390/cancers9020019

4. Ormond A.B., Freeman H.S. Materials 2013, 6(3), 817-840.
https://doi.org/10.3390/ma6030817

5. Hamblin M.R. Curr. Opin. Microbiol. 2016, 33, 67-73.
https://doi.org/10.1016/j.mib.2016.06.008

6. Kustov A.V., Kustova T.V., Belykh D.V., Khudyaeva I.S., Berezin D.B. Dyes and Pigments 2020, 173, 107948.
https://doi.org/10.1016/j.dyepig.2019.107948

7. Osati S., Ali H., Guérin B., van Lier J.E. J. Porphyrins Phthalocyanines 2017, 21, 701-730.
https://doi.org/10.1142/S108842461730004X

8. Glowacka-Sobotta A., Wrotynski M., Kryjewski M., Sobotta L., Mielcarek J. J. Porphyrins Phthalocyanines 2019, 23, 1-10.
https://doi.org/10.1142/S108842461850116X

9. Aggarwal A., Samaroo D., Jovanovic I.R., Singh S., Tuz M.P., Mackiewicz M.R. J. Porphyrins Phthalocyanines 2019, 23, 741-765.
https://doi.org/10.1142/S1088424619300118

10. Singh S., Aggarwal A., N.V.S. Dinesh K. Bhupathiraju, Arianna G., Tiwari K., Drain C.M. Chem. Rev. 2015, 115(18), 10261-10306.
https://doi.org/10.1021/acs.chemrev.5b00244

11. Lovell J.F., Tracy W.B. Liu, Chen J., Zheng G. Chem. Rev. 2010, 110, 2839-2857.
https://doi.org/10.1021/cr900236h

12. Tkachenko N.V., Tauber A.Y., Grandell D., Hynninen P.H., Lemmetyinen H. J. Phys. Chem. A 1999, 103, 3646-3656.
https://doi.org/10.1021/jp983765q

13. Boxer S.G., Closs G.L. J. Am. Chem. Soc. 1976, 98, 5406-5408.
https://doi.org/10.1021/ja00433a066

14. Wasielewski M.R., Svec W.A., Cope B.T. Am. Chem. Soc. 1978, 100, 1961-1962.
https://doi.org/10.1021/ja00474a070

15. Boxer S.G., Bucks R.R. Am. Chem. Soc. 1979, 101, 1883-1885.
https://doi.org/10.1021/ja00501a045

16. Liddell P.A., Barrett D., Makings L.R., Pessiki P.J., Gust D., Moore T.A. Am. Chem. Soc. 1986, 108, 5350-5352.
https://doi.org/10.1021/ja00277a053

17. Wiederrecht G.P., Niemczyk M.P., Svec W.A., Wasielewski M.R. J. Am. Chem. Soc. 1996, 118, 81-88.
https://doi.org/10.1021/ja953159y

18. Kelley R.F., Tauber M.J., Wasielewski M.R. J. Am. Chem. Soc. 2006, 128, 4779-4791.
https://doi.org/10.1021/ja058233j

19. Panda M.K., Ladomenou K., Coutsolelos A.G. Coord. Chem. Rev. 2012, 256, 2601-2627.
https://doi.org/10.1016/j.ccr.2012.04.041

20. Amao Y., Komori T. Biosens. Bioelectron. 2004, 19, 843-847.
https://doi.org/10.1016/j.bios.2003.08.003

21. Amao Y., Yamada Y. Biosens. Bioelectron. 2007, 22, 1561-1565.
https://doi.org/10.1016/j.bios.2006.07.006

22. Alekseev A.S., Tkachenko N.V., Tauber A.Y., Hynninen P.H., Osterbacka R., Stubb H., Lemmetyinen H. Chem. Phys. 2002, 275, 243-251.
https://doi.org/10.1016/S0301-0104(01)00515-8

23. Gryglik D., Miller J.S., Ledakowicz S. Solar Energy 2004, 77, 615-623.
https://doi.org/10.1016/j.solener.2004.03.029

24. Taniguchi M., Lindsey J.S. Chem. Rev. 2017, 117(2), 344-535.
https://doi.org/10.1021/acs.chemrev.5b00696

25. Lindsey J.S. Chem. Rev. 2015, 115(13), 6534-6620.
https://doi.org/10.1021/acs.chemrev.5b00065

26. Montforts F.-P., Gerlach B., Höper F. Chem. Rev. 1994, 94, 327-347.
https://doi.org/10.1021/cr00026a003

27. Ma L., Dolphin D. J. Org. Chem. 1996, 61, 2501-2510.
https://doi.org/10.1021/jo951854i

28. Gerlach B., Brantley S.E., Smith K.M. J. Org. Chem. 1998, 63, 2314-2320.
https://doi.org/10.1021/jo9721608

29. Ma L., Dolphin D. Phytochemistry 1999, 50, 195-202.
https://doi.org/10.1016/S0031-9422(98)00584-6

30. Ma L., Dolphin D. Tetrahedron Asymmetry 1995, 6, 313-316.
https://doi.org/10.1016/0957-4166(95)00001-6

31. Tamiaki H., Kouraba M., Takeda K., Kondo Sh., Tanikaga R. Tetrahedron Asymmetry 1998, 9, 2101-2111.
https://doi.org/10.1016/S0957-4166(98)00199-2

32. Porphyrins: Structure, Properties and Synthesis (Enikolopyan N.S., Ed.) Moscow: Nauka, 1985. 334 p. (in Russ.).

33. Gurinovich G.P., Sevchenko A.N., Solov'ev K.N. Spectroscopy of Chlorophyll and Its Related Compounds. Minsk: Nauka i Tekhnika, 1968. 518 p. (in Russ.).

34. Belykh D.V. Synthesis of Polyfunctional Chlorins Based on Methylpheophorbide a. Syktyvkar: KomiNC UrO RAN, 2012. 164 p. (in Russ.)..

35. Khudyaeva I.S., Shevchenko O.G., Belykh D.V. Russ. Chem. Bull. 2020, 69(4), 742-750.
https://doi.org/10.1007/s11172-020-2827-2

36. Karimov D.R., Makarov V.V., Kruchin S.O., Berezin D.B., Cmirnova N.L., Berezin M.B., Zheltova E.I., Strel'nikov A.I., Kustov A.V. Khimija Rastitel'nogo Syr'ja 2014, 189-196. (in Russ.).
https://doi.org/10.14258/jcprm.201404310

37. Valverde J., This H. J. Agric. Food Chem. 2008, 56, 314-320.
https://doi.org/10.1021/jf070277j

38. Mannina L., Segre A. Grasas y Aceites 2002, 53(1), 22-33.
https://doi.org/10.3989/gya.2002.v53.i1.287

39. Jafari T., Durian G., Rahikainen M., Kortesniemi M., Kangasjärvi S., Sinkkonen J. Phytochem. Lett. 2017, 22, 13-20.
https://doi.org/10.1016/j.phytol.2017.07.015

40. Fujii R., Kita M., Doe M., Iinuma Y., Oka N., Takaesu Y., Taira T., Iha M., Mizoguchi T., Cogdell R.J., Hashimoto H. Photosynth. Res. 2012, 111(1-2), 165-172.
https://doi.org/10.1007/s11120-011-9698-1

41. Guadagno C.R., Della Greca M., De Santo A.V., D'Ambrosio N. Photosynth. Res. 2013, 115(1-2), 115-122.
https://doi.org/10.1007/s11120-013-9833-2

42. Sobolev A.P., Brosio E., Gianferri R., Segre A.L. Magn. Reson. Chem. 2005, 43, 625-638.
https://doi.org/10.1002/mrc.1618

43. Pollesello P., Toffanin R., Erikson O., Kilpeläinen I., Hynninen P.H., Paoletti S., Saris N.-E.L. Anal. Biochem. 1993, 214, 238-244.
https://doi.org/10.1006/abio.1993.1483

44. Schoefs B. J. Chromatogr. A 2004, 1054, 217-226.
https://doi.org/10.1016/j.chroma.2004.05.105

45. Mal'shakova M.V., Pylina Y.I., Belykh D.V. Bioorg. Med. Chem. Lett. 2019, 29, 2064-2069.
https://doi.org/10.1016/j.bmcl.2019.07.019

46. Belykh D.V., Kozlov A.S., Pylina Y.I., Khudyaeva I.S., Benditkis А.S., Krasnovsky A.A. Macroheterocycles 2019, 12(1), 68-74.
https://doi.org/10.6060/mhc190128b

47. Pylina Y.I., Startseva O.M., Rasova E.E., Belykh D.V. Macroheterocycles 2019, 12(2), 165-170.
https://doi.org/10.6060/mhc181219b

48. Belykh D.V., Mazaletskaya L.I., Sheludchenko N.I., Rocheva T.K., Khudyaeva I.S., Buravlev E.V., Shchukina O.V., Chukicheva I.Yu. Molecules 2018, 23, 1718.
https://doi.org/10.3390/molecules23071718

49. Kustov A.V., Belykh D.V., Smirnova N.L., Venediktov E.A., Kudayarova T.V., Kruchin S.O., Khudyaeva I.S., Berezin D.B. Dyes and Pigments 2018, 149, 553-559.
https://doi.org/10.1016/j.dyepig.2017.09.073

50. Otvagin V.F., Nyuchev A.V., Kuzmina N.S., Grishin I.D., Gavryushin A.E., Romanenko Y.V., Koifman O.I., Belykh D.V., Peskova N.N., Shilyagina N.Yu., Balalaeva I.V., Fedorov A.Yu. Eur. J. Med. Chem. 2018, 144, 740-750.
https://doi.org/10.1016/j.ejmech.2017.12.062

51. Nyuchev A.V., Otvagin V.F., Gavryushin A.E., Romanenko Y.I., Koifman O.I., Belykh D.V., Schmalz H.-G., Fedorov A.Yu. Synthesis 2015, 47(23), 3717-3726.
https://doi.org/10.1055/s-0034-1378876

52. Santosh K.Bh., Raja R. Trends in Analytical Chemistry 2012, 35, 5-26.
https://doi.org/10.1016/j.trac.2012.02.007

53. Holzgrabe U. Prog. Nucl. Magn. Reson. Spectrosc. 2010, 57, 229-240.
https://doi.org/10.1016/j.pnmrs.2010.05.001

Attachment	Size
mhc200607b.pdf	964.59 KB

Журнал "Макрогетероциклы"

Navigation

Languages

News

Search

1H NMR Quantitative Determination of Chlorophyll a in the Spirulina Biomass

References: